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Related post: these cultures, very substantial activation occurs. The active principal within the EL-4 supernatants has been previously designated B cell growth factor (BCGF). The properties of BCGF have now been investigated. It is a material which has an approximate molecular weight of 18,000 daltons by neutral gel filtration. Isoelectric focusing reveals that it exists in two forms, one with a pi of approximately 6.4, and the second with a pi of 7.4. Digestion of the B Isordil 30 Mg cell growth factor with neuraminidase yields but a single species with a pi of 9.3. B cell growth factor can be separated Isordil 300 Mg from Interleukin-2 not only by neutral gel filtration and isoelectric focusing, but also by its behavior on phenyl sepharose column chromatography. BCGF binds to phenyl sepharose and is eluted from it with concentrations of approximately 10% ethandiol, substantially less than that required for the elution of Interleukin-2. On sodium dodecylsulphate-polyacryl amide gel electrophoresis, B cell growth factor migrates with a molecular weight of approximately 15,000 daltons. Finally B cell growth factor fails to bind to lines which express receptors for Interleukin-2. Thus, B cell growth factor is an entity clearly independent of Interleukin-2 which appears to be required for the Isordil 60 Mg progress of B cells stimulated with anti-immunoglobulin antibodies to enter the S phase of the cell cycle. Preliminary studies suggest that B cell growth factor is required relatively early in G, rather than at the so-called G-. - G-,. boundary. Attempts to purify B cell growth factor by high pressure liquid chromatography are now in progress as are efforts to obtain reliable in vitro translation from mRNA as a first step in the molecular cloning of the gene which specifies B cell growth factor (Howard and Paul, LI, NIAID; Farrar, Hoffman- LaRoche, Inc. , Nutley, NJ) Activation of Resting B Cells by Non-cognate, Receptor Independent T Cell Interactions . The analysis of the mechanisms by which resting B cells Isordil Tablet are activated has been one of the principal goals of the Laboratory of Immunology scientists. It is generally recognized that at least two distinct pathways exist for this activation process. In one, resting B cells are initially activated as a result of interaction of ligands with their receptors; antigen may interact with antigen binding receptors or anti-immunoglobulin antibodies may act as polyclonal ligands for these receptors. The second major pathway appears to involve the interaction of histocompatibility -restricted helper T cells with B cells through the recognition by the T cell of antigen and class II molecules on B cell surfaces. Such activation Isordil 20 Mg has Isordil 40 Mg been designated cognate activation. During the past year, Laboratory of Immunology scientists have identified a third major pathway of B cell activation which appears to involve the action of T cells but is neither dependent upon a receptor directed signal nor on a cognate interaction. These experiments utilized cloned, antigen specific T cell lines specific for antigens such as the terpolymer of the glutamic acid alanine and tyrosine (GAT) and highly purified populations of resting B cells. It was first demonstrated Isordil 10 Mg that extremely^highly purified populations of resting B cells, containing at least 99% Ig cells, were very effective at stimulating T cell activation as determined by the capacity of T cells to recruit B cells to proliferate. Such highly purified B cells although they activated T cells to cause B cell recruitment, failed to cause T cell proliferation. By contrast, irradiated antigen presenting populations rich in macrophages were excellent stimulants of T cell proliferation, but 20-9 were either inferior to B cells in activating T cells for B cell recruitment or were no better than such populations of B cells. This indicates that the activation of T cells may lead to distinct consequences depending upon the nature of the antigen presenting cell involved in this activation; B cells present antigen to T cells in such a way as to favor the production of factors important in B cell recruitment, whereas, macrophages present antigen to T cells in a way that favors T cell proliferation. A detailed analysis of the B cell recruitment that was caused by T cell activation revealed the following points: 1) Virtually all resting B cells were stimulated to enter the G, phase of the cell cycle by culturing them with a cloned T cell line in the presence of antigen and a major proportion ("^45%) of the B cells that had entered G, were stimulated to enter S phase. This indicated, therefore, that B cells regardless of the specificities of their receptors could be activated by stimulated T cells. 2) There appeared to be no histocompatibility restriction in the activation of the B cells; both syngeneic and allogeneic B cells entered G, and S phases in response to T cell activation if a population of syngeneic antigen presenting cells, either B cells or macrophages, were present to activate the T cells. 3) By varying the concentraton of antigen used for stimulation and the density of the responding cells, this non-histocompatibility restricted T cell recruitment of B cell activation could be made to appear restricted in that only syngeneic cells would be activated. This was interpreted to be due to the greater efficiency of the activation of the B cell which presents antigen to the T cell because of its proximity, than of the activation of allogeneic bystander cells. This result implies that many instances of so-called cognate, histocompatibility restricted T cell-B cell interaction may in fact represent non-cognate interactions dependent upon a T cell activating factor or set of factors capable of acting upon resting B cells Isordil Hydralazine without the need for receptor mediated activation (DeFranco, Ashwell, Schwartz and Paul, LI, NIAID). Measurement of the multispecificity of the combining region of monoclonal anti-DNP antibody by affinity chromatography . An analysis of the binding properties of antibody combining sites as well as the structure of these sites has suggested that they might express the property of multispecificity; that is, that they might bind apparently unrelated ligands through distinct portions of a relatively large combining site. A test of this concept depends upon a simple method of unambiguously measuring the affinity of interaction of a purified monoclonal antibody with a wide range of compounds, both structurally related and not structurally related to the ligand against which the antibody was raised. Laboratory of Immunology scientists have now utilized the quantitative theory of affinity chromatography of Hefcote and DeLisi and extended it so that it could be directly applied to experimental data from the binding of bivalent antibodies to haptens on Buy Isordil affinity columns. The degree of retardation in elution time
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