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1.
AIDS ; 34(2): 189-195, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31634199

RESUMO

OBJECTIVE: CD4CD19 conjugates play an important role in regulating antibody responses and follicular helper T cells development in animal models. However, little is known regarding the characteristic of CD4CD19 conjugates in humans with chronic HIV-1 infection. METHODS: The numbers of CD4CD19 conjugates were counted in 86 HIV-1-infected patients, including 66 typical progressors and 20 complete responders. CD4CD19 conjugates were sorted by flow cytometry and dissociated into CD4 T singlets and CD19 B singlets. The phenotypes of these cells were analyzed in both typical progressors and complete responders, and the levels of HIV-1 DNA in CD4CD19 conjugates were measured in 10 complete responders. RESULTS: We identified CD4CD19 cells as one type of T-B conjugate in peripheral blood, and the numbers and percentages of CD4CD19 conjugates decreased with HIV-1 disease progression. Phenotypic analysis showed CD4CD19 conjugates expressed higher levels of surface CD32. mRNA analysis found that the mRNA levels for CD32b were significantly higher compared with CD32a in CD4CD19 conjugates. Further analysis found that CD4CD19 conjugates expressed higher levels of CCR7 and CXCR5 than CD4 T and CD19 B singlets. A virus infectivity assay showed that CD4CD19 conjugates expressed higher levels of HIV-1-p24 than CD4CD19 cells. CD4CD19 conjugates in lymph node from typical progressors expressed higher levels of HIV-1-p24 than CD4CD19 conjugates in respective peripheral blood. Importantly, CD4CD19 conjugates from complete responders contained higher levels of HIV-1 DNA than total CD4 T cells. CONCLUSION: Our study indicates that CD4CD19 conjugates actively participate in HIV-1 infection and latency, and may serve as a new cellular target to eliminate latency.

2.
Front Immunol ; 10: 2465, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31681335

RESUMO

Background: Treg cells represent important viral reservoirs during chronic HIV infection. CD39 is closely involved in Treg-mediated immunosuppressive effects. However, CD39 expression on nTregs and mTregs and a relationship with HIV DNA levels during HIV infection is still unclear. In this study, we analyzed the distribution of HIV DNA in Treg subsets and the association between HIV DNA and CD39 expression on Treg subsets. Methods: Sixty-two HIV-infected patients with different HIV stages and 14 uninfected individuals were enrolled. nTregs (CD4+CD25+CD127lowCD45RO-) and mTregs (CD4+CD25+CD127lowCD45RO+) were isolated by magnetic selection and flow cytometric sorting. HIV DNA was quantified by real-time polymerase chain reaction (PCR). CD39 expression on nTregs and mTregs was analyzed by flow cytometry. Results: Higher levels of HIV DNA were detected in mTregs than those in nTregs during chronic HIV infection. The frequency of CD39+ nTregs and HIV DNA levels in nTregs were increased in patients with advanced HIV infection. Furthermore, HIV DNA levels in nTregs correlated positively with CD39+ nTreg frequency. CD39+ nTreg frequency was also increased in immune non-responders. Conclusions: mTregs and nTregs are both important reservoirs of virus during chronic HIV infection and HIV DNA levels increase in nTregs in patients with advanced HIV infection. We observed increased frequency of CD39+ nTregs and HIV DNA levels in nTregs in patients with advanced HIV infection. HIV DNA levels in nTregs correlated positively with CD39+ nTreg frequency.

3.
Microbiol Immunol ; 61(6): 239-246, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28500746

RESUMO

To date, there have been no reports characterizing HIV-1 in the semen of Chinese men who have sex with men (MSM) with early infection. In this study, genetic diversity and viral load of HIV-1 in the seminal compartments and blood of Chinese MSM with early HIV-1 infection were examined. Viral load and genetic diversity of HIV-1 in paired samples of semen and blood were analyzed in seven MSM with early HIV-1 infection. HIV-1 RNA and DNA were quantitated by real-time PCR assays. Through sequencing the C2-V5 region of the HIV-1 env gene, the HIV-1 genotype and genetic diversity based on V3 loop amino acid sequences were determined by using Geno2pheno and PSSM programs co-receptor usage. It was found that there was more HIV-1 RNA in seminal plasma than in blood plasma and total, and more 2-LTR circular and integrated HIV-1 DNA in seminal cells than in peripheral blood mononuclear cells from all seven patients with early HIV-infection. There was also greater HIV-1 genetic diversity in seminal than in blood compartments. HIV-1 in plasma displayed higher genetic diversity than in cells from the blood and semen. In addition, V3 loop central motifs, which present some key neutralizing antibody epitopes, varied between blood and semen. Thus, virological characteristics in semen may be more representative when evaluating risk of transmission in persons with early HIV infection.


Assuntos
Variação Genética , Infecções por HIV/sangue , Infecções por HIV/transmissão , Infecções por HIV/virologia , HIV-1/genética , Homossexualidade Masculina , Sêmen/virologia , Carga Viral , Adolescente , Adulto , Motivos de Aminoácidos , Sequência de Aminoácidos , Anticorpos Neutralizantes , Anticorpos Antivirais , Grupo com Ancestrais do Continente Asiático , Contagem de Linfócito CD4 , DNA Viral/análise , Vetores Genéticos , Genótipo , Proteína gp120 do Envelope de HIV , HIV-1/classificação , Humanos , Leucócitos Mononucleares/virologia , Masculino , Fragmentos de Peptídeos , Filogenia , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real , Doenças Sexualmente Transmissíveis/sangue , Doenças Sexualmente Transmissíveis/virologia , Sequências Repetidas Terminais/genética , Adulto Jovem , Produtos do Gene env do Vírus da Imunodeficiência Humana/classificação , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética
4.
Front Immunol ; 8: 1786, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29312314

RESUMO

Background: CXCR5+CD8+ T cells have been demonstrated to play an important role in the control of chronic viral replication; however, the relationship between CXCR5+CD8+ T cells, HIV disease progression, and programmed cell death 1 (PD-1) expression profile on CXCR5+CD8+ T cells during HIV infection remain poorly understood. Methods: We enrolled a total of 101 HIV patients, including 62 typical progressors, 26 complete responders (CRs), and 13 immune non-responders (INRs). Flow cytometric analysis, immunohistochemical staining, and relative function (i.e., cytokine secretion and PD-1 blockade) assays were performed to analyze the properties of CXCR5+CD8+ T cells. Results: HIV-specific CXCR5+CD8+ T cells in the peripheral blood and distribution of CXCR5+CD8+ T cells in the lymph node (LN) were negatively correlated with disease progression during chronic HIV infection. PD-1 was highly expressed on CXCR5+CD8+ T cells and positively associated with peripheral CD4+ T cell counts. Functionally, IFN-γ and TNF-α production of CXCR5+CD8+ T cells were reduced by PD-1 pathway blockade, but the production of IFN-γ and TNF-α from CXCR5-CD8+ T cells increased in response to TCR stimulation. Interestingly, PD-1 expression was constantly retained on CXCR5+CD8+ T cells while significantly decreased on CXCR5-CD8+ T cells after successful antiretroviral treatment in chronic HIV-infected patients. Conclusion: PD-1+CXCR5+CD8+ T cells are functional cytotoxic T cells during chronic HIV infection. PD-1+CXCR5+CD8+ T cells may represent a novel therapeutic strategy for the disease.

5.
World J Gastroenterol ; 20(18): 5519-26, 2014 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-24833882

RESUMO

AIM: To investigate the risk factors for liver-related mortality in chronic hepatitis C (CHC) patients. METHODS: All deceased CHC inpatient data were collected from the Beijing 302 Hospital clinical database, which includes more than 8250 CHC inpatients during the period from 2002 to 2012. The controls were matched to cases by age (± 2 years), sex and date of hospital admission (within the same year). Potential risk factors were included for the evaluation, and odds ratios (OR) and 95%CI were estimated using univariate (unadjusted) and multivariate (adjusted OR, AOR) conditional logistic regression. All statistical tests were two-sided. P values < 0.05 were considered statistically significant. RESULTS: Based on examinations of 144 CHC-related deceased cases and 576 controls, we found that antiviral therapy with interferon-α was associated with a 47% decrease in the risk of hepatic mortality (AOR = 0.53, 95%CI: 0.28-0.99, P = 0.048). Additionally, the initial diagnostic stage of the disease (AOR = 2.89, 95%CI: 1.83-4.56 and P < 0.001 for liver cirrhosis/AOR = 8.82, 95%CI: 3.99-19.53 and P < 0.001 for HCC compared with CHC), diabetes (AOR = 2.35, 95%CI: 1.40-3.95, P = 0.001), hypertension (AOR = 1.76, 95%CI: 1.09-2.82, P = 0.020), alcohol consumption (AOR = 1.73, 95%CI: 1.03-2.81, P = 0.037) and HBsAg positivity (AOR = 22.28, 95%CI: 5.58-89.07, P < 0.001) were associated with a significant increase in the risk of liver-related mortality in CHC patients. CONCLUSION: This study indicates that interferon-α treatment, the stage at the initial diagnosis of the disease and comorbidities are all independent risk factors for liver-related mortality in CHC patients.


Assuntos
Hepatite C Crônica/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Antivirais/uso terapêutico , Causas de Morte , Distribuição de Qui-Quadrado , China/epidemiologia , Comorbidade , Feminino , Hepatite C Crônica/diagnóstico , Hepatite C Crônica/tratamento farmacológico , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Razão de Chances , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Índice de Gravidade de Doença , Fatores de Tempo , Resultado do Tratamento
6.
Mol Med Rep ; 9(3): 831-6, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24430732

RESUMO

Citrobacter rodentium (C. rodentium) infection is a widely used murine model to mimic human enteric bacteria infection and inflammatory bowel disease (IBD). In this model, interleukin (IL)­17A plays critical roles in increasing chemokine and cytokine production in various tissues to recruit innate cells, including monocytes and neutrophils, to the local site of infection. However, the source of IL­17A remains unclear, as the majority of cell types produce IL­17A, including intestinal endothelium cells, innate immune cells and CD4+ T cells in disease development. In the current study, wild­type B6 mice were treated with C. rodentium and the CD4+ Th17 cell subset was observed as being specifically increased in Peyer's patches (PP), but not in mesenteric draining lymph nodes. Furthermore, the research suggested that the differentiation and activation of Th17 cells in PP were dependent on the inflammatory cytokine IL­6, as blocking IL­6 signaling with neutralizing antibodies decreased Th17 cells and resulted in the mice being more susceptible to C. rodentium infection. These results confirmed that the Th17 cell subset was specifically activated in PP and demonstrated that IL­6 is required in Th17 cell activation, which are important to the clinical treatment of IBD.


Assuntos
Infecções por Enterobacteriaceae/imunologia , Regulação da Expressão Gênica , Interleucina-6/metabolismo , Interleucinas/genética , Células Th17/metabolismo , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Neutralizantes/farmacologia , Diferenciação Celular , Citrobacter rodentium/fisiologia , Modelos Animais de Doenças , Infecções por Enterobacteriaceae/mortalidade , Infecções por Enterobacteriaceae/patologia , Feminino , Humanos , Imunoglobulina A/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Interleucina-17/metabolismo , Interleucina-6/imunologia , Interleucinas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Nódulos Linfáticos Agregados/citologia , Taxa de Sobrevida , Células Th17/citologia , Células Th17/efeitos dos fármacos , Perda de Peso
7.
Artigo em Chinês | MEDLINE | ID: mdl-21977585

RESUMO

OBJECTIVE: To construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization (SSH) technique. METHODS: Suppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3.1 (-) -PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. RESULTS: The subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones. CONCLUSION: The obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1T1.


Assuntos
Hepatite B/genética , Hibridização de Ácido Nucleico/métodos , Proteínas Virais/genética , Humanos , Ativação Transcricional/genética
8.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 26(6): 536-8, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-20487644

RESUMO

AIM: To investigate phenotypical and functional characteristics of the CD39(+);FoxP3(+);regulatory T (Treg) cells in humans. METHODS: We collected peripheral blood from 10 healthy volunteers and separated PBMC by using density gradient centrifugation, and then detected the expression of CTLA-4, HLA-DR, CD45RO and Ki-67 on CD39(+);FoxP3(+); and CD39(-);FoxP3(+);Treg cells by using flow cytometric analysis. [(3);H]-thymidine incorporation assay and ELISA were used to monitor the suppressive capacity of CD39(+); FoxP3(+); Treg cells on proliferation and IFN-gamma secretion of Treg-depleted PBMC. RESULTS: The expression of CTLA-4, HLA-DR, CD45RO and Ki-67 was significantly higher in CD39(+);FoxP3(+); Treg cells than those in CD39(-);FoxP3(+); Treg cells. The suppressive capacity of CD39(+);FoxP3(+); Treg cells on proliferation and IFN-gamma secretion of Treg-depleted PBMC was significantly higher than those of FoxP3(+);CD39(-);Treg cells. CONCLUSION: CD39(+);Fxop3(+);Treg subset may play an essential role in immune regulation of Treg, and CD39 can be used as a surface marker to identify the functional Treg cells.


Assuntos
Antígenos CD/metabolismo , Apirase/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Adulto , Antígeno CTLA-4 , Proliferação de Células , Células Cultivadas , Feminino , Antígenos HLA-DR/metabolismo , Humanos , Interferon gama/metabolismo , Antígeno Ki-67/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Masculino , Linfócitos T Reguladores/citologia
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