Mitogenic analysis of murine B-cell heterogeneity.

Abstract

Murine B lymphocytes appear to be heterogeneous from a functional point of view. Thus, those B cells that give rise to antibody-producing cells can be divided into a subset that requires cooperation with T-derived lymphocytes and another subset that is T independent (1-6). Recently, it has been shown that the B-cell population also contains suppressor cells that regulate various functions of T cells (7)(8)(9)(10)(11) and that these suppressor cells seem to be different from antibody-producing cells (10, 11).There is contradictory evidence as to whether the response to different mitogens defines different B-cell subpopulations. In mice, Melchers and Andersson (12) concluded that PPD and lipepelysaccharide (LPS) I stimulate the same cell, whereas Gronowicz and Coutinho (13) The aim of this work was to test directly various alternative hypotheses that could explain the differences in the responsiveness of routine B lymphocytes with regard to various mitogens: (a) One cell possesses the receptors for various mitogens and can be stimulated by all known murine B-cell mitogens separately; (b) the ability to respond to various mitogens depends on the stage of maturation of the B cell; (c) there are different subsets of lymphocytes for different mitogens.We have chosen three well-known B-cell mitogens for this study; LPS (12), Nocardia water-soluble mitogen (NWSM) (18) and DxS (13). We have used the 5-bromo-deoxyuridine (BUdR) suicide technique of Zoschke and Bach (20). This should be a more critical method for ascertaining whether different cells respond to different mitogens by eliminating or inactivating those cells that respond to an initial mitogen.The specific killing by rabbit anti-mouse immunoglobulin serum and by antiIa alloantisera was used to investigate the presence of Ig and Ia on the surface of cells reactive to the above mitogens.1 Abbreviations used in this paper: BUdR, 5-bromo-deoxyuridine; CNRS, Centre National de la Recherche Scientifique; Con A, concanavalin A; DxS, dextran sulfate; LPS, lipopolysaccharide; NWSM, Nocardia water-soluble mitogen. 136J. ExP. MED.

Functional Subsets of Murine and Human B Lymphocyte Cell Lines

“…It remains to be determined whether the fx chain is functional on the surface of cells expressing// only, and whether these cells have already determined which light chain will be synthesized. Several groups have evidence that partially overlapping subsets of normal B cells may respond to several different macromolecular mitogens or PCAs: DxS and poly 1. poly C (Diamantstein et al 1974), endotoxic protein, LPS and PPD (Goodman & Sultzer 1979), LPS or Nocardia mitogen and DxS (Bona et al 1978). Hammerling et al (1978) have used LPS and other agents to induce sIg and other mature markers on bone marrow fractions of mouse pre-B or B lymphocytes.…”

Immune responsiveness of SM/J mice. Cellular characteristics and genetic analysis of hyperresponsiveness to B cell mitogens.

“…B lymphocytes are heterogeneous, and there is evidence based upon sequential stimulation with various different mitogens and upon suicide experiments that subsets of B cells respond optimally to different B cell mitogens (24)(25)(26). However, Andersson et al (27) have postulated that there may also be a B cell subset that expresses all mitogen receptors of a species and therefore responds to all mitogens.…”

Preparation of Antisera Specific for Guinea Pig B Cells

“…The sera The combining site on B cells for our ABS is unknown. ABS produced against murine and chicken B lymphocytes have had activity demonstrated against SMIg [10,11], receptors for the Fc portion of IgG [15] and la antigens [16]. Our ABS is not anti-immunoglobulin and is unlikely to be active against the receptor for Fc as L2C cells do not carry this marker [7; personal observations ].…”

“…Assay of ABS for Anti-Immunoglobulin Activity L2C cells are B lymphocytes and carry readily detectable surface membrane immu noglobulin [9], Some anti-B cell sera in mice have been shown to kill by anti-immunoglobulin activity [10,11], It was there fore important to determine whether our ABS had anti-immunoglobulin activity. Nei ther absorbed nor unabsorbed ABS pro duced any precipitation lines with normal guinea pig scrum in immunoelectrophoretic or double immunodiffusion studies.…”