Gc Protein-derived Macrophage Activating Factor Research Articles

NEW CHALLENGE TO CANCER AND LONGEVITY

“Saisei Mirai” has been focusing on cancer treatments and has developed several new treatments, including GcMAF (Gc protein-derived Macrophage Activating Factor) and oral MAF (Macrophage Activating Factor) which are immunotherapies developed by “Saisei Mirai”. We are collaborating with 6 universities here in Japan to develop new and better treatments mainly for cancer. At the same time, GcMAF and oral MAF showed good effects for many infectious agents, autism spectrum disorder (ASD), chronic fatigue syndrome (CFS), atopic dermatitis, hair loss, multiple sclerosis, autoimmune diseases, injuries and so on. First, I'll explain our cancer treatments.

Some Measures for COVID-19 Including Deep Ultraviolet Light-Emitting Diode (DUV-LED), Gc protein-derived Macrophage-Activating Factor (Gcmaf), and 5-Aminolevulinic Acid (5-ALA)

Several trials for the COVID-19 pandemic are found. Prof. Shunji Nakamura, Tokushima University was awarded the 2014 Nobel prize in Physics for light-emitting diodes (LEDs). Deep ultraviolet LED (DUV-LED) causes SARS-CoV-2 inactivation as 99.9% by 265 nm. Gc protein-derived macrophage-activating factor (GcMAF)-based immunotherapy has a wider application for serious infection, chronic fatigue syndrome, and cancers. Covid center in Naples submitted a phase II report concerning GcMAF and COVID-19 in Feb 2021. 5-aminolevulinic acid (5-ALA) and its metabolite protoporphyrin IX (PpIX) show anti-inflammatory and antiviral effects for Zika, Dengue, influenza A viruses, and SARS-CoV-2. These results hopefully contribute to the research development for COVID-19.

Consumption of an Extremely Biodiverse Probiotic and a Supplement based on Microbial Chondroitin Sulfate is Associated with Very Low Serum Alpha-N-acetylgalactosaminidase (Nagalase) Activity and Decrease of C-reactive Protein Values

We describe the case of a 57-years-old male healthy professional who added to his nutritional plan a novel, extremely biodiverse, probiotic and a supplement based on microbial chondroitin sulfate, vitamin D3 and ultrapure phosphatidylcholine to improve his healthy life expectancy. After three months of such a nutritional regimen, serum alpha-N-acetylgalactosaminidase (nagalase) activity was 0.40 that is a value below the minimum level established for adults (normal values: 0.50-0.95 nMol/mL/min). C-reactive Protein (CRP) was less than 1 (normal values: 0.00-5.00 mg/L), a value significantly lower than that recorded before this experience. At the end of the observation period, serum albumin level was 45 g/L, a value very close to the maximum reference value (normal values: 32.00-46.00 g/L). The Prognostic Inflammatory Nutritional Index (PINI) score was 0.05, a value that is close to the minimum calculated value (normal values: 0.00-1.00) and suggests a very low risk of death for all causes. Observation in silico and experiments in vitro demonstrated that both the probiotic and the supplement formed complexes with human nagalase with an efficiency 100 fold higher than that of purified Gc protein-derived Macrophage Activating Factor (GcMAF). The collection of these observations suggests that the combination of this probiotic and this supplement may lead to overall beneficial effects on health as evidenced by positive changes of indicators of immune system function and healthy aging such as nagalase, CRP, albumin and PINI score.

The Effect of Fucoidan from the Brown Alga Fucus evanescence on the Activity of α-N-Acetylgalactosaminidase of Human Colon Carcinoma Cells.

α-N-acetylgalactosaminidase (EC 3.2.1.49) (alpha-NaGalase) catalyzes the hydrolysis of N-acetamido-2-deoxy-α-d-galactoside residues from non-reducing ends of various complex carbohydrates and glycoconjugates. It is known that human cancer cells express an alpha-NaGalase, which accumulates in the blood plasma of patients. The enzyme deglycosylates the Gc protein-derived macrophage activating factor (GcMAF) and inhibits macrophage activity acting as an immunosuppressor. The high specific activity 0.033 ± 0.002 μmol mg−1 min−1 of the enzyme was found in human colon carcinoma cells DLD-1. The alpha-NaGalase of DLD-1 cells was isolated and biochemical characterized. The enzyme exhibits maximum activity at pH 5.2 and temperature 55 °C. The Km is 2.15 mM, Vmax–0.021 μmol min−1 mL−1, kcat–1.55 min−1 and kcat/Km–0.72 min−1 mM−1 at 37 °C, pH 5.2. The effects of fucoidan from the brown alga Fucus evanescence on the activity of alpha-NaGalase in human colon carcinoma DLD-1 cells and on the biosynthesis of this enzyme were investigated. It was shown that fucoidan did not inhibit free alpha-NaGalase, however, it reduced the expression of the enzyme in the DLD-1 cells at IC50 73 ± 4 μg mL−1.

Is chondroitin sulfate responsible for the biological effects attributed to the GC protein-derived Macrophage Activating Factor (GcMAF)?

We hypothesize that a plasma glycosaminoglycan, chondroitin sulfate, may be responsible for the biological and clinical effects attributed to the Gc protein-derived Macrophage Activating Factor (GcMAF), a protein that is extracted from human blood. Thus, Gc protein binds chondroitin sulfate on the cell surface and such an interaction may occur also in blood, colostrum and milk. This interpretation would solve the inconsistencies encountered in explaining the effects of GcMAF in vitro and in vivo. According to our model, the Gc protein or the GcMAF bind to chondroitin sulfate both on the cell surface and in bodily fluids, and the resulting multimolecular complexes, under the form of oligomers trigger a transmembrane signal or, alternatively, are internalized and convey the signal directly to the nucleus thus eliciting the diverse biological effects observed for both GcMAF and chondroitin sulfate.

Gc Protein-Derived Macrophage Activating Factor (GcMAF) and Autism: Do Clinical Results Require a Novel Interpretation?

Gc Protein-Derived Macrophage Activating Factor (GcMAF) and Autism: Do Clinical Results Require a Novel Interpretation?

Management of Metastatic Colorectal Carcinoma with GcMAF Forte and Thymus Peptides: A Case Report

In the recent years immunotherapy has gained important role in cancer management. Immunostimulating effects of thymus-derived peptides on carcinogenesis have been actively studied in both animals and humans. Gc protein derived macrophage-activating factor (GcMAF) was found to have potent effect on macrophage phagocytic activity. Immunostimulating effects of GcMAF and peptides of thymus can be effectively combined together and with other treatment modalities of cancer. The current case report demonstrates outcomes of combination of GcMAF Forte and HT Thymus (peptides of thymus) in the management of metastatic colorectal adenocarcinoma. This case study suggests that individually prescribed immunotherapy can be a promising alternative in management of advance forms of cancer, especially when options of treatment are limited.

Gc-protein-derived macrophage activating factor counteracts the neuronal damage induced by oxaliplatin.

Oxaliplatin-based regimens are effective in metastasized advanced cancers. However, a major limitation to their widespread use is represented by neurotoxicity that leads to peripheral neuropathy. In this study we evaluated the roles of a proven immunotherapeutic agent [Gc-protein-derived macrophage activating factor (GcMAF)] in preventing or decreasing oxaliplatin-induced neuronal damage and in modulating microglia activation following oxaliplatin-induced damage. The effects of oxaliplatin and of a commercially available formula of GcMAF [oleic acid-GcMAF (OA-GcMAF)] were studied in human neurons (SH-SY5Y cells) and in human microglial cells (C13NJ). Cell density, morphology and viability, as well as production of cAMP and expression of vascular endothelial growth factor (VEGF), markers of neuron regeneration [neuromodulin or growth associated protein-43 (Gap-43)] and markers of microglia activation [ionized calcium binding adaptor molecule 1 (Iba1) and B7-2], were determined. OA-GcMAF reverted the damage inflicted by oxaliplatin on human neurons and preserved their viability. The neuroprotective effect was accompanied by increased intracellular cAMP production, as well as by increased expression of VEGF and neuromodulin. OA-GcMAF did not revert the effects of oxaliplatin on microglial cell viability. However, it increased microglial activation following oxaliplatin-induced damage, resulting in an increased expression of the markers Iba1 and B7-2 without any concomitant increase in cell number. When neurons and microglial cells were co-cultured, the presence of OA-GcMAF significantly counteracted the toxic effects of oxaliplatin. Our results demonstrate that OA-GcMAF, already used in the immunotherapy of advanced cancers, may significantly contribute to neutralizing the neurotoxicity induced by oxaliplatin, at the same time possibly concurring to an integrated anticancer effect. The association between these two powerful anticancer molecules would probably produce the dual effect of reduction of oxaliplatin-induced neurotoxicity, together with possible synergism in the overall anticancer effect.

Retraction. Immunotherapy of HIV-infected patients with Gc protein-derived macrophage activating factor.

Journal of Medical VirologyVolume 86, Issue 11 p. 1998-1998 RetractionFree Access Retraction This article retracts the following: Retracted: Immunotherapy of HIV-infected patients with Gc protein-derived macrophage activating factor (GcMAF) Nobuto Yamamoto, Naofumi Ushijima, Yoshihiko Koga, Volume 81Issue 1Journal of Medical Virology pages: 16-26 First Published online: November 21, 2008 First published: 11 August 2014 https://doi.org/10.1002/jmv.24060AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat This article has been retracted at the request of Editor-in-Chief: Dr. Ari Zuckerman Retraction: Nobuto Yamamoto, Naofumi Ushijima, and Yoshihjima Koga. Immunotherapy of HIV-infected patients with Gc protein-derived macrophage activating factor (GcMAF). J Med Virol 2009;81:16–26. The above article, published online on 21 Nov 2008 Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between Dr. Ari Zuckerman, Editor-in-Chief, Journal of Medical Virology and Wiley Periodicals, Inc. due to irregularities in the documentation for institutional review board approval. REFERENCES Yamamoto N, Ushijima N, Koga Y. 2009. Immunotherapy of HIV-infected patients with Gc protein-derived macrophage activating factor (GcMAF). J Med Virol 81: 16– 26. doi: 10/1002/jmv.21376. Wiley Online LibraryCASPubMedWeb of Science®Google Scholar Volume86, Issue11November 2014Pages 1998-1998 ReferencesRelatedInformation

Gc-protein derived macrophage activating factor (GcMAF) counteracts the neuronal damage induced by oxaliplatin

Oxaliplatin-based regimens are effective in metastasised advanced cancers. However, a major limitation to their use is represented by neurotoxicity leading to peripheral neurophaty (Wolf et al., 2008). In this study we evaluate the effects of an immunotherapeutic agent (Gc protein-derived macrophage activating factor, GcMAF) in preventing oxaliplatin-induced neuronal damage and in restoring microglial activation. The effects of oxaliplatin was studied in human neurons (SH-SY5Y) and microglial cells (c13-nj). Cell density, morphology and viability as well as production of cAMP and expression of vascular endothelial growth factor (VEGF), markers of neuron regeneration and markers of microglia activation were determined. GcMAF reverted the damage inflicted by oxaliplatin on human neurons and preserved their viability; it also increased cAMP production, VEGF and neuromodulin expression. GcMAF did not revert the effects of oxaliplatin on microglial cell viability. However, it induced microglial activation resulting in an increased expression of a specific marker without any increase in cell number. Our results demonstrate that GcMAF may significantly contribute to neutralize the neurotoxicity induced by oxaliplatin, at the same time concurring to an integrated anti-cancer effect.

The in vitro GcMAF effects on endocannabinoid system transcriptionomics, receptor formation, and cell activity of autism-derived macrophages

BackgroundImmune system dysregulation is well-recognized in autism and thought to be part of the etiology of this disorder. The endocannabinoid system is a key regulator of the immune system via the cannabinoid receptor type 2 (CB2R) which is highly expressed on macrophages and microglial cells. We have previously published significant differences in peripheral blood mononuclear cell CB2R gene expression in the autism population. The use of the Gc protein-derived Macrophage Activating Factor (GcMAF), an endogenous glycosylated vitamin D binding protein responsible for macrophage cell activation has demonstrated positive effects in the treatment of autistic children. In this current study, we investigated the in vitro effects of GcMAF treatment on the endocannabinoid system gene expression, as well as cellular activation in blood monocyte-derived macrophages (BMDMs) from autistic patients compared to age-matched healthy developing controls.MethodsTo achieve these goals, we used biomolecular, biochemical and immunocytochemical methods.ResultsGcMAF treatment was able to normalize the observed differences in dysregulated gene expression of the endocannabinoid system of the autism group. GcMAF also down-regulated the over-activation of BMDMs from autistic children.ConclusionsThis study presents the first observations of GcMAF effects on the transcriptionomics of the endocannabinoid system and expression of CB2R protein. These data point to a potential nexus between endocannabinoids, vitamin D and its transporter proteins, and the immune dysregulations observed with autism.

Intra-tumoural nitric oxide release by macrophages activated by Gc-protein-derived Macrophage Activating Factor (GcMAF)

Over past decades, nitric oxide (NO) has emerged as a molecule of interest in cancer treatment because of its tumouricidal properties (Choudhari et al., 2013). Gcprotein- derived Macrophage Activating Factor (GcMAF) induces the synthesis and release of NO by activated macrophages. It was previously demonstrated that molecular complexes of oleic acid (OA) and GcMAF (OA-GcMAF) stimulate macrophage activation in cancer patients (Ward et al., 2014). Here we demonstrate that intratumoural injection of OA-GcMAF leads NO synthesis and release inside the tumour. Under ultrasound guidance, OA-GcMAF was injected into patients harbouring different types of solid tumours; a metastasis from a melanoma, and a metastasis from breast cancer. Intra-tumoural NO synthesis and release was monitored in real-time by power-doppler ultrasonography. One to two minutes after injection, we observed a significant increase in blood flow and in blood vessels diameter, a clear indication of vasodilation due to NO synthesis and release. These observations substantiate the dramatic clinical results previously observed by Ward et al. (2014), and open the way to further investigation in the role of GcMAF as a powerful anticancer agent.

Gc-protein-derived Macrophage Activating Factor (GcMAF) induces ERBB2 shift in human breast cancer

HER2/Neu/ERBB2 is a receptor tyrosine kinase overexpressed in a high percentage of human breast cancers. Gc-protein-derived Macrophage Activating Factor (GcMAF) is a powerful stimulant of the immune system endowed with intrinsic anticancer properties (Pacini et al., 2012). We recently demonstrated that molecular complexes of oleic acid (OA) and GcMAF (OA-GcMAF) show significant therapeutic activity in a variety of tumours (Ward et al., 2014). Here we demonstrate that OAGcMAF eradicates ERBB2 expression in human breast cancer. A biopsy taken before OA-GcMAF treatment showed strong positivity to ERBB2. The patient was then treated with OA-GcMAF administered through subcutaneous injections and with food naturally rich in OA-GcMAF for 3 weeks prior to mastectomy. The subsequent surgery specimen was negative for ERBB2. These results lead to hypothesize: 1. OAGcMAF completely reversed the neoplastic phenotype. 2. OA-GcMAF induced the apoptosis of all ERBB2-positive cancer cells.

GC protein-derived macrophage-activating factor decreases α-N-acetylgalactosaminidase levels in advanced cancer patients

α-N-acetylgalactosaminidase (nagalase) accumulates in the serum of cancer patients and its activity correlates with tumor burden, aggressiveness and clinical disease progression. The administration of GC protein-derived macrophage-activating factor (GcMAF) to cancer patients with elevated levels of nagalase has been associated with a decrease of serum nagalase activity and with significant clinical benefits. Here, we report the results of the administration of GcMAF to a heterogeneous cohort of patients with histologically diverse, advanced neoplasms, generally considered as “incurable” diseases. In most cases, GcMAF therapy was initiated at late stages of tumor progression. As this is an open-label, non-controlled, retrospective analysis, caution must be employed when establishing cause-effect relationships between the administration GcMAF and disease outcome. However, the response to GcMAF was generally robust and some trends emerged. All patients (n = 20) presented with elevated serum nagalase activity, well above normal values. All patients but one showed a significant decrease of serum nagalase activity upon weekly GcMAF injections. Decreased nagalase activity was associated with improved clinical conditions and no adverse side effects were reported. The observations reported here confirm and extend previous results and pave the way to further studies aimed at assessing the precise role and indications for GcMAF-based anticancer immunotherapy.

Gc protein-derived macrophage-activating factor (GcMAF) stimulates cAMP formation in human mononuclear cells and inhibits angiogenesis in chick embryo chorionallantoic membrane assay.

The effects of Gc protein-derived macrophage-activating factor (GcMAF) have been studied in cancer and other conditions where angiogenesis is deregulated. In this study, we demonstrate for the first time that the mitogenic response of human peripheral blood mononuclear cells (PBMCs) to GcMAF was associated with 3'-5'-cyclic adenosine monophosphate (cAMP) formation. The effect was dose dependent, and maximal stimulation was achieved using 0.1 ng/ml. Heparin inhibited the stimulatory effect of GcMAF on PBMCs. In addition, we demonstrate that GcMAF (1 ng/ml) inhibited prostaglandin E(1)- and human breast cancer cell-stimulated angiogenesis in chick embryo chorionallantoic membrane (CAM) assay. Finally, we tested different GcMAF preparations on CAM, and the assay proved to be a reliable, reproducible and inexpensive method to determine the relative potencies of different preparations and their stability; we observed that storage at room temperature for 15 days decreased GcMAF potency by about 50%. These data could prove useful for upcoming clinical trials on GcMAF.

Polymorphism in Vitamin D-Binding Protein as a Genetic Risk Factor in the Pathogenesis of Endometriosis

Context: Previous studies have implicated a deficiency in the inflammatory response in women who develop endometriosis. The specific immunological deficits have not been completely elucidated. Objective: Our objective was to identify differences in protein expression in serum that might shed light on the pathophysiology of endometriosis. Design and Setting: This cross-sectional study of women undergoing laparoscopy between 2003 and 2005 took place at a university medical center. Patients: Patients included consenting women age 18-49 yr undergoing surgery for pain and/or infertility or elective tubal ligation. Women with acute or chronic medical conditions were excluded. Intervention: Blood was collected preoperatively. Main Outcome Measure: Proteomic analysis of serum was done using two-dimensional difference gel electrophoresis. Results: We found 25 protein spots with a significant difference in abundance between women with endometriosis and controls, including acute-phase proteins and complement components. The abundance of vitamin D-binding protein was higher in all endometriosis pools by a factor of approximately 3 compared with the control pool (P < 0.02). Analysis of specific allele products using nano-LC-ESI-MS indicated that it was the GC*2 allele product that was in greater concentration in serum pools, as well as in single validation samples, in women with endometriosis (P = 0.006). In contrast to the GC*1 allele product, which is readily converted to a potent macrophage factor (Gc protein-derived macrophage-activating factor), the GC*2 allele product undergoes practically no such conversion. Conclusions: We speculate that the inability to sufficiently activate macrophages’ phagocytotic function in those carrying the GC*2 polymorphism (more prevalent in endometriosis) may allow endometriotic tissues to implant in the peritoneal cavity. Future studies evaluating specific vitamin D-binding protein polymorphisms as a risk factor for endometriosis in larger populations of women are warranted.

Polymorphism in Vitamin D-Binding Protein as a Genetic Risk Factor in the Pathogenesis of Endometriosis

Context Previous studies have implicated a deficiency in the inflammatory response in women who develop endometriosis. The specific immunological deficits have not been completely elucidated. Objective Our objective was to identify differences in protein expression in serum that might shed light on the pathophysiology of endometriosis. Design and Setting This cross-sectional study of women undergoing laparoscopy between 2003 and 2005 took place at a university medical center. Patients Patients included consenting women age 18-49 yr undergoing surgery for pain and/or infertility or elective tubal ligation. Women with acute or chronic medical conditions were excluded. Intervention Blood was collected preoperatively. Main Outcome Measure Proteomic analysis of serum was done using two-dimensional difference gel electrophoresis. Results We found 25 protein spots with a significant difference in abundance between women with endometriosis and controls, including acute-phase proteins and complement components. The abundance of vitamin D-binding protein was higher in all endometriosis pools by a factor of approximately 3 compared with the control pool (P < 0.02). Analysis of specific allele products using nano-LC-ESI-MS indicated that it was the GC*2 allele product that was in greater concentration in serum pools, as well as in single validation samples, in women with endometriosis (P = 0.006). In contrast to the GC*1 allele product, which is readily converted to a potent macrophage factor (Gc protein-derived macrophage-activating factor), the GC*2 allele product undergoes practically no such conversion. Conclusions We speculate that the inability to sufficiently activate macrophages’ phagocytotic function in those carrying the GC*2 polymorphism (more prevalent in endometriosis) may allow endometriotic tissues to implant in the peritoneal cavity. Future studies evaluating specific vitamin D-binding protein polymorphisms as a risk factor for endometriosis in larger populations of women are warranted.

Polymorphism in Vitamin D-Binding Protein as a Genetic Risk Factor in the Pathogenesis of Endometriosis

Previous studies have implicated a deficiency in the inflammatory response in women who develop endometriosis. The specific immunological deficits have not been completely elucidated. Our objective was to identify differences in protein expression in serum that might shed light on the pathophysiology of endometriosis. This cross-sectional study of women undergoing laparoscopy between 2003 and 2005 took place at a university medical center. Patients included consenting women age 18-49 yr undergoing surgery for pain and/or infertility or elective tubal ligation. Women with acute or chronic medical conditions were excluded. Blood was collected preoperatively. Proteomic analysis of serum was done using two-dimensional difference gel electrophoresis. We found 25 protein spots with a significant difference in abundance between women with endometriosis and controls, including acute-phase proteins and complement components. The abundance of vitamin D-binding protein was higher in all endometriosis pools by a factor of approximately 3 compared with the control pool (P < 0.02). Analysis of specific allele products using nano-scale liquid chromatography-electrospray ionization-mass spectrometry indicated that it was the GC*2 allele product that was in greater concentration in serum pools, as well as in single validation samples, in women with endometriosis (P = 0.006). In contrast to the GC*1 allele product, which is readily converted to a potent macrophage factor (Gc protein-derived macrophage-activating factor), the GC*2 allele product undergoes practically no such conversion. We speculate that the inability to sufficiently activate macrophages' phagocytotic function in those carrying the GC*2 polymorphism (more prevalent in endometriosis) may allow endometriotic tissues to implant in the peritoneal cavity. Future studies evaluating specific vitamin D-binding protein polymorphisms as a risk factor for endometriosis in larger populations of women are warranted.

T.101. Treatment of HIV-Infected Patients with Gc Protein-Derived Macrophage Activating Factor (GcMAF) and Its Coned Derivative (GcMAFc) Eradicates HIV-Infection

T.101. Treatment of HIV-Infected Patients with Gc Protein-Derived Macrophage Activating Factor (GcMAF) and Its Coned Derivative (GcMAFc) Eradicates HIV-Infection

Retracted: Immunotherapy of HIV‐infected patients with Gc protein‐derived macrophage activating factor (GcMAF)

Serum Gc protein (known as vitamin D3-binding protein) is the precursor for the principal macrophage activating factor (MAF). The MAF precursor activity of serum Gc protein of HIV-infected patients was lost or reduced because Gc protein is deglycosylated by alpha-N-acetylgalactosaminidase (Nagalase) secreted from HIV-infected cells. Therefore, macrophages of HIV-infected patients having deglycosylated Gc protein cannot be activated, leading to immunosuppression. Since Nagalase is the intrinsic component of the envelope protein gp120, serum Nagalase activity is the sum of enzyme activities carried by both HIV virions and envelope proteins. These Nagalase carriers were already complexed with anti-HIV immunoglobulin G (IgG) but retained Nagalase activity that is required for infectivity. Stepwise treatment of purified Gc protein with immobilized beta-galactosidase and sialidase generated the most potent macrophage activating factor (termed GcMAF), which produces no side effects in humans. Macrophages activated by administration of 100 ng GcMAF develop a large amount of Fc-receptors as well as an enormous variation of receptors that recognize IgG-bound and unbound HIV virions. Since latently HIV-infected cells are unstable and constantly release HIV virions, the activated macrophages rapidly intercept the released HIV virions to prevent reinfection resulting in exhaustion of infected cells. After less than 18 weekly administrations of 100 ng GcMAF for nonanemic patients, they exhibited low serum Nagalase activities equivalent to healthy controls, indicating eradication of HIV-infection, which was also confirmed by no infectious center formation by provirus inducing agent-treated patient PBMCs. No recurrence occurred and their healthy CD + cell counts were maintained for 7 years.