Studies on the induction of antigen-specific antibody in anti-CD40 cultured human B lymphocytes.
Costimulatory signals provided by T cells are required for B cells to produce specific antibody to T-dependent antigen. We have investigated the suitability of using the CD40 culture system for the proliferation and differentiation of Ag-specific human B cells using cytomegalovirus (CMV) or tetanus toxoid (TT) as antigen. We modified the CD40 culture system (CD32-transfected L cells, anti-CD40, and IL-4) by applying a sequential cytokine stimulation and compared total B-cell cultures with antigen-specific B cells preselected by panning. The detection of specific antibody became possible when antigen-selected B cells were cultured for 7 days in the CD40 system to induce clonal expansion, followed by the addition of IL-2 and IL-10 for an additional 7 days to induce plasma-cell differentiation. We conclude that our initial inability to detect specific antibody in the CD40 system is due to overgrowth of nonspecific B-cell clones and that selection of antigen-specific B cells by panning overcomes this problem. Induction of antigen-specific antibody production was found to be optimal when the initial contact with antigen during panning was limited to between 1 to 24 hours.
Schilizzi
B M
BM
Department of Clinical Immunology, University Hospital, Groningen, The Netherlands.
Harmsen
M C
MC
The
T H
TH
De Leij
L
L
D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D000917:Antibody Formation; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D007378:Interleukins / Q000494:pharmacology; D008213:Lymphocyte Activation; D013154:Spleen / Q000166:cytology; D013745:Tetanus Toxoid / Q000276:immunology
D000907:Antibodies, Bacterial; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019013:CD40 Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007378:Interleukins; D013745:Tetanus Toxoid