title: Proliferation of resting B cells is modulated by CR2 and CR1.
abstract: Absence of the third component of complement, C3, is associated with impaired ability to synthesize antibody, particularly in the presence of limiting antigen [1-9]. The mature B lymphocyte bearing the surface immunoglobulin receptor transduces signals for proliferation and differentiation upon binding of specific antigen. This mature B cell also bears two related membrane proteins, CR2 (the C3d/Epstein-Barr virus receptor) (CD35) [15], which can mediate the binding of ligands to which appropriate cleavage fragments of C3 have become attached [16]. It has been suggested that these receptors play a direct role(s) in B cell activation [17-25]. In light of previous in vivo observations we decided to assess the function of CR2 and CR1 in relation to B cell activation through the membrane IgM receptor. Highly purified splenic B cells were prepared. No contaminating T cells or macrophages were detected by flow cytometric analysis and no proliferative activity was present upon PHA or ConA stimulation of the purified cells. The B cells were separated into low (activated), medium (preactivated) and high density (resting) fractions by Percoll gradient density centrifugation [26]. The responses of the B cell subpopulations to various concentrations of anti mu (DA4.4 monoclonal antibody) [27] were examined for proliferation at 72 h and for IgM/IgG production at 7 days. Low density B cells were maximally stimulated and no concentration of anti-mu was effective in enhancing their responses. High density B cells proliferated to anti-mu in a concentration dependent manner. When substimulatory concentrations of anti-mu were employed, concomitant crosslinking of CR2 (with either of 2 distinct monoclonal antibodies HB-5 [28] or OKB7 [17]) resulted in a 45% enhancement of B cell proliferation above that observed by crosslinking of SIgM alone. In these studies, total IgM and IgG did not increase in the absence of T cells or T cell factors, indicating that terminal differentiation did not occur. In contrast, when a monoclonal antibody to CR1(44D) [29] was employed in an identical experiment, B cell proliferation was completely inhibited. Antibodies to CR2 or CR1 either alone or in crosslinked form did not enhance B cell proliferation. Immune complexes may crosslink the B cell surface in a manner analogous to our model when the immunoglobulin receptor and CR2 are simultaneously engaged. This activation signal may be particularly important in eliciting antibody responses when the quantity of specific antigen or the affinity for antigen is low. The marked inhibition of proliferation induced by CR1 suggests an alternate role for this receptor in modulation of B cell responses.
authors.affiliation: Laboratory of Infectious Diseases, Dana-Farber Cancer Institute, Boston, MA.
authors.forename: J D
authors.initials: JD
authors.lastname: Fingeroth
authors.forename: M E
authors.initials: ME
authors.lastname: Heath
authors.forename: D M
authors.initials: DM
authors.lastname: Ambrosino
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002452:Cell Count; D002455:Cell Division; D002469:Cell Separation; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006684:HLA-DR Antigens / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D051379:Mice; D011951:Receptors, Complement / Q000276:immunology*; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d
chemical_list: D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D005456:Fluorescent Dyes; D006684:HLA-DR Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d