摘要:

AbstractThe activation of murine B cells and macrophages by synthetic lipid A analogues, solubilized with triethylamine and complexed with bovine serum albumin, were investigatedin vitro. The analogues used are nonphosphorylated, C‐1 or C‐4′ monophos‐phorylated and C‐1,4′ diphosphorylated derivatives of β‐1,6‐linked D‐glucosamine disaccharide possessing both ester‐ and amide‐bound fatty acid substituents. They were divided into 4 groups, A, B, C and D in terms of the fatty acid substitution. Ester‐ and amine‐bound fatty acids of the analogues are both tetradecanoic acids (C14) in group A, C14and (R)‐3‐hydroxytetradecanoic acids (C14‐OH) in group B, both C14‐OH in group C and C14‐OH and (R)‐3‐tetradecanoyloxytetradecanoic acids in group D.Mitogenic activity was exhibited in spleen cells from C3H/HeN mice by the C‐1 monophosphorylated analogues in groups A and B, and by the C‐4′ monophosphorylated analogues in groups B, C and D, but not in cells from C3H/HeJ mice. Nonphosphorylated analogues in groups A, B and C, and C‐4′ monophosphorylated analogue in group A showed negative mitogenic activity. Only the nonphosphorylated analogue in group D exhibited mitogenic activity in spleen cells from C3H/HeJ mice as well as those from C3H/HeN mice. None of the other analogues exhibited the activity in C3H/HeJ spleen cells.Polyclonal B cell activation activity was exhibited by the C‐1 monophosphorylated analogues in groups A and B, and by the C‐4′ monophosphorylated analogues in groups C and D. Nonphosphorylated analogues in all groups and C‐4′ monophosphorylated analogues in groups A and B showed negative PBA activity.None of the analogues tested could induce any cytostatic mac

ISSN:0014-2980    

DOI:10.1002/eji.1830140202

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe responses of resting human B lymphocytes to a variety of activation signals were studied. Human tonsillar B lymphocytes were separated according to size by countercurrent elutriation. The small B lymphocytes were then stimulatedin vitrowith various concentrations of anti‐μ antibody in the presence or absence of B cell growth factor (BCGF) or withStaphylococcus aureusCowan strain I (SAC). Cellular volume changes and RNA synthesis were measured over the first 24 h of stimulation and were similar with either 15 μg/ml of anti‐μ, 100 μg/ml of anti‐μ, or SAC. In the subsequent 24 h, however, substantial increases occurred in the amount of RNA synthesis and cell enlargement only in those cultures stimulated with 100 μg/ml of anti‐μ or SAC, but not in the cultures stimulated with 15 μg/ml of anti‐μ. The addition of BCGF to those cultures stimulated with 15 μg/ml of anti‐μ did not alter the increases in cellular volume and RNA synthesis found 24 h after stimulation with anti‐μ alone. However, over the subsequent 24 h, the presence of BCGF in culture enhanced both B cell volume changes and RNA synthesis, when compared to cultures stimulated with 15 μg/ml of anti‐μ alone. In addition, BCGF enhanced DNA synthesis in cultures stimulated with low and high concentrations of anti‐μ. DNA content changes following stimulation with anti‐μ, anti‐μ plus BCGF, and SAC were also measured using propidium iodide staining and flow cytometry. Optimal concentrations of anti‐μ induced 20% of the resting B cells to enter S phase, while optimal concentrations of anti‐μ plus BCGF or SAC induced approximately 40%. Finally, prestimulation of resting B cells for 24 h with a low concentration of anti‐μ, sufficient for cell enlargement but not S phase progression, allowed for rapid entrance of the prestimulated B cells into S phase when a high concentration of anti‐μ or SAC was added. These findings suggest the existence of a control point in the progression of human B cells through the cell cycle. This control point is located in the G1phase of the cycle and is reached 24 to 36

ISSN:0014-2980    

DOI:10.1002/eji.1830140203

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe aim of the present study was to define the cell lineage of mixed lymphocyte culture (MLC)‐induced natural killer (NK) effector cells. Human MLC cells were plated under limiting microculture conditions in the presence of irradiated spleen cells and interleukin 2‐containing supernatant. After 18 days, microcultures were scored for proliferation and for cytolytic activity against specific lymphoblasts and NK‐sensitive K562 target cells. About 1 in 7 and 1 in 5 proliferating microcultures had specific or NK‐like cytolytic activity, respectively. Moreover, several microcultures exhibited dual (specific and NK‐like) cytolytic activity, even when they had been established at relatively low numbers of responding cells/well (0.5–0.25) to ensure a high probability of monoclonality. Direct evidence for the existence of cytolytic effector cells with dual activity was achieved by using clones derived from single MLC T cells by micromanipulation. Out of 26 cytolytic clones so derived, 16 exhibited specific cytolytic activity, whereas 22 lysed K562 target cells. More interestingly, 12 of these 26 clones were active against both types of target cells. Only one of these clones was able to lyse autologous or unrelated target cells. In contrast, all such clones lysed the NK‐sensitive cell lines G11, MOLT‐4, Raji, Daudi, Chang and T‐24. Addition of saturating amounts of B9‐4 monoclonal antibody in the lytic assays resulted in the inhibition of the specific cytolysis, but not the NK‐like activity of clones with dual cytolytic activity. It thus appears that (a) alloreactive cytotoxic T lymphocytes can mediate both specific and NK‐like cytolysis and (b) two independent recognition structures are involve

ISSN:0014-2980    

DOI:10.1002/eji.1830140204

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe antigenic specificities, major histocompatibility complex restrictions and functional properties of influenza virus‐specific proliferative cloned cell lines have been studied. These lines were specific for the H3 hemagglutinin subtype of influenza A viruses. By using a large panel of HLA‐phenotyped antigen‐presenting cells, it was found that the polymorphic structures, defined as DR1 and DR7 molecules, or closely associated structures, function as the restricting elements. We excluded for these lines a possible restricting role of supertypic specificities, known cross‐reacting elements on DR molecules, or products of other loci in known linkage disequilibrium with the HLA‐DR molecules. Such exquisitely restricted clones might be of great help in the class II typing of antigen‐presenting cells. Their specific activity was stable for several months. This has allowed the study of some functional properties of these long‐term‐cultured cloned cell lines: interleukin 2 sensitivity and production, helper function in specific antibody synthesis and ability to stimulate in mixed leuk

ISSN:0014-2980    

DOI:10.1002/eji.1830140205

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe immunogenicity of molecules shed by schistosomula into culture medium (antigens present in schistosomula‐released products, SRP‐A) has been studied. The results obtained show that SRP‐A preferentially induce an IgE response when injected into rats, without the need for adjuvants. Moreover, anti‐SRP‐A IgE is cytotoxicin vitrofor the larvae in the presence of macrophages, eosinophils or platelets, which have previously been demonstrated as being the three efficient killer cells for schistosomula in the presence of specific IgE. Immunofluorescence analysis locates the target antigens at the schistosomulum surface. Among the antigens recognized by anti‐SRP‐A IgE, two molecules of 26 and 22 kDa have been identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by wes

ISSN:0014-2980    

DOI:10.1002/eji.1830140206

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe relationship of la expression and antigen‐presenting function by macrophages has been evaluated. When macrophages are maintained in standard culture media, both Ia antigens and accessory cell function are lost. The reacquisition of these properties follows exposure to an Ia‐inducing lymphokine, for which cDNA‐derived interferon‐γ may substitute. The induction of function is related quantitatively to the level of Ia expression. Moreover, both properties reflect newly expressed Ia determinants, since treatment with anti‐I‐A plus complement at the beginning of culture diminishes neither the subsequent level of Ia expression nor function. Treatment with anti‐Mac‐1 plus complement, however, reduces function commensurate with the effectiveness of macrophage depletion. Finally, we find that fixation of macrophages after exposure to antigen does not inhibit antigen presentation, indicating that metabolic activity, while required for antigen processing, is not necessary

ISSN:0014-2980    

DOI:10.1002/eji.1830140207

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractThe cell‐mediated cytotoxic response against autologous cells modified with the sulfhydryl reagent I‐AED [N‐iodo‐acetyl‐N‐(5‐sulfonic‐1‐naphthyl) ethylene diamine] has been described by Levy, R. B., Shearer, G. M., Richardson, J. C. and Henkart, P. A., [J. Immunol.1981.127:523.]. We have established two H‐2Db‐ and eight H‐2Kb‐restricted, AED ‐specific long‐term cytotoxic T lymphocyte (CTL) clones of C57BL/10 origin. The growth of these clones has been dependent upon presence of both antigen and interleukin 2.Cytotoxicity and proliferation analysis of AED‐specific Kb‐restricted CTL clones with target and stimulator cells from Kbm‐mutant mice demonstrated two categories of clones (A and B) based on different reactivity patterns against hapten‐modified Kbm‐mutant cells. AED‐modified target cells of bm5, bm6 and bm9 origin were lysed by type A clones but not by type B clones, in contrast to AED‐modified bm3, bm8 and bm11 target cells which were lysed by type B but not by type A clones. None of the clones lysed AED‐modified bm1 target cells, but all of them lysed AED‐modified bm4 and bm10 target cells. The restriction fine specificities for both cytolytic and proliferative activities of the analyzed clones were identical.A panel of monoclonal antibodies (mAb) directed against the Kbmolecule was used to inhibit lysis of AED‐modified target cells by CTL clones. mAb which recognize a cluster of allodeterminants located in the second external domain (C1 domain) of the Kbmolecule [Hämmerling, G. J., Rüsch, E., Tada, N., Kimura, S. and Hämmerling, U.,Proc. Natl. Acad. Sci. USA1982.79:4737] only inhibited type B clones, whereas inhibition of both type of clones was observed with mAb specific for allodeterminants clustered in the N‐terminal region of Kb.These findings suggest that different regions on the Kbmolecule serve as self determinants for H‐2‐restricted cytotoxicity. We also demonstrate that allodeterminants recognized by mAb and self determinants involved in H‐2 restriction need not be identical. Our data also support the notion that covalent AED modification of H‐2 molecules is not nec

ISSN:0014-2980    

DOI:10.1002/eji.1830140208

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractWhile T cell proliferation to antigen in the presence of antigen‐presenting cells is well known to be readily inhibited by antibodies directed against Class II major histocompatibility complex (MHC) (Ia/HLA‐DR) products, it has not been possible to inhibit proliferation by antibodies directed against the antigen. Because of the implications of these observations for targets of T cell recognition, this phenomenon was reinvestigated using human T cell clones, recognizing a small (24 amino acid) synthetic peptide (termed p20) derived from the influenza hemagglutinin‐1 molecule. It was found that proliferation of clones to p20 was inhibited efficiently (>90%), using p20 as antigen, and rabbit anti‐p20. Inhibition was possible either by coculturing p20 antigen and antibody to p20 with cloned T cells and antigen‐presenting (E−) cells, or by pulsing antigen‐presenting cells with antigen prior to a brief incubation with antibody before washing the E−cells and using them to stimulate cloned T cells. These results do not indicate why previous attempts had failed, but in view of the different techniques available now (cloned T cells, small synthetic polypeptides, and antibody raised against polypeptide) we investigated the influence of these parameters. It was found that, using cloned T cells, the form of the antigen was of importance, as antibody inhibition of the response to hemagglutinin or whole influenza A was much less apparent. These differences were interpreted as being due to greater access of anti‐p20to p20than to hemagglutinin or influenza. If uncloned T cell lines were used, inhibition was also much harder to detect. This was interpreted as masking of inhibition of the response of some clones in the line by interleukin 2‐

ISSN:0014-2980    

DOI:10.1002/eji.1830140209

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractPrevious work has revealed that the T helper cell (Th) responses to an antigenic determinant of Vλ2315(called λ2.1) is regulated by both H‐2 and non‐H‐2 genes. In the present study this was confirmed and extended to two other determinants, one shared between free λ2315and λ1J558(called λ2.2) and one unique for free λ1J558(called λ1.1). H‐2 genes regulate the responses to the latter determinants, because BALB.B (H‐2b) mice were low responders and BALB/c (H‐2d) mice were high responders. Thus, the H‐2dhaplotype on BALB/c background was associated with high responder status. However, when the H‐2dhaplotype was examined on other genetic backgrounds than BALB/c, the animals could be classified as either intermediate or low responders, depending upon the non‐H‐2 background. This demonstrated that non‐H‐2 genes also influenced Thresponses. Furthermore, C3H‐H‐20, DBA/2 and B10.D2 mice (all H‐2d) responded to only one (λ2.1) but not the other (λ2.2) of two determinants physically linked on the same polypeptide chain (λ2315). This indicated that the non‐H‐2 gene effect is capable of fine discimination,i.e.the non‐H‐2 gene‐mediated low responder phenotype may at least in part be due to failure of recognition of certain antigenic sites, like the H‐2‐linked Ir‐gene defect. F1hybrids responded to the same determinants as their parental strains;e.g., the BALB/c non‐H‐2 background exerted a dominant influence over

ISSN:0014-2980    

DOI:10.1002/eji.1830140210

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY

摘要:

AbstractTo identify major human antibody clonotypes with specificity to N‐acetyl‐D‐glucosamine (GlcNAc), affinity‐purified antibody preparations of different human individuals were analyzed by isoelectric focusing (IEF). A major clonotype (1A) was identified representing 7% of all anti‐GlcNAc antibodies of one donor (no. 371). Anti‐GlcNAc antibodies of donor 371 were used as immunogen to prepare monoclonal anti‐idiotopic antibodies (mAb). Two anti‐idiotopic mAb were specific for the major clonotype 1A, as shown by blotting of the anti‐GlcNAc antibodies after IEF in thin‐layer agarose to nitrocellulose filters and staining with either iodinated antigen or iodinated anti‐idiotopic mAb. The two anti‐idiotopic mAb 13F15 and 10F59 were further analyzed with respect to the antigenic determinant which they recognize. Both were directed to combining site‐related determinants as shown by inhibition studies. Although 13F15 has a 500‐fold higher binding capacity (relative affinity) equal amounts do not inhibit more than 50% of 10F59 binding, suggesting that the two mAb detect two closely related, but not identical, idiotopes in the antigen combining site of clone 1A. Although clonotype 1A is a unique antibody exclusively found in IEF of donor 371, the idiotope 1A.1, defined by mAb 13F15, is a recurrent determinant detectable on different anti‐GlcNAc spect

ISSN:0014-2980    

DOI:10.1002/eji.1830140211

出版商:WILEY‐VCH Verlag GmbH    

年代:1984

数据来源: WILEY