Deoxyspergualin inhibits kappa light chain expression in 70Z/3 pre-B cells by blocking lipopolysaccharide-induced NF-kappa B activation.
Deoxyspergualin (DSG) is a potent immunosuppressive agent that is currently undergoing clinical trials for treatment of transplant rejection, preventive of human anti-mouse Ab response, and blocking autoimmune disease progression. The mechanism of action of DSG appears to be novel, with in vivo activity attributable to the suppression of both humoral and cell-mediated immunity. In this study we investigated the effect of DSG on the induction of lg expression in the 70Z/3 murine pre-B cell line. Treatment of 70Z/3 cells with DSG for 24, 48, or 72 h before LPS or IFN-gamma induction resulted in a time-dependent inhibition of surface lgM expression, with greater than 80% inhibition observed after 72 h of pretreatment. Inhibition of surface expression was specific for lgM, as neither MHC class I nor CD45 (B220) surface expression was affected by DSG pretreatment. Cyclosporin A was ineffective at suppressing surface igM induction. DSG pretreatment results in a 10-fold reduction in LPS- or IFN-gamma-induced kappa L chain protein and mRNA expression. No change was observed in either mu or beta-actin mRNA levels. Analysis of nuclear and cytoplasmic NF-kappa B expression using electrophoretic mobility shift analysis and Western analysis, revealed that DSG blocked LPS-induced NF-kappa B nuclear translocation, but had no effect on cytoplasmic NF-kappa B levels. We conclude that DSG may act to suppress humoral immune responses by blocking the transcriptional activation of kappa L chain expression during certain stages of B cell development.
Tepper
M A
MA
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, WA 98121, USA.
Nadler
S G
SG
Esselstyn
J M
JM
Sterbenz
K G
KG
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D016572:Cyclosporine / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology*; D015395:Histocompatibility Antigens Class I / Q000187:drug effects; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007371:Interferon-gamma / Q000187:drug effects / Q000494:pharmacology; D017493:Leukocyte Common Antigens / Q000187:drug effects; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000096:biosynthesis / Q000187:drug effects*; D011506:Proteins / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D013234:Stem Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism
D006146:Guanidines; D015395:Histocompatibility Antigens Class I; D007145:Immunoglobulin kappa-Chains; D007166:Immunosuppressive Agents; D008070:Lipopolysaccharides; D016328:NF-kappa B; D011506:Proteins; D012333:RNA, Messenger; D007371:Interferon-gamma; D016572:Cyclosporine; D017493:Leukocyte Common Antigens; C037258:gusperimus