摘要:

AbstractThe effects of recombinant rat interferon‐y on class II major histocompatibility complex antigen expressionin vivowere studied by immunohistology in LEW rats after continuous intravenous infusion for three days. Interferon‐γ administration led to a systemic induction of class II molecules in previously negative parenchymal and stromal cells. The induction patterns observed were highly reproducible, but not closely dose dependent within a 25‐fold dose difference tested. However, the effect of interferon infusion differed profoundly in individual cell types, and appeared to be related to the differentiation stage of each cell‐ population. Thus, epithelial cells like duct epithelia, urothelium or basal ear skin keratinocytes as well as endothelia in big vessels were strongly and easily induced for class II antigen expression. Parenchymal cells like cardiomyocytes and hepatocytes showed intermediate reactivity, while capillary endothelia, neurons in the brain, straight proximal kidney tubules or endocrine pancreatic islet cells did not express class II antigens. The induced expression was rapidly lost from most cells within one or two days after interferon withdrawal; the only exception occurred in keratinocytes. Long‐term alterations were, however, still found 14 days after infusion. Interstitial class II‐positive dendritic‐shaped cells were increased in the organs and hepatic Kupffer cells carried class II antigens. On conventional histology all organs appeared perfectly normal at this date. After three days of interferon, cells of an immature myelomonocytic phenotype occluded medium‐sized and small veins in all organs and occurred in granuloma‐like lesions in the liver. Although these cells quickly disappeared after interferon withdrawal they might have been at least partially responsible for single deaths on day three. Our study provides a basis for testing the immunologicalin vivofunction of parenchymal class II antigen expression and its differentiation

ISSN:0014-2980    

DOI:10.1002/eji.1830180502

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractPeripheral blood lymphocytes from a melanoma patient were stimulated with autologous melanoma cells in mixed lymphocyte tumor cultures (MLTC). After three restimulations, the lytic activity of the responder cells directed against the autologous melanoma cells was higher than that against K‐562 and autologous Epstein‐Barr virus‐transformed B cell line (EBV‐B) cells. From these MLTC‐responder cells, we derived specific cytolytic T cell (CTL) clones that lysed the autologous melanoma cells and did not lyse K‐562 or autologous EBV‐B cells. Autologous melanoma clones were found that were resistant to some or all of these CTL clones. The autologous CTL clones recognized at least two different antigens (A, B) on the melanoma cells and three types of melanoma clones could be distinguished (A+B+, A+B−, A−B−). This antigenic heterogeneity of melanoma clones was confirmed by testing the CTL clones in cold target competition and also in antigen‐dependent CTL proliferation assays performed with very small numbers of stimulator cells. The data further indicated an instability of the expression of a melanoma‐associated antigen in the course of a long culture period. Among the melanoma clones that expressed antigen A, one was found to stimulate the proliferation of anti‐A CTL clones much more effectively than the others. This represents a new type of heterogeneity among tumor cells which may be of significance for the elicitation of an autologous ant

ISSN:0014-2980    

DOI:10.1002/eji.1830180503

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractPreviously we studied differential expression of cell surface molecules between the metastatic murine lymphoma ESb and an adhesion variant ESb‐MP. Here we describe the specificity of a monoclonal antibody (12‐15) that showed strong binding to the adhesion variant and weak reactivity against ESb cells. The antibody also reacted to lymphoid but not to macrophage‐derived cell lines and immunoprecipitated a molecule of approx. 60–69 kDa from ESb‐MP cells. N‐terminal sequencing of the' antigen revealed identity to the beta protein of mouse Fcγ receptors. Using monoclonal antibodies against Fcγ receptors (2.4G2 and K9.361) in immunofluorescence assays and cDNA probes specific for alpha, betal and beta2 Fc receptor transcripts in Northern blot experiments the differential expression of Fc receptors in ESb and ESb‐MP cells was confirmed.Biochemical analysis of endoglycosidase F‐treated precipitates revealed that antibody 12‐15 reacted to products of all three transcripts with molecular masses for the protein core of 38.5 kDa (betal), 34 kDa (beta2) and 31 kDa alpha). In addition, an unknown protein of 37 kDa (termed beta3) was identified by antibody 12‐15 which could also be detected in ESb cells and EL4 cells. Antibodies 2.4G2 and K9.361 did not react to the beta 3 chain but reacted to varying extents to the other Fc proteins in macorphage and lymphoid cells. Comparison by peptide mapping of the novel beta3 chain to betal, beta2 and alpha proteins revealed similar, but also distinct peptides. The tissue‐specific reactivity of monoclonal antibody 12‐15 is likely to be due to a carbohydrate epitope associated with all Fcγ receptors in lymphoid but

ISSN:0014-2980    

DOI:10.1002/eji.1830180504

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractPairs of monoclonal antibodies (mAb) defining epitopes T11.1 and T11.2 on the CD 2 molecule are mitogenic for purified human T cells in the presence of a sub‐mitogenic dose of 12‐O‐tetradecanoylphorbol 13‐acetate (TPA). Anti‐CD 28 mAb can substitute for the action of TPA in the anti‐CD 2‐induced proliferative response of resting T cells, whereas each signal alone is unable to mediate this effect. Co‐stimulation by anti‐CD 2 plus anti‐CD 28 mAb is monocyte independent and besides resting T cells also induces strong proliferation of thymocytes and pre‐activated T cells. Modulation of the CD 3‐T cell receptor complex does not inhibit the co‐stimulatory effects of anti‐CD 2 plus anti‐CD 28 mAb. The effect is largely dependent on endogenously produced interleukin 2, since the response is strongly inhibited in the presence of mAb against the 55‐k

ISSN:0014-2980    

DOI:10.1002/eji.1830180505

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractThe processing and presentation of whole irradiatedMycobacterium tuberculosis(Mtbγ) and its purified protein derivative (PPD) by the peripheral blood monocytes from healthy Bacillus Calmette Guérin (BCG)‐vaccinated individuals was investigated. To study processing and presentation as events distinct from T cell recognition and proliferation, monocytes were pulsed with antigens for varying time intervals and fixed. The kinetics of presentation indicate that up to 2 h was required for effective presentation of PPD and 2–4 h for Mtbγ, and that the ability to activate T cells declined as the time interval for which pulsing occurred was increased, so that responses were abolished by 8–10 h. Prefixed monocytes could not present Mtbγ and PPD to T cells indicating that processing was an essential requisite. Lysosomotropic agents chloroquine, monensin, and leupeptin inhibited the presentation of these antigens suggesting the role of lysosomes/endosomes in processing. Furthermore, monocytes incubated with optimal concentration of antigens for different lengths of time released determinants which were still antigenic but circumvented the need for any further processing.Addition of nonprimed syngeneic monocytes, both untreated or paraformaldehyde fixed to cells which had been pulsed and fixed, restored the responses even at the later time periods when responses were not detected. This second interaction of the monocyte with T cells was not major histocompatibility complex restricted in that the addition of monocytes from another donor was equally

ISSN:0014-2980    

DOI:10.1002/eji.1830180506

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractWe reported previously that immature macrophage precursor cells can be isolated from spleen and liver of cyclophosphamide or pyran copolymer‐pretreated mice. We now extended our investigations to livers of normal, untreated specific pathogen‐free mice. Using the response to the macrophage growth factor colony‐stimulating factor‐1 (CSF‐1) and the presence of the mouse macrophage‐specific F4BO antigen as criteria of definition, in the liver of normal mice we could demonstrate macrophage precursor (MϕP) cells by means of proliferation assays and flow cytometric analysis.The amount of MϕP present in the normal liver was significantly increased after administration of pyran copolymer. Also an enhanced proliferative response to CSF‐1 as well as augmented natural killer activity and cytostasis ofCandida albicanswas noted in liver nonparenychymal cells (LNPC) after treatment of bone marrow (BM)‐irradiated, splenectomized mice with pyran copolymer. Since the irradiated BM was actually proven to be silent by assessment of BM number and proliferative capacity and by scoring white blood cells, our findings suggest a response of endogenous liver MϕP under the applied conditions. Further evidence for the presence of endogenous liver hemopoietic cells was obtained from transplantation experiments in which LNPC brought about the survival of lethally irradiated mice. The data point towards a significance of the liver in disposing hemopoietic cells to the organism under impairment of r

ISSN:0014-2980    

DOI:10.1002/eji.1830180507

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractThe antigen receptor of the T lymphocytes is one of the most complex eukaryotic membrane structures studied to date. The T cell receptor (TcR) consists of two disulfide‐linked glycoprotein chains (α/βor γ/δ) and is noncovalently associated with a group of small and invariable CD 3 proteins. Four CD 3 chains have been recognized: two highly homologous glycoproteins CD 3 γ and δ, the more distantly related nonglycosylated CD 3 E chain, and the nonglycosylated CD 3 ζ, the latter being present as a homodimer. The unraveling of the architecture of the TcWCD 3 complex is crucial to our understanding of the processes underlying its assembly, recognition and trans‐membrane signaling. The transmembrane orientation of the TcR chains and of CD 3 γ and CD 3 δ can be directly inferred from their primary structure, based on the presence of concensus N‐linked glycosylation sites N‐terminal of their transmembrane domains. This prediction can not be made, however, for nonglycosylated molecules like the CD 3 E chain. In order to determine the transmembrane orientation of CD 3 E, anti‐peptide antisera directed against the N‐termini of the human and murine CD 3 E chains were generated in rabbits. Both antisera stained intact T cells, demonstrating that the N‐terminus of the CD 3 E chain was located at the outer surface of the plasma membrane. The anti‐human CD 3 E peptide antiserum was found to be mitogenic for peripheral blood T cells, a finding previously reported only for monoclonal anti‐TcW CD 3 reagents. Using a novel transient expression system in murine T lymphocytes, the human CD 3 E chain could be expressed on the surface of CD 3+, but not CD 3−murine T cells, as indicated by fluorescence staining with the anti‐peptide antiserum. This experiment confirmed the specificity of the anti‐peptide antiserum and, perhaps more importantly, indicated that the human CD 3 E chain was correctly assembled in the murine CD 3 complex. Moreover, the anti‐human CD 3 monoclonal antibody UCHT1 was found to stain T cells

ISSN:0014-2980    

DOI:10.1002/eji.1830180508

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractPhorbol esters exert diverse effects on cellular activation and differentiation. CD 7, a differentiation antigen appearing early in T cell ontogeny, may be involved in the activation and differentiation processes. CD 7 was found to be rapidly down‐regulated by 12‐O‐tetradecanoylphorbol l3‐acetate (TPA) from mature T cell surface. The time course of CD 7 down‐regulation was similar to that of other functionally important T cell antigens, CD 3 and CD 4. Within 2 h, TPA at 10 to 30 ng/ml induced a complete down‐regulation of CD 7. Twenty‐four hours later, the reappearance of CD 7 on TPA‐treated cells was observed. This phenomenon was monocyte independent. In contrast, CD 7 expression on thymocytes was resistant to the effect of TPA. In addition, certain leukemic T cells were also resistant to TPA‐induced CD 7 down‐regulation. The mechanism underlying TPA‐induced CD 7 down‐regulation was investigated further. Synthetic diacylglycerol,sn‐l,2‐dioctanoylglycero1, which activates protein kinase C, did not induce down‐regulation of CD 7 on mature T cells. Ionomycin, a calcium ionophore, did not down‐regulate this antigen either. Thus, it is concluded that the processes of protein kinase C activation and/or cytosolic calcium influx are not sufficient for TPA‐induced CD 7 down‐regulation; other pathway

ISSN:0014-2980    

DOI:10.1002/eji.1830180509

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractRecombinant human interleukin 6 (rhIL 6) was injected i.p. into male Wistar rats to investigate its role as a mediator of the acute‐phase response. Hepatic mRNA levels of β‐fibrinogen, α2‐macroglobulin, cysteine proteinase inhibitor, α1‐acid glycoprotein and albumin were measured at different times after the administration of rhIL 6. Maximal increases of mRNA concentrations were observed already 4 h after the injection of rhIL 6 leading to 4.8‐, 19.7‐, 10‐ and 16‐fold stimulations in mRNA levels of β‐fibrinogen, α2‐macroglobulin, cysteine proteinase inhibitor or α1acid glycoprotein, respectively.The rhIL 6‐induced stimulation of acute‐phase protein mRNA was much more rapid than the acute‐phase induction after turpentine, where maximal mRNA levels were found between 16 and 24 h. For all acute‐phase proteins studied, the stimulation of mRNA synthesis was found to be dependent on the dose of rhIL 6 injected. In the case of α2‐macroglobulin mRNA a sex‐specific induction by rhIL 6 was found. Only male rats showed an acute‐phase response, whereas in female rats an acute‐phase reaction of α2‐macroglobulin mRNA was not inducible by IL 6.The increases in mRNA levels of the acute‐phase proteins studied were followed by corresponding changes of the proteins in the serum determined by rocket immunoelectrophoresis. It is concluded that IL 6 represents a potent

ISSN:0014-2980    

DOI:10.1002/eji.1830180510

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY

摘要:

AbstractThe conjugation of two lipophilic p‐oxymethylbenzyl‐3β‐cholestanylsuccinate groups to fully benzylpenicilloylated eicosa‐L‐lysine [BP20Lys20(OSuco)2] greatly potentiated its ability to suppress anti‐BPO IgE antibody formationin vivoin mice. This enhanced tolerogenicity was due to the ability of BPO20Lys20(OSuco)2to induce T suppressor cells (Luescher, I. F. et al.,Eur. J. Immunol.1984.14:68). In the present study the role of adherent cells in the induction of T suppressor cells by BPO20Lys20(OSuco)2was investigated. Low numbers of macrophages pulsedin vitrowith BPO20Lys20(OSuco)2, but not with the hydrophilic analog BP020Lys20‐OH, suppressed anti‐BPO IgE antibody formation upon transfer into syngeneic recipients. This suppression was antigen and IgE specific, and was manifested in naive as well as in previously immunized mice. By cell transfer experiments it was found that the suppression was sensitive to cyclophosphamide. The IgE suppression was not due to carry‐over of BPO20Lys20(OSuco)2by the transferred macrophages since the amount of cell‐associated antigen was approximately 500‐fold too low to account for the observed suppressions.The capability of BPO20Lys20(OSuco)2to elicit anaphylaxis in passively sensitized rats was significantly lower in comparison to the hydrophilic analog BPO21Lys20OH. Adherent cells pulsed with BPO20Lys20(OSuco)2, although strongly suppressing IgE antibody formation, failed to elicit any detectable anaphylaxis. The role of adherent cells in the induction of T cell‐mediated suppression of IgE antibody formation by lipid‐modified antigens is discussed and novel therapeutical concepts to achieve desensitization of drug allergic

ISSN:0014-2980    

DOI:10.1002/eji.1830180511

出版商:WILEY‐VCH Verlag GmbH    

年代:1988

数据来源: WILEY