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1.
Front Immunol ; 9: 2287, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30344522

RESUMO

Recent studies indicated that CXCR5+CD8+ T cells in lymph nodes could eradicate virus-infected target cells. However, in the current study we found that a subset of CXCR5+CD8+ T cells in the germinal centers from human tonsils or lymph nodes are predominately memory cells that express CD45RO and CD27. The involvement of CXCR5+CD8+ T cells in humoral immune responses is suggested by their localization in B cell follicles and by the concomitant expression of costimulatory molecules, including CD40L and ICOS after activation. In addition, CXCR5+CD8+ memory T cells produced significantly higher levels of IL-21, IFN-γ, and IL-4 at mRNA and protein levels compared to CXCR5-CD8+ memory T cells, but IL-21-expressing CXCR5+CD8+ T cells did not express Granzyme B and perforin. When cocultured with sorted B cells, sorted CXCR5+CD8+ T cells promoted the production of antibodies compared to sorted CXCR5-CD8+ T cells. However, fixed CD8+ T cells failed to help B cells and the neutralyzing antibodies against IL-21 or CD40L inhibited the promoting effects of sorted CXCR5+CD8+ T cells on B cells for the production of antibodies. Finally, we found that in the germinal centers of lymph nodes from HIV-infected patients contained more CXCR5+CD8+ T cells compared to normal lymph nodes. Due to their versatile functional capacities, CXCR5+CD8+ T cells are promising candidate cells for immune therapies, particularly when CD4+ T cell help are limited.

2.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 34(4): 289-295, 2018 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-29973317

RESUMO

Objective To investigate the recovery effects of IL-12 on the immune suppression induced by chemotherapeutic medicine in patients and mouse with malignant tumors. Methods Peripheral blood mononuclear cells (PBMCs) from tumor patients with or without chemotherapy and healthy donors were stimulated with or without anti-CD3 antibody plus anti-CD28 antibody in the presence or absent of IL-12. The levels of IFN-γ and TNF-α in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA). The expression of IFN-γ in different subsets of T cells was analyzed by fluorescence activated cell sorter (FACS). Finally, we established the cisplatin toxicity mouse model and measured the levels of IFN-γ and TNF-α by ELISA and FACS. Results PBMCs from the patients with malignant tumors produced significantly lower levels of IFN-γ and TNF-α than PBMCs from healthy donors. The production of IFN-γ and TNF-α was higher in pre-chemotherapeutic patients compared with post-chemotherapeutic patients, whereas IL-12 could remarkably recover the production of IFN-γ and TNF-α in the patients with malignant tumors. FACS showed that IL-12 recovered the expression of IFN-γ by CD4+ and CD8+ T cells in post-chemotherapeutic patients. Finally, the results from the animal studies in vitro and in vivo proved that IL-12 recovered the inhibitory effect of chemotherapeutic drugs on immune function. Conclusion Chemotherapeutics inhibits the immune responses in patients and animals, and IL-12 can recover the suppressive effects of chemotherapeutics on the production of cytokines. Our results indicated that IL-12 might play an important role in the reconstruction of immune function in cancer patients with chemotherapeutics.

3.
Clin Sci (Lond) ; 131(4): 297-308, 2017 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-27923881

RESUMO

The translocator protein (TSPO) ligands affected inflammatory and immune responses. However, the exact effects of TSPO ligands on Th1 responses in vitro and in vivo are still unclear. In the present study, we found that TSPO ligands, FGIN1-27 and Ro5-4864, suppressed the cytokine production in a dose-dependent manner by purified human CD4+ T-cells from peripheral blood mononuclear cells (PBMCs) after stimulation. TSPO ligands inhibited the production of interferon γ (IFN-γ) by memory CD4+ T-cells and the differentiation of naïve CD4+ T-cells into Th1 cells via suppressing the activity of the corresponding transcription factors as indicated by reduced expression of T-bet and down-regulation of STAT1, STAT4 and STAT5 phosphorylation. TSPO ligands suppressed cell proliferation and activation of CD4+ T-cells by the inhibition of TCR signal transduction including membrane proteins: Zap, Lck, Src; cytoplasm proteins: Plcγ1, Slp-76, ERK, JNK and the nucleoproteins: c-Jun and c-Fos. In addition, FGIN1-27 inhibited mixed lymphocyte reactions by human or murine cells. After the transplantation of allogeneic murine skin, injection of FGIN1-27 into mice prevented graft rejection by inhibition of cell infiltration and IFN-γ production. Taken together, our data suggest that TSPO ligands inhibit Th1 cell responses and might be novel therapeutic medicine for the treatment of autoimmune diseases and prevention of transplant rejection.


Assuntos
Rejeição de Enxerto/prevenção & controle , Ácidos Indolacéticos/uso terapêutico , Transplante de Pele , Células Th1/imunologia , Adolescente , Adulto , Animais , Benzodiazepinonas/imunologia , Linfócitos T CD4-Positivos/imunologia , Células Cultivadas , Citocinas/biossíntese , Avaliação Pré-Clínica de Medicamentos/métodos , Feminino , Rejeição de Enxerto/imunologia , Humanos , Ácidos Indolacéticos/imunologia , Ligantes , Ativação Linfocitária/imunologia , Teste de Cultura Mista de Linfócitos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Fosforilação/imunologia , Receptores de GABA/metabolismo , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais/imunologia , Proteínas com Domínio T/metabolismo , Adulto Jovem
4.
Sci Rep ; 6: 30362, 2016 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-27468819

RESUMO

Although CD4(+) T cells are recognized to play an important role in the inflammatory response of nasal polyps (NPs), the biological functions of CD8(+) T cells in polypogenesis remain unclear. In this study, we analyzed cell markers, cytokine expression and transcription factors in IL-21-expressing CD8(+) T cells in polyp tissues of NP patients. The results showed that the majority of IL-21-producing CD8(+) T cells were effector memory cells and they co-expressed IFN-γ. IL-21-expressing CD8(+) T cells in polyp tissues expressed higher CXCR5, PD-1, and ICOS levels than cells in control tissues and showed significantly higher T-bet and Bcl-6 expression levels compared with IL-21(-)CD8(+) T cells. Purified polyp CD8(+) T cells promoted IgG production from isolated polyp B cells in vitro, and recombinant IL-12 modulated the expression of IL-21, IFN-γ and CD40L in purified polyp CD8(+) T cells. Moreover, the percentage of IL-21(+)CD8(+) T cells in polyp tissues was positively correlated with endoscopic and CT scan scores in NP patients. These findings indicated that polyp CD8(+) T cells, by co-expressing IL-21 and IFN-γ and other markers, display a Tfh cell functionality, which is associated with the clinical severity of NP patients.


Assuntos
Linfócitos T CD8-Positivos/metabolismo , Interleucinas/metabolismo , Pólipos Nasais/metabolismo , Adolescente , Adulto , Idoso , Linfócitos B/metabolismo , Ligante de CD40/metabolismo , Estudos de Casos e Controles , Separação Celular , Feminino , Citometria de Fluxo , Humanos , Imunoglobulina G/metabolismo , Interferon gama/metabolismo , Interleucina-17/metabolismo , Masculino , Pessoa de Meia-Idade , Fenótipo , Receptores CXCR5/metabolismo , Proteínas Recombinantes/metabolismo , Adulto Jovem
5.
PLoS One ; 11(3): e0151721, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27031950

RESUMO

Our previous result indicated that memory-like human natural killer (NK) cells from TB pleural fluid cells (PFCs) produced large amounts of IFN-γ in response to Bacille Calmette Guerin (BCG). Furthermore, recent studies have shown that human lymphoid tissues harbored a unique NK cell subset that specialized in production of interleukin (IL)-22, a proinflammatory cytokine that mediates host defense against pathogens. Yet little information was available with regard to the properties of IL-22 production by memory-like human NK cells. In the present study, we found that cytokines IL-15 induced and IL-12 enhanced the levels of IL-22 by NK cells from TB PFCs. In addition, IL-22 but not IL-17 was produced by NK cells from PFCs in response to BCG and M.tb-related Ags. More importantly, the subset of specific IL-22-producing NK cells were distinct from IFN-γ-producing NK cells in PFCs. CD45RO+ or CD45RO- NK cells were sorted, co-cultured with autologous monocytes and stimulated with BCG for the production of IL-22. The result demonstrated that CD45RO+ but not CD45RO- NK cells produced significantly higher level of IL-22. Anti-IL-12Rß1 mAbs (2B10) partially inhibit the expression of IL-22 by NK cells under the culture with BCG. Consistently, BCG specific IL-22-producing NK cells from PFCs expressed CD45ROhighNKG2Dhighgranzyme Bhigh. In conclusion, our data demonstrated that memory-like antigen-specific CD45RO+ NK cells might participate in the recall immune response for M. tb infection via producing IL-22, which display a critical role to fight against M. tb.


Assuntos
Antígenos de Bactérias/imunologia , Interleucina-15/imunologia , Interleucinas/imunologia , Células Matadoras Naturais/imunologia , Tuberculose Pleural/imunologia , Adulto , Idoso , Antígenos de Bactérias/farmacologia , Células Cultivadas , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Memória Imunológica/imunologia , Interferon gama/imunologia , Interferon gama/metabolismo , Interleucina-12/imunologia , Interleucina-12/farmacologia , Interleucina-15/farmacologia , Interleucinas/metabolismo , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/metabolismo , Antígenos Comuns de Leucócito/imunologia , Antígenos Comuns de Leucócito/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Tuberculose Pleural/metabolismo , Adulto Jovem
6.
PLoS One ; 11(1): e0147356, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26785168

RESUMO

In the current study of Mycobacterium tuberculosis (MTB)-specific T and B cells, we found that MTB-specific peptides from early secreted antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) induced the expression of IL-21 predominantly in CD4(+) T cells. A fraction of IL-21-expressing CD4(+) T cells simultaneously expressed Th1 cytokines but did not secrete Th2 or Th17 cytokines, suggesting that MTB-specific IL-21-expressing CD4(+) T cells were different from Th1, Th2 and Th17 subpopulations. The majority of MTB-specific IL-21-expressing CD4(+) T cells co-expressed IFN-γ and IL-21+IFN-γ(+)CD4(+) T cells exhibited obviously polyfunctionality. In addition, MTB-specific IL-21-expressing CD4(+) T cells displayed a CD45RO+CD62Ll(ow)CCR7(low)CD40L(high)ICOS(high) phenotype. Bcl-6-expression was significantly higher in IL-21-expressing CD4(+) T cells than IL-21-CD4(+) T cells. Moreover, IL-12 could up-regulate MTB-specific IL-21 expression, especially the frequency of IL-21(+)IFN-γ+CD4(+) T cells. Taken together, our results demonstrated that MTB-specific IL-21(+)IFN-γ(+)CD4(+) T cells from local sites of tuberculosis (TB) infection could be enhanced by IL-12, which have the features of both Tfh and Th1 cells and may have an important role in local immune responses against TB infection.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Interferon gama/imunologia , Interleucina-12/farmacologia , Interleucinas/imunologia , Mycobacterium tuberculosis/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Tuberculose/imunologia , Adulto , Idoso , Inibidores da Angiogênese/farmacologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/efeitos dos fármacos , Tuberculose/tratamento farmacológico , Tuberculose/microbiologia , Adulto Jovem
7.
Cell Cycle ; 14(21): 3362-72, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26566861

RESUMO

Interleukine-12 is critical for the differentiation of Th1 cells and can improve the development of Th1 cells with Tfh cell features in mouse model. Human effector CD4(+) T cells also exhibit poly-functionality by co-expressing IL-21 and IFN-γ. However, the effects of IL-12 on regulating generation of human IL-21- and IFN-γ-expressing CD4(+) T cells are still incompletely understood. Our studies found that IL-12 but not IL-21 could induce the differentiation of human naive CD4(+) T cells into multi-cytokine expressing CD4(+) T cells in vitro, which co-expressed IL-21 and IFN-γ with or without IL-2 and TNF-α. At early stage of differentiation, addition of excess exogenous IFN-γ could increase the generation of IL-21- and IFN-γ-expressing CD4(+) T cells, furthermore, anti-IFN-γ depressed the percentage of poly-functional CD4(+) T cells. Phenotypically, IL-21(+)IFN-γ(+)CD4(+) T cells exhibited more characteristic features about both of Th1 and Tfh cells than IL-21 or IFN-γ single-expressing CD4(+) T cells. Mechamistically, IL-12 modulated the differentiation of IL-21(+)IFN-γ(+)CD4(+) T cells from naive CD4(+) T cells via the pathways of STAT-1/4, T-bet and BCL(-)6. Different from naive CD4(+) T cells, IL-12 increasing the generation of IL-21(+)IFN-γ(+)CD4(+) T cells from memory CD4(+) T cells was only involved in STAT-4 pathway but not STAT-1. Poly-functional CD4(+) T cells were contributed to generation and progress of varies diseases and our studies provide basic theoretics for the designs of vaccine and therapies of diseases.


Assuntos
Linfócitos T CD4-Positivos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Interferon gama/metabolismo , Interleucina-12/farmacologia , Interleucinas/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Células Cultivadas , Proteínas de Ligação a DNA/metabolismo , Sangue Fetal/citologia , Humanos , Memória Imunológica , Recém-Nascido , Interferon gama/imunologia , Interferon gama/farmacologia , Interleucinas/imunologia , Interleucinas/farmacologia , Cinética , Fenótipo , Proteínas Proto-Oncogênicas c-bcl-6 , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT4/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas com Domínio T/metabolismo , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th1/metabolismo , Regulação para Cima
8.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 31(11): 141-1446, 2015 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-26522348

RESUMO

OBJECTIVE: To investigate the secretion of IL-12p40 and IL-6 by splenocytes, dendritic cells stimulated by different Toll-like receptor (TLR) agonists or in the sera of mice immunized with different TLR agonists, and evaluate the effects of different TLR agonists on the function of T helper (Th) cells, especially the differentiation of Th1 cells. METHODS: Supernatants of splenocytes and dendritic cells stimulated with different TLR agonists for 24 hours or sera of mice immunized with different TLR agonists at different time points were used to determine the levels of IL-12p40 and IL-6 by ELISA. CD4⁺ T cells isolated from the spleens of ovalbumin-T cell receptor (OVA-TCR) transgenic BALB/c (DO11.10) mice were co-cultured with antigen presenting cells (APCs) from congenic BALB/c mice at 1:3 ratio of T:APCs. Cultures were stimulated with OVA peptide or OVA peptide plus different doses of TLR agonists and the supernatants collected at different time points were assayed by ELISA for detecting IFN-γ. RESULTS: Pam3CSK4, R848 and CpG oligodeoxynucleotide (ODN) promoted the production of IL-12p40 and IL-6 by splenocytes and dendritic cells obviously, and induced the expression of IFN-γ in antigen specific CD4⁺ T cells in a time- and dose-dependent manner. Monophosphoryl lipid A from Salmonella minnesota R595 lipopolysaccharide (MPLA-SM) induced low levels of cytokines by splenocytes and couldn't promote the production of IFN-γ by antigen specific CD4⁺ T cells, but increased the expressions of IL-12p40 and IL-6 by DCs. All the sera of mice immunized with the four TLR agonists expressed high levels of IL-12p40 and IL-6. CONCLUSION: Splenocytes, DCs stimulated or sera of mice immunized with different TLR agonists produced different levels of cytokines, which could further affect the differentiation of Th1 cells.


Assuntos
Linfócitos T Auxiliares-Indutores/imunologia , Receptores Toll-Like/agonistas , Animais , Feminino , Interferon gama/biossíntese , Subunidade p40 da Interleucina-12/sangue , Interleucina-6/sangue , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia
9.
Oncotarget ; 6(30): 28633-45, 2015 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-26416419

RESUMO

Natural killer T (NKT) cells from mouse and human play an important role in the immune responses against Mycobacterium tuberculosis. However, the function of CD3(+)TCRvß11(+) NKT cells at the local site of M. tuberculosis infection remains poorly defined. In the present study, we found that after stimulation with M. tuberculosis antigens, NKT cells isolated from tuberculosis (TB) pleural fluid mononuclear cells (PFMCs) produced IL-21 and other cytokines including IFN-γ, TNF-α, IL-2 and IL-17. IL-21-expressing NKT cells in PFMCs displayed effector memory phenotype, expressing CD45RO(high)CD62L(low)CCR7(low). Moreover, NKT cells expressed high levels of CXCR5 and all of IL-21-expressing NKT cells co-expressed CXCR5. The frequency of BCL-6-expression was higher in IL-21-expressing but not in non-IL-21-expressing CD3(+)TCRvß11(+) NKT cells. Sorted CD3(+)TCRvß11(+) NKT cells from PFMCs produced IFN-γ and IL-21 after stimulation, which expressed CD40L. Importantly, CD3(+)TCRvß11(+) NKT cells provided help to B cells for the production of IgG and IgA. Taken together, our data demonstrate that CD3(+)TCRvß11(+) NKT cells from a local site of M. tuberculosis infection produce IL-21, express CXCR5 and CD40L, help B cells to secrete IgG and IgA, and may participate in local immune responses against M. tuberculosis infection.


Assuntos
Antígenos de Bactérias/imunologia , Linfócitos B/imunologia , Imunoglobulina A Secretora/imunologia , Imunoglobulina G/imunologia , Interleucinas/imunologia , Mycobacterium tuberculosis/imunologia , Células T Matadoras Naturais/imunologia , Tuberculose Pleural/imunologia , Adulto , Idoso , Antígenos de Bactérias/metabolismo , Linfócitos B/metabolismo , Linfócitos B/microbiologia , Complexo CD3/imunologia , Ligante de CD40/imunologia , Separação Celular/métodos , Células Cultivadas , Feminino , Citometria de Fluxo , Humanos , Imunoglobulina A Secretora/metabolismo , Imunoglobulina G/metabolismo , Memória Imunológica , Imunofenotipagem , Interleucinas/metabolismo , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/metabolismo , Células T Matadoras Naturais/metabolismo , Células T Matadoras Naturais/microbiologia , Comunicação Parácrina , Fenótipo , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Receptores CXCR5/imunologia , Transdução de Sinais , Tuberculose Pleural/metabolismo , Tuberculose Pleural/microbiologia , Adulto Jovem
10.
Sci Rep ; 5: 12781, 2015 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-26239551

RESUMO

In the previous study, we found that the levels of IL-21 in nasal polyps (NPs) were significantly increased and associated with polyp size and recurrence. However, it is unclear that the cell source of IL-21 and the regulation of IL-21 in NP tissues. In the present study, we isolated the lymphocytes from NP tissues, uncinate tissues and peripheral blood of patients with NPs. The cells were analyzed for cell surface markers, cytokines and transcriptional factors by flow cytometry. The results indicated that CD4(+) T cells were the major IL-21-expressing cells in NP tissues and the majority of IL-21 producing CD4(+) T cells co-expressed IFN-γ or IL-17A. IL-21(+)IFN-γ(+)CD4(+) T cells in NP tissues exhibited the features of both Tfh and Th1 cells which co-expressed significantly higher amount of CXCR5, ICOS, PD-1, Bcl-6 and T-bet than did IL-21(+)IFN-γ(-)CD4(+) T cells (p < 0.05). Treatment of the lymphocytes from NP tissues with IL-12 enhanced the production of IL-21 and IFN-γ, especially the frequency of IL-21(+)IFN(-)γ(+)CD4(+) T cells (p < 0.05). The blockade of IL-12 inhibited the production of IL-21 and IFN-γ (p < 0.05). These findings indicated that IL-12 positively enhanced the generation of IL-21(+)IFN-γ(+)CD4(+) T cells having the features of both Tfh and Th1 cells in NP tissues.


Assuntos
Linfócitos T CD4-Positivos/efeitos dos fármacos , Interferon gama/imunologia , Interleucina-12/farmacologia , Interleucinas/imunologia , Pólipos Nasais/imunologia , Adolescente , Adulto , Idoso , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Estudos de Casos e Controles , Linhagem da Célula/efeitos dos fármacos , Linhagem da Célula/imunologia , Proliferação de Células , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Feminino , Expressão Gênica , Humanos , Proteína Coestimuladora de Linfócitos T Induzíveis/genética , Proteína Coestimuladora de Linfócitos T Induzíveis/imunologia , Interferon gama/genética , Interleucina-17/genética , Interleucina-17/imunologia , Interleucinas/genética , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/patologia , Cultura Primária de Células , Receptor de Morte Celular Programada 1/genética , Receptor de Morte Celular Programada 1/imunologia , Proteínas Proto-Oncogênicas c-bcl-6 , Receptores CXCR5/genética , Receptores CXCR5/imunologia , Proteínas com Domínio T/genética , Proteínas com Domínio T/imunologia
11.
J Biol Chem ; 289(51): 35561-9, 2014 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-25378399

RESUMO

We found that after stimulation for a few hours, memory but not naive CD4(+) T cells produced a large amount of IFN-γ; however, the mechanism of rapid response of memory CD4(+) T cells remains undefined. We compared the expression of transcription factors in resting or activated naive and memory CD4(+) T cells and found that T-bet, but not pSTAT-1 or pSTAT-4, was highly expressed in resting memory CD4(+) T cells and that phenotypic characteristics of T-bet(+)CD4(+) T cells were CD45RA(low)CD62L(low) CCR7(low). After short-term stimulation, purified memory CD4(+) T cells rapidly produced effector cytokines that were closely associated with the pre-existence of T-bet. By contrast, resting naive CD4(+) T cells did not express T-bet, and they produced cytokines only after sustained stimulation. Our further studies indicated that T-bet was expressed in the nuclei of resting memory CD4(+) T cells, which might have important implications for rapid IFN-γ production. Our results indicate that the pre-existence and nuclear mobilization of T-bet in resting memory CD4(+) T cells might be a possible transcriptional mechanism for rapid production of cytokines by human memory CD4(+) T cells.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Citocinas/imunologia , Memória Imunológica/imunologia , Proteínas com Domínio T/imunologia , Transporte Ativo do Núcleo Celular/imunologia , Adulto , Western Blotting , Linfócitos T CD4-Positivos/metabolismo , Núcleo Celular/imunologia , Núcleo Celular/metabolismo , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Feminino , Expressão Gênica/imunologia , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interferon gama/metabolismo , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo , Células Th1/imunologia , Células Th1/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/imunologia , Fatores de Transcrição/metabolismo , Adulto Jovem
12.
J Clin Immunol ; 34(8): 979-90, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25190196

RESUMO

Natural killer T (NKT) cells from mouse and human play a protective role in the immune responses against the infection of Mycobacterium tuberculosis. However, the characteristic of CD3(+)TCRvß11(+) NKT cells at the local site of M. tuberculosis infection remains poorly defined. In the present study, we found that the numbers of CD3(+)TCRvß11(+) NKT cells in pleural fluid mononuclear cells (PFMCs) were significantly lower than those in peripheral blood mononuclear cells (PBMCs). However, CD3(+)TCRvß11(+) NKT cells from PFMCs spontaneously expressed high levels of CD69 and CD25 and effector memory phenotypes of CD45RO(high)CD62L(low)CCR7(low). After stimulation with the antigens of M. tuberculosis, CD3(+)TCRvß11(+) NKT cells from PFMCs produced high levels of IFN-γ. Sorted CD3(+)TCRvß11(+) NKT cells from PFMCs cultured with antigen presenting cells (APCs) produced IFN-γ protein and mRNA. The production of IFN-γ could be completely inhibited by AG490 and Wortmannin. In addition, CD3(+)TCRvß11(+) NKT cells from PFMCs expressed higher levels of Fas (CD95), FasL (CD178) and perforin but lower levels of granzyme B compared with those from PBMCs. Taken together, our data demonstrated for the first time that M. tuberculosis-specific CD3(+)TCRvß11(+) NKT cells participated in the local immune responses against M. tuberculosis through the production of IFN-γ and the secretion of cytolytic molecules.


Assuntos
Memória Imunológica , Mycobacterium tuberculosis/imunologia , Células T Matadoras Naturais/imunologia , Tuberculose Pleural/imunologia , Adulto , Idoso , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interferon gama/sangue , Masculino , Pessoa de Meia-Idade , Complexo Receptor-CD3 de Antígeno de Linfócitos T/sangue
13.
FASEB J ; 28(7): 3238-48, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24675363

RESUMO

The mechanism by which IFN-α regulates the host response to Mycobacterium tuberculosis (M.tb) infection in humans is poorly understood. In the present study, we found that freshly isolated pleural fluid mononuclear cells (PFMCs) from tuberculous pleural effusion but not peripheral blood mononuclear cells (PBMCs) spontaneously expressed IFN-α and IL-1ß in vivo. In addition, exogenous IFN-α significantly inhibited production of IL-1ß in PFMCs after stimulation with Bacillus Calmette-Guérin (BCG). To further evaluate the effect of endogenous IFN-α on BCG-induced IL-1ß production, a neutralizing antibody to IFN-α was added to the cultures of BCG-stimulated PFMCs. As expected, neutralization of IFN-α by antibody significantly enhanced the production of IL-1ß. Notably, we showed that IFN-α inhibited production of IL-1ß through 2 distinct mechanisms: IFN-α signaling, via the STAT1 transcription factor, suppressed caspase-1-dependent IL-1ß maturation, and IFN-α induced the production of IL-10 in a STAT1-dependent manner in which IL-10 reduced the abundance of IL-1ß. In contrast, we found that IFN-α enhanced the production of IFN-γ, and IFN-γ also suppressed IL-1ß production in the PFMCs during BCG stimulation. Our findings demonstrate that IFN-α employs distinct pathways for regulating IL-1ß production and reveal that in the case of M.tb infection, the induction of IFN-α and IFN-γ might be associated with M.tb immune escape and disease progression in infected humans.-Ma, J., Yang, B., Yu, S., Zhang, Y., Zhang, X., Lao, S., Chen, X., Li, B., Wu, C. Tuberculosis antigen-induced expression of IFN-α in tuberculosis patients inhibits production of IL-1ß.


Assuntos
Antígenos de Bactérias/imunologia , Interferon-alfa/genética , Interferon-alfa/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/imunologia , Adulto , Idoso , Caspase 1/genética , Caspase 1/imunologia , Caspase 1/metabolismo , Feminino , Humanos , Interferon-alfa/metabolismo , Interferon gama/genética , Interferon gama/imunologia , Interferon gama/metabolismo , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Mycobacterium bovis/imunologia , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/imunologia , Fator de Transcrição STAT1/metabolismo , Tuberculose/genética , Tuberculose/metabolismo , Adulto Jovem
14.
Tuberculosis (Edinb) ; 94(3): 219-25, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24566284

RESUMO

Increasing evidences in animals and humans suggest that CD8(+) T cells contribute significantly to immune defenses against Mycobacterium tuberculosis (Mtb). In the present study, we found that without any stimulation, CD8(+) T cells in pleural fluid cells (PFCs) expressed significantly higher levels of CD69 than PBMCs from patients with tuberculous pleurisy (TBP). CD8(+)CD69(+) T cells expressed significantly higher levels of CD45RO and HLA-DR and lower levels of CD45RA than CD8(+)CD69(-) T cells, demonstrating that CD8(+)CD69(+) T cells were activated memory cells. Furthermore, we found higher expression of CCR6 and lower expression of CCR7 and CD62L on CD8(+)CD69(+) T cells compared with CD8(+)CD69(-) T cells, suggesting that the expression of CCR6 and reduced expression of CCR7 and CD62L might facilitate the migration of circulating CD8(+)CD69(+) T cells into tuberculous pleural space. Importantly, following stimulation with culture filtrate protein of 10 kDa (CFP10) peptides, CD8(+)CD69(+) T cells but not CD8(+)CD69(-) T cells expressed CD107a/b, IFN-γ and TNF-α, demonstrating that CD8(+)CD69(+) T cells were MTB-specific cells. In addition, the majority of CD8(+)CD69(+) T cells were dominated by polyfunctional T cells. In summary, we demonstrated that CD69 as a useful marker for MTB-specific CD8(+) T cells in PFCs from patients with TBP enabled a direct ex vivo estimation of the quantity, as well as the quality, of MTB-specific CD8(+) responses.


Assuntos
Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Lectinas Tipo C/metabolismo , Tuberculose Pleural/imunologia , Adulto , Idoso , Biomarcadores/metabolismo , Feminino , Humanos , Interferon gama/biossíntese , Antígenos Comuns de Leucócito/metabolismo , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/imunologia , Fator de Necrose Tumoral alfa/biossíntese , Adulto Jovem
15.
Tuberculosis (Edinb) ; 94(2): 123-30, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24406079

RESUMO

We recently reported that pleural fluid mononuclear cells (PFMCs) from tuberculous pleurisy stimulated with Bacillus Calmette-Guérin (BCG) or TB antigens produced high levels of cytokines. However, it was still unclear what mechanism of the PFMCs used to migrate into the pleural fluids in TB infection. In the present study, we found that CD3(+)CD4(+) and CD3(+)CD8(+) T cells from PFMCs expressed significantly high levels of CXCR3 compared to PBMCs. In addition, the levels of CXCL10 (the ligand for CXCR3) in pleural fluids were significantly higher than those in normal serum and cancerous fluids. After stimulation with BCG, PFMCs produced high levels of CXCL10. Importantly, the synthesis of CXCL10 was mainly dependent on the BCG-induced production of IFNs, because the neutralization of endogenous IFN-α or IFN-γ with mAbs significantly reduced the production of CXCL10 from BCG-stimulated PFMCs. In addition, the tubercular pleural fluid (TBPF) or exogenous CXCL10 induced the migration of PFMCs, indicating that IFN-α or IFN-γ modulated the immune response through the expression of CXCL10 to aid the recruitment and selective homing of activated/effector cells to the site of Mycobacterium tuberculosis (M.tb) infection. Taken together, the levels of CXCL10 in pleural fluids were high and BCG-stimulated PFMCs expressed high levels of CXCL10, and CXCL10 induced the migration of PFMCs into the pleural fluids in TB infection.


Assuntos
Quimiocina CXCL10/imunologia , Interferon-alfa/imunologia , Interferon gama/imunologia , Leucócitos Mononucleares/imunologia , Neoplasias Pulmonares/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose Pleural/imunologia , Adulto , Idoso , Antígenos de Bactérias/imunologia , Movimento Celular/imunologia , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Linfócitos T/imunologia , Tuberculose Pleural/tratamento farmacológico
16.
PLoS One ; 8(12): e82196, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24349220

RESUMO

CD8(+) T cells are essential for host defense to Mycobacterium tuberculosis (Mtb) infection and identification of CD8(+) T cell epitopes from Mtb is of importance for the development of effective peptide-based diagnostics and vaccines. We previously demonstrated that the secreted 10-KDa culture filtrate protein (CFP10) from Mtb is a potent CD8(+) T cell antigen but the repertoire and dominance pattern of human CD8 epitopes for CFP10 remained poorly characterized. In the present study, we undertook to define immunodominant CD8 epitopes involved in CFP10 using a panel of CFP10-derived 13-15 amino acid (aa) peptides overlapping by 11 aa. Four peptides in CFP10 were observed to induce significant CD8(+) T cell responses and we further determined the size of the epitopes involved in each individual peptide tested. Four 9 aa CD8 epitopes were finally identified and deleting a single amino acid from the N or C terminus of either peptide markedly reduced IFN-γ production, suggesting that they are minimum of CD8 epitopes. In the individuals tested, each epitope represented a single immunodominant response in CD8(+) T cells. The epitope-specific CD8(+) T cells displayed effector or effector memory phenotypes and could upregulate the expression of CD107a/b upon antigen stimulation. In addition, we found that epitope-specific CD8(+) T cells shared biased usage of T cell receptor (TCR) variable region of ß chain (Vß) 12, 9, 7.2 or Vß4 chains. As judged from HLA-typing results and using bioinformatics technology for prediction of MHC binding affinity, we found that the epitope-specific CD8(+) T cells are all restricted by HLA-B alleles. Our findings suggest that the four epitopes in CFP10 recognized by CD8(+) T cells might be of importance for the development of Mtb peptide-based vaccines and for improved diagnosis of TB in humans.


Assuntos
Alelos , Proteínas de Bactérias/imunologia , Linfócitos T CD8-Positivos/imunologia , Antígenos HLA-B/genética , Epitopos Imunodominantes/imunologia , Mycobacterium tuberculosis/imunologia , Adulto , Idoso , Sequência de Aminoácidos , Linfócitos T CD4-Positivos/imunologia , Epitopos de Linfócito T/imunologia , Feminino , Teste de Histocompatibilidade , Humanos , Memória Imunológica/imunologia , Interferon gama/biossíntese , Proteína 1 de Membrana Associada ao Lisossomo/metabolismo , Proteína 2 de Membrana Associada ao Lisossomo/metabolismo , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Tuberculose Pleural/imunologia , Fator de Necrose Tumoral alfa/biossíntese , Adulto Jovem
17.
Immunol Lett ; 152(2): 96-103, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23727477

RESUMO

IL-27, a member of IL-6/IL-12 cytokine family, plays pro- and anti-inflammatory functions in immune responses. It can promote inflammation by inducing Th1 differentiation and exert the inhibitory effects on Th2 and Th17 mediated immune responses. Moreover, IL-27 suppresses CD28-mediated IL-2 production from mouse naive CD4(+) T cells. In the present study, we demonstrate that IL-27 inhibits the production of IL-22 and induces the expression of IFN-γ in CD4(+) T cells from human umbilical cord blood mononuclear cells (CBMCs) stimulated with anti-CD3 and anti-CD28 in dose-dependent manner. In addition, the suppression of IL-22 is not dependent on the production of IFN-γ and IL-10. Importantly, IL-27 promotes the expression of SOCS1, which could be inhibited by a Jak2/STAT inhibitor, AG490. Importantly, the expression of IL-22 could not be inhibited under the circumstances with the lower expression of SOCS1. Moreover, IL-27 inhibits the production of IL-22 in CD4(+)CD45RA(+) and CD4(+)CD45RO(+) T cells from PBMCs. These data identify that IL-27 may suppress the production of IL-22 by inducing the expression of SOCS1 in human CD4(+) T cells. Furthermore, it demonstrates that IL-27 may be a therapeutic approach in the treatment of IL-22-mediated diseases.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Interleucinas/biossíntese , Interleucinas/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Células Cultivadas , Sangue Fetal , Humanos , Recém-Nascido , Inflamação/imunologia , Interferon gama/biossíntese , Interleucina-10/biossíntese , Antígenos Comuns de Leucócito/metabolismo , Leucócitos Mononucleares/imunologia , Ativação Linfocitária/imunologia , Transdução de Sinais/imunologia , Proteína 1 Supressora da Sinalização de Citocina , Proteínas Supressoras da Sinalização de Citocina/biossíntese , Tirfostinas/farmacologia
18.
PLoS One ; 8(5): e64612, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23741351

RESUMO

IL-21 has pleiotropic effects on innate and adaptive immune response, and plays an important role in the development of autoimmune disease and antitumor activity. It has been reported that IL-21 is produced by CD4(+) T cells and NKT cells. However, the differentiation of IL-21-producing CD4(+) T cells in humans remains largely unclear. In the present study, we showed that cytokines of IL-1ß, IL-6 or IL-21 induced differentiation of human IL-21-producing CD4(+) T cells, and TGF-ß enhanced the effect of inflammatory cytokines on the development of IL-21-producing CD4(+) T cells. Furthermore, we found that the majority of IL-21-producing cells were distinct from Th17 cells and Th1 cells since they did not co-express IL-17 and IFN-γ. TGF-ß significantly inhibited the production of IFN-γ and enhanced the effect of IL-21 on the development of IL-21-producing CD4(+) T cells. In addition, we found that IL-21 inhibited the differentiation of CD4(+) Foxp3(+) T cells induced by TGF-ß. Further study indicated that IL-21 induced phosphorylation of transcriptional factors of STAT1, STAT3 and STAT5, and TGF-ß induced phosphorylation of Smad3 in CD4(+) T cells. Taken together, our data indicated that TGF-ß enhanced IL-21-induced differentiation of IL-21-producing CD4(+) T cells, and the majority of IL-21-producing cells were different from Th17 and Th1 cells. Our results provide a new sight regarding the differentiation of human CD4(+) T cells.


Assuntos
Linfócitos T CD4-Positivos/efeitos dos fármacos , Sangue Fetal/citologia , Interleucinas/farmacologia , Proteína Smad3/genética , Fator de Crescimento Transformador beta/farmacologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Diferenciação Celular/efeitos dos fármacos , Sangue Fetal/imunologia , Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-17/farmacologia , Interleucina-1beta/farmacologia , Interleucina-6/farmacologia , Interleucinas/biossíntese , Cultura Primária de Células , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/imunologia , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/imunologia , Fator de Transcrição STAT5/genética , Fator de Transcrição STAT5/imunologia , Transdução de Sinais , Proteína Smad3/imunologia , Células Th1/citologia , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th17/citologia , Células Th17/efeitos dos fármacos , Células Th17/imunologia
19.
Clin Immunol ; 148(1): 113-23, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23685221

RESUMO

We have previously shown that human memory-like NK cells were persistent in tuberculous pleurisy but it was unclear how NK cells migrated into the pleural fluids. At present, we found that NK cells from TB pleural fluid cells (PFCs) expressed significantly higher levels of CXCR3 and CXCR4 than NK cells from PBMCs. Migration assay demonstrated that IP-10 and SDF-1 induced more migration of NK cells from PFCs than PBMCs. CD45RO(+) or CD45RO(-) NK cells from PFCs were co-cultured with autologous monocytes and stimulated with BCG. The results showed CD45RO(+) but not CD45RO(-) NK cells produced significantly higher levels of IFN-γ, which was IL-12-dependent since anti-IL-12Rß1 mAbs could significantly inhibit the IFN-γ by NK cells. Collectively, our data demonstrated that human Mycobacterium tuberculosis-specific NK cells were migrated into the local site of TB infection mainly via IP-10/CXCR3 and SDF-1/CXCR4 axis, memory-like NK cells might display an important role against M. tuberculosis infection.


Assuntos
Vacina BCG/imunologia , Memória Imunológica/imunologia , Células Matadoras Naturais/imunologia , Mycobacterium tuberculosis/imunologia , Receptores CXCR3/imunologia , Receptores CXCR4/imunologia , Tuberculose Pleural/imunologia , Adolescente , Adulto , Idoso , Vacina BCG/administração & dosagem , Movimento Celular/imunologia , Feminino , Citometria de Fluxo , Humanos , Interferon gama/imunologia , Masculino , Pessoa de Meia-Idade , Receptores CXCR3/biossíntese , Receptores CXCR4/biossíntese , Adulto Jovem
20.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 28(9): 926-9, 2012 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-22980654

RESUMO

AIM: To study the mechanism underlying the IL-12-induced cytotoxic function of NK cells to Jurkat cells. METHODS: NK cells from peripheral blood mononuclear cells (PBMCs) were purified by magnetic sorting and stimulated with or without IL-12. The expression of genes on IL-12-treated and non-IL-12-treated NK cells was analyzed by gene chips and the expression of cytolytic molecules was evaluated by flow cytometry. RESULTS: Seventeen genes were up- (5/17) or down-regulated (12/17) on IL-12-treated NK cells compared with non-IL-12-treated NK cells (fold change≥10). IL-12-induced expression of TRAIL on NK cells mediated the cytotoxicity to Jurkat cells. The expression of TRAIL on subsets of CD56(+);CD16(+); and CD56(-);CD16(+); NK cells significantly increased after the stimulation with IL-12 and Jurkat cells expressed high level of TRAIL receptor 2 (TRAIL-R2). Importantly, the neutralizing mAbs against TRAIL (RIK-2) significantly inhibited the cytotoxicity of NK cells induced by IL-12. CONCLUSION: The expression of TRAIL on human NK cells induced by IL-12 was one of the major mechanisms of cytotoxicity to Jurkat cells.


Assuntos
Citotoxicidade Imunológica , Interleucina-12/farmacologia , Células Matadoras Naturais/imunologia , Ligante Indutor de Apoptose Relacionado a TNF/fisiologia , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células Jurkat , Células Matadoras Naturais/efeitos dos fármacos , Ligante Indutor de Apoptose Relacionado a TNF/genética
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