Sensitivity to methylmercury-induced autoimmune disease in mice correlates with resistance to apoptosis of activated CD4+ lymphocytes.
The sensitivity of splenic lymphoid cells to apoptosis induced by low concentrations of methylmercury (MeHgCl) has been examined in C57BL/6 and SJL mice, which are, respectively, resistant and sensitive to a genetically determined autoimmune disease induced by subtoxic doses of MeHgCl. To determine the implications of subtoxic doses of MeHgCl in the susceptibility of SJL mice to autoimmune disease, Concanavalin A (ConA) stimulated spleen cells from both mouse strains were treated in vitro with MeHgCl concentrations varying between 0.001 and 1.0 microM for 48h. Results have shown that ConA-activated splenic lymphoid cells from SJL mice increased in the presence of low concentrations of MeHgCl while the number of lymphoid cells from C57BL/6 mice rather decreased. Flow cytometric analysis of the cells showing a typical lymphoid forward scatter (FSC)/side scatter (SSC) pattern (region R1), and those characterized by a lower FSC and a higher SSC parameters (region R2), morphologically corresponding to apoptotic cells, revealed that lymphoid cells from C57BL/6 mice suffered a dose-dependent shift from region R1 toward region R2 when treated with concentrations ranging between 0.01 and 1 microM of MeHgCl. However, SJL splenic lymphoid cells cultured in the presence of low concentrations of MeHgCl proved more resistant to apoptosis. The level of apoptosis induced by MeHgCl in both regions was verified by AnnexinV-propidium iodide (PI) and TdT-mediated dUTP nick end labeling (TUNEL) immunolabelings. Phenotyping of lymphoid cells from both mouse strains cultured in the presence of low concentrations of MeHgCl and stimulated with ConA, indicated that CD4+ T cells from SJL mice increased while the corresponding cell subset from C57BL/6 mice became apoptotic. The resistance to apoptosis of ConA-activated lymphoid cells from SJL mice seemed related to an increase of CD4+ cells induced by the lower concentrations of MeHgCl (0.001 and 0.01 microM). However, these SJL cells were sensitive to anti-Fas-mediated apoptosis while residual anti-Fas-resistant cells from C57BL/6 mice were, themselves, sensitive to MeHgCl-induced apoptosis. The in vivo significance of these results has been confirmed by an observed increase in splenic cellularity and in the percentage of activated CD4+ cells from SJL mice. These increases were not observed in C57BL/6 mice.
Pheng
Say-Ry
SR
Département de Médecine du Travail et d'Hygiène du Milieu, Faculté de Médecine, Université de Montréal, Québec, Montréal, Canada.
Auger
Chantal
C
Chakrabarti
Saroj
S
Massicotte
Eric
E
Lamontagne
Lucie
L
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001327:Autoimmune Diseases / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D003208:Concanavalin A / Q000378:metabolism; D053222:Fas Ligand Protein; D005260:Female; D005434:Flow Cytometry; D008562:Membrane Glycoproteins / Q000378:metabolism; D008767:Methylmercury Compounds / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000276:immunology / Q000378:metabolism; D019014:fas Receptor / Q000378:metabolism
D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; D008767:Methylmercury Compounds; D019014:fas Receptor; D003208:Concanavalin A; C004925:methylmercuric chloride