摘要:

AbstractThe technique of two‐dimensional gel electrophoresis has been applied to study the changes in protein patterns occurring in murine T lymphocytes after activation by a T cell mitogen. The protein maps of individual T cell clones obtained by limiting dilution were also compared. Whole cell lysates from [35S]methionine‐labeled cell clones or cell populations were analyzed and radiofluorographs of gels compared.We found that, during the events of differentiation and clonal proliferation, a T cell population undergoes dramatic changes resulting in pronounced changes in the protein pattern. In a typical gel with about 1000 detectable spots, there were about 50 changes from one day to the next. Certain regions of gels were compared using highly exposed films; in these, a much higher proportion of spots was found to vary (47/255 spots).Furthermore, we found that T cell clones obtained by limiting dilution yield a protein pattern that contains both “stable” and “diverse” regions. In the molecular weight region from about 35000–43000, there are 0 to 5 differences when one clone is compared with another. Due to a relatively small number of clones analyzed [16], it is probable that the revealed changes are only a fraction of the diversity which remains to

ISSN:0014-2980    

DOI:10.1002/eji.1830140902

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe technique of two‐dimensional gel electrophoresis has been applied to study the changes in protein patterns occurring in murine B lymphocytes after activation with the synergistic B cell mitogens lipopolysaccharide and dextran sulfate. Whole cell lysates from [35S]methionine‐labeled mitogen‐activated B cells were analyzed and radiofluorographs of the gels compared. We found that a large number of proteins appear and disappear during the 5‐day activation period (74/206 spots of a nonimmunoglobulin region of the gel). The B cell pattern (in the nonimmunoglobulin region) was also compared with that of mitogen‐activated T cells. We identified 19 T cell‐specific and 29 B cell‐specific proteins, the remaining 1000 or so proteins

ISSN:0014-2980    

DOI:10.1002/eji.1830140903

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractNanomolar concentrations of 4β‐phorbol 12,13‐dibutyrate markedly enhanced the redistribution of concanavalin A receptors, the common leukocyte antigen and the Lyt‐3 antigen in human blood lymphocytes, as measured by cap formation. The effect on the lateral mobility of these cell surface molecules was dose dependent and occurred within a few minutes of treatment. 12‐O‐Tetradecanoyl phorbol 13‐acetate, another tumor promoter, was similarly active. 4α‐Phorbol 12,13‐didecanoate, which does not have tumor‐promoting activity, did not enhance cap formation. The effect of various drugs and treatments indicated that the phorbol ester‐enhanced cap formation was energy and temperature dependent and required functional microfilaments. Retinoic acid, an antitumor‐promoting agent, was inhibitory and trifluoperazine, an inhibitor of calmodulin‐dependent processes, had a minor inhibitory effect. Protein secretion and synthesis, extracellular Ca2+/Mg2+and functional microtubules did not seem to be involved. The enhanced capping was inhibited by the alkylating agents tosyl phenylalanyl chloromethyl ketone and tosyl lysyl chloromethyl ketone but not by other protease inhibitors. The effect of various amino acid derivatives suggested the participation of an esterase. A comparative study of dose response, kinetics and sensitivity to drugs indicated a direct correlation between the phorbol 12,13‐dibutyrate‐enhanced redistribution of membrane glycoconjugates and the phorbol ester‐induced binding (adhesion) between human blood lymphocytes, a

ISSN:0014-2980    

DOI:10.1002/eji.1830140904

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractNIMF4‐31.7 is a monoclonal IgM antibody with affinity for terminal galactose residues of membrane glycoproteins, Sepharose and agarose. It is cytotoxic for a subpopulation of murine thymocytes and peripheral lymphocytes, corresponding to the peanut agglutinin‐positive population, but reacts with a majority of cells after neuraminidase treatment. It has revealed a genetic linkage between the Thy‐1 locus and quantitative expression of carbohydrate determinants on the thymocyte mem

ISSN:0014-2980    

DOI:10.1002/eji.1830140905

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractTo elucidate the rapid destruction of allogeneic bone marrow (BM) grafts in nonsensitized rats we have assessed: (a) the 21‐h tissue localization of51Cr‐labeled allogeneicvs. syngeneic BM cells, (b) the ability of allogeneic BM cells to proliferate in the BM of irradiated recipients, (c) the survival of allogeneicvs. syngeneic BM cells in cell‐impermeable diffusion chambers implanted into the peritoneal cavity of rats, and (d) the destruction of allogeneicvs. syngeneic labeled BM cellsin vitroby effector cells from various lympho‐myeloid tissues.Some allogeneic BM cells were destroyed within 24 h after transfer to athymic nude rats, thus demonstrating the thymus independence of the cytotoxicity. Furthermore, allogeneic BM cells also failed to proliferate in the BM of irradiated recipients. However, the survival of allogeneic BM cells was not impaired if they were sheltered from host cells within cell‐impermeable diffusion chambers over a culture period of 4 days. But when the allogeneic BM cells were similarly cultured in hosts presensitized against the BM donor, a substantial reduction in BM cell survival was observed. The effector cells of allogeneic BM cytotoxicity (ABC) were present in the spleen because PVG‐rnu spleen cells were highly effective in lyzing BM cells from the AO, but not the PVG strainin vitroafter only 4 h of culture. Some ABC activityin vitrowas also found for peripheral blood lymphocytes and lymph node cells, but not for BM cells themselves. Taken as a whole these data provide firm evidence that the “natural resistance” to BM allografts is basically different from conventional immune responses, and in most, but not all respects resembles natural

ISSN:0014-2980    

DOI:10.1002/eji.1830140906

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractIn an attempt to identify the site on IgE which binds with high affinity to the Fcϵ receptor (FcϵR) on mast cells, we established monoclonal anti‐IgE antibodies (mAb) by fusion of myeloma cells with rat splenocytes immunized with purified murine IgE mAb. Six individual mAb were found to react with various IgE mAb of different specificities and not with immunoglobulins of other classes. Three different clusters of epitopes on the Fcϵ portion could be detected by antibody competition studies. These antigenic determinants were expressed on the Fcϵ portion and required the two heavy chains in their native conformation. Two groups of mAb and their Fab′ fragments completely inhibited the binding of125I‐labeled IgE to rat basophilic leukemia cells (RBL), and one mAb inhibited the specific IgE binding only partially (55–65%). Likewise, the Fab′ fragments of the purified mAb inhibited the antigen‐mediated, IgE‐dependent, serotonin release of RBL cells. Thesein vitrofindings were confirmed byin vivoexperiments, which demonstrated that the anti‐IgE mAb could specifically block passive cutaneous anaphylaxis reaction when injected i.d., before challenging with the antigen. The differences in blocking reactivity of the various anti‐IgE mAb are discussed in view of heterogeneity in the

ISSN:0014-2980    

DOI:10.1002/eji.1830140907

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractA series of B cell hybridomas was used as a model system to assess quantitatively the role of Ia molecules in antigen presentation to allo‐ or soluble antigen‐reactive T cell clones. These hybrid cell lines were established by fusion between the HGPRT−BALB/c B cell lymphoma M12.4.1 and LPS‐stimulated spleen blasts from B10.BR (H‐2k) mice. Quantitative cellular absorption of appropriate anti‐Ia monoclonal antibodies and flow cytofluorometric analyses revealed that the B cell hybridomas examined herein expressed constitutively a number of surface I‐Akor I‐Ekmolecules that varied in an order of magnitude of 1 to 5. Such quantitative differences could be correlated precisely with (a) the capacity of B cell hybridomas to activate T cell clones to proliferate and/or to produce interleukin 2 in response to Eβkallodeterminant or to poly(Glu60Ala30Tyr10) presented in the context of I‐Akrestriction element, and (b) the amount of monoclonal anti‐I‐Akantibody required to inhibit antigen presentation to T cell clones. The possible implications of these data are discussed in the context of current models of regulation of Ia antigen expression by an

ISSN:0014-2980    

DOI:10.1002/eji.1830140908

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractSequential development of antibody titer, functional avidity, helper and suppressor activities were investigated in rhesus monkeys. These were immunized with a single dose of 0.1 μg to 10 mg of a streptococcal protein antigen (SA) in aluminium hydroxide. The IgG antibody titers followed the classical pattern first established in mice, of high‐dose and low‐dose tolerance with intermediate doses of immunity. This was correlated with a similar pattern of functional avidity of IgG antibodies, as measured by a dissociation assay. Helper and suppressor functions were assayed in parallel by inducing the corresponding factors from monkey lymphocytes in Marbrook flasks and testing the factors which cross the species barrier in cooperative cultures with CBA mouse spleen B cells. A progressive modulation of helper and suppressor activities was elicited by the increasing doses of SA, during the initial 28 days after immunization. Thus, dominant suppressor with minimal helper activity, IgG antibody titer and functional avidity were elicited by 0.1 μg SA. However, 1 or 10 μg SA induced dominant helper with minimal or transient suppressor activity and high IgG antibody titers and functional avidity. Somewhat intermediate responses were elicited by 100 μg SA, but 1 mg and especially 10 mg SA induced dominant suppressor and minimal helper activity, with low IgG antibody titers and functional avidities. When the immune response was established, about 28 days after immunization, the intermediate dose of SA elicited IgG antibodies with high titer and functional avidity, high T cell helper but low suppressor activities. In contrast, both high‐ and low‐dose SA induced partial tolerance, with low IgG antibody titer, functional avidity and T cell helper activity. These studies suggest cyclical development of helper and suppressor functions during the 4 weeks after immunization. The emergence of a dominant helper or suppressor function is antigen dos

ISSN:0014-2980    

DOI:10.1002/eji.1830140909

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractSubcutaneous immunization with hen egg‐white lysozyme (HEL) in complete Freund's adjuvant induces, both in antibody responder and nonresponder mice, a classical delayed‐type hypersensitivity (DTH) reaction evaluated as footpad swelling. This response can be specifically transferred to naive recipients by Lyt‐1+2−T cells and passive transfer is restricted by genes mapping in or to the left of the I‐A region of the H‐2 complex.Fine antigenic specificity analysis shows that HEL‐primed T cells mediating DTH recognize ring‐necked pheasant egg‐white lysozyme, a lysozyme closely related to HEL, but fail to respond to human lysozyme, differing from HEL at 40% amino acid residues. Complete cross‐reactivity between native and denaturated (reduced and carboxymethylated) HEL is exhibited by T cells involved in the DTH response. Subcutaneous injection of HEL coupled to spleen cells is also able to induce antigen‐specific and genetically restricted DTH responses whereas the same cells administered by i.v. or i.p. route induce predominantly suppressor T cell activation. These suppressor T cells specifically inhibit the induction phase of

ISSN:0014-2980    

DOI:10.1002/eji.1830140910

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractCulture supernatant from a monoclonal T cell lymphoma line (LH8‐105) obtained by radiation leukemia virus‐induced transformation of hen egg‐white lysozyme (HEL)‐specific suppressor T lymphocytes is able, when injected into mice, to specifically suppress the delayed‐type hypersensitivity (DTH) reaction induced by HEL. The suppressor T cell factor (TsF) exhibits fine antigenic specificity since it suppresses the DTH response induced by HEL without affecting the DTH response induced by ring‐necked pheasant egg‐white lysozyme (REL), a lysozyme closely related to HEL. Conversely, LH8‐105 TsF is able to suppress the DTH response induced by human lysozyme, distantly related to HEL but sharing a common epitope critical for induction of suppressive activity. The fine antigenic specificity of LH8‐105 TsF for a restricted epitope on the HEL molecule is confirmed by binding to HEL but not to REL immunosorbents. This TsF also bears I‐J determinants, as demonstrated by binding to monoclonal anti‐I‐J immunosorbents, and it suppresses the afferent but not the efferent phase of the DTH response to HEL. The afferent suppression is controlled by genes apparently mapping in the I‐J subregion of the H‐2 complex since I‐J‐incompatible mice are not suppressed by LH8‐105 TsF injection. This inducer‐type TsF induces second‐order effector suppressor T cells only in HEL‐primed mice indicating the primary role of antigen, in association with H‐2 (I‐J) products, in the a

ISSN:0014-2980    

DOI:10.1002/eji.1830140911

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY