Detection and enumeration of circulating HIV-1-specific memory B cells in HIV-1-infected patients.
Memory B cells are long-living cells that circulate throughout the body and differentiate into plasma cells after stimulation by antigens, cytokines, and direct cell-to-cell interaction in lymphoid tissues. For HIV-1-infected patients, we assessed whether in vitro polyclonal B cell activation that induces immunoglobulin secreting cells (SCs) also generates HIV-1-specific resting B cells to synthesize antibodies specific to HIV-1. To this end, highly purified B cells from 10 HIV-1-untreated patients were cultured with or without mouse fibroblastic cells expressing the CD40 ligand in the presence of IL-2 and IL-10. The percentage of immunoglobulin SCs we obtained by using the B cell-CD40L stimulation system was equal to 55% to 98% of the circulating memory B cells. Moreover, the anti-HIV-1 IgG, IgA, or IgM antibody SCs represented 1 x 10-2 to 1 x 10-3 of the total immunoglobulin SCs. The anti-HIV-1-specific antibodies detected in cell culture supernatants were directed to gag-, pol-, and env-encoded viral proteins. We found that in AIDS patients, HIV-1-specific resting memory B cells circulate in the blood and can be quantified by their anti-HIV-1 antibody secretion after strong B cell polyclonal stimulation.
Fondere
Jean-Michel
JM
Laboratoire de Virologie, Hôpital Lapeyronie, Montpellier, France.
Huguet
Marie-France
MF
Macura-Biegun
Anna
A
Baillat
Vincent
V
Ohayon
Viviane
V
Reynes
Jacques
J
Vendrell
Jean-Pierre
JP
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology* / Q000821:virology*; D000918:Antibody Specificity; D015703:Antigens, CD / Q000097:blood; D001402:B-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015483:HIV Antibodies / Q000097:blood; D015497:HIV-1 / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007072:Immunoglobulin D / Q000097:blood; D007156:Immunologic Memory / Q000276:immunology*; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000097:blood; D014766:Viremia / Q000097:blood / Q000276:immunology
D015703:Antigens, CD; D015483:HIV Antibodies; D007072:Immunoglobulin D; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7