Glatiramer acetate induces pro-apoptotic mechanisms involving Bcl-2, Bax and Cyt-c in peripheral lymphocytes from multiple sclerosis patients.
Apoptotic deletion of autoreactive T-cells is defective in patients with multiple sclerosis (MS). Glatiramer acetate (GA) treatment seems to restore apoptosis of detrimental T-cells. We analyzed the mitochondria membrane pro- (Bax) and anti-apoptotic (Bcl- 2) and cytosolic pro-apoptotic (Cyt-c, APAF-1) proteins expression in peripheral lymphocytes from relapsing-remitting (RR) MS patients during GA treatment. Blood samples were collected from 8 healthy controls (HCs) and from 8 RR MS patients prior to and every three months during the 9 months of GA treatment. Peripheral blood mononuclear cells (PBMNCs) Bcl-2, Bax, Cyt-c and APAF-1 were quantified by western blot followed by densitometric scanning and Bax/Bcl-2, cytosolic Cyt-c/Bcl-2 and APAF-1/Bcl-2 ratios were calculated. T-cells were in vitro tested for oxygen consumption by a respirometric analysis. Bax/Bcl-2, cytosolic Cyt-c/Bcl-2 and APAF-1/Bcl-2 ratios in untreated MS patients were significantly (p < 0.05) lower than in HCs. Bax/Bcl-2 ratio increased (p = 0.03) and Cyt-c/Bcl-2 ratio showed a trend to increase during the 9 months of GA treatment in MS patients. A reduction of 58% and 59% in oxygen consumption by PBMNCs was evident after GA treatment in vitro or when GA treated patients' cells were compared with those from HCs, respectively. Our findings suggest that GA exerts a regulatory effect on peripheral T lymphocytes through pro-apoptosis mechanisms involving mitochondria and cytosolic proteins.
Ruggieri
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Dept. of Neurological and Psychiatric Sciences, University of Bari, Policlinico, Piazza Giulio Cesare, Italy. mruggieri@neurol.uniba.it
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D000328:Adult; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D053477:Apoptotic Protease-Activating Factor 1; D015153:Blotting, Western / Q000379:methods; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D016022:Case-Control Studies; D045304:Cytochromes c / Q000378:metabolism; D005260:Female; D000068717:Glatiramer Acetate; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000188:drug therapy / Q000473:pathology*; D010101:Oxygen Consumption / Q000187:drug effects; D010455:Peptides / Q000008:administration & dosage*; D011506:Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012402:Rotenone / Q000494:pharmacology; D018709:Statistics, Nonparametric; D013997:Time Factors; D014475:Uncoupling Agents / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
C506592:APAF1 protein, human; D051017:Apoptosis Regulatory Proteins; D053477:Apoptotic Protease-Activating Factor 1; D007166:Immunosuppressive Agents; D047908:Intracellular Signaling Peptides and Proteins; D010455:Peptides; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D014475:Uncoupling Agents; D051028:bcl-2-Associated X Protein; D012402:Rotenone; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000068717:Glatiramer Acetate; D045304:Cytochromes c