Discrimination between major histocompatibility complex class II DQ and DR locus products in cattle

We have used a panel of anti-major histocompatibility complex (MHC) class II monoclonal antibodies (mAbs) and have assessed their specificity for the products of the individual bovine MHC (BoLA) class II subregions. The mAbs identified two distinct class II molecules by affinity purification and ELISA. Two-dimensional immunoblotting confirmed these data and NH₂-terminal sequencing of the purified class II α chains of one member of each group identified the subregion specificity of the mAbs. The mAbs VPM36, TH22A and TH81A are specific for BoLA DQ, whereas VPM54, TH14B and J11 are specific for BoLA DR. SW73.2 reacts with both MHC subgroups of all cattle tested.     

Two-dimensional gel analysis of a second family of class II molecules by polymorphic HLA-DR4,5, and w9 monoclonal antibodies

Human Ia-like, class II molecules were isolated by immunoprecipitation with monoclonal antibodies from various HLA-D/DR homozygous cell lines and were analyzed by two-dimensional gel electrophoresis. The monoclonal antibody PLM12 reacted with B cells carrying DR4, DR5, DRw6.2, and DRw9 phenotypes, and its reactivity perfectly correlated with the previously defined TB21 (MB3-like) specificity. Class II molecules detected by PLM12 were structurally distinct from those precipitated by the anti-DR monoclonal antibody NC1 on all HLA-DR4, DR5, DRw6.2, and DRw9 homozygous cell lines and showed polymorphism in heavy and light chains among these cell lines. The monoclonal antibodies PLM2 and PLM9 only reacted with B cells carrying DR5 and DRw6.2 and also detected a distinct set of class II molecules from those precipitated by NC1 but identical to those of PLM12. Thus, PLM2 and PLM9 serologically detected a new subtypic antigen of the PLM12-reactive class II molecules. Furthermore, the antibody NC1 precipitated two light chains and one heavy chain from HLA-DRw6.2 homozygous cell line EBV-Sh. The result indicated the presence of three sets of class II molecules: two in a DR family and another carrying the polymorphic determinants detected by PLM2, PLM9, and PLM12 in a second family.     

Analysis of the fine specificities of sheep major histocompatibility complex class II-specific monoclonal antibodies using mouse L-cell transfectants

The fine specificities of two panels of monoclonal antibodies (mAbs) for sheep major histocompatibility complex (MHC) class II molecules were determined using five mouse L-cell transfectaents, each expressing a defined sheep DQ or DR MHC class II A/B gene pair. Using the transfectants in an indirect fluorescence antibody assay, previous immunochemical characterization of the mAbs was confirmed for 16 of 23 mAbs tested. The MHC class II subtype specificity ( DQ or DR ) of each mAb was assigned without interference from the products of other expressed class II loci. This allowed the identification of both cross-locus specificities as well as defining fine specificities of mAbs previously only partially characterized by immunochemical techniques.     

A matrix approach to human class II histocompatibility antigens: Reactions of four monoclonal antibodies with the products of nine haplotypes

Three putative HLA-DC-specific monoclonal antibodies, Genox 3.53, BT3/4 and anti-Leu-10, and the HLA-DR-specific antibody, L243, were compared. Their interactions with molecules from homozygous cell lines expressing DR types 1 through 9 were studied. Indirect radioimmunoassays on 29 cell lines demonstrated that Genox 3.53 reactivity correlated with DR1, 2, 6; BT3/4 reactivity correlated with DR 1, 2, 4, 6, 8; and anti-Leu-10 reactivity correlated with DR1, 2, 4, 5, 6, 8, and 9. In addition, one of six DR3-positive cells and three DR7, DRw10-positive cells reacted with anti-Leu-10 and one of two DR9-positive cells reacted with BT3/4. Binding studies with soluble antigen and competitive radioimmunoassays demonstrated that all three antibodies reacted with the DC1 molecule. Preincubation with BT3/4 blocked anti-Leu-10 binding; Genox 3.53 and L243 did not. Genox 3.53 and L243 were only blocked by themselves. Serial immuno-precipitation showed anti-Leu-10 reacted with non-HLA-DR molecules from cells expressing DR types 1–6, 8 and 9. However, the molecules precipitated by anti-Leu-10 were characteristic class II major histocompatibility complex (MHC) molecules. Their and chains were of lower apparent molecular weight than the DR chains in all haplotypes. They also comigrated with the DC1 molecule precipitated by Genox 3.53. Serial immuno-precipitation also showed that anti-Leu-10 removed all Genox 3.53 reactive molecules from cell lysates, but Genox 3.53 removed only a subset of anti-Leu-10 reactive molecules. These studies show Genox 3.53, BT3/4, and anti-Leu-10 react exclusively with class II MHC molecules that are not HLA-DR, and most likely define different polymorphisms of DC molecules, the human equivalent of mouse I-A products.Abbreviations used in this paper BSA bovine serum albumin - PBS phosphate-buffered saline, pH 7.4 - RIA radioimmunoassays - ¹²⁵I-RAM ¹²⁵I-labeled F(ab)₂ of rabbit anti-mouse IgG - NP40 Nonidet P40 OVA-LB, 0.1% ovalbumin/0.5% NP40, 10mM Tris pH 7.3, 1MM M9Cl₂ 0.5% phenyl methyl sulfonyl fluoride - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - MHC major histocompatibility complex - KD kilodaltons     

Separation of four class II molecules from DR2 and DRw6 homozygous B lymphoid cell lines

Four human class 11 molecules, one FA, one DC1, and two DR-like molecules, were isolated from DR2 and DRw6 homozygous cell lines by means of a variety of monoclonal antibodies and were compared with each other by two-dimensional (2-D) gel electrophoresis. Anti-DR2 or anti-DR3 + 5 + w6 sera immunoprecipitated two distinct light chains (L1 and L2) and one heavy chain (H1) from a DR2 or DRw6 homozygous cell line, respectively. One or both of these two class II molecules were also immunoprecipitated by DR-specific monoclonal antibodies and were considered to constitute a DR family of molecules. Three DC1-specific monoclonal antibodies, SDR4.1, Tu22, and PLM5, immunoprecipitated a set of heavy (H2) and light (L3) chains distinct from those of two DR-like molecules. The heavy chains of the DC1 antigens from DR2 and DRw6 cell lines were indistinguishable, whereas the light chains were structurally distinct from each other. A fourth molecule, FA, was identified by a novel monoclonal antibody and was also detected by two additional antibodies, Tu39 and SG171, that blocked the SB-specific T-cell proliferative response. The FA light chain (L4) was distinct from those of the former three antigens on both cell lines, whereas the FA heavy chain was indistinguishable from the DC1 heavy chain (H2) in this 2-D gel analysis. Thus, four light chains and two heavy chains were isolated from both DR2 and DRw6 homozygous cell lines. A possible gene-antigen organization of the DC-like antigens was also discussed.     

Mechanisms of immunodeficiency in HIV infection and ways of overcoming it]

Though antibodies against HIV-1 appearing in the course of infection are successfully used for the diagnostic purposes, their accumulation on the earlier step leads to: firstly, to the rapid generation of the immunodeficiency by different mechanisms and secondly, to inefficiency of immunotherapy. One of the causes for immunodeficiency seems to be antibodies which are induced in the HIV-infected person by the HIV peptides homologous to the MHC class II molecules by their amino acid sequences. 73% of HIV-1 positive sera are shown to react with human B-lymphoma cells expressing surface class II molecule. The binding is caused by the antibodies preventing the murine monoclonal anti-HLA.DR Ab interaction with B-lymphoma. Three amino acid sequences are identified in both alpha- and beta-chain of the HLA.DR antigen, these sequences being homologous to HIV-1 gp120 or gp42 molecules for 50 to 70%. Using synthetic peptides it was shown that HIV-1-infected persons contain antibodies which cross-react to the homologous peptides of the HIV-1 and of the MHC class II. It is supposed that such antibodies shield the class II molecule on the surface of their own antigen-presenting cell which may lead to immunodeficiency caused by the anti-HIV-1 antibody.     

The MT3 specificity resides on a novel human class II antigen distinct from the HLA-DR antigen and DC-like antigen

The MT3 specificity is closely associated with the HLA-DR4, DR7, and DRw9, and is a supertypic specificity. To determine whether the MT3 specificity resides on a novel class II antigen, the MT3 antigen, DR antigen and the DC-like antigen from the DR4-, DR7- and DRw9-homozygous B lymphoid cell lines were identified and compared with one another by two-dimensional gel electrophoresis using alloantisera. The analysis revealed that each of the three antigens exists as a structurally distinct class II antigen in each cell line. The light chains of the MT3, DR and DC-like antigens are different in charge from one another. The molecular weight of the heavy chains of the MT3 and DR antigens is higher than that of the DC-like antigen. On the other hand, no electrophoretic differences are observed between the heavy chains of the MT3 and DR antigens. These results strongly suggest that the MT3 specificity resides on a light chain of a novel class II antigen distinct from the DR antigen and the DC-like antigen. These observations also support our previous proposition that the MT3 antigen belongs to the fourth group of the human class II antigens.     

The T3/T cell receptor complex: antigenic distinction between the two 20-kd T3 (T3-delta and T3-epsilon) subunits.

Clonally distributed (clonotypic) antigen receptors on human T lymphocytes (alpha and beta chains) are associated with three invariable T3 polypeptide chains (T3 gamma, delta and epsilon), together forming the T3/T cell receptor complex. Monoclonal antibodies prepared against the two 20-kd T3 polypeptide chains demonstrated that T3-delta and T3-epsilon are distinct polypeptide chains. Only one monoclonal antibody (anti-T3-delta chain) reacted with the T cell surface as judged by indirect immunofluorescence, and by its mitogenicity for quiescent peripheral blood lymphocytes. Immunohistological staining and immunoprecipitation experiments showed that both the T3-delta and T3-epsilon chains are T cell-specific. As seen with the anti-alpha/beta chain reagent WT-31, anti-T3-delta chain monoclonal antibodies stained medullary thymocytes more intensely than cortical thymocytes, whereas the difference between the staining of cortical and medullary thymocytes was generally not apparent with anti-T3-epsilon chain antibodies. Because of this specificity and their ability to react with both the denatured and the native forms of each polypeptide chain, these new monoclonal reagents will be useful tools in studies of the biosynthesis and cell surface expression of the T3/T cell receptor complex during normal and malignant thymic differentiation.     

Characterization of the two heavy chains of the T3 complex on the surface of human T lymphocytes

The T3 complex has been defined by a group of monoclonal antibodies which react with all human peripheral blood T lymphocytes and a subpopulation of thymocytes. This membrane structure includes glycoproteins of 44 (alpha), 37 (beta), 25 (gamma), and 20 kDa (delta) as well as a nonglycosylated polypeptide of 20 kDa (epsilon). The characterization of the alpha and beta chains has been of particular interest because they may constitute the T cell receptor for antigen. Here we show that the T3 complex prepared by immunoprecipitation from T lymphocytes of a leukemic patient (Sezary syndrome) displays an unusually strong association of the alpha and beta chains with the 20/25-kDa T3 proteins. The alpha and beta chains (48 and 44 kDa) were co-precipitated by anti-20-kDa T3 monoclonal antibodies as a disulfide-linked 90-kDa heterodimer. A minor 220-kDa multimer composed of proteins similar to the alpha and beta chains was also present in these immunoprecipitates. This multimer could be independently precipitated with a new monoclonal antibody WT-31, which detects the larger polypeptide chains of the T3 complex on all human T lymphocytes. After removal of N-linked oligosaccharides, both the alpha and beta chain were found to have 33-kDa peptide backbones with distinct isoelectric points. Using a monoclonal reagent T40/25, a 90-kDa heterodimer, consisting of 40- and 49-kDa chains with peptide backbones of 34 kDa was found to be T3-associated on the T leukemic cell line HPB-ALL. When the alpha and beta chains from the Sezary patient were compared with the corresponding chains from HPB-ALL by peptide mapping, large differences were observed. Taken together, the data presented here provide strong evidence that the T cell receptor for antigen is part of the T3 complex on the surface of human T lymphocytes.     

Direct identification of class II histocompatibility DR proteins in preparations of human T-cell lymphotropic virus type III

Class II histocompatibility DR antigen alpha and beta chains were isolated from preparations of human T-cell lymphotropic virus type III grown in human H-9 cells. The proteins were purified by reversed-phase high-pressure liquid chromatography and identified by direct N-terminal amino acid sequence analysis of each chain. The purified DR alpha chain had an N-terminal amino acid sequence identical to the known sequence of human DR alpha chain through the first 37 residues. The N-terminal amino acid sequence of the purified DR beta chain was identical to that of human DR4 beta chain. The DR alpha and beta chains appeared to be identical to the p34-36K and p30-32K proteins, respectively, concentrated in immunostimulatory complexes prepared from unfractionated virus and were the major immunogens in these complexes. These proteins represent a ready source of antigens which can cause false-positive enzyme-linked immunosorbent assay reactions in individuals previously exposed to allogenic histocompatibility antigens. The removal of the DR chains from virus preparations by use of available monoclonal antibodies or other means should result in a lower rate of initial false-positive enzyme-linked immunosorbent assay reactions.     

An influenza A virus-specific and HLA-DRw8-restricted T cell clone cross-reacting with a transcomplementation product of the HLA-DR2 and DR4 haplotypes

The clone TA10 is a T3+ T4+ T8- proliferative and cytolytic human T cell clone. This clone has been shown to be specific for the hemagglutinin of influenza A Texas virus and restricted by an HLA class II molecule associated with the DRw8-Dw8.1 phenotype. Here we show that TA10 and all of its subclones can also react with eight HLA-DRw8 negative, Epstein-Barr virus (EBV)-transformed cell lines or phytohemagglutinin blasts in the absence of influenza antigens. All of these cell lines are HLA-DR2/DR4 with a classic DR2 long haplotype. The only nonreactive HLA-DR2/DR4 cell line observed bears a DR2 short haplotype. Only heterozygous HLA-DR2/DR4 but not parental DR2 or DR4 EBV-transformed cell lines can be recognized by TA10, indicating that the cross-reacting determinant is a transcomplementation product between HLA-DR2 and HLA-DR4 haplotypes. DR-specific, but not DQ- or DP-specific monoclonal antibodies, inhibit in the proliferation assay and in the chromium release test both the DRw8-Dw8.1-restricted and the anti-DR2/DR4 reactions. These results show that HLA-DR-restricted, anti-viral human T cell clone can evidence cross-reactivity for allospecific class II molecules of the major histocompatibility complex, and human CTL can recognize transcomplementation products of class II HLA genes. In addition, the results suggest that a beta-chain coded for by an HLA-DR gene and associated with an alpha-chain coded for by a still unidentified but possibly HLA-DQ gene constitute this functional transcomplementation product.     

A novel HLA class II molecule

Molecular evidence has been obtained for a novel monomorphic HLA class II molecule distinct from HLA-DP/DQ/DR using a panel of lymphoblastoid cells which include HLA-loss mutants. The expression of this molecule was investigated using monomorphic affinity-purified mouse monoclonal antibodies (mAbs), including one of the IgG_2a subclass designated EDUA. This antibody reacts strongly in a cell-binding radioimmunoassay with HLA-DR and -DQ loss mutants derived from a lymphoblastoid parental cell. The EDU-1 mAb also reacted with a local panel of homozygous Epstein-Barr virus-transformed cell lines. The reactive molecules were further detected on allostimulated T-cell clones and various leukemic cells including those of myeloid origin which lack surface expression of HLA-DQ molecules. Thus the class II molecule described in this study corresponds to a monomorphic HLA class II determinant expressed on a variety of cells of different origin and HLA phenotypes. Moreover, this antigen structure is distinct from that of HLA-DP/DQ/DR as shown by direct immunoprecipitation, serial immunodepletion experiments, and two-dimensional gel electrophoresis. The molecule could be specified by new class II genes between DP and DQ. An alternative explanation for the genetic basis of the novel molecule is the existence of isotypic associations between alpha and beta chains of various class II molecules (DP, DX, DZ, and DO) but not DR and DQ as the mutant cells tested lack the latter genes.     

Serologic heterogeneity in I-J determinants associated with functionally distinct T cell regulatory factors

Information transfer among regulatory T cell subsets is mediated by biologically active T cell factors. Many of these factors are comprised of two molecules: one that binds antigen, and another that is I-J+ and determines the self recognition capability of the factor (I-J molecule). In the in vitro response to sheep red blood cells, we used three functionally distinct I-J+ factors to study the relationship between polymorphic I-J determinants and the biological activity of these factors. Our study shows that several monoclonal I-J antibodies react with I-J molecules associated with T suppressor-inducer factor (TsiF) and T suppressor-effector factor (TseF), but not with T contrasuppressor inducer factor (TcsiF). In contrast, a different set of monoclonal I-J reagents reacts with TcsiF but not TsiF or TseF. Finally, some monoclonal I-J antibodies distinguish between I-J molecules associated with TsiF and TseF. Thus anti-I-J reagents differentially react with I-J determinants on regulatory factors, and this differential pattern of reactivity correlates with the functional activity of the factors. The possible relationship between I-J heterogeneity and the biological function of I-J molecules in regulation is discussed.     

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.     

HLA-DR7-Specific monoclonal antibodies and a chimpanzee anti-DR7 serum detect different epitopes on the same molecule

We describe here two monoclonal antibodies with HLA-DR7 serologic specificity. The antibodies, SFR16-DR7M, a cytotoxic rat IgM antibody of high affinity, and SFR16-DR7G, a noncytotoxic antibody of the rat IgG 2a class, react with only DR7-positive cells in radioimmunoassay. The cytotoxic activity of SFR16-DR7M correlates completely with the presence of the DR7 specificity, and segregates with the DR7-bearing haplotype in a family. SFR16-DR7M precipitates a class II molecule with the electrophoretic characteristics of DR molecules from LG-10, an HLA-DR7 homozygous cell line. SFR16-DR7G completely inhibits the cytotoxicity of SFR16-DR7M, but only partially inhibits the cytotoxicity of a chimpanzee antiserum with DR7 specificity, Gay/Swei. In binding-inhibition studies, binding of SFR16-DR7M to LG-10 cells is only partially inhibited by the chimpanzee antiserum and vice versa. Both SFR16-DR7M and Gay/Swei reciprocally deplete the same class II molecules from a ³⁵S-methionine-labeled detergent-solubilized membrane preparation of the LG-10 cell line. The chimpanzee serum Gay contains antibodies reactive with epitopes on separated DR7 beta chains, while both SFR16-DR7M and SFR16-DR7G bind only to DR7 alpha-beta complexes. These data suggest that at least two allogeneic epitopes exist which result in the same serologic specificity, and that these epitopes differ in their requirement for alpha-beta complex formation.     

Production of an anti-mouse MHC class II monoclonal antibody with biological activity in transgenic tobacco

To produce a monoclonal antibody specific to a mouse major histocompatibility complex (MHC) class II protein, we synthesized the complementary DNAs for the heavy and light chains of a monoclonal antibody by PCR amplification. These cDNAs were then introduced separately into tobacco plant cells. After performing Northern blot analysis to confirm the expression of each of the chain genes in the transformed plants, we constructed transgenic plants expressing both the heavy and light chains by sexual crossing. The expression of the heavy and light chain genes in the sexually crossed plant was confirmed by Northern and Western blot analyses, respectively. Fluorocytometric analysis showed that the plant-derived antibodies, which we purified using a protein G affinity column, bound specifically to target cells that expressed the cognate MHC class II molecules on their cell surfaces. The results of this study demonstrate that a monoclonal antibody against mouse MHC class II proteins can be expressed in transgenic plants. They also show the specific binding activity of plant-derived antibodies to cognate antigens.     

Analysis of a common-antigen lipopolysaccharide from Pseudomonas aeruginosa.

Lipopolysaccharide isolated from Pseudomonas aeruginosa PAO1 (O5 serotype) was separated into two antigenically distinct fractions. A minor fraction, containing shorter polysaccharide chains, reacted with a monoclonal antibody to a P. aeruginosa common antigen but did not react with antibodies specific to O5-serotype lipopolysaccharide. In contrast, fractions containing long polysaccharide chains reacted only with the O5-specific monoclonal antibodies. The shorter, common-antigen fraction lacked phosphate and contained stoichiometric amounts of sulfate, and the fatty acid composition of this fraction was similar to that of the O-antigen-specific fraction. The lipid A derived from the serotype-specific lipopolysaccharide cross-reacted with monoclonal antibodies against lipid A from Escherichia coli, while the lipid A derived from the common antigen did not react. We propose that many serotypes of P. aeruginosa produce two chemically and antigenically distinct lipopolysaccharide molecules, one of which is a common antigen with a short polysaccharide and a unique core-lipid A structure.     

An antiviral T-Cell clone defines a functional supertypic specificity shared by different HLA-DR molecules from DR2-short,DRw11, and DRwl3 Haplotypes

An influenza virus-specific HLA class IIrestricted human T4⁺ clone (Ij) allows us to define a new functional supertypic HLA class II specificity shared by three different haplotypes. Influenza A virus-infected antigen-presenting cells of these three haplotypes, HLA-DR2 short, DRw11, and DRw13, are able to stimulate Ij cells. The same precise viral specificity is seen in all three cases. Proliferation inhibition experiments using HLA-specific monoclonal antibodies demonstrate that HLA-DR products are involved in all cases. However, according to the DR specificity of the antigen-presenting cell, differential blockings by a series of DR-specific monoclonal antibodies suggest that the functional epitope is shared by different HLA DR molecules. This is confirmed by two-dimensional gel analysis of the HLA DR chains expressed in the three haplotypes.Abbreviations used in this paper APC antigen-presenting cell - EBV Epstein-Barr virus - HA hemagglutinin - HLA human leukocyte antigens - IL-2 interleukin-2 - MHC major histocompatibility complex - MLR mixed lymphocyte reaction - mAb monoclonal antibody - PAGE polyacrylamide gel electrophoresis - PBM peripheral blood mononuclear cells - % RR relative response percent     

Structural relationships between the DR beta 1 and DR beta 2 subunits in DR4, 7, and w9 haplotypes and the DRw53 (MT3) specificity

The class II molecules of DR4, DR7, and DRw9 haplotypes were analyzed by immunoprecipitation, followed by two-dimensional gel electrophoresis and N-terminal amino acid sequencing. By using HLA-DR chain-specific monoclonal antibodies, two distinct DR beta-chains were identified. One beta-chain, designated DR beta 2, had a characteristic acidic mobility. In all three DR types the DR beta 2-chains were indistinguishable by two-dimensional gel electrophoresis and partial N-terminal sequencing. A second DR beta-chain designated beta 1 had a more basic mobility on two-dimensional gel electrophoresis, and differed from the DR beta 2-chains by the consistent presence of phenylalanine at position 18. In contrast to the DR beta 2-chains, the DR beta 1-chains were clearly polymorphic, with specific amino acid sequence differences characteristic of each DR type. The monoclonal antibodies 109d6 and 17-3-3S, recognizing distinct polymorphic epitopes similarly correlated with the DRw53 allospecificity, were found to react with different DR beta-chains. The epitope recognized by monoclonal antibody 109d6 was identified on the DR beta 2-chain in the prototypic DR4, DR7, and DRw9 cell lines. However, the DR7, Dw11, DQw3 cell line BEI was unreactive with antibody 109d6 by either immunofluorescence or immunoprecipitation despite the presence of the DRw53 allodeterminant on this cell line. The other DRw53-like monoclonal antibody, 17-3-3S, reacted with the DR beta 1-rather than the DR beta 2-chain in all DR4 and DR7 cell lines tested, including the cell line BEI. However, antibody 17-3-3S did not react with the DRw53-positive DRw9 cell line ISK. These studies suggest that the DRw53 allospecificity is more complex than previously thought and may comprise a number of distinct epitopes encoded by two different DR beta loci.     

Identification of the components of the murine T cell antigen receptor complex

In addition to the alpha and beta chains of the MHC class II restricted antigen receptor, monoclonal anti-receptor antibodies coprecipitate four polypeptides that appear to be noncovalently associated with the alpha-beta dimer of murine T cells. Included in the murine T cell antigen receptor complex are two glycoproteins of 25 kd (gamma) and 21 kd (delta) and two nonglycosylated polypeptides of 26 kd (epsilon) and 16 kd (zeta). The epsilon chain appears to possess an intrachain disulfide bond and zeta exists in the complex as a disulfide-linked homodimer. The delta chain is phosphorylated on a serine residue in response to T cell activation with antigen. In contrast, both delta and epsilon are phosphorylated in response to treatment of the T cells with phorbol 12-myristate 13-acetate. These polypeptides may play a role in the transduction of the signal(s) in T cell activation.