Focussing human B cell specificity by immunoselection via antigen-presenting cells in vitro.
We present a protocol for in vitro immunization of B cells using monocyte-derived accessory cells (MoAC). MoAC are developed from human peripheral blood monocytes in culture and represent functionally competent inducers of antigen-specific immune responses. Using MoAC, we attempted to immunoselect TT-specific lymphocytes by rosetting. Adherent human MoAC were pulsed with tetanus toxoid (TT) and allowed to form clusters with autologous lymphocytes, followed by removal of non-adherent cells. After one week of culture, a specific anti-TT antibody response emerged on a low background of unspecific Ig. In comparison, cultures which had not been selected for adherent cells produced a high polyclonal background. Our results demonstrate that from peripheral blood cells, previously not a favourable source for in vitro immunization, in a majority of tests antigen-specific B cells could efficiently be immunoselected via adherence to autologous antigen-presenting cells, leading to a high-titre in vitro immunization.
Lenzner
S
S
Department of Immunology, Georg August University, Göttingen, Germany.
Scholtes
U
U
Peters
J H
JH
D000918:Antibody Specificity; D000938:Antigen-Presenting Cells / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002448:Cell Adhesion; D002469:Cell Separation; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009000:Monocytes / Q000166:cytology / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology*
D013745:Tetanus Toxoid