T cell receptor-mediated signs and signals governing T cell development

The developmental fate of T cells is largely controlled by the nature and success of signals mediated by the pre-T cell receptor (TCR) and TCR complexes. These intracellular signals are regulated by cascades of protein tyrosine phosphorylations initiated following ligand binding to the pre-TCR or TCR complexes. The phosphorylation cascades are primarily orchestrated by two distinct families of protein tyrosine kinases (PTKs), the Src- and the Syk/ZAP-70-families. Germline gene targeting experiments, several human immunodeficiencies, and somatic cell mutants have all contributed to our understanding of how these families of kinases coordinate their actions to promote signaling. Upon activation, the PTKs transmit their signals to a number of newly described adaptor proteins including LAT, SLP-76, and vav, among others. The following review combines results derived from different experimental strategies to examine the contributions of the PTKs and the adaptor molecules to pre-TCR and TCR signaling processes.     

T cell recognition of epithelial self

The presentation of self-antigens to circulating T cells is a critical, precipitating event in the induction of autoimmune injury in parenchymal organs. Epithelia expressing these self-antigens are thought to release such moieties for reprocessing by traditional antigen-presenting cells within the lymphoid system. We now demonstrate, however, that some epithelium possess novel functional mechanisms for presenting their own antigens to a responsive, syngeneic T cell repertoire. The presentation of these self-antigens occurs in the context of MHC class II molecules and depends on CD4 associative-recognition determinants. Our findings strongly suggest that organ epithelium may directly activate cell-mediated events to produce autoimmunity through self-recognition.     

Human T cell immunodeficiency

Severe combined immunodeficiency (SCID) is a heterogeneous group of diseases that are invariably fatal in infancy unless treated by hematopoietic stem cell replacement. For many years we have worked to better manage patients affected by SCID through rapid and accurate diagnosis followed by treatment aimed at achieving long-lasting immune reconstitution. By extensive immunological, biochemical, and genetic studies of patient samples, and with the realization of differences between human and murine T cell development, we have successfully been able to identify some of the molecular defects causing SCID. Among these discoveries, we described the first mutated signal transduction protein in T cells (ZAP-70); the first genetic defect leading to SCID and autoimmune phenomena (IL2Rα); and, recently, the critical importance of CD3δ in the development of T cells. Our efforts have significantly advanced the understanding of the role of some of the signal-transducing proteins in T cell maturation and function. This review summarizes several of these discoveries and some of their impact on our understanding of T cells development, function, and homeostasis in humans.     

Human T cell subsets

Several monoclonal antibodies (mAbs) demonstrate further heterogeneity of CD4 and CD8. Within CD4, two major subsets can be identified with helper/inducer and suppressor/inducer functions. It is argued that suppressor/inducers, identified by CD45R mAbs, are immature or virgin T cells and therefore respond poorly to recall antigens. Helper/inducers are CD45R negative but express the UCHL1 antigen, which appears to be part of CD45. These cells respond well to recall antigens and are therefore mature or memory T cells. Several antigens not related to CD45, such as 4B4, Leu8, TQ1 and 9.3, identify functionally distinct subsets rather than stages of T cell maturation.     

T cell antigen recognition at the cell membrane

T cell antigen receptors (TCRs) on the surface of T cells bind specifically to particular peptide bound major histocompatibility complexes (pMHCs) presented on the surface of antigen presenting cells (APCs). This interaction is a key event in T cell antigen recognition and activation. Most studies have used surface plasmon resonance (SPR) to measure the in vitro binding kinetics of TCR-pMHC interactions in solution using purified proteins. However, these measurements are not physiologically precise, as both TCRs and pMHCs are membrane-associated molecules which are regulated by their cellular environments. Recently, single-molecule f?rster resonance energy transfer (FRET) and single-molecule mechanical assays were used to measure the in situ binding kinetics of TCR-pMHC interactions on the surface of live T cells. These studies have provided exciting insights into the biochemical basis of T cell antigen recognition and suggest that TCRs serially engage with a small number of antigens with very fast kinetics in order to maximize TCR signaling and sensitivity.     

T cell receptor crossreactivity as a general property of T cell recognition

TCR recognition of MHC/peptide complexes directs many aspects of T cell biology, including thymic selection, survival of na?ve T cells and differentiation into effector and memory T cells. It was widely thought that TCR recognition is highly specific, with an individual T cell being capable of only recognizing a particular peptide and closely related sequence variants. By considering the structural requirements for peptide binding to MHC molecules and TCR recognition of MHC/peptide complexes, we demonstrated that T cell clones could recognize a number of peptides from different organisms that are remarkably distinct in their primary sequence. These peptides are particularly diverse at those sequence positions buried in pockets of the MHC binding site, while a higher degree of similarity is present at a limited number of peptide residues that create the interface with the TCR. Many examples have now been documented for human and murine T cells, indicating that TCR crossreactivity represents a general feature of TCR recognition.     

Polyspecificity of T cell and B cell receptor recognition

A recent workshop discussed the recognition of multiple distinct ligands by individual T cell and B cell receptors and the implications of this discovery for lymphocyte biology. The workshop recommends general use of the term polyspecificity because it emphasizes two fundamental aspects, the inherent specificity of receptor recognition and the ability to recognize multiple ligands. Many different examples of polyspecificity and the structural mechanisms were discussed, and the group concluded that polyspecificity is a general, inherent feature of TCR and antibody recognition. This review summarizes the relevance of polyspecificity for lymphocyte development, activation and disease processes.     

Differential T cell receptor-mediated signaling in naive and memory CD4 T cells

Naive and memory CD4 T cells differ in cell surface phenotype, function, activation requirements, and modes of regulation. To investigate the molecular bases for the dichotomies between naive and memory CD4 T cells and to understand how the T cell receptor (TCR) directs diverse functional outcomes, we investigated proximal signaling events triggered through the TCR/CD3 complex in naive and memory CD4 T cell subsets isolated on the basis of CD45 isoform expression. Naive CD4 T cells signal through TCR/CD3 similar to unseparated CD4 T cells, producing multiple tyrosine-phosphorylated protein species overall and phosphorylating the T cell-specific ZAP-70 tyrosine kinase which is recruited to the CD3ζ subunit of the TCR. Memory CD4 T cells, however, exhibit a unique pattern of signaling through TCR/CD3. Following stimulation through TCR/CD3, memory CD4 T cells produce fewer species of tyrosine-phosphorylated substrates and fail to phosphorylate ZAP-70, yet unphosphorylated ZAP-70 can associate with the TCR/CD3 complex. Moreover, a 26/28-kDa phosphorylated doublet is associated with CD3ζ in resting and activated memory but not in naive CD4 T cells. Despite these differences in the phosphorylation of ZAP-70 and CD3-associated proteins, the ZAP-70-related kinase, p72^syk, exhibits similar phosphorylation in naive and memory T cell subsets, suggesting that this kinase could function in place of ZAP-70 in memory CD4T cells. These results indicate that proximal signals are differentially coupled to the TCR in naive versus memory CD4 T cells, potentially leading to distinct downstream signaling events and ultimately to the diverse functions elicited by these two CD4 T cell subsets.     

T细胞免疫识别研究进展

<正>免疫识别是疫苗设计的基础。T细胞免疫识别是预防性疫苗、治疗性疫苗(含负调疫苗)发展的基础。通过T细胞受体(T-cellantigen-receptor,TCR)的信号转导的特征是依赖于对表位多肽的限     

T cell recognition of Tn-glycosylated peptide antigens

The mouse hemoglobin-derived decapeptide Hb (67–76), VITAFNEGLK, which binds well to E^k and is non-immunogenic in CBA/J mice, was O-glycosylated with the tumor-associated carbohydrate Tn (α-D -N-acetylgalactosamine, or α-D -GalNAc). Each of the ten positions in the peptide was substituted with serine or threonine having the Tn antigen attached. The complete set of Tn-glycosylated peptides were then studied for binding to E^k and for immunogenicity in CBA/J mice. All of those glycopeptides which had the Tn attached to serine or threonine at a position in the peptide where, according to the crystal structure determinations, the amino acid side chain was oriented downwards into the binding site of the major histocompatibility complex (MHC) molecule, completely lost their capacity for binding to E^k. This was the case for the glycopeptides with Tn attacted at position 68 and 76, which are the major anchor residues and for those with Tn attached at position 71 and 73, which function as secondary anchor residues. Those glycopeptides which had Tn attached to serine or threonine at positions where the side chain pointed away from the binding site maintained their capacity for binding to E^k, except for those with Tn attached at position 70 and 74. Furthermore, some of the MHC-binding glycopeptides were immunogenic. In particular, this was the case for the glycopeptide with Tn attached to the central position 72 in the decapeptide. From previous studies, this is known to be the dominant T cell receptor contact residue of Hb (67–76). The results suggest that T cells may be capable of recognizing epitopes which are partially defined by a small glycan group.     

Human intrathymic T cell differentiation

The human thymus develops early on in fetal gestation with morphologic maturity reached by the beginning of the second trimester. Endodermal epithelial tissue from the third pharyngeal pouch gives rise to TE3+ cortical thymic epithelium while ectodermal epithelial tissue from the third pharyngeal cleft invaginates and splits during development to give rise to A2B5/TE4+ medullary and subcapsular cortical thymic epithelium. Fetal liver CD7+ T cell precursors begin to colonize the thymus between 7 and 8 weeks of fetal gestation, followed by rapid expression on thymocytes of other T lineage surface molecules. Human thymic epithelial cells grown in vitro bind to mature and immature thymocytes via CD2 and CD11a/CD18 (LFA-1) molecules on thymocytes and by CD58 (LFA-3) and CD54 (ICAM-1) molecules on thymic epithelial cells. Thymic epithelial cells produce numerous cytokines including IL1, IL6, G-CSF, M-CSF, and GM-CSF--molecules that likely are important in various stages of thymocyte activation and differentiation. Thymocytes can be activated via several cell surface molecules including CD2, CD3/TCR, and CD28 molecules. Finally, CD7+ CD4-CD8- CD3- thymocytes give rise to T cells of both the TCRab+ and TCR gd+ lineages.     

T cell recognition in chronic beryllium disease

Chronic beryllium disease (CBD) is a granulomatous lung disorder caused by beryllium exposure in the workplace and is characterized by the accumulation of beryllium-specific CD4(+) T cells. Depending on genetic susceptibility and the nature of the exposure, CBD occurs in up to 20% of exposed workers. Genetic susceptibility has been associated with particular HLA-DP alleles, especially those possessing a negatively charged glutamic acid residue at the 69th position of the beta-chain. The mechanism for this association lies in the ability of these HLA-DP molecules to bind and present beryllium to pathogenic CD4(+) T cells. Large numbers of effector memory, beryllium-specific CD4(+) T cells are recruited to the lung of these subjects and secrete Th1-type cytokines upon beryllium recognition. The presence of circulating beryllium-specific CD4(+) T cells directly correlates with the severity of lymphocytic alveolitis. With the presence of a known antigenic stimulus, CBD serves as an important model of immune-mediated, organ destruction. Thus, our findings in CBD have important implications for studies in autoimmune diseases, in particular those with an unknown inciting antigen and an inaccessible target organ.     

HLA-E polymorphisms in hematopoietic stem cell transplantation

Human leukocyte antigen (HLA)-E is an inhibitory ligand of natural killer cells and γ/δ T-cells. Differential expression of HLA-E alleles on the cell surface has been reported to influence outcome of hematopoietic stem cell transplantation (HSCT). We performed HLA-E genotyping in 116 HSCT patients and their HLA-matched unrelated donors. The impact of HLA-E genotypes on patient's overall survival (OS), disease free survival (DFS), cumulative incidences for relapse, transplant-related mortality (TRM) and acute graft vs host disease (aGvHD) was assessed. Neither univariate nor multivariate analysis showed any influence of HLA-E polymorphisms on the investigated endpoints of HSCT in our cohort. We could not confirm any of the previous observations in our cohort and consider it unlikely that HLA-E polymorphisms affect outcome of HSCT.     

Bacterial infections promote T cell recognition of self-glycolipids

Recognition of self is essential for repertoire selection, immune regulation, and autoimmunity and may be a consequence of infection. Self-induced recognition may represent the escape mechanism adopted by pathogens but may also incite autoimmune diseases. Here, we show that bacterial infection may promote activation of T cells reactive to self-glycosphingolipids (self-GSL). CD1+ antigen-presenting cells (APCs) infected with bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, or Mycobacterium bovis-Bacillus Calmette Guerín [BCG]) or treated with the bacterial components lipopolysaccharide, lipoteichoic acid, or Pam3CysSerLys4 (P3CSK4) lipopeptide acquire the capacity to stimulate self-GSL-specific T cells to cytokine release. Immediately after infection, APCs increase the endogenous GSL synthesis and stimulate GSL-specific T cells in a CD1- and T cell receptor (TCR)-dependent manner. This stimulation may contribute to inflammatory responses during bacterial infections and may predispose individuals to autoimmune diseases.     

T cell recognition of cell surface antigens. I. Specificity

Mouse fibroblast monolayers can be used as an immunoadsorbent to separate normal, unsensitized rat lymphocytes with receptors for foreign cell surface antigens. We used this approach to investigate if the specificity of thymus-dependent cellular immune reactions is based on a functional diversity of the T lymphocytes recognizing the relevant antigens. T lymphocyte diversity was demonstrated by the following findings.

• 1) Lymphocytes that are adsorbed to fibroblasts of a given phenotype are relatively restricted in their reactivity to cellular antigen. They strongly react against the adsorbing monolayer type, but their reactivity to another, third party fibroblast type is decreased.
• 2) Lymphocytes that were preabsorbed on one fibroblast type and transferred to fresh monolayer cultures decreased their reactivity to the absorbing fibroblast type, but could fully react against unrelated third party fibroblasts.
• 3) Pretreatment of the lymphocytes with relevant subcellular antigen prevents their specific adherence to fibroblast monolayers, and, thus, recognition of cellular antigen.
• 4) Using mosaic fibroblast monolayers composed of fibroblasts of two unrelated phenotypes, we were able to demonstrate two different lymphocyte populations, each characterized by the capacity to recognize either of the adsorbing fibroblast types.

     

Interleukin-21 receptor-mediated signals control autoreactive T cell infiltration in pancreatic islets

It remains unclear how interleukin-21 receptor (IL-21R) contributes to type 1 diabetes. Here we?have shown that dendritic cells (DCs) in the pancreas required IL-21R not for antigen uptake, but to acquire the chemokine receptor CCR7 and migrate into the?draining lymph node. Consequently, less antigen, major histocompatibility complex (MHC) class II, and CD86 was provided to autoreactive effector cells in?Il21r(-/-) mice, impairing CD4(+) T?cell activation, CD40:CD40L interactions, and pancreatic infiltration by autoreactive T?cells. CD40 crosslinking restored defective CD4(+) cell expansion and CD4 independently expanded autoreactive CD8(+) cells, but CD8(+) cells still required CD4(+) cells to reach the pancreas and induce diabetes. Diabetes induction by transferred T?cells required IL-21R-sufficient host antigen-presenting cells. Transferring IL-21R-sufficient DCs broke diabetes resistance in Il21r(-/-) mice. We conclude that IL-21R controls both antigen transport by DCs and the crucial beacon function of CD4(+) cells for autoreactive CD8(+) cells to reach the islets.     

Superantigen recognition by gammadelta T cells: SEA recognition site for human Vgamma2 T cell receptors

Human gammadelta T cells expressing the Vgamma2Vdelta2 antigen receptors recognize nonpeptide prenyl pyrophosphate and alkylamine antigens. We find that they also recognize staphylococcal enterotoxin A superantigens in a manner distinct from the recognition of nonpeptide antigens. Using chimeric and mutant toxins, SEA amino acid residues 20-27 were shown to be required for gammadelta TCR recognition of SEA. Residues at 200-207 that are critical for specific alphabeta TCR recognition of SEA do not affect gammadelta TCR recognition. SEA residues 20-27 are located in an area contiguous with the binding site of V beta chains. This study defines a superantigen recognition site for a gammadelta T cell receptor and demonstrates the differences between Vgamma2Vdelta2+ T cell recognition of superantigens and nonpeptide antigens.     

Human T cell differentiation antigens characterizing a cytotoxic/suppressor T cell subset

Monoclonal antibodies were raised against the leukemic T cells from a patient with chronic lymphocytic leukemia. Two antibodies, termed T411 and T811, were obtained which were reactive by indirect immunofluorescence only with cells of the T cell lineage. The T411 antibody recognized a polypeptide chain of 100,000 dalton apparent molecular weight which was present on the surface of 94 +/- 4% of peripheral blood T lymphocytes, but only on 20 +/- 8% of thymus cells. The antibody T811 reacted with a surface molecule composed of 2 poly-peptide chains of 32,000 and 34,000 dalton apparent molecular weight, which was expressed only on 25 +/- 10% of blood T lymphocytes and on 90 +/- 4% of thymus cells. Functional analysis of the T811+ and T811- T cell subsets isolated by rosetting with anti-mouse Ig coated ox erythrocytes revealed that both subpopulations were able to mount a proliferative response to allo-antigens, whereas allo-antigen induced cytotoxic cells and their precursors were only found in the T811+ subset. The pokeweed mitogen induced in vitro differentiation of B lymphocytes into immunoglobulin secreting cells was dependent on the presence of the T811- subset, whereas the T811+ T cells efficiently suppressed this differentiation.