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Although the manifestation of inflammatory autodestructive disease is the result of major immunological dysfunction, recent evidence indicates that the immune system attempts to compensate by the production of immunomodulatory autoantibodies. Healthy humans have low levels of naturally occurring autoantibodies directed against the first complementarity-determining region (CDR1) and third framework region (FR3) of their own T-cell receptor (TCR) Vbeta segments, but individuals suffering from rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) can have highly elevated levels of these autoantibodies. We cloned and characterized human anti-TCR monoclonal autoantibodies (mAAbs) from RA and SLE patients. Because of the cross-reactions between distinct CDR1 segments of human TCR Vbeta and corresponding murine homologs, it was possible to show that human mAAbs blocked the capacity of a murine TH1 cell line (DO11.10) to produce IL-2 in response to antigenic stimulation in vitro. These results support the hypothesis that autoantibodies against TCR Vbeta can shut down TH1-mediated inflammatory autodestructive reactions.

The operation of the immune system is a complex orchestration of specific self and non-self-recognition capacities mediated by cells of the innate system acting in coordination with T and B lymphocytes in a series of processes modulated by cytokines. We provide evidence for a natural immunomodulatory system involving autoantibodies directed against a controlling segment of T cell receptor Vbeta chains that downregulate production of stimulatory cytokines balanced by the peptides which in turn upregulate inflammatory activities mediated by TH1-type helper cells. TCR Vbeta-derived peptides effective in retrovirally induced immunosupression could also reverse the effects of immunosenescence in aged mice by restoring the balance of TH1- and TH2-type immunity and the resistance of the animals to cardiac pathology caused by infection with coxsackievirus. An unexpected finding was an adaptive role of the T cells from peptide-treated mice in remodeling damaged hearts by increasing net collagen synthesis by cardiac fibroblasts.

The induction of T helper (TH) lymphocytes by distinct TH ligands results in a differentiation to TH1/TH2 subsets based on their unique pattern of cytokine secretion and effector functions. We hypothesized that the relative proportion of TH1/TH2 directly relates to cardiac fibroblast (CF) function and thereby cardiac extracellular matrix (ECM) composition and cardiac diastolic function in the absence of injury or altered wall stress. We compared the effect of selective TH1 with TH2 inducers on cardiac gene expression, ECM composition, and diastolic function in C57BL/J mice. Twelve weeks after immune modulation, the left ventricular stiffness (β) was significantly increased in the TH1 group and decreased in the TH2 group ( P < 0.01). The TH2 group also demonstrated significantly increased end-diastolic and end-systolic volumes ( P < 0.01). Cardiac gene expression patterns for pro-matrix metalloproteinase (MMP)-9 and -13 were increased by greater than fivefold in the TH2 group and significantly decreased in the TH1 group ( P < 0.05). The total cardiac collagen and cross-linked collagen were significantly increased in the TH1 group and decreased in the TH2 group ( P < 0.01). Coculturing lymphocytes harvested from the treated mice with naive primary CF demonstrated a direct control of the lymphocytes on CF pro-collagen, pro-MMP gene expression, and MMP activity. These results suggest that the TH phenotype differentially affects diastolic function through modulating CF pro-collagen and pro-MMP gene expression, MMP activity, and cardiac collagen cross-linking, resulting in altered ECM composition. Thus modulation of TH lymphocyte function could promote adaptive remodeling in heart failure and postmyocardial infarction.

Viral myocarditis is an important cause of heart failure and cardiomyopathy. Immunosenescence, characterized by a dramatic reduction in immune responsiveness, can increase susceptibility to cardiopathology from viral infections. The T-cell receptor (TCR) Vbeta 8.1 peptide, a 16-mer peptide, has shown immuno-regulating and immunostimulating effects in viral-induced immunodeficiency. In our study, 18-mo-old C57Bl/6 female mice were treated twice with TCR Vbeta8.1 peptide and 10 d before sacrifice were injected ip with coxsackievirus B3. Cardiac histopathology was assessed for lesion severity. Splenocyte cyto-kine production (interleukin-2, -4, -6, interferon-gamma) and heart viral titers were determined. Our data suggest that immunosenescence suppressed both T helper (Th1) and Th2 cytokine production and that treatment with TCR Vbeta8.1 peptide induced cytokine stimulation close to levels seen in young mice. Nontreated aged mice developed some degree of myocarditis (75% mild and 25% severe), whereas only 35% of the peptide-treated aged group developed cardiopathology, with 25% being mild and 10% severe. Heart tissue from nontreated aged mice infected with coxsackievirus had a higher viral titer than hearts of aged mice equally infected but treated with the peptide. In conclusion, TCR Vbeta8.1 peptide induced immunoregulation, and inhibited or reduced coxsackievirus B3-induced cardiopathology in aged mice.

Over the last decade, a number of concerns have arisen related to safety issues that have had an adverse effect on the public's trust, particularly among parents whose children are the primary recipient of the vaccine. Historically, the live attenuated measles virus (MV) vaccine and the combination multivalent measles, mumps, and rubella (MMR) vaccine have had a major impact on the health of children worldwide and have been extremely successful at preventing infectious diseases associated with three childhood viral pathogens. In this report, we describe MV infection, replication, pathogenesis, and immunization. MV is a viral pathogen that exhibits a number of complex processes that can effect its replication, pathogenesis, and the induction of an effective antiviral immune response. We describe the published literature as it relates to MV infection and immunization and report adverse events in an attempt to provide a balanced discussion and an historical perspective of the MMR vaccine and autism.

Infection of people with human immunodeficiency virus (HIV) as well as LP-BM5 infection in mice results in progressive deterioration of the immune system in the majority of untreated hosts. Peptide immunotherapy has been shown to be effective in the stimulation or immunoregulation of T-helper 1 (T(H)1) and T-helper 2 (T(H) 2) response subsets. In murine acquired immunodeficiency syndrome (AIDS), T(H)1 deficiency enables the host to be susceptible to coxsackievirus infection, inducing cardiopathology in a short period. T-cell receptor (TCR) Vbeta8.1 peptide, a 16-mer peptide containing the entire CDR1 segment and part of the FR2 region of human Vbeta8, showed both an immunoregulating and immunostimulating effect in murine AIDS. TCR Vbeta8.1 peptide acts on T cells promoting interleukin-2 production and therefore enhancing a cell-mediated immune response. It retarded development of cardiopathology due to coxsackievirus infection. Retrovirus-infected mice treated with the peptide showed a longer life span than the nontreated, retrovirus-infected animals.

This review considers definitions of the specificity of antibodies including the development of recent concepts of recognition polyspecificity and epitope promiscuity. Using sets of homologous and unrelated peptides derived from the sequences of immunoglobulin and T cell receptor chains we offer operational definitions of cross-reactivity by investigating correlations of either identities in amino acid sequence, or in hydrophobicity/hydrophilicity profiles with degree of binding in enzyme-linked immunosorbent assays. Polyreactivity, or polyspecificity, are terms used to denote binding of a monoclonal antibody or purified antibody preparation to large complex molecules that are structurally unrelated, such as thyroglobulin and DNA. As a first approximation, there is a linear correlation between degree of sequence identity or hydrophobicity/hydrophilicity and antigenic cross-binding. However, catastrophic interchanges of amino acids can occur where changing of one amino acid out of 16 in a synthetic peptide essentially eliminates binding to certain antibodies. An operational definition of epitope promiscuity for peptides is the case where two peptides show little or no identity in amino acid sequence but bind strongly to the same antibody as shown by either direct binding or competitive inhibition. Analysis of antibodies of humans and sharks, the two most divergent species in evolution to express antibodies and the combinatorial immune response, indicates that the capacity for both exquisite specificity and epitope recognition promiscuity are essential conserved features of individual vertebrate antibodies.

Available data suggest that ‘primitive’ antibody-combining sites often include longer than average HCDR3s. Long HCDR3 sequences have been reported in diverse vertebrates, including humans, cattle, camels and sharks. These long HCDR3 segments contain unusual sequence features such as stretches of Gly or Pro residues and multiple Cys residues. We examined how longer than average HCDR3s were accommodated in the V domains of human, murine and camel antibodies with known three-dimensional structures. The main conclusions were that (1) HCDR3s longer than 12 residues should protrude outward from the V domains; (2) descending HCDR3 polypeptides may utilize VL (including LCDR3) constituents as a platform, supporting the protruding segments; (3) intra- and inter-HCDR disulfides are frequently formed to rigidify the structure of HCDR3 or the combining site, and (4) V and C domains were possibly more similar in primordial antibodies than they are in their present day counterparts.

To characterize the binding specificity and light- and heavy-chain variable region usage in monoclonal human autoantibodies (mAAbs) to T-cell receptors, we constructed heterohybridomas from peripheral blood B cells of three rheumatoid arthritis (RA) patients. From a panel of more than 200 heterohybridomas secreting IgM autoantibodies binding to T-cell receptor Vbeta chain first complementarity determining segments (CDR1), we characterized two IgM/lambda molecules from a single patient in detail. These bound to both CDR1 peptide epitopes and intact TCR of recombinant single-chain T-cell receptor constructs, and to T-cell surface TCR. Spectratype analysis using epitopes mimicking a set of 24 Vbeta genes indicated that one molecule bound only a few members of the set, whereas the second showed considerable epitope promiscuity by binding to more than half of the tested CDR1 peptides. Both mAAbs used variants of a Vlambda3 gene that were very similar to one another and to the germline gene. The epitope-promiscuous autoantibody used a V(H)4 gene identical to a germline prototype, while the other incorporated a V(H)3 sequence differing in only a single residue from its germline prototype. The CDR3s of both were large and distinct from each other as well as from the corresponding segments of rheumatoid factors and "cold agglutinins" using the same or related V(H) germline genes. These mAAbs offer models for deciphering the basis of epitope promiscuity, and serve as candidates for direct use in immunomodulation because they are of intrinsic human origin and do not require molecular engineering to adapt them for use in therapy.

Previous studies established that retrovirally infected young mice produced large amounts of autoantibodies to certain T-cell receptor (TCR) peptides whose administration diminished retrovirus-induced immune abnormalities. C57BL/6 young (4 weeks) and old (16 months) female mice were injected with these same synthetic human TCR V beta 8.1 or 5.2 peptides. Administration of these autoantigenic peptides to old mice prevent immunosenescence, such as age-related reduction in splenocyte proliferation and interleukin-2 (IL-2) secretion. TCR V beta peptide injection into young mice had no effect on T- or B-cell mitogenesis and IL-4 production while modifying tumour necrosis factor-alpha (TNF-alpha), IL-6, and interferon-gamma (IFN-gamma) secreted by mitogen-stimulated spleen cells. TCR V beta injection also retarded the excessive production of IL-4, IL-6 and TNF-alpha induced by ageing. These data suggest that immune dysfunction and abnormal cytokine production, induced by the ageing process, were largely prevented by injection of selected TCR V beta CDR1 peptides.

C57BL6 mice were immunized with peptide corresponding to CDR1 of human T cell receptor (TCR) Vs8.1 and immunoglobulin lambdachain MCG after murine retrovirus (LP-BM5) infection. Immunization with the TCR peptide largely prevented the retrovirus-induced reduction in B and T cell proliferation, tissue vitamin E, and Th1 (T helper 1) cytokine (IL-2 and IFN-τ) secretion. The TCR peptide also prevented retroviral stimulation of Th2 cytokine (IL-6 and IL-10) production and lipid peroxidation. Cryptosporidiosis was established in all retrovirus immunosuppressed mice, while non-retrovirus infected mice were refractory to parasite infection. Cryptosporidium parvum colonization of intestinal villi was significantly reduced in immunosuppressed animals that received TCR Vs8.1 peptide immunization. Thus, immune dysfunction with the loss of parasite resistance, induced by murine retrovirus infection, was largely prevented by TCR Vs CDR1 peptide immunization. However, no significant change in survival of the retrovirally infected mice was observed as the lymph nodes continued to expand which induced eventual death from asphyxiation.

Individuals seropositive for human immunodeficiency virus type 1 (HIV) express elevated levels of autoantibodies (AAbs) directed against recombinant T-cell receptors (TCRs) and synthetic peptide epitopes duplicating β chain markers. We performed longitudinal studies of anti-TCR AAbs in HIV-1-infected individuals, making comparisons with uninfected sera and sera from other individuals infected with a nonviral agent. We determined levels of autoantibodies by titration using enzyme-linked immunosorbent assay (ELISA) and developed a means for characterizing “autoantibody CDR recognition spectrotypes” for individual sera. Antibody levels against certain defined synthetic epitopes were substantially elevated in HIV-infected subjects relative to reactivities by control groups. Individual sera showed relatively high AAb levels to a subset of CDR1 peptide epitopes. Two patients who subsequently developed AIDS showed particular reactivity to Vβ2.1, 8.1, 10.1, and 22.1 epitopes. Our results show that production AAbs to TCR Vβ epitopes is a general consequence of HIV infection. The response is individual but shows some restriction and shifts in AAb subpopulations often occur with time.

Autoantibodies (AAbs) directed against particular segments of the variable region of TCR beta chains occur in normal humans and in certain autoimmune diseases, but the factors regulating the appearance of such antibodies are unknown. We report that AAbs binding a peptide determinant corresponding to the CDR1 of the V beta domain are elevated in C57BL/6 mice following infection with the LP-BM5 murine leukemia retrovirus mixture, a treatment used to induce murine AIDS. The elevation of the level of these AAbs is an early event following retroviral infection which corresponds in part to the general polyclonal activation of the B cells, but a selectivity for particular V beta sequences is apparent later. This suggests that the appearance of these antibodies may play a part in the subsequent development of immunodeficiency. Since the antibodies studied are of the IgG isotype, both T cells and B cells are involved in their elaboration.

The fetus is a natural allograft that is protected from immunologic rejection by a complex set of structural and regulatory mechanisms. We determined whether healthy pregnant women differed significantly from healthy non-pregnant controls in their capacity to produce autoantibodies to defined antigenic determinants of the alpha/beta T-cell receptor. Although controls and pregnant women expressed comparable levels of autoantibodies against an intact recombinant T-cell receptor containing the complete V alpha/V beta structures, analysis of comparative reactivity against individual peptide segments of the molecules, indicated enhanced reactivity to regions corresponding to the CDR1 of the alpha chain and to the Fr3 of the variable region of the beta chain. A major difference was noted by increased reactivity of IgG autoantibodies of pregnant women to peptides corresponding to the "switch" region joining the variable and constant domains. This was noted with both the Tcr alpha and beta chains and was directed against highly conserved determinants within these molecules. Antibodies to this region are lacking in the non-pregnant controls. It is possible that autoantibodies directed against conserved regions of the T-cell receptor might function in the suppression of T-cell reactivity of fetal determinants.

A single chain T cell receptor (scTcr) was constructed from the complete V alpha and V beta regions of Jurkat T-cell receptor alpha/beta chain genes using molecular cloning techniques. The recombinant scTcr reacted with a panel of rabbit antisera generated against synthetic 16-mer peptides duplicating the amino acid sequence of Jurkat V alpha and beta chains but not with antisera directed against peptides from the constant domain. Autoantibodies present in sera from systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) patients bound the scTcr in ELISA assays. The recombinant scTcr described here should prove to be a useful reagent with which to study T-cell receptor activity in serological and functional assays.

Although gene segments specifying Igs of all vertebrates show clear homology, their arrangements differ markedly, thereby suggesting that the mechanisms for the generation of diversity and for the regulation of gene expression may be quite distinct. In the sandbar shark, light chain gene segments are distributed as apparently independent clusters consisting of V, J, and C elements that require rearrangement for expression. The usual distance between V and C in the clusters is 3 kb but larger clusters occur. The V, J, and C elements are clearly homologous to those of human λ chains. Shark Igs resemble mammalian IgM in structure and gene similarity. IgM may comprise as much as 50% of serum proteins in the shark. By contrast, IgM in humans comprises less than 5%. Human autoantibodies usually are IgM. These show little dependence on thymic function for expression and tend to increase with age. We have carried out a study of the capacity of Igs of unimmunized sharks and people (normals and patients suffering from autoimmune diseases) to react against a panel of antigens, including those usually considered autoantibodies, such as thyroglobulin and single-stranded DNA. Sharks and humans possess IgM antibodies that react with thyroglobulin and ssDNA. Affinity-purified natural shark antibodies to thyroglobulin or ssDNA constitute small fractions of total IgM. They illustrate extensive cross-reactivity comparable to that shown by polyspecific IgM autoantibodies produced by human B cells (CD5+) that appear early in ontogeny.

Antigen-specific molecules secreted by murine T cell hybrids (TABM) specific for 4-hydroxy 3-nitrophenyl (NP) or azobenzenearsonate (ABA) were purified from ascitic fluid by ion exchange chromatography and/or affinity for antigen. Partial amino acid sequence of reduced Mr 72,000 NP-specific polypeptides and Mr 20,000 peptides prepared by treatment of the ABA-specific immunoprotein with cyanogen bromide was obtained and a septapeptide of the NP-specific TABM shared 3/7 residues with the ABA-specific TABM. Both TABM shared residues present in 95% T cell receptor for antigen (TCR) V alpha subgroup I and 83-96% murine immunoglobulin V kappa Fr3. These results provide evidence that extracellular antigen-specific T cell proteins are soluble analogues of TCR alpha chains and belong to the immunoglobulin supergene family.

Physiologic removal of old and damaged erythrocytes, platelets, and other terminally differentiated cells is initiated by the appearance of an aging antigen that marks them for death by initiating the binding of IgG autoantibody and subsequent removal by phagocytes. We have developed a synthetic aging antigen peptide that blocks binding of IgG to senescent cells in vitro. We hypothesize that the synthetic antigen can be used to prevent cell destruction in diseases such as autoimmune hemolytic anemias and idiopathic thrombocytopenia purpura, and that the antigen itself can be used to manipulate cellular lifespan in vivo.

As cartilaginous fish are the vertebrates most distal from man to produce antibodies, fundamental information regarding conservation and variation of the antigen binding site should be gained by comparing the properties of antibodies directed against the same antigen from the two species. Since monoclonal cell lines cannot be generated using shark B cells, we isolated antigen binding recombinant single chain Fv antibodies (scFv) comprising of the complete variable regions from shark light and heavy chains. Thyroglobulin was used as the selecting antigen as both sharks and humans express natural antibodies to mammalian thyroglobulin in the absence of purposeful immunization. We report that recombinant sandbar shark (Carcharhinus plumbeus) scFvs that bind bovine thyroglobulin consist of heavy chain variable regions (VH) homologous to those of the human VHIII subset and light chain variable regions (VL) homologous to those of the human Vλ6 subgroup. The homology within the frameworks is sufficient to enable the building of three-dimensional models of the shark VH/VL structure using established human structures as templates. In natural antibodies of both species, the major variability lies in the third complementarity determining region (CDR3) of both VH and VL. Copyright © 2005 John Wiley & Sons, Ltd.

In ancestral sharks, a rapid emergence in the evolution of the immune system occurred, giving jawed-vertebrates the necessary components for the combinatorial immune response (CIR). To compare the natural antibody (NAb) repertoires of the most divergent vertebrates with the capacity to produce antibodies, we isolated NAbs to the same set of antigens by affinity chromatography from two species of Carcharhine sharks and from human polyclonal IgG and IgM antibody preparations. The activities of the affinity-purified anti-T-cell receptor (anti-TCR) NAbs were compared with those of monoclonal anti-TCR NAbs that were generated from a systemic lupus erythematosus patient. We report that sharks and humans, representing the evolutionary extremes of vertebrate species sharing the CIR, have NAbs to human TCRs, Igs, the human senescent cell antigen, and to numerous retroviral antigens, indicating that essential features of the combinatorial repertoire and the capacity to recognize the potential universe of antigens is shared among all jawed-vertebrates.

With regard to retroviral mediators of autoimmune/inflammatory diseases, coupled with genetic and environmental factors, it is clear that retroelements, in particular human endogenous retroviruses (HERVs), play a vital and significant role in the etiopathogenesis of such diseases. Because of the genetic diversity of exogenous and endogenous retroviruses, it has been difficult to delineate or identify specific retroviruses associated with specific autoimmune or inflammatory diseases. However, sequencing of the human genome and genome-wide scans of both humans and mice support the hypothesis of a HERV based etiopathogenesis for systemic lupus erythematosus (SLE). Specifically, it is proposed that molecular mimicry between the Gag-related region of (U1)snRNP and HRES-1 initiates the production of cross-reactive auto-antibodies and associated immune complexes. In addition to molecular mimicry, other mechanisms of autoimmunity brought about by HERVs include but are not limited to insertional mutagenesis, superantigen-type activity, cis- or trans-regulation of cellular genes, and immunomodulation by HERV gene products. Continued research in the field of autoimmune disease and HERVs as etiological factors will certainly facilitate the understanding of the very complex disease, SLE, as well as other autoimmune or inflammatory diseases.

The nuclear vitamin D receptor (VDR) mediates the actions of its 1,25-dihydroxyvitamin D(3) ligand to control gene expression in terrestrial vertebrates. Prominent functions of VDR-regulated genes are to promote intestinal absorption of calcium and phosphate for bone mineralization and to potentiate the hair cycle in mammals. We report the cloning of VDR from Petromyzon marinus, an unexpected finding because lampreys lack mineralized tissues and hair. Lamprey VDR (lampVDR) clones were obtained via RT-PCR from larval protospleen tissue and skin and mouth of juveniles. LampVDR expressed in transfected mammalian COS-7 cells bound 1,25-dihydroxyvitamin D(3) with high affinity, and transactivated a reporter gene linked to a vitamin D-responsive element from the human CYP3A4 gene, which encodes a P450 enzyme involved in xenobiotic detoxification. In tests with other vitamin D responsive elements, such as that from the rat osteocalcin gene, lampVDR showed little or no activity. Phylogenetic comparisons with nuclear receptors from other vertebrates revealed that lampVDR is a basal member of the VDR grouping, also closely related to the pregnane X receptors and constitutive androstane receptors. We propose that, in this evolutionarily ancient vertebrate, VDR may function in part, like pregnane X receptors and constitutive androstane receptors, to induce P450 enzymes for xenobiotic detoxification.

Natural autoantibodies (NAAbs) specific for the T-cell receptor (TCR) are present in all human sera, but individuals with rheumatoid arthritis (RA) generally produce higher titres of immunoglobulin M (IgM) isotype autoantibodies (AAbs) against Vbeta TCR epitopes. To investigate possible correlations between the specificity of such AAbs and their role in immunomodulation, we generated seven B-cell hetero-hybridomas, secreting monoclonal IgM NAAbs, from the synovial tissue and peripheral blood of patients with RA. Here we report three anti-TCR monoclonal autoantibodies (mAAbs)--OR2, OR5 and Syn 2H-11--with the ability to bind subsets of murine T cells, including the ovalbumin-specific DO-11.10 clone. These antibodies did not induce apoptosis in vitro, but prevented interleukin-2 (IL-2) production by antigen-specific T cells. These findings suggest an immunomodulatory function for NAAbs to TCR V-region epitopes and serve as the foundation for testing human anti-TCR mAAbs in animal models with the eventual goal of using them as therapeutic agents in human disease.

The classical concept of antibody binding is defined as an exclusive and high-affinity interaction with one epitope. The emerging reality about antibody combing sites, however, is that some can bind unrelated determinants. The studies presented here define this quality as epitope recognition promiscuity by analyzing the capacity of monoclonal human autoantibodies to bind sets of overlapping peptides duplicating the complete structures of T cell receptor (TCR) alpha and beta chains and immunoglobulin lambda chain. We assessed the binding of these monoclonal antibodies (mAbs) to a set of homologous peptides corresponding to the CDR1 segments of human Vbeta gene products, a major epitope used in the selection of the antibodies. We present data on the binding characteristics of four human mAbs selected for the ability to bind TCR epitopes. These mAbs are IgM molecules with VH and VL sequences in germline configuration, but have diverse VH CDR3 regions. These studies aim to characterize the property of epitope promiscuity and show that the relationship between the binding site and its epitope is a complex interaction and unpredictable from antigen sequence alone. Our results support the conclusion that epitope recognition promiscuity is a genuine feature of antibody and TCR recognition.

Although the etiology of systemic lupus erythematosus (SLE) remains unclear, there is substantial circumstantial evidence that the development of SLE is dependent on environmental, genetic, and retroviral factors. SLE patients produce high titer antibodies to various retroviral proteins, including Gag, Env, and Nef of HIV and HTLV, in the absence of overt retroviral infection. We review the factors linking HERVs to SLE and consider the various processes utilized by endogenous retroviruses in the etiopathogenesis of SLE. In particular, we consider the role of HTLV-1-related endogenous sequence (HRES-1) in SLE. We propose that molecular mimicry between HRES-1 and the small ribonucleoprotein complex initiates the production of autoantibodies, leading to immune complex formation, complement fixation, and pathological tissue deposition.

A gelatin sponge model of concomitant tumor immunity was employed in order to examine the clonality of T cells associated with progressing and rejected tumor sites. Here we show that freshly isolated T cells bearing TCR V(beta)1, CDR3 RPGTGN, J(beta)1.1 and TCR V(beta)8, CDR3 GD, J(beta)1.6 predominated progressing and rejected tumor sites. Despite the similarity in T cell populations, the T cells from rejected tumor sites were capable of killing the autologous tumor cells, whereas T cells from progressing tumor sites were not able to do so. The differing cytolytic ability could not be attributed to a difference in TCR zeta chain protein expression levels between both T cell populations. After a 5 day mixed lymphocyte tumor culture the T cells from the progressing tumor site were capable of killing autologous tumor cells, which suggested changes took place within the cell population during in vitro culture. Further TCR analysis revealed T cells bearing TCR V(beta)1, CDR3 RPGTGN, J(beta)1.1 and TCR V(beta)8, CDR3 GD, J(beta)1.6 were not expanded following the in vitro culture. These data suggest that the lack of cytotoxicity of freshly isolated tumor-infiltrating lymphocytes (TIL) was not due to abnormal TCR zeta chain expression or major differences in the TCR V(beta) usage. Additionally, the gain of TIL effector function did not correlate with an expansion of the TCR bearing T cells found to predominate the in vivo response. These data suggest that the predominant TCR V(beta) used by lymphocytes infiltrating regressing or rejected tumors may not represent the tumor reactive T cells that grow in culture or respond to the autologous tumor in vitro.

We propose that the amyloid deposits in senile plaques of Alzheimer's Disease (AD) result from ancient mechanisms in wound-healing and inflammatory processes that preceded the evolution of the inducible combinatorial immune responses characteristic of jawed vertebrates. AD plaques are unlike active plaques in MS, because antibodies, T-cells and, B cells are not conspicuous components of senile plaques or other loci of degeneration. However, senile plaques contain amyloids and other inflammatory proteins of ancient origin that appear to be made by local brain cells, including neurons, astrocytes, and microglia. We describe a highly conversed 16-mer found in pentraxins from mammals and from the horseshoe crab. The senile plaque thus provides a novel opportunity to study primitive features of complement-mediated inflammatory responses in the absence of immunoglobulins.

Murine retrovirus infection causes an aberrant stimulation of several subsets of T helper 2 cells identified by their T cell receptors (TCR). C57BL/6 mice were treated with synthetic peptides based upon different human TCR V beta CDR1 sequences following experimental infection with the murine retrovirus. Previous studies established that retrovirally infected mice produced autoantibodies to certain of these peptides, and their administration after infection diminished many of the cytokine abnormalities induced by the virus. This study determined whether the complete 16-mer synthetic peptides modeling the V beta CDR1/FR3 were required, and whether admixture of autoantigenic peptides synergized immune preservation. Treatment with complete TCR pep beta 3 and pep V beta 5.2 peptide alone and combined largely prevented the retrovirus-induced reduction in B and T cell proliferation and Th1 cytokine secretion while suppressing excessive production of Th2 cytokines, which are stimulated by retrovirus infection. Treatment with overlapping short peptides corresponding to the N-terminal 11-mer and C-terminal 12-mer did not significantly prevent the immune dysfunction in retrovirus-infected mice. These data suggest that immune dysfunction and abnormal cytokine production, induced by murine retrovirus infection, were largely prevented by TCR V beta CDR1 peptides, and the complete CDR1 in association with the five residues from FR2 was required.

Autoantibodies (AAbs) to T cell receptor (TCR) determinants are produced in humans during the course of rheumatoid arthritis and systemic lupus erythematosus as well as in retroviral infections. We have examined the binding specificity of a panel of monoclonal antibodies derived from mutant viable motheaten (mev) mice against several TCR peptide determinants representing the complementary determining region 1 (CDR1) regions of various Vbeta families, and have identified one mAb, UN37-5, that shows high affinity binding with specificity for two CDR1 peptide determinants. The light and heavy chain V genes of UN37-5 were sequenced and compared to known V genes. The UN37-5 V H gene sequence represents the V H J6O6 family and is most similar to a previously reported V H gene derived from a germ line DNA fragment representing a unique J606 family V H gene. This germ line V H gene is also used by previously characterized mev derived mAbs directed against thymocyte and RBC antigens. The UN37-5 V L gene sequence represents the V K 4/5 gene family. It has 87% homology with the V K Ox-1 germline gene. The UN37-5 V K sequence has greater than 95% identity at the amino acid level with VK L chains from IgM hybridomas specific for DNA and the influenza virus hemagglutinin. The specificity of this AAb is determined by the V H CDR3 and a V K chain which has not been utilized in previously reported mev autoantibodies.

Autoantibodies directed against peptide-defined epitopes of T-cell receptors occur in the serum of healthy humans with the levels and isotypic expression dependent upon physiological changes (aging, pregnancy) or upon the development of autoimmune disease. We carried out investigations of autoantibodies against Tcr peptide-defined epitopes in retroviral infections of humans (HIV-1) and mice (LP-BM5 strain of murine leukemia virus) to determine whether infection with these agents disrupted the regulation of the production of these antibodies. Retroviral infection in humans resulted in increased levels of autoantibody production against putative immunoregulatory regions of the Tcr s chain (Vs CDR1 and Fr3), a result reflecting a disruption of regulation. In addition, antigenic mimicry was observed with a cross-reaction shared between the common portion of the V3 neutralizing loop of the HIV-1 gp 120 molecule and the joining segment of T-cell receptors (Js). Infection of mice with the defective retrovirus resulted in the induction of antibodies directed particularly against Vs CDR1 peptide-defined determinants. Analysis of the virally induced response to a set of 8 CDR1 peptide epitopes indicated a selectivity to the process. It was possible to partially reverse aberrant cytokine changes correlated with the onset of murine MAIDS by administration of T-cell receptor peptides in saline. These results show that retroviral infection in humans and mice has a profound dysfunctional effect on the regulation of autoantibodies to T-cell receptors. The function of these autoantibodies in the immunopathogenesis of acquired immunodeficiency remains to be determined.

Normal individuals possess low levels of autoantibodies specific for certain peptide defined regions of T-cell receptor (Tcr) variable regions, particularly CDR1 and Fr3. These regions are predicted to be exposed on the surface of the native molecule and, by analogy and comparison with immunoglobulins, correspond to public idiotype determinants. The anti-Tcr idiotype antibodies appear to be ubiquitous and we propose that they play a role in the regulation of T-cell function. To delineate the parameters of expression of these antibodies, we characterized anti-Tcr antibody activity in normal individuals, in those suffering from the autoimmune diseases rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), and in patients with non-autoimmune arthritis (osteoarthritis) as a disease control. There were significant increases in autoantibody levels in the autoimmune patients. There was also variation in isotype and the particular variable regions recognized IgM autoantibodies directed against a few peptide defined determinants were elevated in RA, whereas SLE patient sera showed high levels of IgG binding to a broad spectrum of Tcr peptides.

Recently it has been observed that administration of intravenous immunoglobulin (IVIG) can have profound effects on a wide variety of diseases related to the dysregulation of the immune system. The mechanisms which explain these activities are poorly understood. Human IVIG and various Cohn plasma fractions contain autoantibodies directed against T cell receptors (Tcr). Previous studies have shown that IVIG contains autoantibodies against T cell receptor peptides. In order to further our understanding of autoantibody specificities, a single chain Tcr (scTcr) was constructed by recombinant DNA techniques from the variable α and variable s chains of the Jurkat cell line. Anti-Tcr autoantibodies were isolated from IVIG and Cohn fractions I + III using a scTcr affinity column. This scTcr affinity purified material reacted with the surfaces of T cells at l0ug/m1 whereas non-purified IVIG did not. Sera from patients with rheumatoid arthritis (RA) as well as serum from patients with systemic lupus erythematosus (SLE) reacted with the scTcr at levels above that of normals.

At some point in their illness, most patients with systemic lupus erythematosus (SLE) develop cold-reactive IgM anti-lymphocyte autoantibodies, which appear to represent a type of natural autoantibody that exhibits a disease-related increase in titer. Increased levels of such autoantibodies are found during flares of SLE disease activity in association with lymphopenia [7, 61], suggesting a causal relationship with T cell depletion and various functional abnormalities of T cells, B cells, and monocytes. Addition of SLE serum or immunoglobulin with anti-lymphocyte autoantibody activity to in vitro cultures of normal peripheral blood mononuclear cells alters their function in ways closely paralleling that of freshly isolated lymphocytes and monocytes from patients with SLE. Mechanisms by which such functional effects could occur in vivo include: (a) elimination of cells by complement-mediated lysis, antibody-dependent, cell-mediated cytotoxicity [15], and/or opsonization; (b) alteration of lymphocyte migration patterns; (c) modulation of surface determinants [63]; (d) agonist, partial agonist, or antagonist effects on cell surface receptors [38, 39, 65]; and (e) interaction with soluble products of activated cells [18]. Thus, autoantibodies to lymphocyte and monocyte surface determinants may constitute one of the pathogenetically significant extrinsic elements that alters cellular immune function in this disorder.

Immunoglobulins and their close relatives, the antigen-specific T-cell receptors, are recognition proteins that express structures which readily serve as self-immunogens. Healthy humans can produce antibodies against variable region-defined recognition structures termed idiotypes, as well as against constant region structures, and the levels of these can increase markedly in autoimmune disease; e.g., rheumatoid factors are autoantibodies directed against a conformational determinant of the gamma heavy chain. More recent analyses employing synthetic peptide technologies and construction of recombinant T-cell receptors document that autoantibodies directed against both variable and constant region markers of the alpha/beta T-cell receptor occur in healthy individuals. Alterations in levels of antibody, usage of IgM or IgG isotypes, and specificity for particular peptide-defined regions vary with natural physiological processes (aging, pregnancy), with artificial allografting, with retroviral infection, and with the inception and progression of autoimmune disease (e.g., rheumatoid arthritis, systemic lupus erythematosus). Two of the major autoimmunogeneic regions of the Tcr alpha/beta are "constitutive" markers inasmuch as all individuals tested produce antibodies against these regions. The most frequently observed autoantibodies are against Tcr V beta CDR1 and Fr3 markers. It is hypothesized that these are normally involved in immunoregulation. Autoantibodies usually are not detected against CDR2 region determinants, or the "private idiotypes" defined by the CDR3 region, or the highly conserved FR4 segment specified by the joining gene segment. However, autoantibodies against the CDR2 of the Tcr alpha chain occur in some SLE patients, and healthy pregnant women produce antibodies against the common peptide determinant expressed by the joining gene and the beginning of the C alpha or C beta domain. Although the precise role of the naturally occurring autoantibodies in immunoregulation remains to be determined, modification of the course of autoimmune diseases in experimental rodent models (experimental allergic encephalomyelitis) has been successfully carried out by immunization with synthetic peptides corresponding to the CDR2 and Fr3/CDR3 segments, and immunization of humans with synthetic V beta CDR2 segments may prove helpful in multiple sclerosis. Moreover, infusion of intravenous immunoglobulins has been successful in the treatment of many autoimmune diseases, including examples where levels of T cells bearing particular V beta gene subsets were elevated. The recent knowledge gained from T-cell receptor structural analysis and antigenic modeling holds promise for determining the roles of particular variable domain structures in antigen recognition MHC-restriction and immunoregulation, and in the development of synthetic and recombinant reagents for modulation of autoimmune and infectious diseases.

Clinically healthy humans as well as patients suffering from various autoimmune diseases produce natural antibodies against a variety of self-components. Such antibodies have been proposed to carry out a physiologic role in maintaining the integrity of self, as well as potentially destructive roles in the generation of autoimmune diseases. Because human autoantigens, particularly membrane proteins, are usually present in extremely small amounts, it is generally impossible to obtain enough to carry out a detailed characterization of the antibodies or the antigenic determinants recognized. To circumvent this difficulty, we developed synthetic autoantigens predicted from the gene sequence of two functionally critical membrane proteins; the band 3 anion transport protein which is found on all cells, and the T-cell receptor (beta chain) which is the antigen-specific receptor on thymus-derived lymphocytes. We have investigated the natural human IgM and IgG antibody responses to peptides selected on the basis of predicted molecular surface exposure and previously known antigenicity, and correlate levels of binding with changes in age and by comparison with autoimmune diseases. We report that the IgM response to synthetic autoantigens tends to be higher than that of IgG molecules, but significant IgG binding occurs to some peptides. This situation is particularly noticeable in comparison of rheumatoid arthritis patients with normal individuals. Distinct peptide portions of individual molecules are recognized differently by the autochthonous immune system as manifested by age dependence of the response and differential levels of IgM and IgG activity. The synthetic autoantigens that tend to generate the highest amounts of natural antibody are those that are either exposed on the surface of the cell (band 3 peptides) or are exposed in the predicted 3-dimensional folding of the molecule (T-cell receptor beta peptides). Rheumatoid arthritis patients tend to give higher IgM reactivities to both band 3 and Tcr beta peptides than do normals, with this effect being less pronounced in the distinct autoimmune disease systemic lupus erythematosus. Studies of normal humans ranging in age from 20 to 90 years suggest two major patterns for the IgM natural antibody response to synthetic peptides giving high response. The first is that the level of IgM reactivity is high early in life and remains high throughout. The second pattern is one in which the reaction is high in younger individuals, but diminishes substantially in the latter decades of life.(ABSTRACT TRUNCATED AT 400 WORDS)

The T-cell receptor V beta subfamily repertoires of synovial and peripheral T cells of 8 rheumatoid arthritis (RA) patients were determined using the polymerase chain reaction. Three normal controls were included. Some of the rheumatoid synovial samples did not express the complete range of V beta families and lacked as many as 6 gene families. However, these patients showed considerable individual variation in expression. Overall, the data do not support preferential T-cell receptor V beta usage in synovial T cells of RA patients either in comparison to their autochthonous peripheral T cells or to peripheral T cells of normal subjects.

An aging antigen, senescent cell antigen, resides on the 911 amino acid membrane protein band 3. It marks cells for removal by initiating specific IgG autoantibody binding (1–22). This appears to be a general physiologic process for removing senescent and damaged cells in mammals and other vertebrates (4). Although the initial studies were done using human erythrocytes as a model, senescent cell antigen occurs on all cells examined (4). The aging antigen itself is generated by the degradation of an important structural and transport membrane molecule, protein band 3 (5). Besides its role in the removal of senescent and damaged cells, senescent cell antigen also appears to be involved in the removal of erythrocytes in clinical hemolytic anemias (7,8), and the removal of malaria-infected erythrocytes (23,24). Oxidation generates senescent cell antigen in situ (6). Neither cross-linking nor hemoglobin appear to play a role. Although storage is the only in vitro model that mimics cellular aging in situ, we have discovered three alterations/mutations of band 3 that permit insight into aging in situ. One mutation with an addition to band 3 has normal or decelerated red cell aging. In contrast, another band 3 alteration with a suspected deletion or substitution that renders band 3 more susceptible to proteolysis, shows accelerated aging. The third alteration which is also more susceptible to proteolysis is associated with neurologic defects.

Retrovirally infected humans and mice showed progressive acquired immunodeficiency accompanied by the production of elevated levels of autoantibodies directed against T-cell receptor variable-domain epitopes. Epitope mapping analyses indicated that a major determinant recognized was defined by a 16-mer peptide containing the entire CDR1 segment and part of the FR2 region of human Vbeta8, and that both species showed reactivity to the same sequence. Either prophylactic or therapeutic administration of this peptide to retrovirus-infected C57/BL/6 mice normalized the balance of T(H)1- and T(H)2-type helper activity and restored the resistance to infection by the opportunistic parasite Cryptosporidium. Administration of the peptide did not generate significantly increased levels of autoantibody, but had a profound effect on T-cell activity as well as other aspects of inflammation, including NK-cell activity. A 16-mer derived from the Jbeta sequence showed similar functional effects on T cells from retrovirus-infected mice. Direct binding of the VbetaCDR1 peptide to recombinant TCR Valpha/Vbeta constructs, as well as to IgM natural autoantibodies, suggests that the cell surface receptor for the peptide is the alpha/beta TCR on T cells and surface IgM in B cells. The Vbeta CDR1 peptide stimulated division of murine splenocytes in vitro, stimulated the production of the T(H)1 cytokine IL-2, and synergized with the T-cell mitogen concanavalin A in proliferation and IL-2 production. These studies indicate that administration of peptides derived from T-cell receptor variable domains to animals immunosuppressed as a result of retroviral infection has a profound immunomodulatory effect enhancing overall T-cell functional capacity, particularly with respect to the cytokine production characteristic of T(H)1-type cells. Our studies are interpreted in the context of other recent investigations of immunomodulatory peptides.

The purpose of this study was to use affinity-purified polyclonal antibodies produced against a synthetic peptide corresponding to the joining (J) region of a human T cell receptor β chain to characterize antigen receptor expression on subpopulations of human lymphocytes. The synthetic peptide used was ANYGYTFGSGTRLTVV, corresponding to the J segment of the human β-chain gene YT35. Biochemical characterization has previously demonstrated binding of anti-J β peptide antibodies to the α β heterodimer and to certain immunoglobulin light chains. Flow cytometric analysis of normal human peripheral blood lymphocytes performed here, using affinity-purified antibodies to the J β peptide, showed expression of the epitope on 50–60% of CD20 (B1)-positive B lymphocytes, and on 40–50% of CD8-positive T lymphocytes. Only background levels were observed on CD4-positive T cells.

Surface immunoglobulins (Igs) of lymphocytes are of considerable interest because these molecules probably function as receptors for antigen, and knowledge of their molecular properties should provide information on the mechanisms of immune differentiation. The density and types of surface Ig on a cell provide markers useful in indicating the class of a lymphocyte and its stage of maturity. Moreover, knowledge of the specificities of the surface Ig of neoplastic lymphocytes might suggest the nature of agents involved in the generation of the disease. Two broad classes of lymphocytes, bone marrow-derived lymphocytes (B cells) active in antibody secretion, and thymus-derived lymphocytes (T cells) which mediate cellular immune reactions, and their subpopulations must be considered with reference to the nature, origin, and function of their surface immunoglobulin. This article analyzes direct and indirect methods for the demonstration of surface Igs and describes certain physicochemical properties of isolated surface Ig molecules. Roles of these surface molecules in recognition of antigen, initiation of all differentiation, and cooperation among lymphocytes and accessory cells are discussed.

Insulin receptors have been demonstrated on activated but not resting T-lymphocytes. We compared insulin binding to several T- and B-cell tissue culture lines transformed by either radiation or lymphotrophic Herpes virus. Two T-cell lines derived from mouse lymphomas, one T-cell derived from the cotton top marmoset and one mouse B-cell line bound, at most, 3% of a tracer dose of [ 125 I]insulin. In contrast, positive B-cell line (1605 L) derived from a leukemic cotton top marmoset, bound 40% of tracer [ 125 I]insulin. The binding was of high affinity (1.9 nM −1 ), displayed analogue specificity characteristic of the insulin receptor and receptor number per cell was 16,600. The data show that a transformed B-lymphocyte binds substantial levels of insulin. The magnitude of such expression might be related to the fact that this B-cell line also expresses Epstein-Barr virus.

The monoclonal murine T cell hybridoma, 51H7D, was previously shown to bind the arsazobenzene hapten and to produce a soluble antigen-binding molecule. In this paper we characterize this antigen-binding immunoprotein for its relationship to known T cell receptors serologically, using antibodies specific for variable region framework, or joining region peptides predicted from gene sequence and by biochemical means. The 51H7D cell expresses a protein with subunit size of approximately 31,000, that reacts antigenically with affinity-purified antibodies directed against synthetic first framework and joining segment peptides, corresponding to the gene sequence of the T cell receptor beta chain, YT35. This molecule does not react with affinity-purified antibodies directed against murine immunoglobulin, framework 1 sequences of alpha and gamma T cell receptors, or with antibodies against synthetic heavy chain joining segments. The subunit of mol. wt. 31,000 can form higher aggregates, notably in the mol. wt range of 60,000-70,000, depending upon extraction conditions. The soluble form of the antigen-binding molecule bears the J beta cross-reactive determinant and occurs predominantly as a charge restricted molecular species of approximate mol. wt 60,000-70,000. The purified molecule has a blocked N-terminus, but quantitative statistical analysis of its amino acid composition indicates a closer relatedness to T cell receptor beta chains and other antigen-binding T cell products, than it has to alpha, gamma or delta TCR chains. No evidence for more than one type of polypeptide chain was found and the polymerization is not dependent upon the formation of disulfide bonds. These studies raise the possibility that antigen-binding soluble T cell molecules might belong to a new family of immunoproteins, that is related to, but distinct from, classical immunoglobulins and alpha beta or gamma delta heterodimers.

Chicken antibodies raised to the (Fab')2 fragment of pplyclonal human IgG recognize a set of configurational determinants, including idiotypic determinants of individual myeloma proteins, formed by certain heavy chain Fd regions in association with κ light chains. Chicken anti-Fab binds, as shown by indirect immunofluorescence, to a surface component of human and marmoset B cells and to some neoplastic T cell lines, as well as to approximately 50% of T cells isolated from human peripheral blood. It does not bind to normal or neoplastic ‘null’ cells. The generation of human alloreactive cytotoxic T cells in vitro is substantially inhibited by incubation with chicken anti-human (Fab')2; whereas chicken anti-mouse (Fab')2 has no effect. Conversely, chicken anti-mouse (Fab')2 completely inhibits the in vitro induction of alloreactive mouse T cells, but does not affect the human mixed lymphocyte reaction. Components bearing Fab-related antigenic determinants were isolated from culture fluids in which two marmoset T cell lines had been grown. When analysed by polyacrylarnide gel etectrophoresis under reducing conditions in sodium dodecyl sulfate-containing buffers, the major Fab-related component from line 70-N2 had an apparent mass of 70,000 dallons and that from line 22-CM-37 had an apparent mass in the range of 44,000–50,000 daltons.

Proteins unique to B16 melanoma cells grown in culture and as solid tumors have been independently described recently. As both proteins have apparent M r of 65,000-70,000, we have more fully compared their biochemical characteristics. The M r 65,000 protein, which is isolated from B16 cells in culture, has an aggregate molecular weight of >200,000 under nonreducing conditions. This M r 65,000 form is shed into the culture medium exclusively and is the predominant molecular species of serum-free culture supernatants. It was found that B700, a major membrane protein of B16 cells in solid tumors, and the M r 65,000 preparation from cultured cells were electrophoretically identical when compared under four separate sets of conditions. These two proteins also share immunologic determinants. The results suggest the identity of these two proteins isolated by completely different approaches.

Immunoglobulin (Ig) released into the medium by monoclonal continuously cultured murine T lymphoma cells of the lines WEHI-22 and WEHI-7 was isolated by serological precipitation or solid-phase immunoadsorption techniques. The intact immunoglobulin had an electrophoretic mobility on sodium dodecyl-sulphate (SDS) containing polyacrylamide gels comparable to that of IgG (mass 150,000). This mobility was significantly faster than that of '7S' IgM of murine B lymphocyte surfaces. The T lymphoma immunoglobulin consisted of a pair of heavy chains linked by disulphide bonds and light chains non-covalently bound to the heavy chains. The isolated heavy chains migrated slightly faster than the mu chains of MOPC 104E IgM. Some, but not all, antisera directed against mu chains of normal mouse serum IgM bound T lymphoma immunoglobulin apparently via a cross-reaction localized to the Fd fragment. These data indicate that immunoglobulin of T lymphoma cells and, presumably normal T lymphocytes, represents an immunoglobulin isotype which is distinct from those immunoglobulins found on the B cell surface.

In vitro investigations were carried out to determine whether lymphocyte-like (small round) cells of the tunicate Pyura stolonifera react to allogeneic cells and mitogens in a manner comparable to that of vertebrate immunocytes. The lymphocyte-like cells possessed receptors for concanavalin A, wheat germ agglutinin and soybean lectin as shown by binding of radioiodinated lectins in scintillation counting and autoradiographic assays. This binding did not induce mitogenesis. Mixtures of cells of P. stolonifera individuals taken from the same or distinct localities did not show enhanced DNA synthesis when assayed at time intervals ranging from 3 to 10 days. The small round cells of this and other tunicates are not completely similar to vertebrate lymphocytes in morphology. Our observations support the concept that these cells are not directly homologous to immunologically competent vertebrate lymphocytes, but may serve as haemopoietic stem cells as proposed by other (Endean, 1954; Freeman, 1970).

Serological studies were carried out to obtain information regarding the relationship of the V H -related determinants expressed by certain permanent in vitro T-cell leukemia lines and corresponding determinants expressed by characterized human serum immunoglobulins. A panel of conventional (goat and rabbit) antisera, produced against various Fab-related fragments of monoclonal human Waldenstrom macroglobulins and polyclonal IgG molecules, bound to certain in vitro T-cell leukemia lines, notably, 70-N2, MT-1, YT4E, and HUT78, as shown by microhemagglutination. Inhibition studies using characterized myeloma proteins to inhibit this agglutination indicated the expression of a restricted V H -related determinant by these T-cell lines. Parallel studies performed using conventional (rabbit) and murine monoclonal/hybridoma antibodies produced against the isolated 68,000-Da V H -related product synthesized by the 70-N2 line showed that the determinant expressed by this molecule was restricted in expression, comprising 2–3% of the normal, polyclonal human Fab pool, and that the determinants found on the other positive T-cell leukemias were cross-reactive rather than identical. The inhibition studies suggest that the determinant resides between residue 22 and the end of the V H region. These results further define the antigenic nature of the V H -related marker found on the surfaces of certain normal and neoplastic T-cell lines.

Data of recent studies based upon three approaches to the problem of the T cell receptor for antigen, namely, use of ‘anti-receptor’ (≡ anti-idiotype) antibodies, utilization of antisera made against circulating antibodies in association with physicochemical characterization procedures, and phylogenetic investigations, support the conclusion that this recognition molecule is an immunoglobulin, but that it represents an isotype distinct from usual circulating antibodies and B cell surface IgM- and IgD-like molecules.