共查询到20条相似文献,搜索用时 31 毫秒
1.
F J Dumont 《Cellular immunology》1987,105(2):355-365
The murine Ly-6A cell surface antigen is normally present on a minor subset of mature T cells. This marker has been shown to become highly expressed on mitogen-activated T cells. We found that expression of Ly-6A is also markedly increased in resting T cells by incubation with IFN-alpha/beta or IFN-gamma. Here, we compared the effect of the immunosuppressant cyclosporine A (CsA) on Ly-6A induction by IFN and concanavalin A (Con A). The augmentation of Ly-6A expression produced by treatment of T cells with IFN-alpha/beta or IFN-gamma was found not to be affected by CsA concentrations up to 2 micrograms/ml. In contrast, at doses as low as 50 ng/ml, CsA prevented the enhancement of Ly-6A expression in Con A-treated T-cell cultures. Culture supernatant transfer experiments were performed to further explore this effect of CsA. It was found that supernatants from Con A-activated T cells enhanced Ly-6A expression in resting T cells. This activity could be neutralized with an anti-IFN-gamma monoclonal antibody. Supernatants from T cells treated with Con A in presence of CsA lacked Ly-6A-enhancing activity. Taken together, these data suggest that the inhibition by CsA of Ly-6A induction in Con A-treated T cells reflects the known inhibitory effects of the drug on IFN-gamma secretion. This may imply the existence in T cells of an autocrine circuit involving IFN-gamma and regulating Ly-6A expression. 相似文献
2.
3.
The discovery of several monoclonal antibodies provided the impetus to revisit the Ly-6 group of antigens. Our serological data point to the existence of at least five separate Ly-6 antigens. They are distinguished by the patterns of their tissue expression as (1) the classical Ly-6 alloantigen of peripheral lymphocytes (Ly-m6.2A), (2) a bone marrow cell-restricted antigen (Ly-m6.2B), (3) an antigen shared by bone marrow cells and peripheral lymphocytes (Lym6.2C, possibly identical with H9/25),(4) an antigen expressed on bone marrow cells, thymocytes, and peripheral lymphocytes (Ly-m6.2D), and (5) an antigen occurring exclusively on lymphoblasts (Ly-m6.IE, similar to Ala-1). ThB is a sixth distinct antigen of the group. The assumption that separate antigens exist is supported by distinctive distribution patterns in normal and neoplastic tissues. The genes controlling Ly-6 antigens are closely linked, as they are transmitted as two haplotypes only. One incidence of a crossover within the Ly-6 region was observed: the Ly-6B.2 alloantigen was expressed in NZB mice, which type Ly-6.1 for other Ly-6 specificities. 相似文献
4.
R G Palfree S Sirlin F J Dumont U H?mmerling 《Journal of immunology (Baltimore, Md. : 1950)》1988,140(1):305-310
The Ly-6C.2 molecule was purified from K36 tumor cells by affinity chromatography and gel filtration. The electrophoretically homogeneous preparation, with m.w. 15,000, was tested with a panel of antibodies that confirmed the presence of the LY-6C.2 epitope. An N-terminal sequence of 39 amino acids was obtained showing 59% homology with the corresponding portion of the Ly-6A.2 polypeptide. Based on the least homologous (29%) 14 amino acid segment, an oligonucleotide probe was constructed, and Ly-6C.2 cDNA was cloned from a BW5147 cDNA library. A 794-base pair cDNA containing the entire coding region had 82% homology with Ly-6A.2 cDNA. The encoded polypeptide sequence of 131 amino acids containing a perfect correlation with the N-terminal sequence data was 63% homologous with that of Ly-6A.2. The greatest homology was in the leader, first 16 N-terminal and last 39 C-terminal amino acids. The latter are likely to be important in determining the attachment of glycophosphatidylinositol. Despite results indicating fewer disulfide constraints in the Ly-6C molecule, the predicted sequence contains 10 cysteine residues nearly perfectly matched with those predicted in Ly-6A. 相似文献
5.
Rat monoclonal antibodies YE3/19.1, defining the murine-activated lymphocyte antigen MALA-1, and D7, detecting an Ly-6 locus-controlled antigen, bound highly purified Ly-6E.1. On western blots of lymphocyte surface proteins which had been solubilized and electrophoretically separated in octylglucoside, they detected bands which comigrated with Ly-6A.2 or Ly-6E.1 antigens. On cells or in an immunoassay they blocked alloantibodies against Ly-6A.2 or Ly-6E.1. The tissue distribution of MALA-1 also correlated with Ly-6A.2 or Ly-6E.1. Upon octylglucoside or sodium dodecyl sulfate-polyacrylamide gel electrophoresis, these antigens displayed similar sizes. Thus, Ly-6A.2 and Ly-6E.1 are most likely products of alternate alleles. Electrophoretic analysis showed a similar size and charge for Ly-6A.2, Ly-6B.2, Ly-613.2, and Ly-27.2. Ly-6C.2 and Ly-28.2 appeared to be identical, and were similar in size to Ly-6A.2, but they differed in charge and in intrachain disulfide constraints. Since Ly-613.2 and Ly-27.2 may represent the same or different epitopes on the Ly-6A.2 molecule, the previously postulated five Ly-6-like antigens that were thought to be separable on the basis of tissue distribution, may represent no more than three separate proteins which can be assigned to one of two distinct categories by electrophoretic mobility in gels containing octylglucoside. 相似文献
6.
Role of Ly-6 in lymphocyte activation. I. Characterization of a monoclonal antibody to a nonpolymorphic Ly-6 specificity 总被引:3,自引:0,他引:3
G Ortega P E Korty E M Shevach T R Malek 《Journal of immunology (Baltimore, Md. : 1950)》1986,137(10):3240-3246
In studies with alloantisera and monoclonal antibodies (mAb) a number of antigenic determinants have been defined that are the products of the Ly-6 locus on murine chromosome 2 and that are expressed primarily on B and T lymphoid cells. It remains controversial whether these antigenic determinants are encoded by a single gene or a multigene complex. We have characterized a new rat mAb, D7, which recognizes a cell surface antigen whose expression on nonactivated peripheral lymphocytes varies from strain to strain. The phenotype of the staining profile, i.e., high or low percentage of D7-positive cells, mapped to the Ly-6 locus as assayed by strain distribution studies, RI lines, and Ly-6 congenic strains. The binding of D7 to Ly-6.1-positive strains could be inhibited by mAb directed to the Ly-6E.1 specificity, whereas D7 could inhibit the binding of mAb specific for Ly-6A.2 to cells from Ly-6.2-positive strains. Coprecipitation studies followed by Western blot analysis confirmed that D7 reacts with both Ly-6E.1- and Ly-6A.2-bearing molecules. The most likely explanation for these findings is that Ly-6A.2 and Ly-6E.1 represent allelic specificities. Further dissection of the complexity of the Ly-6 antigen system and determination of its possible functional importance in lymphocyte activation should be greatly facilitated by the availability of xenogeneic mAb that recognize framework determinants on multiple Ly-6 products. 相似文献
7.
Using immunofluorescence with a monoclonal anti-Ly-6.2 antibody and FACS analysis we have confirmed that the Ly-6.2 antigen is present on approximately 70% of mature T cells and B cells but on few immature lymphocytes. There is a wide range of antigen density among the Ly-6.2+ populations, with the mean density higher on T cells than B cells. Following Con A activation of splenocytes there was a sixfold increase in Ly-6.2 antigen density though approximately 20% of the activated lymphocytes were Ly-6.2?. The increase in Ly-6.2 density was specific since similar density increases did not occur for the closely linked antigens ThB and H . By panning a predominantly T-cell population for Lyt-2-bearing cells, it was found that Lyt-2+ lymphocytes were either negative or dully staining for Ly-6.2. However, activated cells bearing the Lyt-2 antigen were all Ly-6.2 positive. Double-staining experiments showed that T cells which had high Ly-6.2 antigen densities also had high Thy-1 antigen densities. Corticosteroid-resistant thymocytes were highly enriched for Ly-6.2-bearing cells compared to untreated thymocytes and had staining profiles for Ly-6.2 which were similar to peripheral T cells, supporting the idea that steroid treatment selects for a phenotypically mature thymic population. 相似文献
8.
Kazuo Dairiki Shuji Nakamura Shuji Ikegami Madoka Nakamura Takao Fujimori Norikazu Tamaoki Nobuhiko Tada 《Immunogenetics》1989,29(4):235-240
The mouse lymphocyte surface alloantigen, Ly-31, defined by monoclonal antibody N1.10 (IgG2b, k) and controlled by a gene locus closely linked to theAkp-2 locus on chromosome 4, was biochemically investigated. By employing a quantitative immunoassay system, it was found that the Ly-31.1-specific antibody detected an allotypic determinant of mouse alkaline phosphatase. Ly-31.1, i. e., mouse alkaline phosphatase, was expressed predominantly in kidney and bone and was also detected in placenta, lung, and testis. Concerning tumor cell lines, they varied in the amount of antigen present, with both T and B lymphoid lineages selectively possessing the antigen. In normal lymphoid tissues, lesser amounts of antigen were detected. The binding of mouse alkaline phosphatase to Ly-31.1-specific monoclonal antibodies was specific in nature. The Ly-31.1 antigen was immunoprecipitated from the lysates of surface-radiolabeled YAC-1 moloney leukemia cells, and appeared as a single band of about 78 000 under both reduced and nonreduced conditions on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Furthermore, treatment of tumor cell lines with phosphatidylinositol-specific-phospholipase C resulted in the removal of Ly-31 antigen from the cell surface. These results suggest that a gene cluster containing theLy-31 andAkp-2 loci which control the alkaline phosphatase is formed on mouse chromosome 4. The Ly-31 antigen is the first enzyme demonstrated to be a lymphocyte surface alloantigen. 相似文献
9.
A monoclonal antibody that recognizes an Ly-6-linked antigen inhibits the generation of functionally active T cell subsets 总被引:4,自引:0,他引:4
P M Flood D B Murphy M Horowitz K P LeClair F R Smith E Stockert M A Palladino A B DeLeo 《Journal of immunology (Baltimore, Md. : 1950)》1985,135(1):63-72
A monoclonal antibody (mAb) generated against the chemically-induced BALB/c Meth A sarcoma, designated HD42, reacts in cytotoxic tests with Meth A as well as with BALB/c peripheral lymph node cells and mitogen-activated spleen cells. The antigen was detected by FACS analysis on BALB/c spleen and lymph node cells, and by absorption assays on all normal lymphoid cells of BALB/c but not B6 mice. The expression of the antigen was not found on normal adult lung fibroblasts, on brain, nor on an extensive panel of tumors of BALB/c and B6 origin. Because the strain distribution of the antigen is reciprocal to that of Ly-6.2 and is not expressed in congenic C3H.Ly-6b mice, we have tentatively defined it as Ly-6.1 and referred to the mAb as alpha-Ly-6.1. The presence of alpha-Ly-6.1 abrogates both the Con A-induced and the IL 2-dependent proliferative response of normal T cells, whereas the response of normal B cells to LPS remains unaffected. alpha-Ly-6.1 is a potent suppressor of the primary in vitro plaque-forming cell (PFC) response to SRBC. Pretreatment of normal splenic T cells with alpha-Ly-6.1 and complement had no effect on the ability of these cells to generate in vitro either T helper cells (TH) or T suppressor cells (TS) to SRBC. However, addition of antibody in the absence of complement during the generation of TH or TS, or posttreatment of these T cell subsets with antibody and complement after in vitro education, completely removed the functional activity of these cell types. Addition of alpha-Ly-6.1 to MLC suppressed the MLR as well as the generation of cytotoxic lymphocytes (CTL), whereas the presence of the antibody during a cell-mediated lympholysis (CML) had no effect. Therefore, it appears that alpha-Ly-6.1 recognizes an antigen that is important for the generation of TH and TS cell subsets. 相似文献
10.
Dr. Fumio Takei 《Immunogenetics》1982,16(3):201-208
H9/25, an allospecificity encoded by the Ly-6 region, was biochemically characterized. It was sensitive to pepsin and heat treatment, but was resistant to periodate oxidation. Its apparent molecular weight was approximately 12 000 daltons by gel filtration. The antigenic molecule was partially purified by gel filtration and antibody affinity chromatography. The partially purified antigen molecule was radioiodinated, immunoprecipitated with monoclonal antibody H9/25, and analyzed by SDS-polyacrylamide gel electrophoresis. The autoradiograph showed the molecular weight of H9/25 to be approximately 15000 daltons under reducing conditions. These results indicate that H9/25 is a protein with a single polypeptide chain of 12000–15000 daltons molecular weight, and the antigenic specificity is carried by a peptide but not a carbohydrate moiety. 相似文献
11.
The relationship between two alloantigens on mouse lymphocytes, that is Ly-6.2 and H9/25, which have previously been shown to have identical strain distribution patterns, was further investigated. Analysis of 39 (AKR × CBA) × CBA backcross progeny showed no segregation between these two antigens, indicating a close genetic linkage between them. Serological analysis showed that Ly-6.2 and H9/25 are differentially expressed on T-cell hybrid lines. Furthermore, cross-absorption of anti-Ly-6.2 serum with two cell lines revealed a heterogeneity among Ly-6 specificities. Semipurified H9/25 antigen failed to block anti-Ly-6.2 serum while anti-Ly-6.2 serum did not significantly block monoclonal antibody H9/25. These results suggest the presence of multiple allospecificities encoded for by the Ly-6 region. 相似文献
12.
Feeney AJ 《Immunogenetics》1978,7(1):537-543
The antigens Ala-1 and Ly-6 were first thought to be different on the grounds that Ala-1 was present only on activated T and B lymphocytes while Ly-6 was present only on post-thymic T lymphocytes. In this paper, it is shown that Ly-6 is expressed on activated B cells, including PFC and LPS blasts, and that after typing of several recombinant inbred lines,Ala-1 andLy-6 remain genetically inseparable. Based on available data, it is most likely that Ly-6 is in fact Ala-1, although further testing is required to confirm the absence of Ly-6 from nonactivated lymphocytes.Abbreviations used in this paper Con A
concavalin A
- LPS
lipopolysaccharide
- SRBC
sheep red blood cells
- PFC
plaque-forming cells
- RI
recombinant inbred strains
- B6
C57BL/6 mice 相似文献
13.
The Ly-6 alloantigens have been shown to participate in the process of T cell activation based on the ability of anti-Ly-6 mAb to induce IL-2 production and proliferation of T lymphocytes. In the present investigation we have demonstrated that peripheral T lymphocytes from A strain mice exhibited abnormally low proliferative responses after stimulation through Ly-6A/E and Ly-6C molecules when compared to responses of T cells from numerous other mouse strains. The abnormal activation of the Ly-6 pathway of A strain T cells was not due to ineffective FcR cross-linking of the anti-Ly-6 mAb, to inappropriate cellular expression of the Ly-6A/E alloantigen in A strain T cells, or to an active suppressive phenomenon. T lymphocytes from A strain mice proliferated normally when the cells were activated by mAb to Thy-1 or the CD3/TCR complex suggesting that A strain mice did not exhibit a generalized T cell activation defect. Cell separation studies of T cells and accessory cells demonstrated that this defect was quantitative, rather than qualitative, and that it was complex, residing at both the T cell and accessory cell levels. These results suggest that activation of T lymphocytes via the Ly-6 molecule may involve a signaling pathway and/or cell-cell interactions distinct from those required for optimal activation via CD3/TCR. 相似文献
14.
P. Mark Hogarth Bronwyn A. Houlden Susan E. Latham Vivien R. Sutton Ian F. C. McKenzie 《Immunogenetics》1984,20(1):57-69
Three alloantigens encoded by Ly-6-linked genes are defined by monoclonal antibodies. The Ly-27.2 antigen is defined by antibody 5075-19.1, Ly-28.2 by 5075-3.6, -12.1, -16.10 and by 5095-16.6. The strain distribution pattern of these antibodies is the same and identical with Ly-6.2. However the tissue distribution of these antigens is unique and distinguishes these antigens from the Ly-6.2 antigen or any known antigen encoded by Ly-6-linked genes. Ly-27.2 is present on all thymocytes, T cells, and B cells but is absent from bone marrow cells, whereas Ly-28.2 is absent from most thymocytes and is present on a subpopulation of T cells and B cells but is found on 60–70% of bone marrow cells. No recombination between the Ly-6/Ly-27/Ly-28 loci was found in linkage studies using 41 recombinant inbred strains and 57 backcross mice and indicates very close linkage of these genes. In addition, close linkage to 24 minor histocompatibility genes was excluded using the Bailey HW bilineal congenic mice. The data presented indicate that either the Ly-6 complex is composed of a family of tightly linked genes or the antigens are the products of a single gene that undergoes extensive modification during differentiation. 相似文献
15.
The mouse Ly-17.2 alloantigen has recently been defined with both conventional and monoclonal antibodies; it identifies a locus, sited on chromosome 1, the products of which were considered to be specific for B cells. Using another Ly-17.2-specific monoclonal antibody (described herein), the tissue distribution of the Ly-17.2 antigen was shown to extend to a subpopulation of T lymphocytes and to neutrophils. This distribution is remarkably similar to that of the Fc receptor for immunoglobulin. Indeed, we now demonstrate that the Ly-17 locus codes for a polymorphism of the Fc receptor, a conclusion based upon (a) an identical tissue distribution of Ly-17.2 and FcR on both normal and tumor tissue; (b) specific inhibition of EA rosette formation by F(ab)2 fragments of anti-Ly-17.2; (c) inhibition of the binding of the 2AG2 monoclonal rat antimouse Fc receptor antibody by Ly-17.2 antibody; (d) precipitation of an identical series of molecules by our Ly-17.2-specific antibody and by the recognized Fc receptor-specific antibody (2.4G2); and (e) the demonstration by coprecipitation that the Ly-17.2 specificity is present on Fc receptor molecules. The studies suggest that the xenogeneic monoclonal antibody (2.4G2) which recognizes an invariant site on the FcR molecule and the polymorphic site are closely associated. In addition, the studies firmly map a gene coding for or regulating the expression of the FcR to chromosome 1.Abbreviations used in this paper Ig
immunoglobulin
- FcR
receptor for the Fc portion of Ig
- TNP
trinitrophenyl
- Fab
antigen-binding fragment
- pA
Protein A
- SDS-PAGE
sodium dodecyl sulfatepolyacrylamide gel electrophoresis
- PBS
phosphate-buffered saline
- BSA
bovine serum albumin
- SAMIg
sheep antimouse Ig
- SRBC
sheep red blood cells
- C
complement
- FITC
fluorescein isothiocyanate
- CNBr
cyanogen bromide
- EA
antibody-sensitized erythrocytes 相似文献
16.
BACKGROUND: The Ly-6 family has many members, including Ly-6C and Ly-6G. A previous study suggested that the anti-Ly-6G antibody, RB6-8C5, may react with Ly-6Chi murine bone marrow (BM) cells. This finding has been interpreted as cross-reactivity of RB6-8C5 with the Ly-6C antigen, and has been generalized to many hematopoietic cell types, using the terminology Ly-6G/C. The present study was undertaken to determine whether anti-Ly-6G antibodies truly cross-react with the Ly-6C antigen on multiple hematopoietic cell types. METHODS: Splenocytes, thymocytes, and BM cells obtained from Ly-6.1 and Ly-6.2 strains of mice were stained with a variety of antibodies to Ly-6C and Ly-6G. Flow cytometric analysis was performed on these populations. RESULTS: Evaluation of anti-Ly-6C and anti-Ly-6G staining showed only Ly-6C expression and no Ly-6G expression on subsets of splenic T and B cells and thymocytes from Ly-6.1 and Ly-6.2 mice. Bone marrow cells were identified that express both Ly-6G and Ly-6C; no Ly-6G+Ly-6C- populations were seen. CONCLUSIONS: Multiple Ly-6C+ hematopoietic cell populations were identified that do not stain with anti-Ly-6G antibodies. This calls into question the use of the Ly-6G/C nomenclature and suggests that epitopes recognized by anti-Ly-6G antibodies should simply be designated Ly-6G. 相似文献
17.
Most hemopoietic cells express one or more members of the Ly-6 supergene family of small glycosylphosphatidylinositol-linked proteins. Although levels of Ly-6 proteins vary with stages of differentiation and activation, their function largely remains unknown. To ascertain whether ligands for Ly-6 proteins exist, chimeric proteins were constructed in which Ly-6E, Ly-6C, and Ly-6I were fused to the murine IgM heavy chain. These chimeras specifically stained both developing and mature B lymphocytes, as assessed by flow cytometry. Analysis of variants of the CH27 B cell lymphoma revealed that Ly-6A/E and Ly-6I recognized different molecules. CH27 cells with low levels of Ly-6A/E ligand activity also lost expression of CD22, and cells transfected with CD22 gained the ability to bind the Ly-6A/E chimera and, to a lesser extent, the Ly-6C and Ly-6I chimeric proteins. As many mature B cells coexpress Ly-6A/E and CD22, the function of Ly-6 molecules may be to associate with other membrane proteins, possibly concentrating these ligands in lipid rafts, rather than acting directly as cell:cell adhesion molecules. 相似文献
18.
A mouse monoclonal cell line (20-1.5) was produced by the cell fusion method and the antibody secreted by this line defined the Ly-9.2 specificity — the reciprocal specificity to that previously identified as the Lgp 100 or the T100 molecule. Although most concentrated on lymph-node cells, the antigen is also found on thymocytes, spleen and bone-marrow cells as well as liver and brain tissue. The monoclonal antibody precipitates a 100000 molecular weight moiety from thymocytes. The antigenic specificities appear to be highly immunogeneic and antibodies to these specificities contaminate many antisera. These sera are noncytotoxic as is the case with the monoclonal antibody even though it is of the IgG2a subclass. As with T100 or Lgp 100, theLy-9 locus appears to be linked to theH-25 locus. 相似文献
19.
Dr. Arpi Matossian-Rogers P. Rogers J. A. Ledbetter L. A. Herzenberg 《Immunogenetics》1982,15(6):591-599
Various murine tumor lines were screened by FACS analysis for the surface antigens ThB and Ly-6.2. Positive cell lines were used for immunoprecipitation studies. A monoclonal ThB-specific antibody immunoprecipitated a unique acidic protein of approximately 16 000 daltons from several positive tumors and from concanavalin A (Con-A) and LPS activated splenic lymphocytes. Monoclonal Ly-6.2-specific antibody was used to immunoprecipitate a 33 500 dalton protein that was shown to exist in four similarly sized forms with different basic charges. In the course of these studies, the apparent molecular weight of the surface antigen T 30, immunoprecipitated with a monoclonal T 30-specific antibody from the cell line EL4, was found to be approximately 25 000 daltons. 相似文献
20.
N Hollander 《Journal of immunology (Baltimore, Md. : 1950)》1987,139(2):437-442
IL 2 production by T cell variants, which lack the Thy-1 or Ly-1 surface glycoproteins, was studied. Cross-linking of the Thy-1 molecule resulted in IL 2 production by the EL4 thymoma and by a T cell hybridoma, suggesting that Thy-1 may play a role in T lymphocyte triggering. To further study the functional role of this molecule, Thy-1-negative variants were selected and analyzed for IL 2 production in response to phorbol-12-myristate-13-acetate (PMA) or to Con A. It was demonstrated that in spite of their failure to express Thy-1, the Thy-1-negative clones were capable of IL 2 production. These results indicated that although Thy-1 cross-linking triggers cell activation, a signal provided by Thy-1 is not indispensable for cell activation by mitogens. The T cell tumor line LBRM331A5 responds synergistically to IL 1 and PHA by releasing IL 2. It was demonstrated that anti Ly-1 monoclonal antibodies and PHA co-stimulated LBRM331A5 cells, as did IL 1 plus PHA. Thus, anti Ly-1 antibodies mimic the effect of IL 1, suggesting a role for Ly-1 antigen in T cell activation, perhaps by serving as an IL 1 receptor or as an associated molecule. To further study the functional role of Ly-1 and its relation to IL 1 receptor, Ly-1-negative variants of the LBRM331A5 cell line were selected and analyzed for IL 2 production in response to PHA plus IL 1. It was demonstrated that the Ly-1-negative clones were capable of IL 2 production as efficiently as Ly-1-positive clones. These results indicate that the Ly-1 and IL 1 receptor are distinct molecules, which are involved in different activation pathways. 相似文献