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The many effects of GcMAF on cancer.

Abstract: Multifaceted immunotherapeutic effects of vitamin D-binding protein-derived macrophage activating factor (GcMAF) on human breast cancer cells

Thyer L**, Gulisano M*  Published at the 5th Immunotherapeutics & Immunomonitoring Conference January 31st 2013 San Diego, CA, USA. And presented to the International PMTC Cancer Conference at the University of Sharjah UAE on the 1st February 2013.

Vitamin D-binding protein-derived macrophage activating factor (GcMAF) is a powerful stimulant of the immune system that has been proposed as an immunotherapeutic agent in the treatment a variety of conditions ranging from cancer to neurological disorders.

We previously demonstrated that GcMAF added to human breast cancer cells exerts multiple effects that concur to its antitumor potential; in fact, GcMAF inhibited cancer cell proliferation and metastatic potential, reverted the neoplastic phenotype and inhibited cancer cell-induced angiogenesis, a critical step in cancer progression.

In the present study, we describe for the first time the effects of GcMAF on the activation of macrophages that were added to a culture of the human breast cancer cell line MCF-7 (HPA Culture Collection). MCF-7 cells, incubated for 2 days, showed the anarchic morphology typical of carcinoma cells. They grouped in disordered cell masses extending above the cell monolayer that appeared as large lumps of cells protruding upwards.

Macrophages (cell line Raw 264.7, HPA Culture Collection) were activated by culturing them in the presence of 100 ng/ml GcMAF (Immuno Biotech Ltd) for 72 h prior to addition to the MCF-7 cell culture. The macrophages were added at a ratio of 1:1 to the MCF-7 cells. The cells were then allowed to settle for 1 h before time lapse photography. Photography was taken over a 60 h period using an Olympus CK2 microscope and a GXCAM-3 with NCH Debut capture software.


Results – Analysis of film

Un-stimulated macrophages appeared as round spherical cells that refract light. Once activated by GcMAF, macrophages sent out cytoplasmic protrusions that we interpreted as if the cells were trying to seek out and make contact with MCF-7 cancer cells. After about 60 h, it could be observed that the irregular growth of the breast carcinoma cells was arrested and the large protruding cell biomass was reduced. Carcinoma cells were phagocytised by the activated macrophages.

Taken together, these results support and reinforce the hypothesis that GcMAF is a molecule endowed with multiple biological activities relating to its antitumor properties. It inhibits human breast cancer cell proliferation and metastatic potential; it reverts the neoplastic phenotype; it inhibits cancer cell-induced angiogenesis; and it stimulates tumoricidal macrophages that phagocytise cancer cells. It is foreseeable that GcMAF will soon become part of an integrated immunotherapy approach to breast cancer.  (END)

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