Reconciling views on T cell receptor germline bias for MHC

Whether MHC restriction by the T cell receptor (TCR) is a product of evolutionary pressures leading to germline-encoded 'rules of engagement' remains avidly debated. Structural results derived from analysis of TCR-peptide-MHC complexes appear to support a model of physical specificity between TCR germline V regions and MHC. Yet, some recent evidence suggests that thymic selection, and co-receptors may have misled us into thinking the TCR is exclusively MHC-specific, when in fact, TCRs can robustly engage non-MHC ligands when given the chance. Here, I propose that seemingly contradictory data and hypotheses for, and against, germline bias are, in fact, compatible and can be reconciled into a unifying model.     

NK receptor interactions with MHC class I molecules in pregnancy

Both HLA class I molecules and their receptors on Natural Killer cells, the KIR molecules, are highly polymorphic. It is generally believed that this variation is driven in response to the role of these receptors and counter-receptors in resistance to disease. Uterine NK cells are the major maternal leukocyte population present within the decidua, and they express KIR2D receptors for HLA-C, the only polymorphic class I molecule on trophoblast. Genetic and functional data suggest that the maternal KIR/fetal HLA-C interaction in pregnancy may affect the delivery of an optimal blood supply to mother and fetus. The drive for novelty in HLA-C and KIR2D allelic diversity may relate not only to survival from infections but also to reproductive success.     

Aggregation of MHC class I molecules on a CD8+ alphabeta T cell clone specifically inhibits non-antigen-specific lysis of target cells

MHC class I molecules are target molecules recognized by TCR or NK receptors encoded in the NK gene cluster or leukocyte receptor cluster. We show that aggregation of MHC class I molecules by specific monoclonal antibodies on cytotoxic T cells, inhibits the anti-CD94 redirected lysis of P815. This inhibition is not the consequence of apoptosis or anergy of the cytotoxic T lymphocytes. In contrast, aggregation of MHC class I molecules does not inhibit either the anti-CD3 redirected cytotoxicity or the CD94-triggered up-regulation of CD25 molecules of the same T cell clone. MHC class I ligand molecules expressed by antigen presenting cells and/or T lymphocytes could therefore be able to modulate nonspecific cytotoxicity upon interaction with MHC class I molecules expressed by effector cytotoxic T lymphocytes.     

Expression of inhibitory receptors for MHC class I molecules on T cells

Inhibitory receptors (IRs) specific for MHC class I molecules and originally described on natural killer (NK) cells are also expressed on a fraction of peripheral T cells. The presence of these receptors on T cells is poorly understood. In this review, the different antigen specificities described to date for IR+ T cells and the expression pattern of these receptors on T cells are analyzed. This analysis indicates that the population of T cells defined by IR expression is heterogeneous and that different IRs (or families of IRs) may play different roles in T-cell biology.     

Mutant MHC class I molecules define interactions between components of the peptide-loading complex

Class I histocompatibility molecules, consisting of a heavy chain, beta2-microglobulin and peptide, are assembled in the endoplasmic reticulum (ER) with the assistance of several molecular chaperones and accessory proteins. Peptide binding occurs when assembling class I molecules associate with a loading complex consisting of the transporter associated with antigen processing (TAP) peptide transporter, tapasin, ERp57 and calreticulin (CRT)/calnexin. To assess the physical organization of this complex, we generated a series of mutants in the murine H-2Dd heavy chain and assessed their association with components of the complex. Seven mutations, clustered between amino acids 122 and 136 in the heavy chain alpha2 domain plus one mutation at position 222 in the alpha3 domain, resulted in loss of interaction with tapasin. Association with TAP was always lost simultaneously, supporting the view that tapasin acts as an obligatory bridge between class I molecules and TAP. Compared with previous studies on the HLA-A2 molecule, some differences in points of tapasin interaction were observed. Failure of the H-2Dd mutants to bind tapasin resulted in low cell-surface expression and altered intracellular transport. Most mutants retained a substantial degree of peptide loading, consistent with the view that although tapasin may promote peptide binding to class I, it is not required. A surprising observation was that all mutants lacking tapasin interaction retained normal association with CRT. This contrasts with previous observations on other class I molecules and, combined with differences in tapasin interaction, suggests that the organization of the ER peptide-loading complex can vary depending on the specific class I molecule examined.     

Cytotoxic T lymphocyte recognition of hybrid MHC class I molecules

We have utilized a group of MHC class I genes produced by in vitro recombination between Dp and Dd to study recognition of MHC class I molecules by cytolytic T cells (CTLs). Both polyclonal allo-specific and H-2-restricted CTLs require that alpha 1 and alpha 2 of the target class I molecule be derived from the same haplotype for efficient killing. By using T-cell lines we showed that within the bulk population there must exist a fraction of T cells which can recognize epitopes in alpha 1 or alpha 2. Critical residues for T-cell recognition have been identified using these chimeric genes.     

Inhibition of T cell activation by blockade of MHC class II molecules

Autoimmune diseases result from the activation of self-reactive T cells induced by autoantigens or by foreign antigens cross-reactive with an autoantigen. A striking characteristic of autoimmune diseases is the increased frequency of certain HLA alleles in affected individuals. Moreover, as demonstrated for example in rheumatoid arthritis and insulin-dependent diabetes mellitus, class II alleles positively associated with autoimmune diseases share amino acid residues in the hypervariable HLA regions involved in peptide binding. Therefore, it is likely that disease-associated HLA class II molecules have the capacity to bind the autoantigen and present it to T cells, thereby inducing and maintaining, under appropriate conditions, the autoimmune disease. The data reviewed here demonstrate MHC-selective inhibition of antigen-induced T cell responses in vivo by parenterally administered soluble, MHC-binding peptide competitors, under conditions in which the competitor is not immunogenic. This suggests the feasibility of a therapeutic approach based on MHC blockade in the treatment of HLA-linked autoimmune diseases.     

Regulation of T cell function by NK cell receptors for classical MHC class I molecules

Inhibitory receptors for MHC class I molecules were initially characterised on NK cells. Human and mouse NK cell receptors (NKRs) are also expressed on T cells, predominantly on a subset of memory-phenotype CD8(+) T cells. This review focuses on the precise determination of interactions between NKRs and MHC class I, as well as on the unexpected in vivo function of NKRs on T cells.     

The Fanconi anemia DNA damage repair pathway in the spotlight for germline predisposition to colorectal cancer

Colorectal cancer (CRC) is one of the most common neoplasms in the world. Fanconi anemia (FA) is a very rare genetic disease causing bone marrow failure, congenital growth abnormalities and cancer predisposition. The comprehensive FA DNA damage repair pathway requires the collaboration of 53 proteins and it is necessary to restore genome integrity by efficiently repairing damaged DNA. A link between FA genes in breast and ovarian cancer germline predisposition has been previously suggested. We selected 74 CRC patients from 40 unrelated Spanish families with strong CRC aggregation compatible with an autosomal dominant pattern of inheritance and without mutations in known hereditary CRC genes and performed germline DNA whole-exome sequencing with the aim of finding new candidate germline predisposition variants. After sequencing and data analysis, variant prioritization selected only those very rare alterations, producing a putative loss of function and located in genes with a role compatible with cancer. We detected an enrichment for variants in FA DNA damage repair pathway genes in our familial CRC cohort as 6 families carried heterozygous, rare, potentially pathogenic variants located in BRCA2/FANCD1, BRIP1/FANCJ, FANCC, FANCE and REV3L/POLZ. In conclusion, the FA DNA damage repair pathway may play an important role in the inherited predisposition to CRC.     

Structural basis of T cell recognition of peptides bound to MHC molecules

Helper T lymphocytes play a critical role in immune system activation following recognition of MHC class II-bound peptide ligands (pMHCII). These CD4 T cells stimulate B cell antibody production and cytolytic T cell generation. Until recently, the structural basis of coordinate T cell receptor (TCR) and CD4 co-receptor interaction with a given pMHCII was unknown. Here we review current structural data on specific pMHCII recognition by T cells and compare TCR and co-receptor docking to pMHCI versus pMHCII ligands. The implications of these findings for thymic selection, helper versus cytolytic T cell recognition and alloreactivity are discussed.     

An autonomous CDR3delta is sufficient for recognition of the nonclassical MHC class I molecules T10 and T22 by gammadelta T cells

It remains unclear whether gammadelta T cell antigen receptors (TCRs) detect antigens in a way similar to antibodies or alphabeta TCRs. Here we show that reactivity between the G8 and KN6 gammadelta TCRs and the major histocompatibility complex class Ib molecule T22 could be recapitulated, with retention of wild-type ligand affinity, in an alphabeta TCR after grafting of a G8 or KN6 complementarity-determining region 3-delta (CDR3delta) loop in place of the CDR3alpha loop of an alphabeta TCR. We also found that a shared sequence motif in CDR3delta loops of all T22-reactive gammadelta TCRs bound T22 in energetically distinct ways, and that T10(d), which bound G8 with weak affinity, was converted into a high-affinity ligand by a single point mutation. Our results demonstrate unprecedented autonomy of a single CDR3 loop in antigen recognition.     

Antigenic peptide interaction with MHC molecules: implications for the design of artificial vaccines

Both helper and cytotoxic T cells see primarily a limited number of immunodominant sites on a protein. The reasons for immunodominance are many. We have explored primarily the roles of antigen processing and binding to histocompatibility molecules, as well as the structural features of the antigenic peptide. This information has led to the identification and characterization of sites stimulating helper or cytotoxic T cells specific for antigens from malaria or HIV, and ways to couple and immunize with these, that may be useful for vaccine development. We also observed a striking concordance between helper and cytotoxic T cell sites.     

Accessory proteins that control the assembly of MHC molecules with peptides

The stable assembly of Major Histocompatibility Complex (MHC) molecules with peptides is controlled by a number of cofactors, including proteins with general housekeeping functions and proteins with dedicated functions in MHC assembly. Recent work in my laboratory has focused on two chaperones, tapasin (tpn) and DM, that play critical roles in the loading of peptides onto MHC class I and MHC class II molecules, respectively. Tapasin is a transmembrane protein that tethers empty class I molecules in the endoplasmic reticulum to the transporter associated with antigen processing. DM is a peptide exchange factor that binds with empty and peptide-loaded class II molecules in endosomal and lysosomal compartments. Although a number of different functions for tapasin and DM have been proposed, emerging evidence suggests that both of these chaperones retain unstable MHC molecules in peptide-loading compartments until they bind with high-affinity peptides. These cofactors therefore promote the surface expression of long-lived MHC-peptide complexes.     

两种抑制MHCⅡ类分子表达方法的比较

目的：评价cⅡTA（MHC classⅡ transactivator，CⅡTA）基因突变体和反义RNA对MHCⅡ类分子的抑制效率和胞内稳定性，为改造MHC分子奠定基础。方法：用PCR、酶切及连接技术，构建不含起始密码子的peDNA3mCⅡTA2，含起始密码子的peDNA3mCⅡTA3突变体及cⅡTA基因的反义RNA-pcDNA3／CⅡTAJ亘。用脂质体转染法，将突变体、反义RNA及空载体pcDNA3转入PIEC细胞和123细胞中。持续四个月，流式细胞术观察它们对PIEC／123细胞株MHCⅡ类（sLA-DR）分子的诱导性和组成性表达的影响。结果：转染pcDNA3、pcDNA3mCⅡTA2后，PIEC／123细胞SLA-DR的表达没有受到抑制，转染peDNA3mCⅡTA3后，9／20（45．0％）的PIEc细胞、10／20（50．O％）的L23细胞SLA-DR的表达受到明显抑制，抑制率高达90％以上；转染pcDNA3mCIITA＆后，7／15（46．7％）PIEC细胞、5／15（33．3％）123细胞sLA-DR的表达受到明显抑制，抑制率高达85％以上。持续检测4个月，转染peDNA3mCⅡTA3的PIEC／123细胞SLA-DR的表达受抑率均在90％以上。转染反义RNA的PIEC／123细胞在第65／45天。SLA-DR表达抑制率降至10％以下。结论：构建成功的CⅡTA突变体和反义RNA均能有效抑制sLA-DR表达，但CⅡTA突变体稳定存在于胞内的时间长，构建突变体是和用CⅡTA抑制MHCⅡ类分子的好方法。