• Phoenix Rising needs funds to operate: please consider donating to support PR

GcMAF for XMRV--Gc protein-derived macrophage activating factor--anyone taking it?

serg1942

Senior Member
Messages
421
Likes
772
Location
Spain
Hi moonwalker,

I have been reading and trying hard to understand Dr. Yamamoto's papers on HIV and cancer, and it's clear to me what he is stating in the above text:

This is an old paper, and he was discovering that both cancer and HIV infected patients had high Nagalase, and that this substance inhibited the activation of macrophages. Nagalase inhibits the formation of MAF (macrophage activation factor) by deglycosilating the Serum Gc protein (precursor of MAF).

So, by administering GcMAF, we are actually bypassing the inhibition of Gc protein, and therefore activating macrophagues, both in cancer and in HIV infected patients.

Hope this helps,
Sergio
 
Messages
20
Likes
0
Hi Sergio! Thank you so much for explaining it to me, but I knew that already :)
I'm a bit skeptic it's really all that simple in practice. I'd like to meet one person who cured their virus using gcmaf.
 

mojoey

Senior Member
Messages
1,213
Likes
74
Gc PROTEIN-DERIVED MACROPHAGE ACTIVATING FACTOR (GcMAF) STIMULATES
ACTIVATION AND PROLIFERATION OF HUMAN CIRCULATING MONOCYTES
M. Ruggiero*, S. Pacini**, N. Yamamoto
*Department of Experimental Pathology and Oncology, University of Firenze, Italy
**Department of Anatomy, Histology and Forensic Medicine, University of Firenze, Italy
Division of Molecular Immunology and Immunotherapy, Socrates Institute for Therapeutic Immunology, Philadelphia, PA, USA
THPE0051
Background. Vitamin D binding protein-macrophage activating factor (GcMAF) is a powerful stimulator of the immune system. Its effects were studied in conditions
where macrophage/monocyte function is deficient, from HIV infection to cancer (Transl Oncol 1:65-72, 2008; J Med Virol 81:16-26, 2009), whereas the effects on
monocytes from healthy subjects have not been studied. Thus, here we report the results obtained challenging human monocytes from healthy subjects with GcMAF:
we demonstrate that the individual degree of responsiveness is dependent on vitamin D receptor (VDR) gene polymorphisms. In addition, since the signal transduction
pathway of GcMAF is not fully understood, we studied the effects of GcMAF on the formation of intracellular cAMP. Finally, we studied the effects of GcMAF on normal
and cancer cell-induced angiogenesis in the chick embryo chorioallantoic membrane (CAM) assay.
Materials and Methods. GcMAF was prepared by Prof. N. Yamamoto. Peripheral blood monocytes were isolated from healthy subjects using Ficoll-Paque gradient
centrifugation. 100oeL of cells from donors harbouring different VDR polymorphisms (identified by BsmI and FokI restriction enzymes; see Adv Chronic Kidney Dis
15:186-90, 2008) were cultured with GcMAF for different time intervals (30 min - 96 h). Each condition was replicated in quadruplicate and each subject served as
internal control. Cell proliferation and viability were assessed by the cellular reduction of tetrazolium salts to colored formazans and measured as optical density
(Leuk Lymph 44:1957-62, 2003). cAMP was measured using the Cayman Chemical cAMP assay. CAM assay was performed as described in J Environ Pathol Toxicol
Oncol 28:85-8, 2009.
Results. Normal murine and human monocytes, incubated with 10 pg GcMAF/ml for 30 min and cultured for 3-6 h, were highly activated and produced 30-fold
increased superoxide generation (Table 1).
In fact, subjects harbouring homozygous “bb/FF” genotypes showed the highest response and 100 pg GcMAF/ml stimulated monocyte proliferation to an extent
comparable to that achieved by the highest concentration of lipopolysaccharide (1 μg/ml), taken as positive control. Heterozygous subjects (“Bb/Ff”) showed a smaller,
but still significant, response, whereas “BB/ff” homozygous did not respond.
In subjects harbouring “bb/FF” genotypes, GcMAF sustained cell viability for about 98 h
whereas un-stimulated cells were no longer viable after 48 h, as if, in those subjects, GcMAF had rescued monocytes from apoptosis
(Cell Death Dis 1, e30; doi:10.1038/cddis.2010.8, 2010).
In preliminary experiments with healthy donors, however, we had noticed that individual responses
to GcMAF significantly varied. Therefore, donors were selected for VDR genotypes and we
observed that the “b” and “F” alleles of the VDR gene were associated with the highest responses
in terms of cAMP formation and proliferation (Tables 2 and 3).
Chronic HIV infection is associated with deregulated angiogenesis possibly resulting in some of the pathological processes that occur in AIDS patients
(Angiogenesis 5:141-51, 2002). In the past, it was demonstrated that GcMAF inhibited growth factor-induced cell proliferation, chemotaxis, and tube formation in vitro
by using cultured endothelial cells and in vivo by using a mouse cornea micropocket assay (J Natl Cancer Inst 94:1311-19, 2002).
Its effects on CAM assay (i.e. on an entire developing embryo), however, had not been studied. Table 4, shows
that 1 ng GcMAF/ml (i.e. a concentration 10-fold higher than that required to stimulate monocytes) completely
inhibited the angiogenesis induced by prostaglandin E2 or by a human breast cancer cell line, MCF-7.
GcMAF alone did not modify basal angiogenesis or chick embryo viability and development.
Acknowledgements: this study was subsidized by grants from the University of Firenze to M.R. and S.P.
Author for correspondence: Prof. Marco Ruggiero, MD, PhD: marco.ruggiero@unifi.it
Monocytes activated with 10-100 pg GcMAF/ml developed a large amount of Fc-receptors as
well as significant variation of receptors that recognize IgG-bound and unbound HIV virions.
Thus, monocyes/macrophages activated by GcMAF preferentially phagocytize IgG-bound HIV
virions.
Thus, monocyes/macrophages activated by GcMAF preferentially phagocytize IgG-bound HIV
virions.
Figure 1
Discussion. These results elucidate the cellular and molecular mechanisms through which stimulation of the immune system leads to eradication of HIV
(J Med Virol 81:16-26, 2009; Figs 1, 2 and 3). In fact, we demonstrate that GcMAF has a potent mitogenic activity in vitro and these data are consistent with the
observation that intravenous administration of GcMAF increased the systemic cell counts of the activated macrophages to >200-fold. In addition, we demonstrate
for the first time that the response of human monocytes to GcMAF is dependent on VDR gene polymorphisms.
It is worth noting that the alleles “b” and “F” are also
associated with the highest sensitivity to vitamin D; a convergence of the vitamin D and GcMAF signalling pathways can thus be hypothesized. Thus, domain I of
GcMAF complexes with vitamin D and then binds to monocytes. However, gene-cloned domain III alone fully activates monocytes in a manner identical to entire
GcMAF. Whatever the case, these results can prove instrumental in identifying those HIV-positive subjects that could benefit the most from GcMAF treatment.
Figure 2 Figure 3
 

mojoey

Senior Member
Messages
1,213
Likes
74
Following the above, does anyone know what labs provide the VDR bsm/fok genotyping tests to predict our response to gcmaf?
 

garcia

Aristocrat Extraordinaire
Messages
975
Likes
247
Location
UK
Following the above, does anyone know what labs provide the VDR bsm/fok genotyping tests to predict our response to gcmaf?
Hi Joey,
I know Amy Yasko tests VDR fok as part of the genetic testing she offers (as well as VDR Taq). But I don't know where to get tested for VDR bsm. Any ideas?

Also do you know if mutations in other parts of VDR, e.g. VDR Taq (which I have) would have an influence on GcMAF?

many thanks,
garcia.
 

mojoey

Senior Member
Messages
1,213
Likes
74
I've done Yasko testing, but I'm not sure if the SNPs are applicable to the alleles tested above. if anyone can answer that, I think we'd all really appreciate it.
 

mojoey

Senior Member
Messages
1,213
Likes
74
I emailed Dr. Ruggiero to ask him about commercial labs. I wanted to ask Dr. Yamamoto again but he usually keeps me on the phone for a few hours so I'm gonna have to have a really good day for that.
 

garcia

Aristocrat Extraordinaire
Messages
975
Likes
247
Location
UK
I've done Yasko testing, but I'm not sure if the SNPs are applicable to the alleles tested above. if anyone can answer that, I think we'd all really appreciate it.
Yes the VDR Fok is the same.
 

mojoey

Senior Member
Messages
1,213
Likes
74
Just looked it up and yeah you're right. Yasko panel did include bsm/taq but don't think the cluster is relevant here (in fact, I'm not sure what SNPs for a gene cluster even means)

My Fok is +/- so that's Ff. Now just need to find a lab that tests for Bsm
 

aquariusgirl

Senior Member
Messages
1,649
Likes
616
yasko lab tests for 3 VDR receptors:
Fok
Taq
BSM/Taq

Can't remember if all 3 were in her original DNA test or if 1 or more were added later.

If they were part of the extended panel, you can buy the add-on as a separate test. Unknow the price.
 

garcia

Aristocrat Extraordinaire
Messages
975
Likes
247
Location
UK
yasko lab tests for 3 VDR receptors:
Fok
Taq
BSM/Taq

Can't remember if all 3 were in her original DNA test or if 1 or more were added later.

If they were part of the extended panel, you can buy the add-on as a separate test. Unknow the price.
I only got tested for Fok and Taq.
I am ++ for Taq and -- for Fok.

Aquariusgirl do you know why is the last one called "BSM/Taq" and not "BSM"? Also how did you find out this is now offered? Did you have one done recently? I had one done in 2008 & it was absent then. Also the sample test they give doesn't seem to mention it.

Many thanks,
garcia.
 

mojoey

Senior Member
Messages
1,213
Likes
74
Genova Diagnostics doesn't offer the Bsm test either. Argh.

Dr. Ruggiero said he could help me with this, but won't be able to until September because he's out of the country. I emailed Dr. Pacini as well.
 

aquariusgirl

Senior Member
Messages
1,649
Likes
616
garcia... holisticheal tells me bsm/taq tracks with taq.....ie they should be the same ...if you're hetero for one.. you're hetero for both etc.

teh girl in the office could not explain why this is.. she says they are separate SNPs...

haven't had time to google for an explanation
 

mojoey

Senior Member
Messages
1,213
Likes
74
I haven't found anything about bsm/taq cluster on google besides Yasko. We're gonna need explanations from non-Yasko sources for this. The last explanation you posted from Yasko's office just sounds very suspect....I haven't read anything about bsm and taq being identically expressed.
 

Jenny

Senior Member
Messages
1,386
Likes
555
Location
Dorset
I had VDR tests through Sciona Cellf - it was free as I was part of a study, so not sure how much it costs.

I don't fully understand the results though - they were:

VDR Taq1 - positive - they say this is also called VDR variation Fok 1 2590 T>C

VDR Bsml - negative - this is also called VDR Bsm1 58980 G>A, VDR T Bsml C

VDR Fokl - positive - this is also called VDR Taq1 60058 T>C, VDR T Taq1 C

Anyone know how these results relate to VDR bb, FF etc, or being homo or heterozygous?

Jenny
 

Overstressed

Senior Member
Messages
406
Likes
140
Location
Belgium
Hi Jenny,

I think you might be on the good site concerning GcMaf - I read it as bb/FF, but I'm not sure, maybe other members can clarify ?

OS.
 

mojoey

Senior Member
Messages
1,213
Likes
74
I'm also very interested what you find out Jenny. I've been looking for a relatively affordable way to get this testing done, and Ruggiero still hasn't gotten back to me.

thanks for posting!
joey