RESUMO
Background: Natural Killer T (NKT) cells are CD1d-restricted innate-like T cells that can rapidly release stored cytokines upon recognition of lipid antigens. In mice, type I NKT cells seem to promote liver inflammation, whereas type II NKT cells seem to restrict hepatitis. Here, we aimed at characterizing the role of human type I and type II NKT in patients with autoimmune hepatitis (AIH). Methods: NKT cells were analyzed by flow cytometry in peripheral blood and liver of AIH patients and control groups. α-galactosylceramide-loaded or sulfatide-loaded tetramers were used to detect type I or II NKT cells, respectively. Hepatic CD1d was stained by in situ-hybridization of liver biopsies. Results and Conclusions: Type II NKT cells were more prevalent in human peripheral blood and liver than type I NKT cells. In AIH patients, the frequency of sulfatide-reactive type II NKT cells was significantly increased in peripheral blood (0.11% of peripheral blood leukocytes) and liver (3.78% of intrahepatic leukocytes) compared to healthy individuals (0.05% and 1.82%) and patients with drug-induced liver injury (0.06% and 2.03%; p < 0.05). Intrahepatic type II NKT cells of AIH patients had a different cytokine profile than healthy subjects with an increased frequency of TNFα (77.8% vs. 59.1%, p < 0.05), decreased IFNγ (32.7% vs. 63.0%, p < 0.05) and a complete lack of IL-4 expressing cells (0% vs. 2.1%, p < 0.05). T cells in portal tracts expressed significantly more CD1d-RNA in AIH livers compared to controls. This study supports that in contrast to their assumed protective role in mice, human intrahepatic, sulfatide-reactive type II NKT cells displayed a proinflammatory cytokine profile in patients with AIH. Infiltrating T cells in portal areas of AIH patients overexpressed CD1d and could thereby activate type II NKT cells.
Assuntos
Hepatite Autoimune/imunologia , Fígado/imunologia , Sulfoglicoesfingolipídeos/imunologia , Adulto , Idoso , Antígenos CD1d/análise , Citocinas/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Células T Matadoras Naturais/imunologia , Fenótipo , Receptores de Quimiocinas/sangueRESUMO
We report the first 2 cases of hepatosplenic T-cell lymphoma in adolescents diagnosed with autoimmune hepatitis under long-term treatment with azathioprine. Patients presented with fatigue, hepatosplenomegaly, and pancytopenia. The diagnosis could be confirmed performing biopsies of liver and spleen or bone marrow, which demonstrated infiltration of neoplastic T lymphocytes with the typical phenotype with both markers, CD56(+) and TCRγδ(+) Thus, these cases emphasize the need to constantly reevaluate the administered dose and duration of thiopurines for autoimmune hepatitis, especially in adolescents.
Assuntos
Azatioprina/efeitos adversos , Hepatite Autoimune/tratamento farmacológico , Imunossupressores/efeitos adversos , Neoplasias Hepáticas/induzido quimicamente , Linfoma de Células T/induzido quimicamente , Neoplasias Esplênicas/induzido quimicamente , Adolescente , Azatioprina/uso terapêutico , Feminino , Hepatite Autoimune/complicações , Humanos , Imunossupressores/uso terapêutico , Neoplasias Hepáticas/diagnóstico , Linfoma de Células T/diagnóstico , Masculino , Neoplasias Esplênicas/diagnósticoRESUMO
BACKGROUND & AIMS: Hepatic ischemic reperfusion injury (IRI) is a major complication of liver transplantation and resectional hepatic surgeries. Natural killer T (NKT) cells predominate in liver, where they recognize lipid antigens bound to CD1d molecules. Type I NKT cells use a semi-invariant T-cell receptor and react with α-galactosylceramide; type II NKT cells use diverse T-cell receptors. Some type II NKT cells recognize the self-glycolipid sulfatide. It is not clear whether or how these distinct NKT cell subsets mediate hepatocellular damage after IRI. METHODS: We examined the roles of type I and type II NKT cells in mice with partial hepatic, warm ischemia, and reperfusion injury. RESULTS: Mice that lack type I NKT cells (Jα18-/-) were protected from hepatic IRI, indicated by reduced hepatocellular necrosis and serum levels of alanine aminotransferase. Sulfatide-mediated activation of type II NKT cells reduced interferon-γ secretion by type I NKT cells and prevented IRI. Protection from hepatic IRI by sulfatide-mediated inactivation of type I NKT cells was associated with significant reductions in hepatic recruitment of myeloid cell subsets, especially the CD11b(+)Gr-1(int), Gr-1(-), and NK cells. CONCLUSIONS: In mice, subsets of NKT cells have opposing roles in hepatic IRI: type I NKT cells promote injury whereas sulfatide-reactive type II NKT cells protect against injury. CD1d activation of NKT cells is conserved from mice to human beings, so strategies to modify these processes might be developed to treat patients with hepatic reperfusion injury.
Assuntos
Fígado/irrigação sanguínea , Fígado/patologia , Células T Matadoras Naturais/efeitos dos fármacos , Traumatismo por Reperfusão/prevenção & controle , Sulfoglicoesfingolipídeos/uso terapêutico , Animais , Antígeno CD11b/análise , Antígeno CD11b/imunologia , Feminino , Interferon gama/imunologia , Fígado/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Células Mieloides/imunologia , Células T Matadoras Naturais/imunologia , Necrose/tratamento farmacológico , Necrose/imunologia , Receptores de Quimiocinas/análise , Receptores de Quimiocinas/imunologia , Traumatismo por Reperfusão/imunologia , Traumatismo por Reperfusão/patologiaRESUMO
TCR-mediated recognition of beta-linked self-glycolipids bound to CD1d is poorly understood. Here, we have characterized the TCR repertoire of a CD1d-restricted type II NKT cell subset reactive to sulfatide involved in the regulation of autoimmunity and antitumor immunity. The sulfatide/CD1d-tetramer(+) cells isolated from naïve mice show an oligoclonal TCR repertoire with predominant usage of the Valpha3/Valpha1-Jalpha7/Jalpha9 and Vbeta8.1/Vbeta3.1-Jbeta2.7 gene segments. The CDR3 regions of both the alpha- and beta-chains are encoded by either germline or nongermline gene segments of limited lengths containing several conserved residues. Presence of dominant clonotypes with limited TCR gene usage for both TCR alpha- and beta-chains in type II NKT cells reflects specific antigen recognition not found in the type I NKT cells but similar to the MHC-restricted T cells. Although potential CD1d-binding tyrosine residues in the CDR2beta region are conserved between most type I and type II NKT TCRs, CDR 1alpha and 3alpha regions differ significantly between the two subsets. Collectively, the TCR repertoire of sulfatide-reactive type II NKT cells exhibits features of both antigen-specific conventional T cells and innate-like cells, and these findings provide important clues to the recognition of beta-linked glycolipids by CD1d-restricted T cells in general.
Assuntos
Glicolipídeos/imunologia , Células T Matadoras Naturais/imunologia , Receptores de Antígenos de Linfócitos T/metabolismo , Sequência de Aminoácidos , Animais , Antígenos CD1d/metabolismo , Autoimunidade , Sequência de Bases , Regiões Determinantes de Complementaridade , Primers do DNA/genética , Genes Codificadores dos Receptores de Linfócitos T , Glicolipídeos/química , Imunidade Inata , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , Células T Matadoras Naturais/classificação , Sulfotransferases/deficiência , Sulfotransferases/genéticaRESUMO
Natural killer T (NKT) cells generally recognize lipid-antigens presented in the context of the MHC class I-like molecule CD1d. CD1d-restricted NKT cells consist of two broad subsets: Type I, which express an invariant T cell receptor (TCR) and type II, which utilize diverse TCR gene segments. A major type II NKT subset has been shown to recognize a self-glycolipid, sulfatide. Both subsets play important roles in autoimmune diseases, tumor surveillance, and infectious diseases. While type I NKT cells protect from tumor growth by enhancing tumor surveillance, type II NKT cells may suppress anti-tumor immune responses. In a murine autoimmune hepatitis model, type I NKT cells contribute to pathogenesis, whereas activation of sulfatide-reactive type II NKT cells protects from disease. Sulfatide-mediated activation of type II NKT cells results in modification of dendritic cells and induction of anergy in type I NKT cells. Elucidation of this novel pathway of cross-regulation among NKT cell subsets will provide tools for intervention in autoimmune diseases and for designing strategies for effective anti-tumor immunity.
Assuntos
Antígenos/imunologia , Imunidade/imunologia , Células T Matadoras Naturais/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Autoimunidade/imunologia , Humanos , Neoplasias/imunologiaRESUMO
Gastrin-induced release of calcitonin from medullary thyroid carcinomas (MTC) is based on the expression of the cholecystokinin(2)-receptor (CCK(2)R) in these tumors. Recently, we have shown that the CCK(2)R is expressed not only in MTC but also in C-cells within the normal thyroid gland. The functions of the CCK(2)R in MTC and C-cells are largely unknown. We therefore explored the effects of gastrin-induced CCK(2)R stimulation in the highly differentiated MTC cell line, TT. CCK(2)R expression in TT-cells is detectable by RT-PCR as well as immunocytochemistry. Stimulation of the CCK(2)R by gastrin induces immediate release of calcitonin from TT-cells. Moreover, quantitative (LightCycler) RT-PCR demonstrates that gastrin stimulates transcription of the calcitonin and chromogranin A genes in TT-cells. TT-cell proliferation, assessed by counting of viable cells and (3)H-thymidine uptake, is markedly increased by gastrin. This effect is inhibited by the CCK(2)R-specific antagonist L-365,260. Our findings suggest physiological functions for the CCK(2)R in calcitonin-secretion and gene expression as well as a pathophysiological role in MTC proliferation. CCK(2)R antagonists might have therapeutic potential in these tumors.