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Related post: also be isolated by separating cells on the Fluorescence Activated Cell Sorter [FACS] according to the amounts of IgM, IgD, and total Ig they express on their membranes. In general, B lymphocytes responsive to anti-p bear relatively small amounts of IgM, intermediate to large amounts of IgD, and large amounts of Ig, Thus, the anti-p responsive B lymphocyte has a high ratio of IgD to IgM. It is interesting in this regard, that the B lymphocyte defect caused by xid is associated with absence of proliferative responses to anti-p and with the absence or diminished numbers of B lymphocytes which have a high ratio of IgD to IgM. Our current studies of this phenomenon are centered around three areas. We have produced a series of monoclonal rat anti-p antibodies through fusion of rat antibody producing cells with the BALB/c myeloma NS-1 . We Buy Meclizine are now in the process of characterizing the specificity of these monoclonal antibodies and determining whether they are capable of stimulating proliferative responses. A second area of emphasis is that of producing cloned lines of B lymphocytes reactive to anti-p (see Section C). Such cells should provide ideal materials for the study of the cell biology of B lymphocyte activation by anti-p. It is the study of early events in B cell activation which constitutes the third major area in which study has recently Buy Meclizine Online begun. B. B Lymphocyte Ontogeny, Heterogeneity and Activation 1. CBA/N Mice and the xid Gene Our general approach to the study of the functional diversity of B lymphocytes has been based on the use of genetic systems. We have placed 22-lff ZOl-AI-00030-12 LI particular emphasis on the x-1 inked genetic defect of CBA/N mice which is characterized by a profound but discrete defect in B lymphocyte function. This defect involves the failure of CBA/N mice and mice with CBA/N x-chromo- somes to respond to certain thymus-independent (TI) antigens. These include hapten derivatives of Ficoll, type III pneumococcal polysaccharide, and levan and dextran and their hapten conjugates. In Where To Buy Meclizine contrast, mice with the CBA/N de- fect respond to another group of TI antigens, which includes the TNP deriva- tives of Brucella abortus (TNP-BA), lipopolysaccharide (LPS) and Nocardia water soluble mitogen (NWSM). On the basis of the distinctive responsiveness of CBA/N mice to these two sets of TI antigens, we proposed that they should be separately designated. TI antigens to which CBA/N respond are designated TI-1 antigens while those to which CBA/N mice do not respond are TI-2 antigens. In general, neonatal mice are unresponsive to TI-2 antigens but do respond to TI-1 antigens and alloanti-6 antibodies block in vitro responses to TI-2 Purchase Meclizine but not TI-1 antigens. Finally, TI-1 antigens tend to be powerful polyclonal B lymphocyte activators whereas TI-2 antigens are, for the most part, inactive in this regard. We have recently mapped the gene responsible for the CBA/N defect. This gene, termed x-1 inked immunodeficiency gene ( xid ) , is found on the x-chromo- some equidistant from the Ta (Tabby) and hyp (hypophosphatemia) genes. 2. The Lyb5 and Lyb7 Differentiation Antigens Our working hypothesis is that the genetic defect of mice homozygous or hemizygous for the xid gene involves the deletion of a subpopulation of B lymphocytes. These deleted cells may either be mature members of a single B lymphocyte lineage or members of one of two distinct lineages of B lymphocytes, With this in mind, we produced antisera which identify the subpopulation(s) of B lymphocytes absent in mice with the xid determined defect. Although a variety of immunization procedures were tried, the one which was successful involved Purchase Meclizine Online immunization of C57BL/6 mice with DBA/2 spleen cells. This serum, after absorption with DBA/2 thymocytes and spleen cells from defective (CBA/N X DBA/2)F-, male mice, contains two types of antibodies, anti-Lyb5.1 and anti- Lyb7.1, which recognize antigenic determinants expressed on (DBA/2xCBA/N)F-, male B lymphocytes but not on cells from the defective (CBA/NxDBA/2)F, male. The determinants recognized by these antibodies are each determined by an independently segregating gene. The Lyb5 gene is not linked to the H-2 , Igh , Mis , or Lyb4 loci. The Lyb7 gene is also unlinked to the H-2 , Mis, and Lyb4 loci, but it is loosely linked to the Igh gene complex being located approxi- mately 20 to 25 centimorgans from the IgGg locus. The Lyb5 antigen is detected by the aoility of anti-Lyb5 antisera to lyse B lymphocytes in the presence of complement. By contrast, anti-Lyb7 antisera does not lyse B lymphocytes but it does inhibit the in vitro response of B ymphocytes to TI-2 but not TI-1 Meclizine Buy antigens. This result is particularly pro- vocative because it indicates that a membrane molecule encoded by a gene linked to, but Meclizine Online separate from, the Igh gene complex has an important role in regulating B lymphocyte activation. 22-19 ZOl-AI-00030-12 LI We decided to search for monoclonal antibodies directed at markers of mouse B lymphocyte subpopulations, both for the intrinsic value of any such antibodies, and in the hope that we might find one which identified Lyb5 or Lyb7. To do so, we immunized Lewis rats with DBA/2 spleen cells or DBA/2 B lymphocytes, fused the rat cells to NS-1 and screened, using the FACS, for monoclonal antibodies that bound B cells, or subsets of them. Through this approach, we have obtained several Generic Meclizine anti-B cell antibodies, most of which bind all B cells. In some instances, these antibodies identify antigens expressed to different extents on B cell subsets, as indicated by the finding that cells from mice with the xid determined defect may express more or less antigen than control cells. Of these, three are of special interest. They^are 17C9 which recognizes the ThB antigen; 14D10 which recognizes most Ig cells and another group of Ig~ cells which may include B_cell Order Meclizine precursors, and 14G8, which appears to be Buy Cheap Meclizine principally found on Lyb5 B cells. We are now in the process of a more precise deter- mination of the cell types upon which these cells are expressed and in efforts to purify and to functionally test these cells. 3. Responses of xid Mice to Polysaccharide Determinants The study of the xid determined B lymphocyte defect of CBA/N mice has been a major interest of Order Meclizine Online our laboratory for a considerable period of time. We have recently extended our study of this genetic model in several directions in an effort to gain greater insight into the nature
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