A new thymocyte membrane differentiation antigen for mouse and rat

A new thymocyte membrane-brain antigenic system is defined for the mouse and rat. Monoclonal antibody NIM-M129 recognizes a membrane antigen that has a similar size to Thy-1 (20–23 kd) but entirely different tissue distribution. It is absent from cortisone-resistant thymocytes, murine peripheral T cells, T-cell blasts, and T-cell lymphomas, or rat bone marrow. However, the determinant exhibits a quantitative difference in expression on thymocytes of Thy-1 ^a (low) and Thy-1 ^b (high) mice, with intermediate values for their F₁ progeny. Backcross and F₂ segregation tests between CBA and CBA-Thy-1^a congenic lines established firm genetic linkage between Thy-1 and the level of epitope expression on the thymocyte cell surface.     

Intermolecular complexes between three human CD1 molecules on normal thymus cells

The first cluster of differentiation (CD1) defines at least three distinct human thymic cell-surface differentiation antigens-CD1a, CD1b, and CD1c. We looked for structural homology of the three CD1 heavy chains at their peptide level by two-dimensional peptide maps. We show here that the CD1a M _r 49 000 heavy chain and the CD1b M _r 45 000 heavy chain appear to be more homologous to each other than to the CD1c M _r 43 000 heavy chain and that only one tyrosil peptide is common to the three heavy chains. Study of the CD1 heavy chains from several individuals reveals a very limited polymorphism of these molecules. We also demonstrate here that CD1a or CD1a-like molecules and other CD1 molecules can form intermolecular complexes on the surface of normal thymus cells. Molecules that are structurally very similar to CD1a molecules are associated noncovalently either with CD1c molecules or with CD1b molecules, and only CD 1a molecules can associate covalently with CD8 molecules. In contrast, we could not find these intermolecular complexes on the surface of leukemic T-cell lines in culture.Abbreviations used in this paper CD cluster of differentiation - mAb monoclonal antibody - MHC major histocompatibility complex - NP-40 Nonidet P 40 - B2m beta-2 microglobulin - PMSF phenylmethylsulfonyl fluoride - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - TL thymus leukemia     

Analysis of T-T lymphocytes and T-accessory cell interactions using cloned T cells: MHC I restricted cloned T cells activate MHC II restricted T helper cells

We evaluated the role of molecules of the major histocompatibility complex (MHC) involved in the cellular interactions of two T-cell clones by testing the effect of monoclonal antibodies on the responses of the clones in vitro. The two T-cell clones used in the study are specific for minor histocompatibility antigens and restricted to the H-2K^k. In the absence of exogenous IL-2 the clones require the presence of Ia⁺, Thy-1⁻ accessory cells and of Thy-1⁺, Lyt-1⁺2⁻ cells in the irradiated spleen cell suspension used as stimulator. It is also necessary that both the accessory cells and the T cells in the stimulator cell populations are recognized specifically by the clones. Monoclonal antibodies specific for the H-2K product inhibited the lytic effector function of the cytolytic clone. These antibodies when added to cultures of stimulator cells and clones inhibited also the proliferation of this clone and of a nonlytic clone. When antigen recognition was measured by the increase in sensitivity of the clones to IL-2 while confronted with uv-irradiated stimulator cells, both clones were blocked efficiently by anti-H-2K antibodies. Thus, these results suggest that the interaction of monoclonal antibodies with the restricting H-2K molecule is sufficient to block the recognition signal, a prerequisite for proliferation. In contrast, monoclonal antibodies specific for A_αA_β and/or E_αE_β had no effect on cytolysis or on restricted recognition. However, they inhibited the proliferative responses as efficiently as the H-2K specific antibodies. Inhibition by class II-specific antibodies was not abolished when stimulator cell populations were depleted of Lyt-2⁺ cells. The blocking effect, however, was reversed by the addition of IL-2. No inhibition was obtained with antibody specific for E_αE_β when B10.A(4R) spleen cells, which do not express E_αE_β, or when B10.A(4R) accessory cells, which were reconstituted with (BALB/c X B10.A(4R)) F1 T cells, were used as stimulators. Stimulator cells heterozygous for H-2 could be inhibited by antibodies to the parental haplotype not encoded in the clones (H-2K^d). These and previous results suggest that H-2K-restricted minor histocompatibility antigen-specific recognition transmits an activating signal to the clones and to the stimulator cells. The clones probably are induced to express more IL-2 receptors. The stimulator T cells seem to interact through A_αA_β and E_αE_β molecules with syngeneic accessory cells. This interaction results in IL-2 production by the stimulator T cells and thus in the proliferation of the clones.     

Evidence for a gene extinguishing cell-surface expression of the Thy-1 antigen

Hybrids between the Thy-1^– murine myeloma S194 and the Thy-l⁺ lymphoma BW5147 (Thy-l⁺) express neither the Thy-1.1 nor the Thy-1.2 antigen on their cell-surface. Subclones isolated from Thy-1^– clones express both Thy-1.1 and Thy-1.2 antigens in amounts similar to those present on wild-type Thy-1⁺ lymphomas, demonstrating that the Thy-1^– hybrids retain all the genes necessary for Thy-1 expression. The results are consistent with the idea that myeloma cells have a functional gene which acts to extinguish cell-surface Thy-1 expression in hybrids. The exact mechanism by which the gene acts to produce the Thy-1^– phenotype remains to be determined.     

Structural Studies of T200 Glycoprotein and the Il-2 Receptor

Abstract

Immunological analysis of the cell surface of hematopoietic cells has led to the identification of many different cell membrane molecules, some of which have well-defined functions as receptors. In general, however, the role of most lymphocyte cell surface molecules remains ill-defined even in cases in which antibody inhibition studies have given some insight into the biological processes in which they participate. Here we describe molecular and biochemical studies of T200 glycoprotein (leukocyte-common antigen) and the IL-2 receptor which illustrate the kinds of approaches that can be currently used to characterize individual molecules. T200 glycoprotein is a large M_r glycoprotein found exclusively on leukocytes. However, the exact M_r varies in a cell-type-specific fashion and this property is conserved between different species. Comparison of the rat, mouse and human cDNA sequences show that the large cytoplasmic portion of the molecule is well-conserved, approximately 90%, whereas the exterior portion is only about 50% homologous. Cell-type-specific differences in the primary sequence of the molecule have been identified in the N-terminal portion of the molecules. In contrast to T200, the function of the IL-2 receptor is well-known. The interaction of IL-2 with its receptor provides a growth signal that determines the magnitude and duration of T-cell responses. Limited proteolysis studies provide the first direct biochemical evidence that the external region of the IL-2 receptor consists of two independent domains. ¹²⁵I-labeled IL-2 has been chemically crosslinked to the receptor and proteolytic cleavage of the crosslinked product indicates that IL-2 is selectively bound to the N-terminal domain of the receptor.     

Structural differences between insulin and somatomedin-C/insulin-like growth factor-1 receptors revealed by autoantibodies to the insulin receptor

Polyadenylated RNA prepared from first trimester human placenta was translated in a membrane-free cell-free system derived from wheat germ. Analysis of the [³⁵S]methionine-labeled products by SDS-polyacrylamide electrophoresis demonstrated two proteins with apparent M_rs of 14,500 and 16,000 that were specifically immunoprecipitated by antiserum to reduced and carboxylated bovine LH_α, and two different proteins with apparent M_rs of 18,500 and 21,000 that were specifically immunoprecipitated by antiserum to hCG_β. None of these products was sensitive to cleavage by endoglycosidase H, whereas the M_r 21,000 product precipitated by antisera to bovine LH_α and to hCG_α from translations supplemented by canine pancreatic microsomes was processed to a product with M_r 13,000 by endoglycosidase H. We suggest that the two forms of the α and β subunit precursors could arise from the translation of two distinct mRNAs encoding each subunit.     

The synthesis and turnover of 5′-nucleotidase in primary cultured hepatocytes

The synthesis and degradation of 5′-nucleotidase has been studied in rat hepatocytes. Primary cultures of rat hepatocytes were established with the cells showing evidence of polarity after 24–36 h in culture. After a 30 h lag period 5′-nucleotidase activity increased to a plateau level similar to the activity found in whole liver. The half life of the enzyme after reaching the plateau of activity was 22.8 h. Pulse-chase biosynthetic labelling studies of 5′-nucleotidase in the cultured hepatocytes using [³⁵S]methionine showed that the 5′-nucleotidase monomer was synthesised as an M_r 67 000 form which was converted to the mature M_r 72 000 form. [³⁵S]Methionine labelling studies in the presence of tunicamycin showed that the unglycosylated protein monomer was an M_r 57 000 form. The immature M_r 67 000 form of 5′-nucleotidase was sensitive to endoglycosidase H, whereas the mature form was sensitive only to endoglycosidase F. The data presented are consistent with 5′-nucleotidase in a polarised cell being synthesised and processed like other membrane glycoproteins, in contrast to earlier reports.     

Depolymerization of hyaluronan by sonication

High molecular weight hyaluronan (M _r 400 000) obtained from human umbilical cord was depolymerized by sonication for 10 h into small molecules and finally into molecules of constant size (M _r 11 000). The molecular size of the depolymerized hyaluronan was unaltered even under different conditions of sonication. After sonication, the main sugar residues at the reducing and non-reducing termini of depolymerized hyaluronan wereN-acetylglucosamine (86%) and glucuronic acid (98%), respectively. Hyaluronans derived from rooster comb (M _r 1×10⁶) andStreptococcus zooepidemicus (M _r 1.2×10⁶) were deploymerized into molecules of different but characteristic sizes by sonication. On the other hand, neither chondroitin sulfate nor glycogen was depolymerized by sonication. These results suggest that high molecular weight hyaluronan may have some weak linkages related toN-acetylglucosamine in the chain, which are extremely sensitive to sonication. At present, however, the nature of these linkages is still unclear.Abbreviations HA hyaluronan - PA 2-aminopyridine     

A mutant lymphoma cell line with a defectiveThy-1 glycoprotein gene

We characterized a mutant T -cell lymphoma line selected for the inability to express the Thy-1 glycoprotein. This cell line is a member of the D complementation class of Thy-1^– somatic cell mutants, and it lacks detectable cell-surface Thy-1.1 glycoprotein and detectable cytoplasmic Thy-1 mRNA. Southern blot analysis using a number of probes isolated from the clonedThy-1.2 gene demonstrated that, in the mutant, one copy of theThy-1 gene is absent from the genome and the other has undergone rearrangement. This rearrangement results from a deletion of the 5 portion of the gene removing the first two alternate exons and promoters and a portion of the second intron. The deletion breakpoint within the mutantThy-1 gene was localized to within 400 nucleotides by Southern blot analysis. The breakpoint is near two classes of mouse repetitive elements-a mouse B1-family repetitive element and a simple repetitive sequence-suggesting a mechanism of rearrangement leading to the mutation. Southern blot analysis demonstrated that two closely linked molecular markers on chromosome 9 are unaltered, demonstrating that the deletion in this mutant cell line is subchromosomal.     

Close linkage between mouse genes determining the two forms of complement component C6 and component C7, and cis action of a C6 Regulatory Gene

Two forms of mouse complement component C6, with molecular weights (M _rs) of 90 and 100 kilodaltons (kd), are present in the sera from certain inbred strains such as the CBA strain; other strains, such as the BALB/c and DBA/2 strains, have only the 90 kd C6A form. The present work was undertaken to determine whether the two M _r forms were the products of genes coding at separate loci. We screened sera from mice from a number of inbred strains by isoelectric focusing and found one strain, AKR, exhibiting allotypic structural variations of C6 forms. To distinguish the various types, we designated the 90 kd types from CBA and AKR mice C6A1 and C6A2, respectively, and the corresponding 100 kd types C6B 1 and C6B2, respectively. Mice possessing only one M _r form were all typed as C6A1. Results of breeding experiments strongly suggested that the two M _r forms of C6 are coded for at two closely linked loci. Sera from a number of inbred strains were also screened for a complement C7 polymorphism by means of isoelectric focusing and functional overlay. C7 from all strains, excepting the AKR strain, produced identical C7 band patterns. AKR C7 produced a unique band pattern, and results of breeding experiments with AKR and BALB/c mice showed the C6 and C7 loci to be closely linked. In addition, we identified a regulatory gene for C6 production. The gene apparently requires androgen to facilitate C6 production in the majority of strains. In these strains C6 activity is virtually absent from female sera. However, we observed moderate levels of C6 activity in sera from IS/Cam females, indicating that, in this strain, male physiological androgen levels are not necessary for C6 production. IS/Cam possess one form of circulating C6 which appears identical with BALB/c C6A1, and therefore IS/Cam mice differ from AKR mice at both the C6 structural and regulatory loci. These two strains were thus suitable for use in breeding experiments to determine the manner of action of the regulatory gene. Results showed that it acted in a cis manner.Abbreviations used in this paper M _r molecular weight - kd kilodaltons - SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - IEF isoelectric focusing - Slp sex-limited protein - MHC major histocompatibility complex     

Sequential activation and loss of the pre-B cell Thy-1 gene in T-cell x pre-B cell somatic hybrids

In somatic cell hybrids between the pseudodiploid Thy-1^– Abelson-leukemia-virus-induced pre-B cell lymphoma RAW 253.1 and the Thy-1⁺ T-cell lymphoma, AKR1 (Thy-1⁺), all cells express the Thy-1 allele of the T-cell parent but most hybrid cells do not express the Thy-1 allele of the pre-B cell lymphoma parent. The Thy-1 allele of the pre-B cell parent, however, is spontaneously activated in a minor proportion of hybrid cells. By sorting for cells expressing the Thy-1 allele of the pre-B cell parent, derivative clones in which 100% of cells express both parental Thy-1 alleles can be isolated. Revertants with a phenotype identical with that of the original hybrid cell line can be isolated from these derivatives by sorting for nonexpression of the Thy-1 allele of the pre-B cell parent. These first-generation revertant cell lines have lost one copy of the Thy-1 gene derived from the pre-B cell lymphoma parent. By a further cycle of sorting, derivatives in which 100% of cells express both parental Thy-1 alleles can again be obtained. Second-generation revertants isolated by sorting these Thy-1⁺ hybrid cells for nonexpression of the Thy-1 allele of the pre-B cell parent no longer contain a normal copy of the pre-B cell Thy-1 allele and this surface antigen is no longer expressed by any cells in the population. These results are consistent with a mechanism that sequentially activates each copy of the Thy-1 gene derived from the pre-B cell lymphoma parent. Hybrids between the class D Thy-1 ^– mutant, AKR1(Thy-1^–d), in which the 5 region of the Thy-1 structural gene has been deleted, and RAW 253.1 cannot be activated to express either Thy-1 allele. This result indicates that a sequence upstream of exon 2 of the active Thy-1 allele is critical for the initial activation event.     

A Thy-1 − mutant defining a gene acting in trans position to regulate cell-surface Thy-1 glycoprotein expression and Thy-1 messenger RNA content

The Thy-1 glycoprotein is a differentiation antigen which exhibits tissue-specific regulation. A mutant of a Thy-1.1⁺ T-cell lymphoma has been isolated which does not express Thy-1 glycoprotein on the cell surface and does not accumulate Thy-1 mRNA in the cytoplasm. Hybrids between the mutant and a Thy-1.2⁺ T-cell lymphoma express 20–30-fold lower levels of Thy-1 glycoprotein on their cell surface compared to wild-type T-cell lymphomas, and they have correspondingly low levels of cytoplasmic Thy-1 mRNA. A revertant of one hybrid was isolated which expressed wild-type levels of both Thy-1 alleles on its surface and contained correspondingly increased levels of Thy-1 mRNA. A Thy-1⁺ revertant of the Thy-1 ^– mutant was isolated by cell sorting. A second generation Thy-1 ^– mutant could be isolated from this revertant which also did not accumulate Thy-1 mRNA and which behaved in a way similar to the first generation mutant when hybridized to a Thy-1.2⁺ lymphoma. No changes in the structure or copy number of the Thy-1 structural gene could be detected in this series of mutants and revertants. These properties are consistent with a mutation in one (or more) gene(s) which acts in trans position to regulate Thy-1 glycoprotein expression.     

Structure and expression of glycoproteins controlled by the Qa-1 a allele

The alloantigen controlled by the Qa-1 ^a allele is a glycoprotein that exists in two forms. The first, an intracellular molecule of apparent M_r of 44 000 daltons, appears to be a kinetic precursor of the second, a cell-surface molecule with an apparent size of 47 000 daltons. The intracellular form of Qa-1 is distinct from that of the TL glycoprotein in two ways: (1) its polypeptide backbone is approximately 5000 daltons shorter, and (2) it possesses three sites of high-mannose carbohydrate attachment, while TL has only one. In the cell-surface form of Qa-1, all three carbohydrate chains are processed to structures that resist endoglycosidase H digestion, presumably complex-type oligosaccharides. Concomitant with these late carbohydrate-processing steps is the formation of stable complexes between Qa-1 and ₂-microglobulin. The timing of this association provides a further contrast between Qa-1 and TL, which is associated with ₂-microglobulin shortly after its synthesis. The Qa-1 glycoproteins have been identified genetically by their synthesis in B6-TL⁺ (Qa-1 ^a /Tla ^a ) splenocytes but not in splenocytes of congenic 136K1 and B6.K2 (Qa-I ^b /Tla ^b ) mice, and by their absence from the products of BALB/c (Qa-I ^vb /Tla ^c ) splenocytes. The cells synthesizing Qa-1 are at least as prevalent in Ig⁺ spleen-cell populations as in T-cell-enriched splenic Ig^– populations. Thus, active Qa-1 synthesis appears to take place at a high rate in normal splenic B cells without mitogenic stimulation.Abbreviations used in this paper EDTA disodium ethylenediaminetetraacetate - Endo H endo--N-acetylglucosaminidase H - FCS heat-inactivated fetal bovine serum - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - Ig immunoglobulin - PBS Dulbecco's phosphate-buffered saline - SDS sodium dodecyl sulfate - TL thymus leukemia antigen     

I-Region genetic restrictions imposed upon the recognition of KLH by murine T-cell clones

Data presented in this paper show that the recognition of keyhole limpet hemocyanin by murine T-cell clones is restricted by products of the I region. These data have been obtained by genetic mapping studies as well as by the use of monoclonal la-specific antibodies which inhibit the ability of antigen-presenting cells to effectively present antigen to such T-cell clones. Use of heterozygous antigen-presenting cells derived from crosses between B6.C-H-2 ^bm12 and B10.A(4R) mice have allowed us to show that both trans-complementing I-A products are used for restriction of recognition of KLH. These data were confirmed using monoclonal Ia antibodies to inhibition recognition of KLH by the same T-cell clones. Thus, we have shown that there exist hybrid molecules formed by free combinatorial association of products encoded within the I-A subregion which restrict the recognition of soluble antigen. Additionally, we have shown that the molecule formed by complementation between the alpha chain encoded within the I-E region and a beta chain encoded within the A region (A_e) can function effectively in presenting KLH to certain murine T-cell clones. These results support the hypothesis that the recognition of individual antigenic epitopes within large multideterminant antigens is under the control of Ir genes.     

Interactions between MHC-encoded products and cloned T-cells. I. Fine specificity of induction of proliferation and lysis

To study the interactions between T cells and class I MHC products, we developed in vitro a T-cell line reactive to H-2K^b stimulating cells and derived T-cell clones from it. Although the T-cell line could proliferate in the absence of exogeneous T-cell growth factors when stimulated with H-2K^b spleen cells, each of the derived T-cell clones required both H-2K^b stimulating cells and an external source of T-cell growth factor for its propagation. Each of the T-cell clones was also cytolysic for H-2K^b target cells. Such T-cell clones allowed the comparison of the antigenic requirements for proliferation and cytolysis. By using H-2K ^b mutant mice, we found that while the original anti-H-2K^b T-cell line reacted with each of the six mutants tested, the individual T-cell clones could be distinguished in terms of their reactivity pattern. Similar fine specificity patterns were found when H-2K ^b mutant cells were used as stimulating or target cells for any given T-cell clone. Each of the three monoclonal H-2K^b-specific antibodies reacting with different epitopes of the H-2K^b molecule totally inhibited H-2K^b-induced proliferation and lysis by the T-cell clones. Further blocking studies involved use of Fab antibody fragments and definition of their reactivity on cells from the H-2K ^b mutants. We concluded that: (1) blocking with a monoclonal antibody does not prove identity of alloantigens recognized by the T-cells and the antibody; (2) a monoclonal antibody could either block or not block H-2K^b-CTL interactions depending on structural variations of the H-2K^b molecule not affecting the CTL-H-2K^b functional interaction; (3) blocking one type of H-2K^b-T-cell interaction (induction of proliferation) always affects the other type (cytolysis).Abbreviations used in this paper MHC major histocompatibility complex - CTL cytotoxic - T lymphocytes - Th T helper cells - PMA 4-phorbol 12-myristate 13-acetate - Con A Concanavalin A - LPS E. coli lipopolysaccharide - SCA Con A stimulated rat spleen-cells supernatant - SBD B6 anti-DBA/2 mixed lymphocyte culture supernatant - TCGF T-cell growth factors - IL-2 interleukin 2 - mAb monoclonal antibody - FCS fetal calf serum - PBS phosphate buffered saline - C complement     

A human homolog of the mouse CD8 molecule,Lyt-3: genomic sequence and expression

The CD8 antigen is a marker for those cytotoxic T cells that recognize antigen in the context of class I major histocompatibility antigens (MHC) and has now been identified in many species. In rodents the CD8 antigen is a heterodimer of two distinct chains, Lyt-2 and Lyt-3 in the mouse and OX-8 M _r 32 000 and 37 000 chains in the rat. Human CD8 has consistently been described as a homodimer/homomultimer on mature T cells made up of one chain homologous to the Lyt-2 and OX-8 M _r 32 000 chains. This paper identifies a human equivalent of the second rodent CD8 chain (Lyt-3 and OX-8 M _r 37 000 chains) at the genomic level and shows that this gene is transcribed in human thymocytes and in some acute leukemic T-cell lines. The existence of a human Lyt-3 homolog raises the possibility that human CD8, like mouse CD8, may exist as a heterodimer.     

Interactions between the Thy-1 and T-cell antigen receptor pathways in the activation of cytotoxic T cells: evidence from synergistic effects, loss variants, and anti-CD8 antibody-mediated inhibition

The relationship between activation of cytolytic T-lymphocyte (CTL) clones via the T-cell receptor (Ti) or the Thy-1 molecule was investigated. Anti-Ti and anti-Thy-1 monoclonal antibodies (mAb) can activate CTL clones to secrete interferon-gamma (IFN-gamma). Suboptimal doses of anti-Ti and anti-Thy-1 mAb, as well as suboptimal doses of two different anti-Thy-1 mAb, can synergize to activate T-cell clones. The addition of phorbol myristic acetate (PMA), which is not stimulatory by itself, can enhance the synergistic effect of mAb on IFN-gamma production. Although the Ti and Thy-1 molecules were not found associated at the cell surface, the results presented here indicate that these molecules are functionally associated. Use of Ti loss variants of a CTL clone confirms that Thy-1-mediated signaling is not an alternative to, but is dependent on the Ti-mediated activation pathway. Additionally, use of anti-Lyt-2/3 mAb, previously described as interfering with class I MHC-Ti binding and/or activation and, in some cases, with anti-Ti-mediated activation revealed that anti-Thy-1 mAb-mediated activation was also greatly reduced by the presence of Lyt-2/3-specific mAb. Thus the interaction between Thy-1 and Ti might also involve Lyt-2 (Lyt-3) molecules.     

Biochemical complexity of serum HLA class I molecules

Human serum was found to contain a variety of class I-like molecules by Western blotting with anti-class I heavy chain reagents: major bands usually are observed around M _r 44 000, 40 000, and 35 000–37 000. HLA-A24-positive individuals are distinguished by higher serum levels of M _r 44 000 and 40 000 class I-like molecules than those found in HLA-A24-negative individuals. The M _r 44 000 serum molecules are probably intact class I molecules that have been shed from the cell membrane, because they contain both a transmembrane segment (TM), as deduced from detergent-binding experiments, and a cytoplasmic tail (CT), as inferred from reactivity with an antipeptide serum specific for the cytoplasmic domain of class I antigens (RaCT). The M _r 35 000 and 37 000 molecules contain neither a TM nor a CT region and therefore are probably proteolytic breakdown products of cellular and/or serum M _r 44 000 molecules, although the existence of Q10-like molecules in man cannot be ruled out. The M _r 40 000 molecules do not contain a TM region. M _r 40 000 molecules reactive with the RaCT serum were found in the minority (2/13) of sera tested. We conclude that alternative splicing resulting in a precise excision of the TM exon plays a minor role in the generation of serum HLA class I antigens.Abbreviations used in this paper B2m beta-2 microglobulin - BSA bovine serum albumin - EBV-BLCL Epstein-Barr virus-transformed B lymphoblastoid cell line - mAb monoclonal antibody - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - TCA trichloroacetic acid     

Modulation of T-cell antigen receptor on lymphocyte membrane

A mouse monoclonal antibody, Mo. Ab. 108.45, detecting cell-surface determinants associated with the T-cell receptor for alloantigen was produced by immunizing mice with an alloreactive human T-cell clone and fusing the splenocytes with the NS₁ plasmocytoma. This Mo. Ab. (1) reacts with the immunizing T-cell clone but not with autologous or allogeneic lymphocytes, lymphoblasts or monocytes; (2) stimulates the proliferation of the immunizing T cells in the absence of the alloantigen; (3) inhibits the response to the specific stimulator; and (4) precipitates a disulfied linked heterodimer composed of two distinct glycoproteins of molecular weights 40 000 and 46 000. The receptor molecule detected by Mo. Ab. 108.45 modulates on the surface of the cells, reaching the highest levels 5 days following exposure to the specific stimulator. The receptor-associated molecule detected by Mo. Ab. 108.45 was expressed by T-cell clones obtained independently in two different mixed lymphocyte cultures between the same responder and stimulator.