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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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