Receptor/ligand avidity determines the capacity of Ly49 inhibitory receptors to interfere with T-cell receptor-mediated activation

The specificity and the relative affinity of many Ly49 receptors for major histocompatibility complex class I ligands have been studied in detail in various adhesion and binding assays. However, how the level of cell surface expression of a given Ly49 receptor and its ligand affinity influence the strength of the inhibition signal is not well documented. To address this issue, we developed a series of human Jurkat T-cell transfectants expressing the whole range of Ly49A and Ly49C levels found in vivo on natural killer and T cells and evaluated their capacity to alter superantigen-induced NF-AT activation and interleukin-2 production. We show that the strength of the inhibition induced by Ly49A/H-2Dd interaction correlates with Ly49A density up to a certain level after which increasing expression does not further inhibit significantly the T-cell receptor-induced activation. This system also represents a valuable tool for the determination of the relative strength of the inhibitory signals of Ly49 receptors following their interactions with different ligands. Even at high levels of expression there was no evidence that engagement of Ly49A with H-2b class I molecules provided an inhibitory signal. Moreover, we showed that functional inhibitory interactions of Ly49C with H-2b class I molecules were only the result of H-2Kb and that H-2d represent lower affinity ligands for Ly49C than H-2b. Therefore, depending on the relative affinity of Ly49 receptors for their ligands, the modulation of their expression level will be determinant for the functional outcome of activated T cells.     

Molecular basis of cross-reactivity among allergen-specific human T cells: T-cell receptor V alpha gene usage and epitope structure.

Cross-reactivities between the major grass pollen allergens, at the level of T-cell recognition was examined employing several Lolium perenne I (Lol p I)-specific human T-cell clones. Nine of these Lol p I-specific T-cell clones exhibited cross-recognition of the recombinant Poa pratensis IX (Poa p IX) allergen, rKBG7.2, indicating that these two major antigens of a grass pollen share T-cell epitopes. Furthermore, proliferative responses of two other T-cell clones demonstrated that individual allergens of diverse grass pollens also possess common T-cell epitopes. Examination of the T-cell receptor (TcR) V alpha genes of these T-cell clones indicated that these cloned cells utilized distinct J alpha genes and that nine out of 10 clones possessed V alpha 13 gene. Furthermore, sequence comparisons of several allergenic molecules indicated that this cross-reactivity may be due to the presence of epitope(s) with structure(s) similar to the major T-cell epitope of Poa p IX allergens. Taken together, these results suggest for the first time that the major grass pollen allergens share cross-reacting T-cell epitope(s), and that this cross-reactivity is due to the structural homologies among allergens and restricted usage of TcR V alpha genes.     

A human suppressor T-cell factor that inhibits T-cell replication by interaction with the IgM-Fc receptor (CD7)

We have previously described the induction of human suppressor T cells from fresh peripheral blood lymphocytes of a kidney transplant recipient by in vitro stimulation with an autologous irradiated antidonor CTL line (EE-1) grown from a biopsy of the patient's own renal allograft. The induced T cells (designated TsEE) were shown to inhibit the in vitro generation of proliferative and cytotoxic responses of autologous T cells and nonautologous T cells that shared HLA-B7 with TsEE cells. Stimulation of TsEE cells by the autologous irradiated inducer line (EE-1) produced soluble factors (designated TsEEF) that similarly inhibited autologous and nonautologous T-cell responses to alloantigens and mitogens, but in a non-HLA-restricted manner. In this study, we examined the functional interaction of TsEEF with various cells surface receptors. TsEEF specifically inhibited the proliferation of stimulated and transformed T cells expressing CD7, a putative receptor for IgM-Fc (FcRmu). Blocking or capping of CD7-FcRmu determinants on responder T cells by pretreatment with IgM or anti-CD7 monoclonal antibodies (3A1, HuLy-m2) abrogated TsEEF activity. Conversely, pretreatment of T cells with TsEEF significantly reduced their binding of IgM and HuLy-m2. TsEEF was demonstrated not to be IgM or IgG, and its activity was not removed by preabsorption with IgM or IgG; however, its activity could be competitively inhibited by coculture with IgM. By cocapping experiments and studies utilizing CD7- (Hut-78) and CD7+ (HSB, Molt-4) T-cell lines. TsEEF activity did not appear to involve interactions with other T-cell or non-T-cell surface receptors. These findings suggest a novel role for FcRmu-CD7 T-cell surface receptors in binding certain soluble T-cell factors that result in the inhibition of T-cell replication.     

Binding energetics of T-cell receptors: correlation with immunological consequences.

Recent crystal structures of the T-cell receptor (TCR) engaged with peptide–major histocompatibility complex (MHC) ligands suggest the TCR binds to such ligands in a conserved orientation, with key Vα residues contacting the α helices. TCR mutagenesis has permitted energy measurements of these interactions. Here, Thomas Manning and David Kranz review binding energetics data that might explain the inherent reactivity of TCRs with MHC products.     

T-cell antigen receptors: fact and artefact

Much has been written to support the view that T-cell antigen-recognition molecules share idiotypes with immunoglobulin and are encoded in the same V_H genes. Here Jacques Miller and his colleagues disbute this view and argue instead, from recent evidence, that the genes for T-cell receptors are entirely different from Ig genes.     

Extraordinary cross-reactivity of an autoimmune T-cell receptor recognizing specific peptides both on autologous and on allogeneic HLA class II molecules

A T-cell receptor's (TCR) recognition of a human leukocyte antigen (HLA)-peptide complex (pHLA) is normally described as being restricted by the HLA molecule and specific for the peptide. This is, however, not always true. Several TCRs have been described, which cross-react with other peptides bound to the restricting HLA molecule. This phenomenon has been considered a variant of molecular mimicry and is suggested to be one of the mechanisms behind autoimmunity. The positive selection of T cells in the thymus imposes low-affinity recognition of the TCRs toward self-pHLA, which increases the probability of the TCR to be promiscuous by nature, and further implies that the T-cell repertoire contains TCRs prone to be autoreactive and thus able to induce autoimmunity. We present an autoimmune TCR showing extreme cross-reactivity to several pHLA comprising both own HLA class II restriction element and allogeneic HLA class II restriction elements in complex with both self-derived and microbially derived peptides. The existence of such a significant cross-reactivity in the context of distinct HLA-DR molecules might be more common among autoimmune TCRs than previously anticipated and potentially reveals a new way of designing altered peptide ligands for therapeutic use.     

Coevolution of T-cell receptors with MHC and non-MHC ligands

The structure and amino acid diversity of the T-cell receptor (TCR), similar in nature to that of Fab portions of antibodies, would suggest that these proteins have a nearly infinite capacity to recognize antigen. Yet all currently defined native T cells expressing an α and β chain in their TCR can only sense antigen when presented in the context of a major histocompatibility complex (MHC) molecule. This MHC molecule can be one of many that exist in vertebrates, presenting small peptide fragments, lipid molecules, or small molecule metabolites. Here we review the pattern of TCR recognition of MHC molecules throughout a broad sampling of species and T-cell lineages and also touch upon T cells that do not appear to require MHC presentation for their surveillance function. We review the diversity of MHC molecules and information on the corresponding T-cell lineages identified in divergent species. We also discuss TCRs with structural domains unlike that of conventional TCRs of mouse and human. By presenting this broad view of TCR sequence, structure, domain organization, and function, we seek to explore how this receptor has evolved across time and been selected for alternative antigen-recognition capabilities in divergent lineages.     

Characterization of murine complement receptor type 2 and its immunological cross-reactivity with type 1 receptor.

We have previously demonstrated that some mAbs prepared against mouse complement receptor type 1 (CR1) bind a 150,000 Mr protein in addition to the 190,000 Mr CR1 protein. We now identify the 150,000 Mr murine protein as complement receptor type 2 (CR2), since: (i) one of the monoclonal antibodies that bind this protein inhibits rosette formation between mouse B cells and C3d-bearing sheep erythrocytes; (ii) as is known for human CR2, this protein is present on B lymphocytes but not T lymphocytes; and (iii) this protein must have affinity for C3b, since it has weak factor I cofactor activity. In addition, this protein resembles the 145,000 Mr human CR2 molecule in size. Since four of the five mAbs that were produced by immunization with CR1 also bound CR2, and they bind to different CR1 epitopes, it seems that murine CR1 and CR2 share multiple epitopes. Injection of mice with one of the CR1-CR2 cross-reactive mAbs almost eliminated both CR1 and CR2 expression, but did not decrease B cell numbers or the expression of B cell IgM, Ia, or B220 antigens. In contrast, injection of mice with a non-cross-reactive anti-CR1 antibody only modulated CR1 expression. These antibodies should thus provide useful tools for the study of the in vivo roles of B cell complement receptors.     

Antigen-induced regulation of T-cell motility,interaction with antigen-presenting cells and activation through endogenous thrombospondin-1 and its receptors

Antigen recognition reduces T-cell motility, and induces prolonged contact with antigen-presenting cells and activation through mechanisms that remain unclear. Here we show that the T-cell receptor (TCR) and CD28 regulate T-cell motility, contact with antigen-presenting cells and activation through endogenous thrombospondin-1 (TSP-1) and its receptors low-density lipoprotein receptor-related protein 1 (LRP1), calreticulin and CD47. Antigen stimulation induced a prominent up-regulation of TSP-1 expression, and transiently increased and subsequently decreased LRP1 expression whereas calreticulin was unaffected. This antigen-induced TSP-1/LRP1 response down-regulated a motogenic mechanism directed by LRP1-mediated processing of TSP-1 in cis within the same plasma membrane while promoting contact with antigen-presenting cells and activation through cis interaction of the C-terminal domain of TSP-1 with CD47 in response to N-terminal TSP-1 triggering by calreticulin. The antigen-induced TSP-1/LRP1 response maintained a reduced but significant motility level in activated cells. Blocking CD28 co-stimulation abrogated LRP1 and TSP-1 expression and motility. TCR/CD3 ligation alone enhanced TSP-1 expression whereas CD28 ligation alone enhanced LRP1 expression. Silencing of TSP-1 inhibited T-cell conjugation to antigen-presenting cells and T helper type 1 (Th1) and Th2 cytokine responses. The Th1 response enhanced motility and increased TSP-1 expression through interleukin-2, whereas the Th2 response weakened motility and reduced LRP1 expression through interleukin-4. Ligation of the TCR and CD28 therefore elicits a TSP-1/LRP1 response that stimulates prolonged contact with antigen-presenting cells and, although down-regulating motility, maintains a significant motility level to allow serial contacts and activation. Th1 and Th2 cytokine responses differentially regulate T-cell expression of TSP-1 and LRP1 and motility.     

T-cell receptor retrogenic mice: a rapid, flexible alternative to T-cell receptor transgenic mice

The T-cell receptor (TCR) is unique in its complexity. It determines not only positive (life) and negative (death) selection in the thymus, but also mediates proliferation, anergy, differentiation, cytotoxicity and cytokine production in the periphery. Through its association with six CD3 signalling chains (εγ, δε and ζζ), the TCR is capable of recognizing an extensive variety of antigenic peptides, from both pathogens and self-antigens, and translating these interactions into multiple signalling pathways that mediate diverse T-cell developmental and functional responses. The analysis of TCR biology has been revolutionized by the development of TCR transgenic mice, which express a single clonotypic T-cell population, with diverse specificities and genetic backgrounds. However, they are time consuming to generate and characterize, limiting the analysis of large numbers of TCR over a short period of time in multiple genetic backgrounds. The recent development of TCR retrogenic technology resolves these limitations and could in time have a similarly important impact on our understanding of T-cell development and function. In this review, we will discuss the advantages and limitations of retrogenic technology compared with the generation and use of TCR transgenic mice for studying all aspects of T-cell biology.     

The half-life of the T-cell receptor/peptide-major histocompatibility complex interaction can modulate T-cell activation in response to bacterial challenge

T-cell activation results from engagement of the T-cell receptor (TCR) by cognate peptide-major histocompatibility complex (pMHC) complexes on the surface of antigen-presenting cells (APC). Previous studies have provided evidence supporting the notion that the half-life of the TCR/pMHC interaction and the density of pMHC on the APC are two parameters that can influence T-cell activation. However, whether the half-life of the TCR/pMHC interaction can modulate T-cell activation in response to a pathogen challenge remains unknown. To approach this question, we generated strains of bacteria expressing variants of the ovalbumin (OVA) antigen, carrying point mutations in the SIINFEKL sequence. When bound to H-2K(b), this peptide is the cognate ligand for the OT-I TCR. Variants of the H-2K(b)/SIINFEKL bind to the OT-I TCR with distinct half-lives. Here we show that dendritic cells (DCs) infected with bacteria expressing OVA variants were incapable of activating OT-I T cells when the half-life of the TCR/H-2K(b)/OVA interaction was excessively short. Consistent with these data, T-cell activation was only observed in mice infected with bacteria expressing OVA variants that bound to OT-I with a half-life above a certain threshold. Considered together, our data suggest that the half-life of TCR/pMHC interaction can significantly modulate T-cell activation in vivo, as well as influence recognition of antigens expressed by bacteria. These observations underscore the importance of the TCR/pMHC half-life on the clearance of pathogens.     

CD8 T-cell ability to exert immunodomination correlates with T-cell receptor: epitope association rate.

When presented alone, H7 a and HY antigens elicit CD8 T-cell responses of similar amplitude, but H7 a totally abrogates the response to HY when both antigens are presented on the same antigen-presenting cell. We found that H7a- and HY-specific T-cell precursors had similar frequencies in nonimmune mice and expressed similar levels of CD5. The H7a -specific CD8 T-cell repertoire harvested at the time of primary response showed highly restricted T-cell receptor (TCR) diversity. Furthermore, T cells specific for H7a and HY expressed equivalent levels of CD8 and TCR and displayed similar tetramer decay rates. The key difference was that anti-H7a T cells exhibited a much more rapid TCR:epitope on-rate than anti-HY T cells. Coupled with evidence that primed CD8 T cells limit the duration of antigen presentation by killing or inactivating antigen-presenting cells, our data support a novel and simple model for immunodomination: the main feature of T cells that exert immunodomination is that, compared with other T cells, they are functionally primed after a shorter duration of antigen presentation.     

Lack of cross-reactivity of a monoclonal antibody against the neurotransmitter site of Torpedo nicotinic acetylcholine receptor with muscle receptors from several sources

A monoclonal antibody raised against Torpedo nicotinic acetylcholine receptor has been used to study the neurotransmitter binding site of the acetylcholine receptor from several sources. When tested on Torpedo receptor, this monoclonal antibody inhibited binding of -bungarotoxin to 50% of the available sites. The failure to complexity inhibit binding of the toxin is attributed to the orientation of the determinant for the monoclonal antibody on the receptor molecule. This monoclonal antibody bound very poorly to the acetylcholine receptor from muscle tissues. This suggests either a modification or a reduced accesibility of the determinant to the monoclonal antibody.     

Sequential activity of gamma-delta T-cell receptor bearing lymphocytes, specific T-cell factors, and alpha-beta T-cell receptor bearing T-cells: a hypothesis

The cell-mediated immune response against cell-bound antigens are biphasic responses. The 'classical' components of cell-mediated immunity such as delayed type hypersensitivity (DTH) and cytotoxic T-cells, are preceded by antigen-specific T-cell factor production. These antigen-specific T-cell factors can be detected in the serum 1-2 days after immunization, which suggests that these antigen-specific T-cell factors play an important role in the initiation of cell-mediated immune responses. Factor-producing lymphocytes can be detected already 1 day after immunization at the site of the antigen challenge, and at first 4-5 days after immunization in the peripheral lymphoid organs. The phenotype of these factor-producing lymphocytes is Thy-1+, CD3+, L3T4 (CD4)-, Lyt2 (CD8)-, whereas the T-lymphocytes inducing the DTH response are Thy 1+, CD3+, L3T4 (CD4)+, Lyt2 (CD8)-. This suggests that these factor-producing lymphocytes are gamma-delta bearing T-lymphocytes. gamma-delta TCR bearing T-cells can develop independently of the thymus, and are present in athymic nude mice. Immunization of athymic (nude) mice induces antigen-specific T-cell factor production as well. This suggests that the gamma-delta bearing T lymphocytes present in nude mice can react to cell-bound antigens and produce these factors.     

Investigation of antigen cross-reactivity of Mycobacterium leprae-reactive murine T-cell lines and clones.

Inguinal lymph node lymphocytes from BALB/c mice immunized intradermally with 10(8) 60Co-irradiated Mycobacterium leprae were cloned by limiting dilution culture. In general, cloned T-cell lines exhibited helper type activity producing interleukin-2, macrophage activation factor and gamma-interferon and lines were further characterized in terms of their cross-reactivities with other species of mycobacteria. M. leprae clones derived after a period of in vitro restimulation were found to cross-react with other species of mycobacteria probably recognizing non-specific or closely related common cell wall associated mycobacterial determinants. On the other hand, lines established by cloning directly from immune mice appeared more M. leprae-specific, exhibiting antigen-dependent lymphokine production and proliferation in vitro.