ISOLATION OF IA-LIKE ANTIGEN-BEARING CELLS FROM HUMAN PERIPHERAL LYMPHOCYTES THROUGH THE USE OF A MONOCLONAL-ANTIBODY TO FRAMEWORK DETERMINANTS OF IA-LIKE ANTIGENS

作     者：INDIVERI, F NG, AK RUSSO, C QUARANTA, V PELLEGRINO, MA FERRONE, S 

作者机构：SCRIPPS CLIN & RES FDN RES INST DEPT MOLEC IMMUNOL LA JOLLA CA 92037 USA 

出 版 物：《JOURNAL OF IMMUNOLOGICAL METHODS》 (免疫法杂志)

年 卷 期：1980年第39卷第4期

页      面：343-354页

核心收录：

学科分类：0710[理学-生物学] 1001[医学-基础医学(可授医学、理学学位)] 10[医学] 

基　　金：NCI NIH HHS [CA 16069, CA 16071] Funding Source: Medline NIAID NIH HHS [AI13154] Funding Source: Medline 

主　　题：ADCC antibody dependent cellular cytotoxicity AET 2-aminoethylisothiouronium bromide AET-E erythrocytes treated with AET C3 third component of complement C3d conversion product of C3 E erythrocytes EA antibody coated erythrocytes EAC1-3 mo EA coated with C1–C3 from mouse serum deficient in the fifth component of complement EAγ erythrocytes sensitized with IgG antibodies EAμ erythrocytes sensitized with IgM antibodies GoE goat erythrocytes MbIg membrane bound immunoglobulin MEM minimal essential medium MLR mixed lymphocyte reaction MoAb monoclonal antibody NK natural killer PBL peripheral blood lymphocytes PWM pokeweed mitogen ShE sheep erythrocytes Tγ T cells which express receptor for the Fc fragment of IgG Tμ T cells which express receptor for the Fc fragment of IgM 

摘      要：A non-complement fixing monoclonal antibody (Q2/70) to framework determinants of human Ia-like antigens was used to develop a method for isolating Ia-like antigen bearing, i.e., Ia-like (+) cells and cells lacking these antigens, i.e., Ia-like (-), from human peripheral blood lymphocytes (PBL). The method was based on PBL sensitization with the antibody Q2/70, followed by rosetting with sheep (ShE) coated with purified rabbit anti-mouse Ig antibodies, differential centrifugation on a Ficoll-Hypaque gradient, and finally recovery of Ia-like (+) and Ia-like (-) cells from the bottom and the interface of the gradient, respectively. Marker analysis of the 2 cell subpopulations showed that the majority of the bottom cell fraction were Ia-like (+) and carried B cell markers such as membrane bound Ig (MbIg) and C3 receptors. The majority of the interface cell fraction were Ia-like (-) and carried T cell markers such as receptors for 2-aminoethylisothiouronium treated sheep erythrocytes (AETShE) and goat erythrocytes (GoE). Serological and functional studies showed that the Ia-like (+) cells: were useful targets in complement mediated cytotoxicity assays for HLA-DR typing;served as stimulator but not as responder in unidirectional mixed lymphocyte reactions (MLR);did not display lytic activity in natural killer (NK) cell cytotoxicity and in antibody dependent cellular cytotoxicity (ADCC);and proliferated in response to pokeweed mitogen (PWM) stimulation in the presence of helper T cells. The Ia-like (-) cells responded to but failed to stimulate allogeneic lymphocytes in the MLR;were highly active in NK and ADCC assays;and, provided helper activity in PWM stimulation of purified B cells.