Vitamin D Binding Protein

Manifestations of the disease

We need to change our paradigm from the existing one that symptoms are directly due to the pathogen or insult to the understanding that the symptoms and in fact clinical markers are due to the immune response. So instead of believing shingles erupts or herpes erupts or cystitis symptoms erupt when our immunity is depressed, we see these eruptions as an immune activation.

Of course, if there are factors suppressing immune function then these reactions will be ineffective long term. But our routine approach of suppressing symptoms when faced with an “acute illness” is likely promoting chronic problems.

The immune response in autism

GcMAF/VDTP is produced in all higher order animals and is a glycoprotein that the body both needs and expects.  Therefore it produces no side effects in humans and any symptoms are the result of an activated immune system – the manifestation of the disease. 

Read more: Manifestations of the disease

What is Vitamin D Binding Protein (VDBP)?

 

Vitamin D Binding Protein is a multi-functional plasma protein with many important functions. The most well known one, and the reason for its name, is the transport of vitamin D metabolites .  Because VDBP is the primary transporter of vitamin D, it has a role in maintaining the total levels of vitamin D in the body and in regulating the amounts of free (unbound) vitamin D available for specific tissues and cell types to utilise.

Other functions of VDBP are to control bone development, binding of fatty acids, sequestration of actin and a range of less-defined roles in modulating immune and inflammatory responses.  The actin removal is considered to be a very important role for VDBP, actins being toxic  and released into the body following cell death.

Two amino acids on this protein chain have been shown to be very strongly associated with the activation of macrophages.  In some publications, the vitamin D binding protein is shown to respond to enzymes released by inflammation and this is often referred to as MAF (Macrophage Activating Factor).  Studies show that this isoform not only activates macrophages, it turns them off when no longer needed, so it is an immune system regulator.

Studies in laboratory cultures have shown that Calcitriol (Vitamin D bound with VDBP) can significantly increase the number of dopamine neurons.

Vitamin D Binding Protein is otherwise know as:

  • VDBP
  • Gc-Protein
  • Glycoprotein
  • Transport protein
  • Gc-Globulin
  • GcMAF

Sources of Vitamin D Binding Protein

VDBP is a protein that occurs naturally in all higher order animals and is often produced by the first milk of mothers (colostrum) that helps boost the immunity of their newborns.

  • Human & animal plasma
  • Collostrum
  • Pro-biotic yoghurts
  • Bacular expression
  • Laboratory synthesis

Factors that influence VDBP levels

Liver disease decreases levels of VDBP. Chronic liver disease will decrease levels less than acute liver failure. Trauma and surgery will decrease Vitamin DBP. Septic infections will consume Vitamin DBP faster than production can be increased.

Normal VDBP levels in serum are 350-500 mg/l. Levels of VDBP less than 80 mg/l yield positive and negative mortality predictive values of 85% and 43% respectively. Survivors had levels greater than 102 mg/l.

http://www.saisei-mirai.or.jp/gan/macrophage_eng.html

 

Therapeutic potential of vitamin D-binding protein

 

Vitamin D-binding protein (DBP) is a multi-functional plasma protein with many important functions. These include transport of vitamin D metabolites, control of bone development, binding of fatty acids, sequestration of actin and a range of less-defined roles in modulating immune and inflammatory responses. Exploitation of the unique properties of DBP could enable the development of important therapeutic agents for the treatment of a variety of diseases.

http://www.ncbi.nlm.nih.gov/pubmed/15245906

 

New perspectives on the vitamin D binding protein

 

The serum vitamin D binding protein (DBP), also known as GC-globulin, is a multifunctional protein known for its role in the transport of vitamin D metabolites. DBP also binds fatty acids and actin monomers, preventing their polymerization that could be detrimental in the circulatory system.

DBP may have immune functions independent of its role as a transporter of vitamin D.

Because of the abundance of DBP, many aspects of its basic biochemistry were quickly established. Other features of vitamin D action, particularly transcriptional mechanisms of regulation, received greater focus and early interest in DBP centred on its value as a tool for population genetics because of its intriguing genetic variations. Nonetheless, knowledge of DBP mechanisms in physiology was obtained, and functions beyond vitamin D ligand binding were identified.

With the recent increased attention regarding the benefits of vitamin D (bone health and immunological regulation), there has been a resurgence of interest in DBP. Because DBP is the primary transporter of vitamin D, it has a role in maintaining the total levels of vitamin D for the organism and in regulating the amounts of free (unbound) vitamin D available for specific tissues and cell types to utilize.

This review will describe the findings on the basic biochemistry and molecular biology of DBP, the studies that elucidated its biological functions and highlight these results in light of the current renewed interest in vitamin D and human health, as well as the debate over what constitutes sufficient levels of vitamin D.

http://www.ncbi.nlm.nih.gov/pubmed/22528806

Safety pharmacology, toxicology and pharmacokinetic assessment of VDBP.

 

VDBP is an important protein of the plasma actin-scavenger system. As such, it has been shown to bind free actin and prevent hypercoagulation and shock in patients with massive actin release resulting from severe tissue injuries.

Treatment of such patients with VDBP could therefore potentially be life-saving.

The VDBP formulation was shown to be stable for at least 4 years with full retention of actin-binding capacity. In vitro studies did not reveal activation of the kallikrein system or the complement system and cellular studies showed no toxic effects on a variety of human cell lines. In vivo studies showed no acute toxic effects in mice, rats or guinea pigs upon intravenous infusion.

A 14-day local tolerance study in rabbits showed no adverse effects, and 14-day toxicity studies in rats and horses did not show any unwanted reactions.

A 14-day toxicology study in beagle dogs, formation of antibodies was seen and in the end of the study period, three out of four dogs showed clinical immunological reactions, which could be ascribed to the formation of antibodies.

The half-life, T, for human VDBP was 12 hr in rats, 16 hr in horses and 30 hr in dogs. The safety profile of plasma-derived VDBP is concluded to be consistent to that required for use in man.

http://www.ncbi.nlm.nih.gov/pubmed/20560927


Autism & MAF as a potential immune development protocol

Initial Observations of elevated Nagalase activity associated with Autism and observed reductions from GcMAF

Full paper is available here

Abstract & Background

Autism spectrum disorders (ASD) are developmental disorders affecting 1:88 children, and which appear to be associated with a variety of complex immune dysregulations including autoimmunity. The enzyme, alpha-N-acetylgalactosaminidase (Nagalase) deglycosylates serum Gc protein (vitamin D3 – binding protein) rendering it incapable of activating macrophage defenses. Increased Nagalase activity has been associated with a variety of malignancies, immune disorders, viral and bacterial infections. 

Macrophage activating factor (GcMAF) has been repeatedly published as an intervention to lower serum Nagalase activity for a variety of cancer and HIV patients. GcMAF is a naturally occurring protein with well-established safety and therapeutic benefits supported by numerous human studies. 

Methods

Initially, parents of 40 individuals with ASD sought testing for Nagalase serum activity as part of an evaluation of immune dysregulation. Nagalase enzyme activity measurement was performed by the European Laboratory of Nutrients (ELN), Bunnik, the Netherlands, using an end-point enzymatic assay of a chromogenic substrate. Some parents of patients with elevated Nagalase activity opted for weekly GcMAF injections. GcMAF is purified from human serum obtained from the American Red Cross using 25-hydroxyvitamin D3-Sepharose high affinity chromatography. The protein is then further diluted to obtain therapeutically appropriate levels for patients based on their clinical presentations.

Results

Individuals with ASD (32 males and 8 females, n = 40, ages: 1 year 4 months - 21 years 2 months) had initial and post treatment assessment of Nagalase activity. Dosing of GcMAF was recommended based on previously reported response curves adjusted by the treating clinician for age, weight, and Nagalase levels. The average pre-treatment Nagalase activity of the autism group was 1.93 nmol/min/mg of substrate. This was well above the laboratory reported normal range of ,0.95 nmol/min/mg. For the ASD group the average level at the time of second testing was 1.03 nmol/min/mg, reflecting an average reduction of 0.90 nmol/min/mg (P , 0.0001). Apart from the likely immunological benefits of lowering the Nagalase activity of these individuals, uncontrolled observations of GcMAF therapy indicated substantial improvements in language, socialization and cognition. No significant side-effects were reported during the course of injections.

Conclusions

In this first report of Nagalase activity in patients with autism, it appears that most individuals have substantially higher levels than the expected healthy ranges. Although Nagalase is a nonspecific marker of immune dysregulation, its observed levels in autism may have both etiological and therapeutic significance. Importantly, this is also the first report of reduction of Nagalase activity in an autism population with GcMAF injections. 

 

Dr Bradstreet Initial Observatons of Elevated Nagalase Activity

Associated with Autism and Observed Reductions from GcMAF

             

Nagalase dataset for pre-post GcMAF therapy with iCGl response per subject.

           

 

 

 

Nagalase

Nagalase

 

Nagalase

 

Gender

Age 

Pre Result

Post Result

Days Between

Difference

 iCGl *

Male

6

0.90

0.47

76

-0.43

Considerable Improvement (4)

Male

21

1.00

0.44

79

-0.56

Very Considerable Improvement (5)

Male

7

1.30

0.72

143

-0.58

Moderate Improvement (3)

Male

10

2.20

1.30

200

-0.90

Slight Improvement (2)

Male

7

1.90

0.76

112

-1.14

Considerable Improvement (4)

Female

11

1.90

1.00

112

-0.90

Very Considerable Improvement (5)

Female

9

1.90

1.20

111

-0.70

Very Considerable Improvement (5)

Male

4

1.70

1.10

112

-0.60

Considerable Improvement (4)

Male

4

1.00

0.76

116

-0.24

Moderate Improvement (3)

Male

6

1.20

0.79

69

-0.41

Considerable Improvement (4)

Female

5

1.66

0.40

76

-1.26

Very Slight Deterioration (-1)

Male

12

1.69

0.47

88

-1.22

Very Slight Improvement (1)

Male

12

7.80

4.40

56

-3.40

Moderate Improvement (3)

Male

7

1.50

0.90

60

-0.60

Considerable Improvement (4)

Female

12

1.98

0.81

104

-1.17

Moderate Improvement (3)

Male

1

1.50

1.00

95

-0.50

Considerable Improvement (4)

Male

6

2.60

2.50

95

-0.10

Slight Improvement (2)

Male

1

2.80

1.80

111

-1.00

State Unchanged (0)

Male

18

1.30

0.92

69

-0.38

Moderate Improvement (3)

Male

4

3.00

1.00

87

-2.00

Very Considerable Improvement (5)

Male

17

1.20

0.80

128

-0.40

Considerable Improvement (4)

Male

4

1.60

1.10

53

-0.50

Moderate Improvement (3)

Female

5

0.92

0.62

103

-0.30

State Unchanged (0)

Male

11

1.00

0.90

88

-0.10

Moderate Improvement (3)

Male

5

1.40

0.81

105

-0.59

Moderate Improvement (3)

Female

5

3.90

1.60

126

-2.30

Very Considerable Improvement (5)

Female

4

1.10

0.61

89

-0.49

Slight Improvement (2)

Male

18

4.00

1.40

70

-2.60

Considerable Improvement (4)

Male

3

2.60

1.40

81

-1.20

Moderate Improvement (3)

Male

5

1.20

0.96

184

-0.24

Moderate Improvement (3)

Male

9

1.79

0.57

95

-1.22

Slight Improvement (2)

Female

18

1.90

1.20

159

-0.70

State Unchanged (0)

Male

16

1.82

0.62

77

-1.20

Slight Improvement (2)

Male

11

2.90

0.93

82

-0.97

Moderate Improvement (3)

Male

7

1.73

0.51

38

-1.22

Considerable Improvement (4)

Male

7

2.90

1.20

75

-1.70

Very Considerable Improvement (5)

Male

10

1.20

0.82

103

-0.38

Very Slight Improvement (1)

Male

8

1.10

0.68

103

-0.42

Slight Improvement (2)

Male

4

1.00

0.89

117

-0.11

Slight Improvement (2)

Male

4

1.20

0.91

138

-0.29

Very Considerable Improvement (5)

 


Read more: Initial Observations of elevated Nagalase activity associated with Autism and observed reductions from GcMAF

Immune dysfunction in autism

Autism spectrum disorders (ASD) are developmental disorders affecting 1:88 children, and which appear to be associated with a variety of complex immune dysregulations including autoimmunity. The enzyme, alpha-N-acetylgalactosaminidase (Nagalase) deglycosylates Vitamin D Binding Protein (VDBP) rendering it incapable of activating macrophage defenses. 

Increased Nagalase activity has been associated with a variety of malignancies, immune disorders and viral infections. Macrophage activating factor (MAF) has been repeatedly published as an intervention to lower serum Nagalase activity for a variety of cancer and HIV patients. 

MAF is a naturally occurring protein with well-established safety and therapeutic benefit(s) supported by numerous human studies.

Uncontrolled observations of MAF therapy in Autism indicated substantial improvements in language, socialization and cognition. During this time a reduction in Nagalase levels was recorded.  Although Nagalase is a nonspecific marker of immune dysregulation, its observed levels in autism may have both etiological and therapeutic significance.

Autism, once a rare disorder, it is now approaching epidemic, if not pandemic, proportions.

While no consensus exists, this trend in autism is at least suggestive of an infective pathogen. Within this context, various organisms have been postulated to be involved, including: gastrointestinal infections, Polyomaviruses, Chlamydophila, Bornaviruses, Paramyxoviruses, and Borrelia burgdorferi. While any of these may contribute to a small percentage of autism cases, it seems unlikely that any of them indi¬vidually represents the origin of this epidemic.

Despite this uncertainty, growing evidence supports significant immune dysfunction, including autoimmunity, in autism.

One possible explanation for the pattern of immune dysregulation and observed in autism spectrum disorders (ASD) could be persistence of active pathogens, perhaps from the perinatal or a subsequent period of child development.

Nagalase has been published as a biomarker associated with various types of cancer, systemic lupus erythematosus (SLE), influenza, and human immunodeficiency virus infection (HIV). It  appears to be an important indica¬tor of secondary immune dysregulation.

Nagalase is an enzyme that deglycosylates the Vitamin D binding protein (VDBP), rendering it inca¬pable of immune activation and thereby preventing its regulation of macrophage activation.    It is noteworthy that vitamin D deficiency, either in pregnancy or during postnatal development, is an apparent risk factor for autism.The impact of Nagalase on VDBP transportation of vitamin D is not known. However, vitamin D deficiency is a known risk factor for autoimmunity.

Initial Observations of Elevated Nagalase Activity Associated with Autism and Observed Reductions from GC Protein—Macrophage Activating Factor Injections

Autism spectrum disorders (ASD) are developmental disorders affecting 1:88 children, and which appear to be associated with a variety of complex immune dysregulations including autoimmunity. The enzyme, alpha-N-acetylgalactosaminidase (Nagalase) deglycosylates serum Gc protein (vitamin D3 – binding protein) rendering it incapable of activating macrophage defenses. Increased Nagalase activity has been associated with a variety of malignancies, immune disorders and viral infections. Macrophage activating factor (GcMAF) has been repeatedly published as an intervention to lower serum Nagalase activity for a variety of cancer and HIV patients. GcMAF is a naturally occurring protein with well-established safety and therapeutic benefit(s) supported by numerous human studies.

Methods: Initially, parents of 40 individuals with ASD sought testing for Nagalase serum activity as part of an evaluation of immune dysregulation. Nagalase enzyme activity measurement was performed by the European Laboratory of Nutrients (ELN), Bunnik, the Netherlands, using an end-point enzymatic assay of a chromogenic substrate. Some parents of patients with elevated Nagalase activity opted for weekly GcMAF injections provided by Immuno Biotech Ltd., Guernsey UK (www.gcmaf.eu). GcMAF is purified from human serum obtained from the American Red Cross using 25-hydroxyvitamin D3-Sepharose high affinity chromatography. The protein is then further diluted to obtain therapeutically appropriate levels for patients based on their clinical presentations.

Results: Individuals with ASD (32 males and 8 females, n = 40, ages: 1 year 4 months - 21 years 2 months) had initial and post treatment assessment of Nagalase activity. Dosing of GcMAF was recommended based on previously reported response curves adjusted by the treating clinician for age, weight, and Nagalase levels. The average pre-treatment Nagalase activity of the autism group was 1.93 nmol/min/mg of substrate. This was well above the laboratory reported normal range of <0.95 nmol/min/mg. For the ASD group the average level at the time of second testing was 1.03 nmol/min/mg, reflecting an average reduction of 0.90 nmol/min/mg (P < 0.0001). Apart from the likely immunological benefits of lowering the Nagalase activity of these individuals, uncontrolled observations of GcMAF therapy indicated substantial improvements in language, socialization and cognition. No significant side-effects were reported during the course of injections.

Conclusions: In this first report of Nagalase activity in patients with autism, it appears that most individuals have substantially higher levels than the expected healthy ranges. Although Nagalase is a nonspecific marker of immune dysregulation, its observed levels in autism may have both etiological and therapeutic significance. Importantly, this is also the first report of reduction of Nagalase activity in an autism population with GcMAF injections.

http://www.la-press.com/initial-observations-of-elevated-alpha-n-acetylgalactosaminidase-activ-article-a3450

Autistic disorder and viral infections

Autistic disorder (autism) is a behaviorally defined developmental disorder with a wide range of behaviors. Although the etiology of autism is unknown, data suggest that autism results from multiple etiologies with both genetic and environmental contributions, which may explain the spectrum of behaviors seen in this disorder.

One proposed etiology for autism is viral infection very early in development. The mechanism, by which viral infection may lead to autism, be it through direct infection of the central nervous system (CNS), through infection elsewhere in the body acting as a trigger for disease in the CNS, through alteration of the immune response of the mother or offspring, or through a combination of these, is not yet known. Animal models in which early viral infection results in behavioral changes later in life include the influenza virus model in pregnant mice and the Borna disease virus model in newborn Lewis rats.

Many studies over the years have presented evidence both for and against the association of autism with various viral infections. The best association to date has been made between congenital rubella and autism; however, members of the herpes virus family may also have a role in autism.

Recently, controversy has arisen as to the involvement of measles virus and/or the measles, mumps, rubella (MMR) vaccine in the development of autism.

Biological assays lend support to the association between measles virus or MMR and autism whereas epidemiologic studies show no association between MMR and autism.

Further research is needed to clarify both the mechanisms whereby viral infection early in development may lead to autism and the possible involvement of the MMR vaccine in the development of autism.

J Neurovirol. 2005 Feb;11(1):1-10.

Libbey JE, Sweeten TL, McMahon WM, Fujinami RS.

Department of Neurology, University of Utah, Salt Lake City, Utah 84132-2305, USA.

More on Viral Persistence in a Subgroup of Autism

First published April 12 2011

Let’s talk about the link between our fellow travelers (all the bacteria, viruses and other organisms residing with us – actually mostly in us). Biological psychiatry is just coming to terms with the microbiome/microbiota, and its influence on the function of the brain. As we collectively try to understand the influence of this unseen influence on the human condition, we cannot forget the influence of our shared viral travelers.  The obvious viruses, herpes as an example, exist with us from our first encounter until the end of our days. But many viruses are capable of lifelong cohabitation with humans.

The most obvious viruses are; Human Papilloma (HPV), Herpes Simplex (HSV), Cytomegalovirus (CMV), Hepatitis (Hep B and C as well as others), Human Immunodeficiency (HIV), Epstein Barr (EBV – mononucleosis), Varicella (chickenpox), Measles, Polio, SV40, BK and JC. XMRV is a potential new addition to this list, and despite its controversy I believe it is a true human pathogen.  My belief in XMRV as a necessary member of the list is based on both PCR and anti-XMRV antibodies, not merely XMRV PCR (genetic) material. 

Some of these have a potential legacy with autism.  We discussed this in an earlier post. But this viral community interfaces with the bacteria community in complex and incompletely understood ways. 

Read more: More on Viral Persistence in a Subgroup of Autism

Autism and Macrophage Stimulation – 2 years of learning.

1st published January 2013

A brief summary of MAF

MAF is a sugar protein that acts as a Macrophage Activating Factor (MAF) in the immune system.  It is part of the Active Vitamin D Molecule (1,25-dihydroxyvitamin D3), produced in the liver, is completely natural and the body expects it. Over 57 research papers have been published that demonstrate GcMAF activates Macrophages in both in vivo and in vitro studies

Macrophages – The ‘Big Eaters’

Macrophages function in both non-specific defense (innate immunity) as well as help initiate specific defence mechanisms (adaptive immunity) of vertebrate animals. Their role is to phagocytise (engulf and then digest) cellular debris and pathogens, either as stationary or as mobile cells. They also stimulate lymphocytes and other immune cells to respond to pathogens. They are specialized phagocytic cells that attack foreign substances, infectious microbes and cancer cells through destruction and ingestion.

Nagalase – collapsing the immune system.

In order to defeat the immune system and be able to grow and spread unchecked, cancers and many viruses secrete an enzyme called Nagalase which prevents the human body from producing its own MAF.  At a biological level the human body is constantly under attack so there is an excepted 'normal' level at 0.65 nmol/mg.  Levels above 0.65 nmol/mg indicate the immune system may have been compromised and the macrophages are not activated. 

85% of autistic children are found to have high levels of Nagalase.

Nagalase’s only purpose is to destroy GcMAF and this it does with great precision, time and time again with no other collateral damage. In this simple way Nagalase has been responsible for billions of deaths in human history.

MAF therapy

MAF is simply a protein replacement therapy, giving the body the protein it should be producing and expects. 

Dr. Yamamoto’s three studies showed that incredibly small weekly doses (100 billionths of a gram—an amount that is invisible to the naked eye) of GcMAF had cured early metastatic breast, prostate, and colon cancers in 100% of (non anaemic) patients. In a fourth paper, he used the same treatment to cure 100% of non anaemic HIV-infected patients.

More recently Dr Bradstreet & Dr Antonucci have performed MAF therapy on over 2000 autistic children with encouraging results.  Over 85% have responded favourably to MAF and 15% of these have lost the label of autism altogether

Read more: Autism and Macrophage Stimulation – 2 years of learning.

Debunking the lamestream view of autism

First published April 4th 2012

With more than one in a hundred children in the UK succumbing to autism the medical establishment is still no nearer to discovering the causes of, or providing any successful treatment for, this dreaded illness.   There is, however, growing hope of recovery at any age.

The causes Autism?

The static theory - The mainstream view of autism is that it is a “static” encephalopathy with its roots in the womb or in genetic causes.  There is yet no hope of successful treatment as the brain and nervous system are considered to be permanently damaged.

The viral theory - That the onset of autism can be attributed to a particular illness or vaccine and that person “regressed” into autism.  The virus has compromised the immune system allowing the virus to stay locked in the body for years, even life! As Dr. Bradstreet notes in December 2011 - “For years I have observed that even the most substantially effected children have moments where they are profoundly better – perhaps only for mere fleeting periods of time.  This alone implies there is some interference with neuro-processing which is not always present at the same level.  Therefore, it is conceivable that this interfering substance is amenable to biomedical therapy. “

It has now been proven that over 80% Autistic Children are deficient in Vitamin D and have elevated levels of the destructive enzyme 'Nagalase'

Read more: Debunking the lamestream view of autism

Low methylation levels associated with autism

 

The methylation cycle, is a complex biochemical system that is very important as it occurs within every cell of our body and takes place billions of times every second. It is responsible for many reactions within the body including aiding the repair to your DNA on a daily basis, it also manages homocysteine. Homocysteine is an amino acid, when it is not regulated properly it becomes elevated and can cause various adverse reactions including damage to inner arterial lining (endothelium) which can result in cardiovascular disease. Methylation also helps recycle molecules needed for detoxification (Glutathione). Without these significant activities happening the body cannot regulate or react properly and the knock on effect can have numerous health implications. To keep methylation running smoothly you need optimal levels of bioavailable B vitamins. Without enough of these B vitamins methylation breaks down and the results can be catastrophic. 

Low methylation levels are not only associated with depression but also cardiovascular disease, birth defects, downs syndrome, miscarriages, osteoporosis, diabetes, cancer, behavioural disorders, Autism, dementia, fibromyalgia, chronic fatigue syndrome, stroke and some serious health problems, some of which can be debilitating and/or life threatening.

Read more: Low methylation levels associated with autism

Gluten and Casein - negative effects in Autism

Gluten is a protein found in wheat, rye, barley and oats. When you eat, the food in your stomach gets broken down, the body utilises the good bits and the other bits get flushed out as waste matter. With autistic children foods containing gluten do not break down fully. The broken down pieces are called peptides and these peptides are small enough to pass through the wall of the stomach rather than being fully processed. Children and adults with autism usually have something called Leaky gut, this means that the wall of the stomach is damaged and these peptides are small enough to pass through the wall into the bloodstream and the central nervous system including the brain, causing various adverse reactions.
 
When the protein gluten gets broken down, it becomes the peptide known as gluteomorphin. Most people are aware of the highly addictive drug Morphine and this peptide is very similar in its effects on the brain and body of the autistic person.

Casein which is the protein found in dairy products. This protein breaks down to the peptide Caseomorphin but again like gluten, does not fully break down and passes through the lining of the stomach into the bloodstream, contributing to the adverse reactions listed below.
 
Gluten & Casein foods are also inflammatory , where you have inflammation you have a perfect environment for disease to flourish.
 
Parents of autistic children have observed the following reactions when their children have eaten foods containing gluten and/or casein.

Read more: Gluten and Casein - negative effects in Autism

Parasites & Autism

A parasite is a creature that lives off another living creature consuming all its nutrients and causing them to become very sick.  Parasites live everywhere, but they particularly thrive in warm, moist areas. There are various parasite, liver flukes, Tapeworm, Ropeworm, Roundworm, Hookworm & pinworm and also bacterial parasites like Giardia just to name a few.

Parasites can lay tens of thousands of eggs each day! Around the full moon the parasites which reside in the intestine move to the stomach to breed.

Parasites are very calculating and thrive when the immune system is under stress and cannot fight them off, the gut then becomes inflamed resulting in a perfect environment for parasites to flourish.

Parasites are very common around the world. 

A few ways that Parasites can spread:- from animals to humans; in water, this includes bath water, swimming water & drinking water; by eating under cooked meat, Pork is the worst; raw fish - Sushi; unwashed fruit and vegetables; poor hygiene (not washing hands after visiting the toilet)  soil; feces; urine; saliva, including mouthing of toys in children; sandboxes; nappy changing tables; nurserys & schools; toilets; shopping trolleys; kitchen utensils and worktops; We may even be born with them; Some spread through the air, through insect bites, if the insect is a carrier.

The symptoms of parasites can vary and testing for them is almost impossible as they do not show up on routine blood test and stool test produces a lot of false negatives because Laboratories only test for 40 or 50 out of literally thousands of varieties of parasites.

Read more: Parasites & Autism

Autism and the leaky gut

To understand leaky gut and how this impedes your health, you first need to fully comprehend the role of the gut, what happens inside it and the implications this has on overall health.
 
Your immune system and your gut are linked in every respect. Roughly eighty percent of your immune tissue is occupied within your digestive system.
 
The digestive system is a complex network of various functions, when one of these functions becomes imbalanced it can create a knock on effect within other areas of the body, causing extreme health issues.
 
The digestive system comprises of cells, proteins, tissues and organs which work together in a intricate way to defend the body against any harmful pathogens including bad bacteria, viruses, infectious diseases, parasites and toxins. In fact the gut mucosa connects with the greatest inhabitants of immune cells in the body. These are also known as gastrointestinal immune cells, which come from the lymphoid section of the immune system. Their aim is to produce lymphocyte cells which attack harmful pathogens that invade the body. These lymphatic cells also group themselves together, these collections are known as Peyers Patches.

Read more: Autism and the leaky gut

Toxins in the home

Environmental experts have established that many households contain approximately 62 toxic chemicals and although manufacturers state that there are only small amounts present and unlikely to cause any concern, if you are constantly being exposed to these toxins day in day out then the build up over a period of time can be considerable. With children who have autism already struggle to detoxify this can be detrimental for them. Ingredients in common household products have been linked to asthma, cancer, hormone disturbances, reproductive issues and neurotoxicity.

These are some of the most common toxic chemicals found in household products:-

PHTHALATES

These have been widely used since the 1950's. They are used in the plastic industry to make the plastic softer so when it is bent it does not become brittle and crack, for example food packaging or storage and drink bottles. Because they are not chemically bound to the plastic, this allows the phthalates to be continuously released into the food, drink or environment. They are also used in many fragranced household products,  for example, air fresheners & soaps but also if the manufacturers are stating "fragrance" on the label then there will be a good chance that Phthalates will be an ingredient but due to proprietary laws, companies do not have to disclose what’s in their scents so you most probably will not see phthalates listed on the label.

Read more: Toxins in the home

Ethics in Autism Care

Due to the particular “social” status of an autistic patient, ethics in approaching autism and its treatments and research gain much consideration. There are many problems to be faced by relatives of autistic people, with consequences on the quality of life of all members of the family. Recent evidences indicate a negative impact on the employment of the parents of children with autism, with an increasing percentage of mothers that had to leave work or reduce it. The burden of care and surveillance amounts to an average for the family to 17.1 hours/day [5].

Autism is a disorder that persists throughout the entire existence, so that, autistic people need, throughout their life, continuous protection, assistance, as well as, prolonged specialized services and opportunities for independent adult life by the family. Psychological intervention in the clinical management of ASDs is a priority.

Once a child is diagnosed of autism, many hours per week are requested to perform an optimal treatment and structured behavioural and educational intervention, often at overwhelming expense to families. Applied Behaviour Analysis (ABA), a program that uses a one-on-one teaching approach that reinforces the practice of various skills, is now recognized as the most effective psychological treatment. ABA programs are usually performed in a child’s home under the supervision of a behavioural psychologist. However, ABA programs can be very expensive and have not been widely adopted by school or healthcare systems. The family also takes care of this aspect.

Unfortunately, too many autistic children and adolescents are still excluded from education system, and cannot achieve any form of assistance outside of parental care. This fact could strongly influence the onset of their adolescence and adulthood. The loss of an appropriate perspective of a dignified adulthood, together with a prospective of marginalization, increases the stress and the sense of guilt and helplessness of the family.

Read more: Ethics in Autism Care

Support for Immune Causes of Autism

Genome-wide expression studies in Autism spectrum disorder, Rett syndrome, and Down syndrome.

Abstract

Though different in their aetiology, autism spectrum disorder (ASD), Rett syndrome (RTT) and Down syndrome (DS) are three neurodevelopmental disorders sharing significant clinical and neuropathological overlaps.

Genome-wide expression studies are reviewed and available datasets from post-mortem brains reanalyzed to identify genes and gene pathways dysregulated in all three disorders. Our results surprisingly converge upon immune, and not neurodevelopmental genes, as the most consistently shared abnormality in genome-wide expression patterns.

A dysregulated immune response, accompanied by enhanced oxidative stress and abnormal mitochondrial metabolism seemingly represents the common molecular underpinning of these neurodevelopmental disorders. This conclusion may be important for the definition of pharmacological therapies able to ameliorate clinical symptoms across these disorders.

Lintas C, Sacco R, Persico AM.

Laboratory of Molecular Psychiatry and Neurogenetics, University “Campus Bio-Medico”, Rome, Italy

Department of Experimental Neurosciences, I.R.C.C.S. “Fondazione Santa Lucia”, Rome, Italy

Neurobiol Dis. 2010 Dec 2. [Epub ahead of print]

http://www.sciencedirect.com/science/article/pii/S0969996110003852

Common Sense About Autism Spectrum Disorders

 

Autism has a number of causes - which means you can do something to help reverse it and improve the life of your child.

This report will teach you what the various causes are of autism or autism spectrum disorder. There are at least 13 different causes or conditions that lead to the development of autism. A particular case of autism is not caused by all these conditions, but many of the causes will be involved in the development of autism or an autism spectrum disorder in your child.

We call this common sense because what goes on all too often in the medical field regarding autism has nothing to do with common sense. All too often you are told that autism is mostly caused by genetics, possibly with some environmental causes too. And even worse, that there is little to do about it, or to prevent it. It seems that just about everyone in the standard medical field says, with little evidence to back it up, that vaccines have nothing to do with autism. All this flies in the face of common sense. Why?

A great number of children have significantly improved or even been cured by taking vigorous actions with supplements and therapies. This would not and could not happen if autism was primarily genetic, if there is nothing you can do since it’s genetic. Unfortunately, because drugs are ineffective in dealing with autism, and may even help cause it, the traditional medical world all to often says there is nothing to do.

This is not the case. Many many MDs and health professionals have been able to successfully help autistic children improve, sometimes greatly. If this can happen with even one autistic child or adult, it can happen with any person with autism.

It is a basic tenant of the scientific method that any hypotheses, such as autism is primarily genetic and as a consequence there is nothing to do about it, is disproved when even one event proves it to be false. The hundreds of children who have recovered from autism each disprove or falsify the theory that there is nothing you can do about autism.

Even though that theory has effectively been disproved many times, it persists because, unfortunately, modern medical treatments are anything but scientific. Drugs are a big, money making business. You and your children are victims, as is everyone using non steroidal anti-inflammatory drugs, steroids, statins, osteoporosis drugs, and just about any drug there is on the market.

To put it simply, drugs don’t work well and have many harmful side effects because they are modified from nature so that they can be patented. Our bodies are designed to only use nutrients that are natural. Anything not natural, like drugs, and genetically modified foods, chemicals and the whole onslaught of manmade toxins we have created, has toxic side effects in the body.

Again, to put it simply, an autistic child has been over-exposed to these toxins. There is no way further toxins, in the form of drugs, are going to correct this issue. And they don’t. Which is why the mainstream medical profession is so ineffective in dealing with autism. It can’t possibly work. So rather than admit that their approach is stupid, unscientific and backwards, they claim that autism is primarily genetic and that there is little or nothing you can do to help your child.

As easily thousands of children with autism have improved over the years, with some becoming normal again, to say that nothing can be done, other than giving a few drugs to help with the symptoms, lacks common sense, and is a barbaric way to treat, or should we say, not treat, someone with autism or autism spectrum disorders.

Just because your doctor knows of little to do, don’t feel that there is nothing you can do to improve autism or autism spectrum disorders. Too many people with autism have been improved as the underlying causes are dealt with, for this to be true. Your child will get better too, if you take significant and appropriate action. There is no way they couldn’t. This is just plain common sense.

This report is going to help you identify what the best actions to take are, what needs to be done to improve your child. We start out by covering the causes of autism.

Read more: Common Sense About Autism Spectrum Disorders

Autism treatment strategies – before you take GcMAF

 

Whilst many parents are keen to see their child get onto GcMAF immediately, it is not recommended until the diet and digestion is addressed, the body detoxed of heavy metals and parasites.  

The list below is from the Autism Research Institute, showing the order in which interventions are normally completed.  It has been shown that GcMAF can increase behavioural issues, and this is likely due to an overload of toxins following pathogen destruction by an improved immune system.  Detoxification first is likely to minimize this reaction.

  • Improved diet
  • Find out and remove food sensitivities
  • GFCF diet
  • Vitamin and mineral supplements
  • High Dose vitamin B6 and magnesium
  • Essential fatty acids
  • Gut treatments
  • - Antifungals
  • - Probiotics
  • - Digestive enzymes
  • Amino Acids
  • Carnitine
  • Melatonin
  • Thyroid supplements
  • Sulfation
  • Methylation/glutathione/oxidative stress
  • Immune system regulation including GcMAF
  • Hyperbaric oxygen therapy

As with anything, working closely with a qualified practitioner will enable you to understand whether these interventions are beneficial or not, using regular bloodwork and medical testing.  Everyone is different, and these guidelines above are not intended to be medical advice. 

Starting MAF - what to expect with autism and other coinfections

this is an extract from Dr Bradstreet Paper which you can view online here.

“During the first few weeks of treatment, 3 of 40 patients (7.5%) experienced low to moderate rise in body temperature, typically occur-ring 24 to 48 hours after the GcMAF injection and lasting less than 24 hours. 

Parents were instructed to use ibuprofen only if the temperature exceeded 102° F (approximately 39°C), and two were treated during the first few weeks. By the second month, no patients experienced significant febrile events. 

Interestingly, during the first 3 weeks, 6 of 40 patients (15%) were observed to have rashes compatible with viral exan¬themas (generally on the trunk and in fine papules more commonly than maculae). Petechiae were not observed. These rashes could represent the manifesta¬tion of latent or persistent viral infections interacting with activated macrophages”

 

 


Nagalase

Nagalase the basics

Nagalase occurs naturally in the intestines where it breaks down glyocproteins in our food.

Nagalase is also produced in quantities by cancers, viruses & bacteria to break down a specific glycoprotein in our blood - Vitamin D Binding (Glyco)Protein.  Nagalase breaks VDBP at a triage of sugars that are required to activate our immune cells, thus elevated levels of Nagalase in the blood can lead to immune-suppression then cancers and pathogens grow unchecked. 

Because it is an enzyme—a catalyst—Nagalase performs this malicious ritual over and over and over again, and each time it comes away unscathed and unchanged. One Nagalase molecule can thus destroy a huge quantity of Vitamin D Binding Protein molecules. 

Nagalase has no natural enemies. No bodily process, no drug, no treatment could outsmart this diabolical killer. Until Dr. Nobuto Yamamoto researched and published Nagalase, we had no idea as to the actual cause of the immune shutdown that allows cancers and viruses grow unchecked.

Nagalase is a very sensitive marker and can indicate cancer cells before they can be seen on any current scans, tests, or biopsies.  Thus it is not recognised in mainstream medicine as a marker, as they could only tell you that you have the beginnings of a cancer, but they could not give any medication for this.  At this very early stage, a change in lifestyle and the addition of MAFActive may be all it takes to reverse the symptoms

Using Nagalase to screen for cancer & viruses

In the (hopefully not-too-distant) future, once Nagalase testing and GcMAF are available, all people at risk of cancer—i.e., everyone over the age of 40—will get an annual Nagalase test along with their other routine blood tests (complete blood count, comprehensive metabolic panel, vitamin D, lipid panel, etc.).

Nagalase could become our standard marker for early cancer detection. Those with an elevated Nagalase (after ruling out a viral cause) will be treated “presumptively” (i.e., we know it’s in there even though we can’t actually see it) with GcMAF and/or other alternative cancer therapies.

Follow up Nagalase testing will document the patient’s progress. Once the cancer is gone, as documented by a return of Nagalase to baseline, subsequent testing will provide an early warning if cancer starts growing again.

Testing for Nagalase

The test measures the activity of  alfa-N-acetylgalactosaminidase (nagalase) in blood.  Nagalase is an extracellular matrix-degrading enzyme that is (increased) secreted by cancerous cells in the process of tumour invasion. It also is an intrisic component of envelope protein of various virions, such as  HIV and  the influenza virus. It is secreted from virus-infected cells..1,3,4

Nagalase deglycosylates the vitamin D3-binding protein DBP (in humans better known as Gc-protein). Gc-protein is the precursor for the major macrophage-activating factor (MAF). Gc-protein carries one trisaccharide consisting of N-acetylgalactosamine with dibranched galactose and sialic acid termini. By the deglycosylation the  (complete) trisaccharide is removed from the Gc-protein. The glycosylated Gc-protein cannot be converted to MAF anymore.

Normally MAF is produced from Gc-protein by sequential removal of the galactose and sialic acid termini from this protein by respectively beta-galactosidase of inflammation- primed B lymphocytes and sialidase of T lymphocytes, with N-acetylgalactosamine as the remaining sugar. Macrophage-activation for phagocytosis and antigen presentation is the first step in the immune development cascade. Lost precursor  activity leads to immunosuppression.

Increased activity of nagalase has been detected in the blood of patients with  a wide variety of cancers, like cancer of the prostate, breast, colon, lung, oesophagus, stomach, liver, pancreas, kidney, bladder, testis, uterus and ovary, mesothelioma, melanoma, fibrosarcoma, glioblastoma, neuroblastoma and various leukeamias.1,3,4  For various types of tumors various levels of nagalase activity were found.7 It seems likely that secretory capacity of individual tumor tissue varies among tumor types depending upon tumor size, staging, and the degree of malignancy or invasiveness.7  Increased nagalase activity has not been detected in the blood of healthy humans.1

It has been established that the nagalase activity is directly proportional to viable tumour burden..1,2  Studies correlating nagalse levels with tumour burden suggest that the measurement of this enzyme can diagnose the presence of cancerous lesions below levels detectable by other diagnostic means.1 In research a day after surgical removal of primary tumours from cancer patients nagalase activity suddenly decreased to near the tumour-free control level, suggesting that the half-life value of nagalse is less than 24 hous.1,6 The short half-life of nagalase is valuable for prognosis of the disease during various therapies.1,5

Read more: Testing for Nagalase

Using Nagalase to track effectiveness of cancer & viral therapy

The more cancer cells present in the body, the more Nagalase they generate. Thus, Nagalase is the perfect marker for determining response to cancer therapies because, in a given patient, tumour burden will always be proportional to the Nagalase level.

In cancer patients, a declining Nagalase level reflects a reduction in total tumour “load” or “burden.” We would expect a lower Nagalase after chemo, radiation, or surgery because these treatments all reduce tumour burden. Having used these therapies does not mean the disease is gone, however. A very low (baseline) Nagalase level would indicate the cancer has been cured.

If there is even a remote chance of metastatic disease—as indicated by an elevated Nagalase level—it would be wise to use GcMAF to activate the immune system so it can find and polish off any remaining cancer cells. Repeat testing with declining levels would indicate that the treatment is working.

Pathogenic significance of Nagalase activity found in the hemagglutinin of influenza virus

Serum vitamin D3-binding protein (Gc protein) is the precursor for the principal macrophage activating factor (MAF). The precursor activity of serum Gc protein was reduced in all influenza virus-infected patients.

These patient sera contained α-N-acetylgalactosaminidase (Nagalase) that deglycosylates Gc protein. Deglycosylated Gc protein cannot be converted to MAF, thus it loses the MAF precursor activity, leading to immunosuppression. An influenza virus stock contained a large amount of Nagalase activity. A sucrose gradient centrifugation analysis of the virus stock showed that the profile of Nagalase activity corresponds to that of hemagglutinating activity.

When these gradient fractions were treated with 0.01% trypsin for 30 min, the Nagalase activity of each fraction increased significantly, suggesting that the Nagalase activity resides on an outer envelope protein of the influenza virion and is enhanced by the proteolytic process. After disruption of influenza virions with sodium deoxycholate, fractionation of the envelope proteins with mannose-specific lectin affinity column along with electrophoretic analysis of the Nagalase peak fraction revealed that Nagalase is the intrinsic component of the hemagglutinin (HA).

Cloned HA protein exhibited Nagalase activity only if treated with trypsin. Since both fusion capacity and Nagalase activity of HA protein are expressed by proteolytic cleavage, Nagalase activity appears to be an enzymatic basis for the fusion process. Thus, Nagalase plays dual roles in regulating both infectivity and immunosuppression.

http://www.sciencedirect.com/science/article/pii/S1286457905000560

Nagalase is remarkably efficient

 

Because it is an enzyme—a catalyst—Nagalase performs this malicious ritual over and over and over again, and each time it comes away unscathed and unchanged. One Nagalase molecule can thus destroy a huge quantity of Vitamin D Binding Protein molecules.

Nagalase has no natural enemies. No bodily process, no drug, no treatment could outsmart this diabolical killer. Until Dr. Nobuto Yamamoto researched and published Nagalase, we had no idea as to the actual cause of the immune shutdown that allows cancers and viruses grow unchecked.

 

 

Why is Nagalase important?

 

  1. Nagalase causes immunodeficiency. Nagalase blocks production of MAF, thus preventing the immune system from doing its job. Without an active immune system, cancer and viral infections can grow unchecked.
  2. As an extremely sensitive marker for all cancers, Nagalase provides a powerful system for early detection.
  3. Serial Nagalase testing provides a reliable and accurate method for tracking the results of any therapeutic regimen for cancer, AIDS, or other chronic viral infection.

Virus particles also make Nagalase. Their goal is the same as that of the cancer cells: survival by incapacitating their number one enemy: the immune system.

Nagalase’s behaviour toward us has been reprehensible and disastrous. Working in cahoots with cancer and HIV—not shy about getting into bed with our mortal enemies—Nagalase can rightfully claim direct responsibility for billions of human deaths. And it would just as soon add you to the list.

Nagalase is a “genocidal murderer” and yet so few people have heard of it and even fewer laboratories (6 at the last count) offer a test to measure the levels of Nagalase in the blood.

Rising Nagalase levels indicate a cancer or virus is growing and spreading. Conversely, Nagalase levels will decrease if the cancer or infection is being effectively destroyed.

Any treatment that lowers cancer cell (or viral numbers) will lower Nagalase levels.
Nagalase will, for example, always drop after surgery (whether or not the entire tumour was removed). Chemotherapy and radiation also reduce Nagalase levels. So does MAF.

If, after these treatments, the depressed level begins to rise again, this is the warning sign that the cancer was not completely removed, and/or that metastatic disease is hiding out somewhere. With viral infections, increasing Nagalase levels indicate return of the infection.

Consecutive rising Nagalase levels are therefore a red flag, warning us it may be time to entertain new treatment options. Conversely, if levels are going down, stay the course: the cancer or virus is going away.

Nagalase – A Near-Universal Cancer Marker?

Even though further clarification is required regarding the structure and generation of GcMAF, Yamamoto’s body of work has given rise to two key propositions that may have extraordinary importance if sustained by future research: namely, that serum nagalase can serve as a virtually universal marker for cancer, and that parenteral administration of pre-formed GcMAF can exert a profound immunostimulant effect in cancer patients, enabling the effective control and eradication of many cancers when they are in a micrometastatic or very early nascent form.

In 1996, Yamamoto and colleagues published a study in which they assayed serum nagalase in 20 patients with a wide range of cancers, as well as in 5 healthy subjects.10 Whereas the serum nagalase activity in the healthy subjects averaged 0.23 (nmoles/mg/min), it ranged in the cancer patients from a low of 0.64 to a high of 5.21. In several patients who were receiving radiation therapy, the serum nagalase activity declined progressively, coming close to the normal range in some. Conversely, the ability of Gc protein derived from the patients to give rise to GcMAF activity in vitro (assessed by stimulation of superoxide production by macrophages) tended to be low in cancer patients, and vary inversely with serum nagalase; this precursor activity increased progressively during radiation therapy.

A year later, Yamamoto published a study confirming these findings in 36 patients with oral cancer; whereas serum nagalase averaged 0.29 in twelve healthy controls, this activity was above 1 in all except one of the cancer patients (0.74), and was above 6 in four of them.11 Once again, serum precursor activity for GcMAF generation tended to be subnormal and correlated inversely with serum nagalase in the cancer patients. Surgical excision of primary tumors or metastasized nodes was followed by a rapid decline in serum nagalase, coming close to the normal range in 3 patients.

Yamamoto’s assessment of serum nagalase in patients with micrometastatic breast, colorectal, and prostate cancers appears to provide further confirmation for the utility of nagalase as a cancer marker; these findings are discussed below.

Read more: Nagalase – A Near-Universal Cancer Marker?


Macrophages

Swollen lymph nodes - immune reaction

As your immune system develops so new immune cells are created in your lymph nodes this is why we feel tenderness and swelling in our glands.

The explosive increase in the number of lymphocytes, both B cells and T cells, from just a few to millions in the presence of an infection was discovered in the 1950s. The process, called clonal expansion, is what gives the adaptive immune system its extraordinary might and specificity. You can tell that clonal expansion is occurring when you feel tender bumps (swollen lymph nodes) in your neck or other areas.

When lymphocytes multiply during clonal expansion, some of them are destined to live on as memory T and B cells. These clones are a subset of the expanded number of T and B cells that develop from your first exposure to a germ, and they protect you against subsequent attacks by the same germ.

Read more: Swollen lymph nodes - immune reaction

Macrophages - Your Awesome Killing Machine

Macrophages are big and smart white blood cells that chase, capture, engulf, and digest intruders. They trap and phagocytize (literally, “eat”) their enemies. They can multiply rapidly when necessary. However, they’re naturally indolent and need to be activated by VDBP.

Opsonin “super glue” helps them stick to their prey. Their electron-driven free radical death ray (AKA “oxidative burst”) blasts holes in microbes and cancer cells. Once a microbe or cancer cell has been phagocytized by a macro, it is encapsulated inside a “phagolysosome” (the intracellular “death chamber”), where it is then killed (if it isn’t dead already), and then dissected down into its component parts, which are then recycled.

Here’s how it works. When it isn’t swimming in the blood stream, a macrophage can slowly “walk” through tissues using self-generated stumpy little (one micron) “legs” (about ten of them sprout at a time). The macrophage ambles over to and snuggles up alongside a “foreign invader” (e.g., cancer cell or virion), quickly identifies it as foe, sprays it with membrane-frying free radical-laden bursts, grabs, engulfs, smothers, kills, and digests it. If the enemy is further away, or trying to escape, the macrophage chases after it, extrudes a cluster of long thin sticky spaghetti-like tentacles that wrap around and ensnare the fugitive cell, clutching it in an unbreakable strangle hold.

In a process known as phagocytosis, the macrophage draws in its victim, engulfs and smothers it, then encases it in a small bubble-like cyst (called a phagolysosome) inside its cytoplasm. The phagolysosome then secretes a cocktail of corrosive free radicals and enzymes that rapidly digest its victim down into its component parts (amino acids, nucleic acids, fatty acids, etc.). The macrophage then spits out these pieces into the intercellular “soup.”  

Because the remnants of viruses and cancer cells are fundamental cellular building blocks, the body quickly recycles them using the “spare parts” to build brand new healthy cells.

Macrophages and their oxidative bursts

A powerful weapon possessed by a Macrophage weapon is the “oxidative burst” (also widely known as the “respiratory burst”). An enzyme (called NADPH oxidase) stationed in the Macrophage’s outer membrane sprays out a beam of highly reactive free electrons, like bullets from a machine gun.

The NADPH gun emits a particle beam that blast tumour cells and microbes to smithereens. The electrons in the beam emerge one at a time, but they really really don’t want to be “free,” so—as fast as they possibly can—they snatch another electron to form a stable pair (we are talking nanoseconds here). A chain reaction of electron-snatchings triggered by the oxidative burst literally vaporizes molecules in the outer wall of a cancer cell or viral capsid, ripping holes in it.

Now the membrane that held the victim together literally falls apart, spilling out its contents. Without an intact outer membrane, a cancer cell can’t survive for very long. Oxidative bursts don’t happen all of the time. That would be a waste of firepower. The “trigger” that turns it on is the perceived proximity of a “foe,” a cancer cell, HIV virus, hepatitis virus, or a bacterium. When a macro comes into immediate contact with “enemy,” then—and only then—does it turn on the electron death beam.

There are lots of oxygen (O2) molecules everywhere in our bodies. (We need plenty of oxygen and glucose, the “fuels” from which we generate the “energy” that drives all of the cellular chemical reactions that make life possible.) When released, most of the electrons in the death ray beam crash into one of these omnipresent oxygen molecules, from which they quickly grab the electron they need to make a stable pair. The oxygen molecule now is missing one of its electrons, and is thus transformed into the violently corrosive free radical known as “superoxide” (O2-). Now superoxide is the one wanting an electron, and it will destroy anything in its path to get one. That “anything” would be the virus, bacterium, or cancer cell our macro has grabbed with its pseudopod. Suddenly the invader finds itself with a huge hole in its outer membrane. It’ll die soon.

The free electrons and superoxides also trigger chain reactions forming other reactive free radical species. One of these is the hydroxyl ion (OH-). This is hydrogen peroxide, just like the stuff that comes out of that brown bottle, but 33 times as potent—a locally generated intercellular dose. Perfect for frying microbes and tumor cells.

By oxidizing omnipresent chlorine atoms, the electron beam also generates noxious hypochlorous acid (HClO), which can poke a hole in an enemy membrane in nothing flat. Now we have a toxic soup of free radical oxidizing agents that can do tremendous local damage to our enemies.

MAF Activated Macrophages and the “Oxidative Burst”
Only MAF activated macrophages are going to deliver oxidative bursts that are potent enough to be effective. If Nagalase from viruses or cancer cells has put the macrophages to sleep, the oxidative burst degenerates into a piddly potato gun that’s not going to hurt anybody. Firepower—or lack thereof—is what we are talking about here.  Activated macrophages fire the atomic equivalent of millions of rounds a second and never have to pause to reload.

Macrophage phagolysosome execution (and dismantling) chamber

If, somehow, a microbe or cancer cell has survived the oxidative burst and phagocytosis, it will not survive the death chamber. Once eaten, internalized, and embedded in the macrophage’s cytoplasm, the enemy is imprisoned in a round cyst-like bubble inside the macrophage (called a phagolysosome) into which are squirted all sorts of digestive enzymes and many more rounds of oxidative burst, just for good measure. Pretty things do not happen inside of phagolysosomes. If the cancer cell or microbe is not already dead, the phagolysosome “death chamber” will certainly polish it off. (“Phago” means “to eat.” “Lyso” means “to dissolve.” “Some” means “sack” or “bag.”)

Once the dismembering process is complete, the phagolysosome slides over and makes contact with the outer cell membrane, merges with it, then disgorges the now harmless breakdown products (nucleic acids, fatty acids, amino acids, etc.) out into the extracellular fluid. They are then taken up by nearby cells and recycled into new body parts.
The ecologically-minded among us should find the efficiency of this process commendable. Nothing is wasted. Scary toxic bad guys are killed, dismantled, and transformed into spare parts for the good guys: us.A sophisticated communication system

Macrophage exponential self-cloning: the ultimate weapon.


Last, but definitely not least, Macrophages—if outgunned—play the population card: they multiply rapidly. When they find themselves in an area of high cancer cell or viral particle density, they don’t have to call up the draft to get more troops; they simply clone themselves, which they can do on very short notice. More Macrophages automatically translates into more of all the other weapons enumerated above. But, again, this multiplication process occurs only in activated Macrophages.

Vitamin D Binding Protein Activation

Without VDBP, Macrophages languish. In the presence of Vitamin D Binding Protein, their activity level increases exponentially. Once activated, Macrophages multiply rapidly and attack ferociously.

Macrophages – Communicating with the immune system

Immune cells—macrophages and lymphocytes— carry on a constant blather, like a huge town hall chat room where everybody is talking at once. However, since the talking is a release of “messenger molecules” and the listening is done by protein receptors, immune cells can actually listen while they are talking!! No need to complain about being interrupted! It’s weird, and foreign to us humans, but this simultaneous talking and listening makes for a far faster exchange of messages than if you had to stop and listen every time the other guy was talking (like we humans usually do).

There is so much activity, what with the constant molecular chatter coupled with a madhouse of cellular scrambling to grab and kill enemy cells as rapidly as possible, that the casual observer might get the impression of chaos. But she would be sadly mistaken. There are no wasted efforts here. Like a Beethoven symphony, everything is extremely well-organized and perfectly coordinated.

The chemical chatter among macrophages and other immune cells is so rapid and efficient that it would make a sophisticated military communications system look like a bunch of kids with tin can phones. Macrophages release clouds of messenger molecules (cytokines, interferons, leukotrienes, and other small molecules)—at rates of up to thousands of molecules per second per cell. Each molecule carries a specific request or command. Like “Bring me this,” or “We need some of that over there,” or “Kill everything that looks like this.” “We need an inflammatory response over here.” Or “We don’t need to do that anymore.” They discuss what the enemy looks like and how aggressive he is. They tell each other how hard to work. They label targets for other cells to identify and kill. They talk about where the enemy is hiding. They discuss current enemy strategy and how best to outmaneuver it.

Feeding your immune cells

The immune system consists of a finely orchestrated, complex collection of tissues and cells that protect your body from allergens, bacteria, viruses, and other potentially harmful organisms, collectively known as antigens. Skin and the membranes that line entrances to the body -- nasal passages, eyes, and respiratory and digestive tracts -- are the first line of defense, providing a physical barrier against invaders. Internally, specialized white blood cells fight antigens that make it past the skin: T-lymphocytes continuously patrol the body in search of antigens; B-lymphocytes manufacture antibodies, special blood proteins that neutralize or destroy germs; and neutrophils and macrophages scavenge antigens from the blood for delivery to the lymphatic system, which disposes of them. To work smoothly, these cells depend on you feeding them....this is what your immune cells need.

selenium which helps white blood cells produce the proteins they need to clear out viruses. 

zinc (pumpkin seeds)which is important in the development of white blood cells. - See more at: http://www.sunwarrior.com/news/immune-boosting-foods/#sthash.5pdCQkzR.dpuf

vitamin A, a component of healthy skin. The skin is the first line of defense against infection. Vitamin A is also important to T-Cells and natural killer cells. - See more at: http://www.sunwarrior.com/news/immune-boosting-foods/#sthash.5pdCQkzR.dpuf

glutamine, an amino acid that is used by immune cells during times of stress, inflammation, and infection, especially by lymphocytes, macrophages, and neutrophils. - See more at: http://www.sunwarrior.com/news/immune-boosting-foods/#sthash.5pdCQkzR.dpuf

glutathione, an antioxidant that strengthens the immune system. - See more at: http://www.sunwarrior.com/news/immune-boosting-foods/#sthash.5pdCQkzR.dpuf

allicin, which fights bacterial infections and cancer - See more at: http://www.sunwarrior.com/news/immune-boosting-foods/#sthash.5pdCQkzR.dpuf

Lentinan, a type of beta glucan found in shiitake mushrooms, is believed to reduce tumor activity and lessen the side effects of cancer treatment.5 Researchers at Teikyo University’s Biotechnology Research Center in Kawasaki, Japan, showed that lentinan has anti-tumor properties, suppressing the formation and development of tumors.

beta glucan (PGG glucan) that enhances the function of macrophages and neutrophils. Researchers looked at beta glucan’s ability to work in partnership with antibiotics to decrease mortality of the rats. “Results of these studies demonstrated that prophylaxis with PGG glucan in combination with antibiotics provided enhanced protection against lethal challenge with Escherichia coli or Staphylococcus aureus as compared with the use of antibiotics alone,”

The Immune Response: Clonal Selection

The inflammatory response stimulates neutrophils and macrophages to migrate to a site of infection. This animation can be used to demonstrate to students the microorganism–macrophage interaction that leads to antibody synthesis and immune memory.

The macrophage phagocytizes the microorganism, killing it and breaking its macromolecules into fragments. These fragments, in conjunction with the major histocompatibility complex type II (MHC II), are displayed on the macrophage cell surface. This presentation triggers CD4+ T-helper lymphocytes to combine with the presented antigen and activate antibody synthesis by differentiated B-cells called plasma cells. Instead of differentiating into plasma cells, other B lymphocytes remain committed and ready for the next contact with this specific antigen.

Macrophages and T-helper cells release, and are activated by, secreted products called cytokines. Cytokines are small proteins that serve a variety of functions and include interleukins, interferons, colony stimulating factors, tumor necrosis factors, and growth factors. Some cytokines positively regulate the immune response by stimulating growth and maturation of target cells; other cytokines negatively regulate the immune response through a series of inhibition reactions.


Cancer & Macrophage Activation

The fundamentals of cancer

With all the scientific papers and scientific explanations, it is easy to get lost in a fog of information.  Despite the millions of pounds and dollars spent on ‘cancer research’ over the years, scientists are still no closer to giving a definitive reason as to what causes cancer and because they are looking so closely at the minute details, they seem to be missing the point about how to cure cancer.   There are many people who have suffered from, and survived, cancer.  Some of them have been through the medical route and endured painful chemo and radiation therapy.  Others have been told that their cancer is too difficult to treat or too far progressed to be able to be saved with conventional medicine, and were told to go home and put their affairs in order.  

Those who survive this have done so by listening to alternatives, thinking about and learning from their own body and trying remedies that are considered ‘quackery’.

The causes of cancer.

Cancer cells are not aliens.  They are your own cells that have mutated due to some environmental factor that has put them under enormous stress.  They have mutated to survive, because that is the main driving force behind every living thing, the instinct to live.  Cancer is a symptom of the disease or an environmental stress, and removing the disease is the key to preventing the spread of the cancer.  

Read more: The fundamentals of cancer

Macrophage activation as an Immunotherapeutic approach to cancers

 

Macrophage activation is an immunotherapeutic approach that reinforces the immune system and helps it to fight cancer; it also has a direct effect on cancer cells in vitro and therefore it can be proposed as an effective anticancer approach. 

Macrophage activation's best effects are observed when it is administered in early stages of cancer when there is not yet the need of conventional therapies. However, several Doctors in the world have observed significant effects also in advanced cancers and after other conventional treatments such as surgery, radio- or chemo-therapy.

Yamamoto’s three cancer studies showed that incredibly small weekly doses (100 billionths of a gram—an amount that is invisible to the naked eye) of Macrophage activation had cured early metastatic breast, prostate, and colon cancers in 100% of (nonanemic) patients. In a fourth paper, he used the same treatment to cure 100% of nonanemic HIV-infected patients.

 For the three cancer studies, Dr. Yamamoto had chosen patients who had recently received the standard mainstream triad of surgery, chemo, and radiation. Despite these treatments, every patient had evidence of metastatic disease, which means that despite the best efforts of conventional medicine, their cancers were out of control and still growing. Their prognoses were poor at best. Nevertheless, this patient group had one thing in common: their tumor mass (also known as tumor burden) had been drastically reduced by the therapies they had received, and this in turn dramatically increased the likelihood that Macrophage activation would remove the remaining cancer cells.

We have not observed adverse effects in such a context and we feel safe to state that it can be considered in many cases as a valid complementary approach to other anticancer therapies.

Read more: Macrophage activation as an Immunotherapeutic approach to cancers


Cancer & MAF Research Papers

Inhibitory effect of vitamin D-binding protein-derived macrophage activating factor on DMBA-induced hamster cheek pouch carcinogenesis and its derived carcinoma cell line.

 

A study on MAF in hamsters infected with squamous cell cancer.

All 15 hamsters in the control group died within 20 weeks.The other group were infected and administered MAF, although the doses, source and intervals are not stated.2 of the 14 hamsters in this group did not develop any tumours.

The other 12 showed a significant delay in developing tumours and none died within the 20 weeks that they were observed.MAF was administered to the 5 hamsters in the control group at week 13, and tumour growth was slightly suppressed, with all dying from tumour burden.

However, the mean survival time was significantly extended.

Oncol Lett. 2011 Jul;2(4):685-691. Epub 2011 May 13. PubMed PMID: 22848250; PubMed Central PMCID: PMC3406437.

Vitamin D-binding protein-derived MAF, and prostate cancer.

 

Summary: MAF was lost or reduced in prostate cancer patients due to high levels of nagalase.Macrophages activated by MAF develop a considerable variation of receptors that recognize malignant cell surfaces.16 nonanemic prostate cancer patients received weekly administration of 100 nanograms of MAF , during which time their nagalase levels reduced.After 14 to 25 weeks all 16 patients had low nagalase levels equivalent to those of healthy people, indicating the patients were cancer free.No recurrence occurred for 7 years.

Cancer Immunol Immunother. 2012 Dec;61(12):2377-8. doi: 10.1007/s00262-012-1310-9. Epub 2012 Jun 28. PubMed PMID: 22740161.

Effects of vitamin D-binding protein-derived macrophage-activating factor on human breast cancer cells.

 

Searching for additional therapeutic tools to fight breast cancer, we investigated the effects of vitamin D-binding protein-derived macrophage activating factor (DBP-MAF, also known as GcMAF) on a human breast cancer cell line (MCF-7).

MATERIALS AND METHODS:

The effects of DBP-MAF on proliferation, morphology, vimentin expression and angiogenesis were studied by cell proliferation assay, phase-contrast microscopy, immunohistochemistry and western blotting, and chorioallantoic membrane (CAM) assay.

RESULTS:

DBP-MAF inhibited human breast cancer cell proliferation and cancer cell-stimulated angiogenesis. MCF-7 cells treated with DBP-MAF predominantly grew in monolayer and appeared to be well adherent to each other and to the well surface. Exposure to DBP-MAF significantly reduced vimentin expression, indicating a reversal of the epithelial/mesenchymal transition, a hallmark of human breast cancer progression.

CONCLUSION:

These results are consistent with the hypothesis that the known anticancer efficacy of DBP-MAF can be ascribed to different biological properties of the molecule that include inhibition of tumour-induced angiogenesis and direct inhibition of cancer cell proliferation, migration and metastatic potential.

Anticancer Res. 2012 Jan;32(1):45-52. PubMed PMID: 22213287.

MAF is associated with the formation of chemical messenger pathways

 

The effects of MAF have been studied in cancer and other conditions where blood supply conditions are deregulated.This study demonstrates that the division of human blood mononuclear cells (immune system cells) when treated with MAF was associated with the formation of chemical messenger pathways.The effect was dose dependant, but maximum stimulation was achieved at 0.1ng/ml.Heparin inhibited the formation of chemical messenger pathways.In addition, MAF at that dose inhibited the formation of prostate and breast cancer cell blood pathways in laboratory tests.They tested different MAF preparations in this type of test (CAM test) and this test proved to be a reliable, reproducible and inexpensive method of determining the potencies of different preparations and their stability. 

Cancer Immunol Immunother. 2011 Apr;60(4):479-85. doi: 10.1007/s00262-010-0953-7. Epub 2010 Dec 14. PubMed PMID: 21170647.

Vitamin D binding protein-macrophage activating factor directly inhibits proliferation, migration, and uPAR expression of prostate cancer cells

 

MAF shows a direct and potent effect on prostate tumour cells in the absence of macrophages.  It demonstrates a reduction in proliferation as well as metastic clones of these cells.Studies show that this is not due to the death of the cancer cells, but due to the reduction of multiplication.  MAF also prevents the migration of prostate cancer cells in the lab.It stopped the prostate cancer cells from producing a receptor, and evidence shows that this receptor correlates with metastasis.It concludes that MAF has strong inhibitory activity even in the absence of macrophages.

Gregory KJ, Zhao B, Bielenberg DR, Dridi S, Wu J, Jiang W, Huang B, Pirie-Shepherd S, Fannon M

 

Glycosylation status of vitamin D binding protein in cancer patients

 

Based on earlier studies, it appears that vitamin D binding protein (DBP) is significantly or completely deglycosylated in cancer patients, thus destroying the molecular precursor of the immunologically important MAF.  This investigation was to directly investigate the volume of the three sugar molecules of serum derived DBP in various cancers. This study showed that there was no significant depletion of the trisaccharide in the 56 patients relative to healthy controls.

This suggests that the hypotheses regarding the structural or molecular origins of GcMAF being a trisaccharide should be reconsidered.

PubMed PMID: 19642159

Immunotherapy for Prostate Cancer with Gc Protein-Derived Macrophage-Activating Factor

 

Serum Gc protein (known as vitamin D(3)-binding protein) is the precursor for the principal macrophage-activating factor (MAF). The MAF precursor activity of serum Gc protein of prostate cancer patients was lost or reduced because Gc protein was deglycosylated by serum alpha-N-acetylgalactosaminidase (Nagalase) secreted from cancerous cells.

Therefore, macrophages of prostate cancer patients having deglycosylated Gc protein cannot be activated, leading to immunosuppression. Stepwise treatment of purified Gc protein with immobilized beta-galactosidase and sialidase generated the most potent MAF (termed GcMAF) ever discovered, which produces no adverse effect in humans. Macrophages activated by GcMAF develop a considerable variation of receptors that recognize the abnormality in malignant cell surface and are highly tumoricidal. Sixteen nonanemic prostate cancer patients received weekly administration of 100 ng of GcMAF.

As the MAF precursor activity increased, their serum Nagalase activity decreased. Because serum Nagalase activity is proportional to tumor burden, the entire time course analysis for GcMAF therapy was monitored by measuring the serum Nagalase activity. After 14 to 25 weekly administrations of GcMAF (100 ng/week), all 16 patients had very low serum Nagalase levels equivalent to those of healthy control values, indicating that these patients are tumor-free. No recurrence occurred for 7 years.

http://www.ncbi.nlm.nih.gov/pubmed/18633461

Immunotherapy of metastatic colorectal cancer with vitamin D-binding protein-derived macrophage-activating factor

 

Serum vitamin D binding protein (Gc protein) is the precursor for the principal macrophage-activating factor (MAF). The MAF precursor activity of serum Gc protein of colorectal cancer patients was lost or reduced because Gc protein is deglycosylated by serum alpha-N-acetylgalactosaminidase (Nagalase) secreted from cancerous cells.

Deglycosylated Gc protein cannot be converted to MAF, leading to immunosuppression. Stepwise treatment of purified Gc protein with immobilized beta-galactosidase and sialidase generated the most potent macrophage-activating factor (GcMAF) ever discovered, but it produces no side effect in humans. Macrophages treated with GcMAF (100 microg/ml) develop an enormous variation of receptors and are highly tumoricidal to a variety of cancers indiscriminately. Administration of 100 nanogram (ng)/ human maximally activates systemic macrophages that can kill cancerous cells.

Since the half-life of the activated macrophages is approximately 6 days, 100 ng GcMAF was administered weekly to eight nonanemic colorectal cancer patients who had previously received tumor-resection but still carried significant amounts of metastatic tumor cells. As GcMAF therapy progressed, the MAF precursor activities of all patients increased and conversely their serum Nagalase activities decreased. Since serum Nagalase is proportional to tumor burden, serum Nagalase activity was used as a prognostic index for time course analysis of GcMAF therapy.

After 32-50 weekly administrations of 100 ng GcMAF, all colorectal cancer patients exhibited healthy control levels of the serum Nagalase activity, indicating eradication of metastatic tumor cells. During 7 years after the completion of GcMAF therapy, their serum Nagalase activity did not increase, indicating no recurrence of cancer, which was also supported by the annual CT scans of these patients.

http://www.ncbi.nlm.nih.gov/pubmed/18058096

Immunotherapy of metastatic breast cancer patients with vitamin D-binding protein-derived macrophage activating factor

Serum vitamin D3-binding protein (Gc protein) is the precursor for the principal macrophage activating factor (MAF). The MAF precursor activity of serum Gc protein of breast cancer patients was lost or reduced because Gc protein was deglycosylated by serum alpha-N-acetylgalactosaminidase (Nagalase) secreted from cancerous cells. Patient serum Nagalase activity is proportional to tumor burden. The deglycosylated Gc protein cannot be converted to MAF, resulting in no macrophage activation and immunosuppression.

Stepwise incubation of purified Gc protein with immobilized beta-galactosidase and sialidase generated probably the most potent macrophage activating factor (termed GcMAF) ever discovered, which produces no adverse effect in humans. Macrophages treated in vitro with GcMAF (100 pg/ml) are highly tumoricidal to mammary adenocarcinomas. Efficacy of GcMAF for treatment of metastatic breast cancer was investigated with 16 nonanemic patients who received weekly administration of GcMAF (100 ng). As GcMAF therapy progresses, the MAF precursor activity of patient Gc protein increased with a concomitant decrease in serum Nagalase. Because of proportionality of serum Nagalase activity to tumor burden, the time course progress of GcMAF therapy was assessed by serum Nagalase activity as a prognostic index. These patients had the initial Nagalase activities ranging from 2.32 to 6.28 nmole/min/mg protein.

After about 16-22 administrations (approximately 3.5-5 months) of GcMAF, these patients had insignificantly low serum enzyme levels equivalent to healthy control enzyme levels, ranging from 0.38 to 0.63 nmole/min/mg protein, indicating eradication of the tumors. This therapeutic procedure resulted in no recurrence for more than 4 years.

http://www.ncbi.nlm.nih.gov/pubmed/17935130

Pancreatic carcinogenesis: apoptosis and angiogenesis.

Summary:

Cell death and the creation of blood supplies to the cells (angiogenesis) are processes that are changed with cancers.Several reports show that a tumor suppressor gene that is in pancreatic cancer and related to malignancy can induce cell death and reduce the blood supply.This study has discovered two new angiogenesis inhibitors- one of them being GcMAF, the other aaAT-III.These molecules were able to regress tumours in immunodeficient mice, and show potent inhibition of circulatory cell proliferation.The angiogenesis inhibitors induced tumour dormancy in the animal model

Abstract

Apoptosis and angiogenesis are critical biologic processes that are altered during carcinogenesis. Both apoptosis and angiogenesis may play an important role in pancreatic carcinogenesis. Despite numerous advances in the diagnosis and treatment of pancreatic cancer, its prognosis remains dismal and a new therapeutic approach is much needed. Recent research has revealed that apoptosis and angiogenesis are closely interrelated. Several reports show that a tumor suppresser gene that is expressed in pancreatic carcinoma and related to malignant potential can induce apoptosis and also inhibit angiogenesis. At present, it is generally accepted that tumor growth in cancers, including pancreatic cancer, depends on angiogenesis. We have identified 2 new angiogenesis inhibitors from a conditioned medium of human pancreatic carcinoma cell line (BxPC-3): antiangiogenic antithrombin III (aaAT-III) and vitamin D binding protein-macrophage activating factor (DBP-maf). These molecules were able to regress tumors in severe combined immunodeficiency disease (SCID) mice, demonstrating potent inhibition of endothelial cell proliferation. Moreover, the angiogenesis inhibitors induced tumor dormancy in the animal model. These results suggest that antiangiogenic therapy using angiogenesis inhibitors may become a new strategy for treatment of pancreatic cancer in the near future.

http://www.ncbi.nlm.nih.gov/pubmed/15084979

Effect of salivary gland adenocarcinoma cell-derived alpha-N-acetylgalactosaminidase on the bioactivity of macrophage activating factor

Summary:

To study the effects of nagalase on the bioactivity of GcMAF.A cancer cell line with high levels of nagalase, produced by the human salivary gland (HSG), was studied.GcMAF prepared enzymatically was able to activate macrophages.However, GcMAF treated with nagalase did not do this.Thus, salivary gland cancer in this study was able to produce large quantities of nagalase, which inactivates GcMAF produced from GcProtein, resulting in reduced phagocytic activity.This study suggests that HSG nagalase acts as an immunodeficiency factor in cancer patients.

Abstract

The aim of this study was to clarify the effects of alpha-N-acetylgalactosaminidase (alpha-NaGalase) produced by human salivary gland adenocarcinoma (SGA) cells on the bioactivity of macrophage-activating factor (GcMAF). High exo-alpha-NaGalase activity was detected in the SGA cell line HSG. HSG alpha-NaGalase had both exo- and endo-enzyme activities, cleaving the Gal-GalNAc and GalNAc residues linked to Thr/Ser but not releasing the [NeuAc2-6]GalNac residue. Furthermore, GcMAF enzymatically prepared from the Gc protein enhanced the superoxide-generation capacity and phagocytic activity of monocytes/macrophages. However, GcMAF treated with purified alpha-NaGalase did not exhibit these effects. Thus, HSG possesses the capacity to produce larger quantities of alpha-NaGalase, which inactivates GcMAF produced from Gc protein, resulting in reduced phagocytic activity and superoxide-generation capacity of monocytes/macrophages. The present data strongly suggest that HSG alpha-NaGalase acts as an immunodeficiency factor in cancer patients.

 

Vitamin D binding protein-macrophage activating factor (DBP-maf) inhibits angiogenesis and tumor growth in mice

Summary

Daily administration of GcMAF is able to slow down solid tumour growth (4ng/kg/4 days).At higher doses, GcMAF causes tumour regression (4ng/kg/day).The injections were into the body cavity (similar to IV in humans).The data suggests that GcMAF prevents the growth of blood supplies to tumours, and stimulates the macrophages to attack both the outside and the tumour cell compartment of a cancer.

Abstract

We have isolated a selectively deglycosylated form of vitamin D binding protein (DBP-maf) generated from systemically available DBP by a human pancreatic cancer cell line. DBP-maf is antiproliferative for endothelial cells and antiangiogenic in the chorioallantoic membrane assay. DBP-maf administered daily was able to potently inhibit the growth of human pancreatic cancer in immune compromised mice (T/C=0.09). At higher doses, DBP-maf caused tumor regression. Histological examination revealed that treated tumors had a higher number of infiltrating macrophages as well as reduced microvessel density, and increased levels of apoptosis relative to untreated tumors. Taken together, these data suggest that DBP-maf is an antiangiogenic molecule that can act directly on endothelium as well as stimulate macrophages to attack both the endothelial and tumor cell compartment of a growing malignancy.

PMID: 12659668 [PubMed - indexed for MEDLINE] PMCID: PMC150212

Effects of vitamin D(3)-binding protein-derived macrophage activating factor (GcMAF) on angiogenesis

Summary

As with other results, GcMAF inhibits cancer cell proliferation, independent of tissue origin, so it is not cancer specific.The ability of GcMAF to prevent blood supply creation to cancers may be regulated by the CD36 receptor.

BACKGROUND:

The vitamin D(3)-binding protein (Gc protein)-derived macrophage activating factor (GcMAF) activates tumoricidal macrophages against a variety of cancers indiscriminately. We investigated whether GcMAF also acts as an antiangiogenic factor on endothelial cells.

METHODS:

The effects of GcMAF on angiogenic growth factor-induced cell proliferation, chemotaxis, and tube formation were examined in vitro by using cultured endothelial cells (murine IBE cells, porcine PAE cells, and human umbilical vein endothelial cells [HUVECs]) and in vivo by using a mouse cornea micropocket assay. Blocking monoclonal antibodies to CD36, a receptor for the antiangiogenic factor thrombospondin-1, which is also a possible receptor for GcMAF, were used to investigate the mechanism of GcMAF action.

RESULTS:

GcMAF inhibited the endothelial cell proliferation, chemotaxis, and tube formation that were all stimulated by fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor-A, or angiopoietin 2. FGF-2-induced neovascularization in murine cornea was also inhibited by GcMAF. Monoclonal antibodies against murine and human CD36 receptor blocked the antiangiogenic action of GcMAF on the angiogenic factor stimulation of endothelial cell chemotaxis.

CONCLUSIONS:

In addition to its ability to activate tumoricidal macrophages, GcMAF has direct antiangiogenic effects on endothelial cells independent of tissue origin. The antiangiogenic effects of GcMAF may be mediated through the CD36 receptor.

Anti tumor effect of vitamin D-binding protein-derived macrophage activating factor on Ehrlich ascites tumor-bearing mice.

Summary

Administration of GcMAF to cancer bearing mice, to ascertain doses and intervals, using survival time and nagalase levels to record the effects.

The control group survived around 13 days.

1 administration of 100pg (picogram)/mouse was given to 8 mice at the same time as tumour transplantation, 7 survived around 21 days, 1 more than 60.

2 administrations of GcMAF, at day 0 and at day 4 after transplantation, 6 survived about 31 days, 2 more than 60 days.

3 administrations of GcMAF in 4 day intervals after transplantation, 6 of the eight survived more than 60 days, with a nagalase level as low as healthy control group during the survival period.No mention of the survival time of the other 2 mice.

Conclusion is that the administration of small amounts of GcMAF caused protracted macrophage activation.

http://www.ncbi.nlm.nih.gov/pubmed/9893164

Immunotherapy of BALB/c mice bearing Ehrlich ascites tumor with vitamin D-binding protein-derived macrophage activating factor

Summary

Administration of GcMAF and cdMAF to cancer bearing mice, to ascertain doses and intervals, using survival time and nagalase levels to record the effects.The readings of nagalase increased as the tumours grew.

The control group survived around 16 days.

1. administration ofGcMAF 100pg/mouse gave a survival time of 35 days

2. administrations of GcMAF, at day 0 and at day 4 after transplantation, survival increased to more than 50 days.

3. Mice that received GcMAF at days 4 and 8 after transplantation survived up to 32 days.

4.  administrations of GcMAF or cdMAF in 4 day intervals after transplantation, showed an extended survival of 90 days, with a nagalase level as low as healthy control group during the survival period.

This seems to be a poorly reported study, as it gives no indication of the performances of the different MAFs in relation to each other.

http://www.ncbi.nlm.nih.gov/pubmed/9187119

Prognostic utility of serum alpha-N-acetylgalactosaminidase and immunosuppression resulted from deglycosylation of serum Gc protein in oral cancer patients.

Summary

46 oral cancer patients were tested for Gcprotein.22% had greatly reduced gcprotein, 61% was moderately reduced, 17% were equivalent to healthy controls.Patients with low gcprotein had high nagalase.Those with high Gcprotein had low nagalase.Thus there is an inverse relationship.Surgical removal of tumours results in a subtle decrease in nagalase.Tests in mice show that nagalase is directly proportional to tumour burden.Hence it could be used as a diagnostic

Abstract

Vitamin D3-binding protein (Gc protein), a serum glycoprotein, is the precursor for the macrophage activating factor.

Cancer patient sera contain alpha-N-acetylgalactosaminidase that deglycosylates Gc protein. Deglycosylated Gc protein cannot be converted to macrophage activating factor, leading to immunosuppression.

Of 46 oral cancer patients with squamous cell carcinoma, approximately 22% had greatly reduced precursor activities. The precursor activity of approximately 61% of these patients was moderately reduced. The remaining patients (17%) had precursor activities equivalent to those of healthy humans. Patients with low precursor activity of serum Gc protein had high serum alpha-N-acetylgalactosaminidase activity.

In contrast, patients with high precursor activity had low serum alpha-N-acetylgalactosaminidase activity.

Thus, levels of serum alpha-N-acetylgalactosaminidase of individual patients have an inverse correlation with precursor activities of their serum Gc protein. Surgical removal of tumors resulted in a subtle decrease in serum alpha-N-acetylgalactosaminidase activity with concomitant increase in the precursor activity of serum Gc protein. Serum enzyme analysis of nude mice transplanted with a human oral squamous carcinoma cell line revealed that serum alpha-N-acetylgalactosaminidase activity is directly proportional to tumor burden. Thus, alpha-N-acetylgalactosaminidase activity in patient bloodstream can serve as a diagnostic/prognostic index.

http://www.ncbi.nlm.nih.gov/pubmed/9000571

Macrophage-directed immunotherapy as adjuvant to photodynamic therapy of cancer.

Summary

DBPMAF was made using mice serum (as opposed to GcMAF from human serum), as can mice make antibodies to human GcMAF. The photodynamic therapy used in conjunction with GcMAF in mice squamous cell carcinomas resulted in a 100% recovery rate.

Abstract

The effect of Photofrin-based photodynamic therapy (PDT) and adjuvant treatment with serum vitamin D3-binding protein-derived macrophage-activating factor (DBPMAF) was examined using a mouse SCCVII tumour model (squamous cell carcinoma). The results show that DBPMAF can markedly enhance the curative effect of PDT.

The most effective DBPMAF therapy consisted of a combination of intraperitoneal and peritumoral injections (50 and 0.5 ng kg-1 respectively) administered on days 0, 4, 8 and 12 after PDT. Used with a PDT treatment curative to 25% of the treated tumours, this DBPMAF regimen boosted the cures to 100%. The DBPMAF therapy alone showed no notable effect on the growth of SCCVII tumour. The PDT-induced immunosuppression, assessed by the evaluation of delayed-type contact hypersensitivity response in treated mice, was greatly reduced with the combined DBPMAF treatment.

These observations suggest that the activation of macrophages in PDT-treated mice by adjuvant immunotherapy has a synergistic effect on tumour cures. As PDT not only reduces tumour burden but also induces inflammation, it is proposed that recruitment of the activated macrophages to the inflamed tumour lesions is the major factor for the complete eradication of tumours.

http://www.ncbi.nlm.nih.gov/pubmed/9010027

Deglycosylation of serum vitamin D3-binding protein leads to immunosuppression in cancer patients.

Summary

Blood monocytes / macrophages of 52 cancer patients were incubated with 100pg/ml of GcMAF.The monocytes / macrophages were efficiently activated.Naturally occurring GcMAF was severely reduced in about 25% of the patients, and moderately reduced in 45%.This was found to be the result of higher than normal levels of nagalase.The source of the nagalase appeared to be the cancerous cells as radiation therapy to reduce the cancer also reduced the level of nagalase.Thus both the levels of GcMAF and the levels of nagalase in the blood can serve as diagnostic indices.

Abstract

Serum vitamin D3-binding protein (Gc protein) can be converted by beta-galactosidase of B cells and sialidase of T cells to a potent macrophage activating factor, a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor of the macrophage activating factor (MAF). Treatment of Gc protein with immobilized beta-galactosidase and sialidase generates an extremely high titered MAF, Gc-MAF. When peripheral blood monocytes/macrophages of 52 patients bearing various types of cancer were incubated with 100 pg/ml of GcMAF, the monocytes/macrophages of all patients were efficiently activated. However, the MAF precursor activity of patient plasma Gc protein was found to be severely reduced in about 25% of this patient population. About 45% of the patients had moderately reduced MAF precursor activities. Loss of the precursor activity was found to be due to deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase detected in the patient's bloodstream. The source of the enzyme appeared to be cancerous cells. Radiation therapy decreased plasma alpha-N-acetylgalactosaminidase activity with concomitant increase of precursor activity. This implies that radiation therapy decreases the number of cancerous cells capable of secreting alpha-N-acetylgalactosaminidase. Both alpha-N-acetylgalactosaminidase activity and MAF precursor activity of Gc protein in patient bloodstream can serve as diagnostic and prognostic indices.

http://www.ncbi.nlm.nih.gov/pubmed/?term=D3-binding+protein+leads+to+immunosuppression+in+cancer+patients

Tumor cell alpha-N-acetylgalactosaminidase activity and its involvement in GcMAF-related macrophage activation

Summary

Nagalase causes the suppression of GcMAF.This is a study of the types of nagalase from several tumor cell lines.It concludes that nagalase derived from tumours is different to that from normal Chang liver cells (a type of laboratory cell line).Tumour derived nagalase decreases the potency of GcMAF activation

Abstract

Alpha-N-acetyl galactosaminidase (alpha-NaGalase) has been reported to accumulate in serum of cancer patients and be responsible for deglycosylation of Gc protein, which is a precursor of GcMAF-mediated macrophage activation cascade, finally leading to immunosuppression in advanced cancer patients.

We studied the biochemical characterization of alpha-NaGalase from several human tumor cell lines. We also examined its effect on the potency of GcMAF to activate mouse peritoneal macrophage to produce superoxide in GcMAF-mediated macrophage activation cascade. The specific activity of alpha-NaGalases from human colon tumor cell line HCT116, human hepatoma cell line HepG2, and normal human liver cells (Chang liver cell line) were evaluated using two types of substrates; GalNAc-alpha-PNP (exo-type substrate) and Gal-beta-GalNAc-alpha-PNP (endo-type substrate).

Tumor-derived alpha-NaGalase having higher activity than normal alpha-NaGalase, had higher substrate specificity to the exo-type substrate than to the endo-type substrate, and still maintained its activity at pH 7. GcMAF enhance superoxide production in mouse macrophage, and pre-treatment of GcMAF with tumor cell lysate reduce the activity. We conclude that tumor-derived alpha-NaGalase is different in biochemical characterization compared to normal alpha-NaGalase from normal Chang liver cells. In addition, tumor cell-derived alpha-NaGalase decreases the potency of GcMAF on macrophage activation.

http://www.sciencedirect.com/science/article/pii/S1095643301005220