TCRγδ Cells Research Articles

Epicutaneous (EC) immunization with myelin basic protein (MBP) induces TCRαβ + CD4 + CD8 + double positive suppressor cells that protect from experimental autoimmune encephalomyelitis (EAE)

Multiple sclerosis (MS) is a central nervous system (CNS) chronic inflammatory autoimmune disease with limited treatment modalities. Oral tolerance is one of the experimental methods that protects from autoimmune diseases. However, this method failed to be therapeutic in clinical trials. In our previous work we found that epicutaneous (EC) immunization with protein antigen induced a state of profound immunosuppression that inhibited inflammatory response in contact sensitivity, in experimental autoimmune encephalomyelitis (EAE) and in allogeneic skin graft rejection. In our current work, we precisely determined the phenotype of EC induced T suppressor (Ts) cells that reduce the progress of EAE. Employing TCRδ−/−, CD1d−/− mice, we showed that EC induced Ts cells do not belong either to the population of TCRγδ cells or CD1d restricted NKT cells. Moreover, we noticed that a lack of CD1d−/− restricted NKT lymphocytes resulted in the induction of much stronger suppression of EAE than in wild type mice. This might suggest that NKT cells could interfere with the induction of Ts cells. Using β 2m−/− mice, negative selection and positive selection of EC induced Ts cells, we showed that Ts cells protecting from EAE belong to the population of TCRαβ + CD4 + CD8 + double positive lymphocytes.

Dendritic Cells: More Than Just Adaptive Immunity Inducers?

Dendritic cells (DC) are well known for their capacity to induce immune responses and there is also accumulating evidence of their ability to interact with various cell types of the innate system, such as NK, NKT or TCRγδ cells. These interactions are bi-directional, mediated by soluble or cell surface molecules and have been mainly described in the context of immune responses to infectious agents and tumors. NK, NKT or TCRγδ cells induce the maturation of DC, as shown by the increased expression of CD86, IL12 production and priming of T cell responses. On the other hand, mature DC have the ability to activate NK, NKT or TCRγδ cells for sustained innate immune responses and activated NK cells may kill immature DC. In addition, DC and NK or TCRγδ cells share similar functions such as cytotoxic and antitumor activity, interferon production and antigen presentation capacity.

Maximum Immunobioactivity of Murine Small Intestinal Intraepithelial Lymphocytes Resides in a Subpopulation of CD43+ T Cells

CD43 has been linked to many function-associated T cell activities. Using mAbs that recognize two different CD43 determinants, we show that, although mouse small intestinal intraepithelial lymphocytes (IELs) expressed the CD43 core molecule reactive with mAb R2/60, only about one-half of the total IELs-including some but not all of the TCRalphabeta and TCRgammadelta cells-expressed the CD43 S7(-) reactive determinant. CD43 S7(+) IELs secreted more IL-2, IL-4, IL-10, IL-17, and IFN-gamma following anti-CD3 stimulation, and were >4-fold more cytotoxic in fresh isolates and >16-fold more cytotoxic after anti-CD3 stimulation, than S7(-) IELs. S7(+) but not S7(-) IELs from the ileum of IL-10(-/-) mice spontaneously produced IFN-gamma. In vivo BrdU uptake by IELs in non-Ag-primed mice was greatest in the S7(+) population, indicating that significantly more S7(+) IELs than S7(-) IELs undergo cell expansion under normal homeostatic conditions. DNA microarray analyses showed that S7(+) IELs expressed higher levels of genes associated with activated T cells, whereas S7(-) IELs expressed genes used in the regulation of NK cells. These findings define two functionally distinct populations of IELs based on CD43 expression independent of TCR class, and they identify a subset of IELs that may serve as a target to better control intestinal inflammation.

Distinct requirements for IL-7 in development of TCR gamma delta cells during fetal and adult life.

TCRgammadelta-transgenic IL-7(-/-) mice were generated to determine whether T cells containing productively rearranged TCRgammadelta genes have additional requirements for IL-7 within the thymus or peripheral lymphoid tissues. Differences in developmental requirements for IL-7 by TCRgammadelta cells were noted and were linked to derivation from fetal- vs adult-type precursors in the thymus. Although TCRgammadelta cells are absent from IL-7(-/-) mice, TCRgammadelta cells were restored to the thymus and periphery by expression of TCRgammadelta transgenes. Endogenous TCRgamma chains were expressed by IL-7(+/-) but not IL-7(-/-) TCRgammadelta-transgenic mice, providing direct support for findings that IL-7 is necessary for rearrangement and expression of TCRgamma genes. The number of TCRgammadelta thymocytes was 10-fold reduced in TCRgammadelta-transgenic IL-7(-/-) embryos; however, adult TCRgammadelta-transgenic IL-7(-/-) or IL-7(+/-) mice had similar numbers of fetal thymus-derived TCRgammadelta cells in their skin. Thus, fetal TCRgammadelta cells required IL-7 for TCR rearrangement, but not for proliferation or survival in the periphery. In contrast, the numbers of TCRgammadelta cells in other tissues of TCRgammadelta-transgenic IL-7(-/-) mice were not completely restored. Moreover, coincident with the transition from the first to second wave of T cell precursors maturing in neonatal thymus, thymus cellularity of TCRgammadelta-transgenic IL-7(-/-) mice dropped significantly. These data indicated that in addition to TCRVgamma gene rearrangement, TCRgammadelta cells differentiating from late fetal liver or adult bone marrow precursors have additional requirements for IL-7. BrdU incorporation studies indicated that although IL-7 was not required for TCRgammadelta cell proliferation, it was required to prolong the life span of mature TCRgammadelta cells.

Human T cell receptor gammadelta cells recognize endogenous mevalonate metabolites in tumor cells.

T lymphocytes expressing the T cell receptor (TCR)-γδ recognize unknown antigens on tumor cells. Here we identify metabolites of the mevalonate pathway as the tumor ligands that activate TCR-γδ cells. In tumor cells, blockade of hydroxy-methylglutaryl-CoA reductase (HMGR), the rate limiting enzyme of the mevalonate pathway, prevents both accumulation of mevalonate metabolites and recognition by TCR-γδ cells. When metabolite accumulation is induced by overexpressing HMGR or by treatment with nitrogen-containing bisphosphonate drugs, tumor cells derived from many tissues acquire the capacity to stimulate the same TCR-γδ population. Accumulation of mevalonate metabolites in tumor cells is a powerful danger signal that activates the immune response and may represent a novel target of tumor immunotherapy.

Circulating T lymphocyte subsets in coeliac disease (CoD) patients and healthy family members.

Increased proportions of circulating antigen-primed CD45RO+ TCR gammadelta cells have been found in untreated CoD patients. As certain immunological features are now found in both CoD and healthy persons carrying the HLA DQ2 heterodimer, we sought to establish whether healthy members of the families of CoD patients who are positive for HLA DQ2 and also have increased densities of TCR gammadelta intraepithelial lymphocytes (IEL) in their small bowel mucosa have elevated levels of circulating TCR gammadelta memory cells. Peripheral blood T cells were analysed by flow cytometry in 22 patients with CoD and 16 healthy family members. Untreated CoD patients had higher percentages of circulating CD45RO+ TCR gammadelta cells and CD45RO+ Vdelta1+ cells than healthy family members. On the other hand, the amount of circulating Vdelta1+ lymphocytes was lower in patients with CoD compared with healthy family members. In contrast, no differences were found between HLA DQ2+ and HLA DQ2- healthy family members in respect of circulating TCR gammadelta cell subsets. The change in circulating TCR gammadelta cell subsets found in patients with CoD is thus a consequence of an ongoing immunological process which diminishes on a gluten-free diet rather than a phenomenon directly caused by DQ2. These changes in peripheral blood are not found in healthy individuals who have the same HLA alleles DQA1*0501 and DQB1*0201 encoding the HLA DQ2 and who also have increased densities of TCR gammadelta IEL in their otherwise normal jejunal mucosa.

Control of T cell development in vivo by subdomains within the IL-7 receptor alpha-chain cytoplasmic tail.

IL-7/IL-7R signaling functions in both growth and differentiation during T cell development. In this study, we examined the extent these activities were controlled by signaling associated with distinct IL-7R alpha cytoplasmic domains by transgenic expression of wild-type or cytoplasmic deletion mutants of IL-7R alpha in the thymi of IL-7R alpha(-/-) mice. We show an essential requirement for the tyrosine-containing carboxyl-terminal T domain in restoring thymic cellularity, pro-/pre-T cell progression, and survival. In contrast, the functional differentiation of TCR alpha beta cells and the development of TCR gamma delta cells are partially independent of the T domain. Thus, separate cytoplasmic domains of the IL-7R alpha chain differentially control distinct functions during T cell development, whereas normal IL-7R-dependent thymic development requires the integrated activity of all these domains.

Enterocyte expression of interleukin 7 induces development of gammadelta T cells and Peyer's patches.

The intestinal mucosa is suggested to support extrathymic T cell development, particularly for T cell receptor (TCR)-gammadelta intraepithelial lymphocytes (IELs). TCR-gammadelta cell development requires interleukin (IL)-7; IL-7(-/)- or IL-7 receptor(-/)- mice lack TCR-gammadelta cells. Using the intestinal fatty acid binding protein (iFABP) promoter, we reinstated expression of IL-7 to mature enterocytes of IL-7(-/)- mice (iFABP-IL7). In iFABP-IL7 mice, TCR-gammadelta IELs were restored, as were cryptopatches and Peyer's patches. TCR-gammadelta cells remained absent from all other tissues. Likewise, T cell development in thymus and B cell maturation in the bone marrow and spleen retained the IL-7(-/)- phenotype. Thus, IL-7 expression by enterocytes was sufficient for extrathymic development of TCR-gammadelta cells in situ within the intestinal epithelium and was crucial for organization of mucosal lymphoid tissue.

IL-2Rβ/IL-7Rα Doubly Deficient Mice Recapitulate the Thymic and Intraepithelial Lymphocyte (IEL) Developmental Defects of γc−/− Mice: Roles for Both IL-2 and IL-15 in CD8αα IEL Development

AbstractIL-7Rα-chain-deficient (IL-7Rα−/−) and common γ chain-deficient (γc−/−) mice both exhibit abnormal thymic and intestinal intraepithelial lymphocyte (IEL) development, but the developmental inhibition is not equivalent. In this report, we assessed whether the defects in T cell development associated with γc−/− mice were due to currently defined γc-dependent cytokines by cross-breeding IL-7Rα−/− mice to mice lacking either IL-2, IL-4, or IL-2Rβ. IL-2/IL-7Rα and IL-4/IL-7Rα double knockout (DKO) mice demonstrated equivalent thymic development to IL-7Rα−/− mice, whereas IL-2Rβ/IL-7Rα DKO mice, which lack IL-2, IL-7, and IL-15 signaling, displayed thymic T cell defects identical to γc−/− mice. Collectively, these data indicate that of the γc-dependent cytokines, only IL-7 and IL-15 contribute to the progression and production of thymic T cells. In the IEL, IL-7Rα−/− mice selectively lack CD8αα TCRγδ cells, whereas IL-2Rβ−/− mice show a significant reduction in all CD8αα cells. IL-2−/− and IL-2/IL-7Rα DKO mice demonstrated a reduction in CD8αα IELs to nearly the same extent as IL-2Rβ−/− mice, indicating that IL-2 functions in CD8αα IEL development. Moreover, IL-2Rβ/IL-7Rα DKO mice lacked nearly all TCR-bearing IEL, again recapitulating the phenotype of γc−/− mice. Thus, these data point to the importance of IL-2, IL-7, and IL-15 as the γc-dependent cytokines essential for IEL development.

TCRγδ cells and viruses

T-cell receptor gammadelta cells (TCRgammadelta) are often found in increased numbers during the course of several viral infections in humans. Although these findings suggest an important role for this unique subset, their precise function has not been ascertained. Recent studies in murine models of both RNA and DNA virus infections have begun to shed new light on the potential function for TCRgammadelta cells in antiviral immunity. It is clear that TCRgammadelta cells participate in the immune response to human immunodeficiency virus (HIV), influenza, Sendai, coxsackie, vaccinia, vesicular stomatitis virus (VSV), and herpes simplex virus-1 (HSV-1) viral infections since they become activated and home to the sites of viral replication. In this review we will summarize current efforts to dissect the role of TCRgammadelta cells in these disease settings, emphasizing the effector functions utilized, the TCR repertoire, and the antigens recognized. Particular focus will be placed on HSV-1 infections where we have begun to address these issues and have shown that TCRgammadelta cells are sufficient for protection from lethal infection and are able to recognize the herpes virus antigen glycoprotein I.

Pneumocystis carinii pneumonia in mutant mice deficient in both TCRalphabeta and TCRgammadelta cells: cytokine and antibody responses.

Resistance to Pneumocystis carinii is achieved through cell-mediated and humoral immunity, but the interplay between these two systems in the immunocompetent host is not fully understood. TCRbetaxdelta-/- double-mutant mice deficient of all T cell populations naturally acquired P. carinii pneumonia with lethal consequences. Moribund mutants displayed numbers of pulmonary pathogens comparable to RAG-1-/- mice lacking all functional T and B lymphocytes. Pulmonary lavage cells of diseased TCRbetaxdelta-/- mutants secreted proinflammatory cytokines tumor necrosis factor-alpha, interleukin (IL)-12, and interferon-gamma, but not IL-4, -5, or -10. Serum immunoglobulin levels of both healthy and diseased mice were significantly reduced compared with immunocompetent animals. Secreted antibodies were mainly IgM, which also bound P. carinii. Mutants completely lacked IgG1, emphasizing strict T cell dependence of immunoglobulin switching to this isotype. Other IgG subclasses were strongly reduced and did not bind P. carinii. These results suggest that T cells are crucial for generation of antibodies against P. carinii relevant to resistance.

Circulating TCR gammadelta cells in the patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a disorder with a wide range of immunological abnormalities. The results of the studies undertaken in the last decade indicated that SLE pathogenesis was mainly connected with the breakdown of the activation control of B and T cells, generating humoral or cell-mediated responses against several self-antigens of affected cells. The last studies demonstrate that the role of gammadelta T lymphocytes in autoimmune diseases can be especially important. Flow cytometry techniques were used to investigate the number and percentage of TCR gammadelta T cells and their most frequent subtypes in peripheral blood of 32 patients with SLE and 16 healthy volunteers. We also correlated TCR gammadelta cells number with the level of T CD3+, T CD4+, T CD8+, and NK (CD16) cells (cytometric measurements) and SLE activity (on the basis of clinical investigations). Our studies were preliminary attempts to evaluate the role of that minor T cell subpopulation in SLE. Absolute numbers of cells expressing gammadelta TCR in most SLE blood specimens were significantly lower than in the control group (P<0.006). However, since the level of total T cell population was also decreased in the case of SLE, the mean values of the percentage gammadelta T cells of pan T lymphocytes were almost the same in both analysed populations (7.1% vs 6.3%, respectively). In contrast to Vdelta2+ and Vgamma9+ subtypes of pan gammadelta T cells, Vdelta3+ T cells number was higher in SLE patients (20 x 10 cells/microl) than in healthy control group (2 x 2 cells/microl) (P=0.001). However, we found no differences between the numbers of pan gammadelta T lymphocytes and studied their subtypes in the patients with active and inactive disease. These cell subpopulations were doubled in the treated patients with immunosuppressive agents in comparison with untreated ones; however, data were not statistically significant. Our study indicated that Vdelta3+ subtype of gammadelta T cells seems to be involved in SLE pathogenesis; however, we accept the idea that the autoimmunity does not develop from a single abnormality, but rather from a number of different events.

HSV-1 Glycoprotein I-Reactive TCRγδ Cells Directly Recognize the Peptide Backbone in a Conformationally Dependent Manner

Abstract Despite the description of numerous antigenic ligands recognized by TCRγδ cells, detailed information concerning the structural nature of these antigenic epitopes is lacking. In addition, the recent descriptions of human TCRγδ cells recognizing mycobacterium-derived low m.w. lipid molecules confirms that the spectrum and nature of biologic structures that are capable of being recognized by TCRγδ cells are unclear. We have previously described a murine TCRγδ cell clone, TgI4.4, that is reactive to herpes simplex virus (HSV)-1 glycoprotein I (gI). Unlike TCRαβ-mediated, MHC-restricted Ag recognition but similar to Ig Ag recognition, TgI4.4 recognizes purified gI directly, in the absence of Ag processing or presentation. Since gI is a complex glycoprotein, the nature of the antigenic epitope was investigated. First, gI recognition by TgI4.4 is conformationally dependent, as revealed by denaturation and proteolytic experiments. Secondly, the epitope recognized by TgI4.4 was mapped to the amino terminus by using insertion mutants of gI. Lastly, TgI4.4 recognizes the gI protein directly since completely deglycosylated forms of gI are efficiently recognized. Therefore, TCRγδ cells are capable of recognizing a variety of molecular structures, including proteins. The ability of TgI4.4 to recognize a nonglycosylated form of gI suggests that HSV-1 recognition by TCRγδ cells in vivo is not limited by cell-specific glycosylation patterns or glycosylation-dependent conformational influences.

The Role of IL-7 in Thymic and Extrathymic Development of TCRγδ Cells

AbstractIL-7-deficient (IL-7−/−) mice have reduced numbers of B and TCRαβ cells, but lack mature TCRγδ cells. Although most T cell development occurs in the thymus, some intestinal intraepithelial lymphocytes (IEL), including TCRγδ cells, can develop extrathymically. Epithelial cells in both thymus and intestine synthesize IL-7, suggesting that TCRγδ cell development could occur in either site. To evaluate the role of thymic IL-7 in development of TCRγδ cells, newborn TCRβ-deficient (TCRβ−/−) thymi were grafted to IL-7−/− mice. Donor- and host-derived TCRγδ cells were recovered from thymus grafts, spleen, and IEL. However, when IL-7−/− thymi were grafted to TCRβ−/− mice, no development of graft-derived TCRγδ cells occurred, indicating that extrathymic IL-7 did not support TCRγδ IEL generation from newborn thymic precursors. In contrast, TCRγδ IEL development occurred efficiently in adult, thymectomized, irradiated C57BL/6J mice reconstituted with IL-7−/− bone marrow. This demonstrated that extrathymic development of TCRγδ IEL required extrathymic IL-7 production. Thus, intrathymic IL-7 was required for development of thymic TCRγδ cells, while peripheral IL-7 was sufficient for development of extrathymic TCRγδ IEL.

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Phenotypic and functional maturation of TCR γδ cells in the human thymus

Activation of natural killer cells by heat-killed Listeria monocytogenes requires additional signals from lymphoid cells

Regulatory and protective functions have been attributed to murine natural killer (NK) cells in a number of infectious diseases including listeriosis. We have developed an in vitro model to study parameters underlying the activation of naive NK cells using heat-killed Listeria monocytogenes (HKL) as stimulator. Independent from expression of the cell surface marker NK1.1, NK cells lysed YAC-1 cells after in vitro stimulation with HKL or HKL + Interleukin (IL)-2, but not medium or IL-2 alone. In contrast, NK cells from severely immunocompromised SCID or RAG-1 − − mutant mice failed to respond to HKL alone, but required exogenous IL-2. Using single-gene-disruption mutant mice, we show that NK-cell activation can be supported by either T-cell receptor (TCR)αβ cells, TCRγδ cells, MHC class I or MHC class II gene products. We conclude from these data that recognition of listerial components alone is insufficient for activation of naive NK cells, and that additional costimulatory signals are necessary. These can be provided by various lymphoid cells and appear to be cytokines.

Development of TCRγδ iIELs

Interest has focused recently on the development and function of T and B lymphocytes in the mucosal surface, particularly on a population of intestinal intraepithelial lymphocytes (iIELs) that develop by thymic and extrathymic pathways in the mucosa (for review see ref 1). Our approach to studying TCRγδ iIEL development has been to utilize G8 transgenic (Tg) mice expressing an antigen-specific, developmentally-regulated, γδTCR. These mice have provided a useful tool to dissect T-cell development of intestinal TCRγδ cells. In this review, we summarize our findings of studies using this model system to dissect the complex events that regulate TCRγδ iIEL development and functional activity. The data suggest a maturational scheme for TCRγδ iIELs wherein the majority of TCRγδ iIELs migrate to the gut in an immature state where they develop in response to TCR ligation. As TCRγδ iIELs develop, surface and functional phenotypes change in a discernable pattern. These findings provide an important first step in analysing TCRγδ development, maturation and differentiation in normal animals

TCR gamma delta cells: mysterious cells of the immune system.

Gamma delta cells participate in pathogenic infections and autoimmune conditions, yet, almost a decade after their discovery, little is known regarding their TCR repertoire or effector functions. Unlike MHC-restricted antigen recognition employed by TCR alpha beta cells, TCR gamma delta cells can recognize whole unprocessed antigens in an MHC- independent manner. The nature of positive and negative selection used to shape the repertoire of TCR gamma delta cells is unclear, especially in the nonlymphoid tissues where these cells predominate. While TCR gamma delta cells express an activated phenotype and are present in pathological conditions, their roles in immunological protein is unknown. This review will focus on our efforts to study these issues of TCR gamma delta biology.