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AIM: To investigate the possibility of simultaneously ex vivo generating cytomegalovirus (CMV) pp65 and Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTL) from human umbilical cord blood (CB). METHODS: Mononuclear cell derived from CB (CBMC) was used to construct EBV-transformed B-lymphoblastoid cell lines (BLCL). Then BLCL were transduced with a recombinant retrovirus encoding pp65, the immunodominant CMV polypeptide. CBMC from the same CB donor were stimulated with pp65-expressing BLCL (BLCLpp65) weekly for 5~6 weeks. Chromium release assays (CRA) were performed to detect the specific cytotoxicity of the CTL against EBV and CMV. RESULTS: Western blot analysis and immunocytochemistry confirmed that BLCLpp65 could simultaneously express CMVpp65 and EBV antigen. CRA results showed that the generated CTL possessed specific cytotoxic against EBV and CMV, and the cytotoxicity was mediated by CD8+ CTL. CONCLUSION: BLCLpp65 can be used as antigen-presenting cells to stimulate expansion of EBV and CMV specific CTL simultaneously from the predominantly native T cell population in CB. 相似文献
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WU Qi CHEN Xiao-qian WANG Tian-yang CHEN Jing-feng PAN Xiao-dong WU Cun-zao XIA Peng CHEN Bi-cheng YANG Yi-rong 《园艺学报》2014,30(2):302-307
AIM:To investigate the association of killer cell immunoglobulin-like receptor (KIR) genotype and cytomegalovirus (CMV)-BK virus (BKV) reactivation/infection in the first year after renal transplantation. METHODS:KIR genotypes were analyzed by PCR with sequence-specific primers (PCR-SSP) in 48 renal transplant recipients and KIR genotypes were grouped into AA if they contained only the canonical group A haplotype genes. The genotype containing additional KIR genes was referred to BX, as it contained at least 1 group B haplotype. Real-time quantitative PCR for CMV-BKV DNA was performed to test the viral load. The association of KIR genotype and CMV-BKV infection and the effect of KIR genotype on renal function were analyzed. RESULTS:No obvious difference in the concentration of immunosuppressant and the occurrence of CMV infection/reactivation in the first year after renal transportation was observed between KIR-AA and KIR-BX genotype groups. However, there was a significantly negative correlation between KIR-BX genotype and the cumulative incidence of BKV infection/reactivation. The average concentration of serum creatinine over 1~12 months after operation in KIR-AA genotype group was lower than that in KIR-BX genotype group. The levels of blood urea nitrogen and uric acid showed no obvious difference between the 2 groups. CONCLUSION:There is a significantly negative correlation between KIR-BX genotype and the cumulative incidence of BKV infection/reactivation, but not CMV infection/reactivation in the first year after renal transplantation. 相似文献
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[目的]建立人α干扰素（interferon-α,IFN-α）转基因小鼠,为研究IFN-α在抗病毒中的作用提供模型动物。[方法]将人IFN-α基因插入CMV启动子下游,构建转基因表达载体,通过原核显微注射法建立IFN-α转基因小鼠。[结果]通过特异性引物PCR法和South-ern杂交法检测出4只阳性转基因小鼠。分离4个转基因鼠系F1代PCR阳性小鼠血液中的淋巴细胞进行RT-PCR检测,其中3个鼠系为阳性。收集阳性小鼠血清,用ELISA方法和微量板染色测定法检测出3个转基因鼠系均有IFN-α表达。[结论]成功建立了CMV启动子启动的表达人IFN-α基因转基因小鼠,为研究干扰素抗病毒机制及对其他动物进行抗病毒感染基因工程育种研究奠定了基础。 相似文献
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AIM: To investigate the effects of simvastatin on the expression of Toll-like receptor 2 (TLR-2), interferon-γ (IFN-γ) and monocyte chemoattractant protein-1 (MCP-1) in lung tissues of mice with mouse cytomegalovirus (MCMV) pneumonia and to explore the possible mechanism. METHODS: Male BALB/c mice (6~8 weeks old, n=40) were randomly divided into 5 groups: normal control (NC) group, MCMV infection group, simvastatin group 1 (SMV1 group), simvastatin group 2 (SMV2 group), and simvastatin group 3 (SMV3 group). The mice in SMV1, SMV2 and SMV3 groups were gavaged with simvastatin (50 mg·kg-1·d-1 for 7 d) 7 d before, on the same day of and 3 d after intraperitoneal injection of MCMV, while the mice in normal control group and MCMV infection group were gavaged with the same volume of normal saline. HE staining was used to observe the pathological changes of lung tissues in mice. Total tissue protein was extracted from the lung homogenates to detect the expression of TLR-2 by Western blot and immunohistochemical staining. Real-time PCR was used to analyse the content of MCMV DNA. The levels of IFN-γ and MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with NC group, the pathological changes of the lung tissues of the mice in MCMV group showed alveolar interstitial edema, alveolar wall widening and a large number of inflammatory cells. The expression of TLR-2 in the lung tissues of the mice in model group was increased significantly. The content of MCMV DNA was increased, and the expression of IFN-γ and MCP-1 was also increased significantly. Compared with the mice in MCMV group, the pathological changes of the lung tissues of simvastatin groups showed that the inflammatory cells were decreased. The expression of TLR-2 was down-regulated. The content of MCMV DNA was decreased, and the levels of IFN-γ and MCP-1 were also decreased significantly. At the same time, the expression of TLR-2 and the content of MCMV DNA in SMV1 group were less than those in SMV2 and SMV3 groups (P<0.05), and no statistically significant difference between SMV2 and SMV3 groups was observed. CONCLUSION: Simvastatin down-regulates the TLR-2 signaling pathway, and reduces the expression of TLR-2 and replication of MCMV DNA, thus attenuating the pathological damage of the lung tissue. Early intervention with simvastatin plays an important role in preventing the infection of MCMV and reducing the inflammation. 相似文献
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G.V Quinnan F.A Ennis 《Comparative immunology, microbiology and infectious diseases》1980,3(3):283-290
Cell-mediated immunity is of critical significance in the host response to cytomegalovirus infection. This fact is evidenced by numerous reports of disease in immunosuppressed hosts and by other clinical and experimental observations. Methodologies are now available for elucidation of the specific role of the various facets of cell-mediated immunity in normal and susceptible individuals during different stages of infection. Most progress has been made recently using assays of lymphocyte transformation and cell-mediated immune lysis, but other areas also deserve specific attention. A comprehensive picture of the function of cellular immunity should greatly advance understanding of disease pathogenesis as well as direct the development of future models for immunotherapy. 相似文献
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AIM: To investigate the effect of cytomegalovirus (CMV) infection on the immunological functions of dendritic cells (DC) derived from monocytes. METHODS: RT-PCR assay was used to detect the mRNA expression of CMV immediate early antigen (IEA) and glyceraldehyde phosphate dehydrogenase (GAPDH) genes in immature and mature dendritic cell (cmv-imDC, cmv-mDC) infected by 50-folds median tissue culture infective dose (TCID50) of CMV. The expression of early antigen (EA) in cmv-imDC and cmv-mDC was analyzed by indirect immunofluorescence assay. CMV late antigen pp65 was determined by flow cytometry. The allogeneic stimulating capacity of cmv-DC was assayed by mixed lymphocyte reaction (MLR) with BrdU incorporation. RESULTS: The expression of IE mRNA in cmv-mDC was lower than that in cmv-imDCs at 12 h after infection (0.102±0.020 and 0.862±0.124, respectively, P<0.05). EA, primarily localized in nucleus, was found in cmv-imDC (62.32±14.20)% and cmv-mDC (10.78±3.04)% at 24 h (P<0.01). pp65 positive cells in cmv-imDC and cmv-mDC at 72 h were 4.86% and 0.82%, respectively. Compared with untreated mDC, cmv-imDC showed depressed antigen presentation even after stimulated with maturation signal factor LPS (both P<0.05), while cmv-mDC had weaker stimulating capacity only when DC/T cell ratio was 1∶1 (P<0.05). CONCLUSION: CMV efficiently infectes and replicates in imDC. CMV suppresses the antigen presenting capacity of cmv-DC. 相似文献
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