中华微生物学和免疫学杂志最新文献

{"title":"Safety of yellow fever vaccine strain (Tiantan strain) in China and WHO vaccine strain 17D-213","authors":"Ling Wang, Na Li, Enyue Fang, Y. Yin, Yuhua Li","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.003","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.003","url":null,"abstract":"Objective \u0000To study the intracerebral pathogenicity of the yellow fever vaccine strains of Tiantan strain used in China and WHO vaccine strain 17D-213 in mice. \u0000 \u0000 \u0000Methods \u0000Mice of different ages and strains were intracerebrally injected with same amount of Tiantan strain and 17D-213 strain. The death and survival of mice were observed and recorded. The LD50/ml and half survival time of the two vaccine strains were compared and analyzed. \u0000 \u0000 \u0000Results \u0000There was no difference in LD50/ml between the Tiantan strain and 17D-213 strain when used through intracerebral injection in one-day-old suckling mice, 7-9 g mice or 12-14 g mice. Moreover, no significant difference in survival trend was found in 7-9 g mice or 12-14 g mice injected with the two vaccine strains. However, the two strains had statistically different influences on the survival trend of one-day-old suckling mice. The half survival time of the Tiantan strain was 11 d, while that of the WHO vaccine strain 17D-213 was 6 d. Excepting in NIH mice, no significant differences in LD50/ml were detected between the same amount of two strains in BALB/c, KM, ICR or C57 mice. \u0000 \u0000 \u0000Conclusions \u0000The yellow fever Tiantan vaccine strain and WHO vaccine strain 17D-213 have no significant difference in the intracerebral pathogenicity in mice of different ages and strains with good safety. \u0000 \u0000 \u0000Key words: \u0000Yellow fever; Vaccine strain; Intracerebral pathogenicity; Safety","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"657-661"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45706735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gut microbiota composition and diversity in the elderly supported by institution-based care and home-based care","authors":"Huiping Qiu, Yan-li Ren, Hui Xu, S. Yao, M. Hong, Qin Yang, Zhi Chen","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.007","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.007","url":null,"abstract":"Objective \u0000To investigate the gut microbiota diversity between the elderly supported by institution-based care and home-based care. \u0000 \u0000 \u0000Methods \u0000Fresh stool samples were collected from 18 aged persons supported by institution-based care (G1 group), 20 aged persons with home-based care (G2 group) and 20 middle-aged and young adults (G3 group). The V3-V4 hypervariable region of 16S rDNA was amplified and sequenced by next generation sequencing technology. Operational taxonomic units (OTUs) were analyzed by QIIME analysis platform for species annotation, diversity analysis, and inter-group difference analysis. Statistical analysis was performed using RStudio software. \u0000 \u0000 \u0000Results \u0000The top 6 microbiological taxa in the three groups were Firmicute, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria and Verrucomicrobia. The abundance of the Firmicute in the G1 and G2 groups showed significant differences [(61.47±5.58)% vs (76.55±3.64)%, P 0.05). Results of the NMDS analysis showed that the intra-group differences were greater than inter-group differences in G1 and G2 groups. \u0000 \u0000 \u0000Conclusions \u0000No significant difference in the diversity of gut microbiota was detected between the elderly supported by institution-based care and home-based care, but there were differences in the composition of the predominant gut microbiota. \u0000 \u0000 \u0000Key words: \u0000Institution-based care; Home-based care; Gut microbiota; 16S rRNA","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"680-685"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69858344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epidemiological and genetic characteristics of EV71 strains circulating in Jiangsu Province from 2009 to 2018","authors":"H. Fan, Ji Hong, X. Huo, Jianli Hu, X. Qi","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.008","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.008","url":null,"abstract":"Objective \u0000To analyze the molecular epidemiology, genetic variations and evolution of enterovirus 71 (EV71) strains isolated in Jiangsu Province from 2009 to 2018. \u0000 \u0000 \u0000Methods \u0000Statistical methods were used to analyze the data about epidemiological characteristics and results of pathogen detection in cases with EV71 infection in Jiangsu Province from 2009 to 2018. The complete VP1 sequences of 80 EV71 strains were amplified and sequenced for analysis of diversity and phylogenesis. \u0000 \u0000 \u0000Results \u0000A total of 41 858 enterovirus-positive hand, foot and mouth disease cases were reported in Jiangsu Province from 2009 to 2018. EV71 was the predominant pathogen, accounting for 36.52%, and responsible for most of the severe cases. However, the percentage of EV71 among all pathogens gradually decreased over time. EV71 infection reached the peak in April to June and mainly occurred in children aged six months to five years old with higher incidence in males than in females. In terms of regional distribution, EV71 infections were characterized by area clustering in Jiangsu Province, mainly detected in Nanjing, Suzhou, Wuxi and Lianyungang. The 80 EV71 isolates belonged to C4a genotype. Nucleotide differences between them and three vaccine strains (H07, FY23 and FY7VP5)were 0.6%-5.5%, 0.8%-5.7% and 1.9%-6.9% and amino acid difference were 0-1.4%, 0.3%-2.0% and 0.3%-2.0%, respectively. Amino acid mutations in the epitopes of the 80 EV71 strains did not marked by years or regions. \u0000 \u0000 \u0000Conclusions \u0000EV71 strains showed obvious epidemiological characteristics in time, population and regional distribution in Jiangsu Province from 2009 to 2018.All of the 80 EV71 isolates belonged to C4a subgenotype. The nucleotide sequences between them and the vaccine strains varied greatly, but the homology of amino acids was relatively high, indicating the existence of some synonymous mutations and no risk of antigenic drift. This study would provide reference for EV71 vaccination in Jiangsu Province. \u0000 \u0000 \u0000Key words: \u0000Hand, foot and mouth disease; Enterovirus 71; Epidemiology; Genetic characteristics","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"686-692"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41671652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application potential of bacterial flagellin in treatment of autoimmune rheumatic diseases with Treg epitope peptides","authors":"Mengmeng Li, Xiao Liu, Jian Yin, Zhen Wang, Yaqun Liu, F. Qian, Huji Xu","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.012","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.012","url":null,"abstract":"Immunization with regulatory T cell (Treg) epitope peptides to activate and induce Tregs, by which to suppress pathological autoimmune responses and reconstitute a new homeostasis, is a promising therapeutic regimen for autoimmune rheumatic diseases. However, it is usually hard to induce potent peptide-specific immune responses in vivo with small molecular peptides. Bacterial flagellin is one of the agonists triggering innate immune responses. When used as carrier, it shows strong adjuvant activity to its conjugated antigens. In some particular situations, bacterial flagellin can also activate and induce Tregs. Thus if Treg epitope peptides are covalently conjugated to a bacterial flagellin, the conjugates should be able to effectively enhance the Treg-based immune responses via flagellin itself and the adjuvanticity of flagellin to Treg epitope peptides, and thereby enhance the immunotherapeutic effects on autoimmune rheumatic diseases. \u0000 \u0000 \u0000Key words: \u0000Bacterial flagellin; Treg epitope; Autoimmune rheumatic disease; Conjugate; Vaccine efficacy","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"710-714"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45445842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic characteristics of coxsackievirus A16 isolated in Shenzhen from 2016 to 2017","authors":"X. Yao, Weiqi Wang, Long Chen, Hong Yang, J. Meng, Hong Pan, Hai-long Zhang, Hong-Yu Zhang, Renli Zhang, Yaqing He","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.002","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.002","url":null,"abstract":"Objective \u0000To investigate the genetic characteristics of VP1 genes carried by coxsackievirus A16 strains isolated from cases of hand foot and mouth disease (HFMD) in Shenzhen during 2016 to 2017. \u0000 \u0000 \u0000Methods \u0000Fecal and anal swab specimens were collected from patients with mild HFMD in four sentinel hospitals and the Institute of Pathogen Biology, Shenzhen Center for Disease Control and Prevention, China during 2016 to 2017. All specimens were tested for CVA16 viral RNA using real-time RT-PCR. The VP1 genes of 51 randomly selected CVA16 strains were amplified by RT-PCR and then sequenced using TaKaRa Biomedical Technology (Dalian). Bioinformatics software, including Mega6.02, BioEdit and DNAStar, was used for comparison and analysis of the VP1 genes. \u0000 \u0000 \u0000Results \u0000CVA16 strains in Shenzhen during 2016 to 2017 mainly belonged to B1a and B1b subtypes as well as an emerging subtype B3. The epidemic of B1b subtype was found in both 2016 (28 strains) and 2017 (19 strains), while the B1a subtype (two strains) was only detected in 2017. Two B3 subtype strains were detected in 2017. The strains of B1b subtype were closely related to the strains isolated in Shanghai (JQ314149), Wenzhou (KP289416) and Beijing (KU254598), while the B1a subtype strains were closely related to the strains isolated in Kunming (JQ316639) and Tailand (GQ184139). The B3 subtype strain was an emerging CVA16 epidemic strain in mainland China. Further comparison of the CVA16 epidemic strains in Shenzhen area during 2016 to 2017 with the CVA16 strains causing severe neurological symptoms showed that two amino acid mutations (S14N and M23L) were found in VP1 protein. \u0000 \u0000 \u0000Conclusions \u0000The epidemic strains of CVA16 were B1b subtype in Shenzhen area in 2016. However, B1a, B1b and the emerging B3 subtype strains were prevalent in 2017. Compared with the CVA16 strains causing severe neurological symptoms, the CVA16 strains circulating in Shenzhen during 2016 to 2017 carried two amino acid mutations inVP1 protein. \u0000 \u0000 \u0000Key words: \u0000Coxsackievirus A16; Hand, foot and mouth disease; Phylogenetic analysis","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"652-656"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44234869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vitro antibacterial activity of triclosan in combination with different antibacterial agents against triclosan-resistant multidrug-resistant Acinetobacter baumannii","authors":"Ye Xu, Yizhi Zhang, Chunquan Xu, Siqin Zhang, Xiucai Zhang, Wenya Xu","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.006","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.006","url":null,"abstract":"Objective \u0000To investigate the in vitro antibacterial activity of triclosan combined with different antibacterial agents against triclosan-resistant multidrug-resistant Acinetobacter baumannii (A.baumannii). \u0000 \u0000 \u0000Methods \u0000A total of 626 A. baumannii strains were collected from the First Affiliated Hospital of Wenzhou Medical University from 2016 to 2017. The sensitivity of these A. baumannii strains to common antibiotics was detected by VITEK 2-compact automatical microbiological analyzer and the minimum inhibitory concentrations (MIC) of triclosan were detected by agar dilution method. Checkerboard method was used to detect the changes in MIC values of triclosan against 16 triclosan-resistant multidrug-resistant A. baumannii strains after it was used in combination with four external ointments, including gentamicin, erythromycin, chloramphenicol and kanamycin, and three common antibiotics of imipenem, meropenem and ciprofloxacin, respectively. Fractional inhibitory concentration index (FICI) was used to evaluate the joint bacteriostatic effects. \u0000 \u0000 \u0000Results \u0000Among the 626 A. baumannii strains, 17 were resistant to triclosan with a drug resistance rate of 2.7% (17/626). These triclosan-resistant strains had high MIC values for ciprofloxacin, imipenem, ceftazidime and other commonly used clinical antibiotic and most of them were multidrug-resistant. After triclosan was used in combination with seven different antibacterial drugs, the MIC values of all drugs decreased to various degrees compared with those when they were used alone. Triclosan in combination with gentamicin, chloramphenicol and ciprofloxacin showed synergistic effects on 62.5%, 56.25% and 62.5% of the 16 strains and additive effects on 37.5%, 43.75% and 37.5%, respectively. When it was used in combination with erythromycin, kanamycin, imipenem and meropenem, synergistic effects on 37.5%, 25%, 12.5% and 12.5%, additive effects on 37.5%, 56.25%, 62.5% and 62.5%, and indifferent effects on 25%, 18.75%, 25% and 25% of the strains were detected, respectively. No antagonistic effect was found between triclosan and any of the above antibiotics. \u0000 \u0000 \u0000Conclusions \u0000Triclosan combined with gentamicin, chloramphenicol and ciprofloxacin had better in vitro antibacterial effects against the triclosan-resistant multidrug-resistant A. baumannii strains in this study with synergistic and additive effects. Some indifferent effects were found between triclosan and kanamycin, erythromycin, imipenem and meropenem, but no antagonistic effects were detected. \u0000 \u0000 \u0000Key words: \u0000Triclosan; Acinetobacter baumannii; Multidrug resistance; In vitro antibacterial activity; Drug combination","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"674-679"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48895643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Roles of innate lymphoid cells in tumor immunity and their clinical significance","authors":"C. Chai, Xi Ma, Bang Xin, Yang-an Wen","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.013","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.013","url":null,"abstract":"Innate lymphoid cells (ILCs) are a recently characterized family of immune cells that have critical roles in innate immunity, immune regulation, maintenance of tissue homeostasis, and tissue repair and remodeling. Besides the conventional innate lymphocytes including NK cells and lymphoid tissue-inducer cells, the ILC family can be categorized into three groups, ILC1s, ILC2s and ILC3s. These non-cytotoxic ILC subsets have been identified to confer a diverse array of functions in oncogenesis and metastasis, immune surveillance, and antitumor immunity. In this review, we summarized the emerging findings in recent years regarding the roles of ILCs in immuno-oncology, and highlighted their potentials in immunotherapeutic approaches to tumors. \u0000 \u0000 \u0000Key words: \u0000Innate lymphoid cell; Immuno-oncology; Immunomodulation","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"715-719"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49408595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epidemiological characteristics of norovirus outbreaks in Shenzhen during 2005 to 2017","authors":"Zhen Zhang, Yuan Li, Hailong Zhang, Yan Lu, S. Mei, Jia Du, Xu Xie","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.009","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.009","url":null,"abstract":"Objective \u0000To investigate the epidemiological characteristics of norovirus outbreaks in Shenzhen during 2005 to 2017 in order to provide reference for disease control and prevention. \u0000 \u0000 \u0000Methods \u0000Monitoring data of norovirus outbreaks in Shenzhen from January 1, 2005 to December 31, 2017 were collected from Shenzhen Communicable Disease Information System and China Information System for Disease Control and Prevention. Descriptive epidemiological methods were used for data analysis. \u0000 \u0000 \u0000Results \u0000From January 2005 to December 2017, 346 norovirus outbreaks (five or more cases in one community within one week) were reported in Shenzhen, of which 6.36% (22/346) were public health emergency events. Fewer outbreaks were reported during 2006 to 2013 and they were mainly caused by GⅡ.4 genotype, but the number increased sharply since 2014 with 57.80% (200/346) occurred in 2016—2017 and the epidemic genotype changed from GⅡ.4 to GⅡ.17 and GⅡ.2. The outbreaks peaked during November to March (76.88%, 266/346). There were 63.87% (221/346) reported in urban areas, 67.05% (232/346) in nurseries and 23.70% (82/346) in primary/middle schools. Among the 22 public health emergency events, 40.91% (10/22) were caused by person-to-person contacts, 40.91% (10/22) by foodborne transmission and 13.64% (3/22) by waterborne transmission. Moreover, 75.80% (238/314) of the outbreaks in nurseries and primary/middle schools were confined to one classroom and most were due to contact transmission. \u0000 \u0000 \u0000Conclusions \u0000Norovirus outbreaks increased obviously since 2014, which might be related to the changes of the predominant genotype from GⅡ.4 to GⅡ.17 and GⅡ.2. It is necessary to strengthen a comprehensive prevention and control in key units such as nurseries and primary/middle schools in winter and spring. \u0000 \u0000 \u0000Key words: \u0000Norovirus; Outbreak; Public health emergency; Genotype","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"693-697"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42366351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of interferon inducible protein 16 (IFI16) on viral protein and pro-inflammatory cytokine expression in human T-cell leukemia virus type 1 (HTLV-1)-positive T cells","authors":"Yuhan Cui, Yuhe Guan, Yue Liu, Ge Zhang, Fan Chen, Mengmeng Chen, Jing-Jing Sun, X. Ren, Bo Yang, Jie Wang","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.005","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.005","url":null,"abstract":"Objective \u0000To investigate the effects of interferon inducible protein 16 (IFI16), a cytosolic DNA sensor, on the expression of human T-cell leukemia virus type 1 (HTLV-1) proteins and pro-inflammatory cytokines in adult HTLV-1-positive T cells. \u0000 \u0000 \u0000Methods \u0000IFI16 expression in different HTLV-1-positive T cell lines was detected by immunoblot assay. Specific siRNA targeting the IFI16 gene was constructed and the gene silencing efficiency was detected by immunoblot assay. Expression of HTLV-1 Tax protein at mRNA and protein levels was respectively detected by real-time PCR and immunoblot assay after knocking down the expression of IFI16 in HTLV-1-positive T cells with siRNA. Expression of interferon (IFN)-α, IFN-γ, tumor necrosis factor (TNF)-α, Tax and Env were detected by real-time PCR. \u0000 \u0000 \u0000Results \u0000Compared with the HTLV-1-negative T cell line Jurkat, IFI16 expression was enhanced in the HTLV-1-positive T cell lines MT2, MT4 and C8166. Tax expression was increased, while that of IFN-α, IFN-γ and TNF-α was decreased in MT2 and MT4 cells after silencing the expression of IFI16 with siRNA. \u0000 \u0000 \u0000Conclusions \u0000IFI16 expression was increased in HTLV-1-positive MT2 and MT4 cells. Meanwhile, IFI16 promoted the production of interferon and pro-inflammatory cytokines and inhibited the expression of HTLV-1 proteins. \u0000 \u0000 \u0000Key words: \u0000Interferon inducible protein 16 (IFI16); Human T-cell leukemia lymphoma virus type 1 (HTLV-1); Interferon; T lymphocyte; TNF-α","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"668-673"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43131532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recombinase-mediated in vitro rapid construction of replication-competent human adenovirus type 14 vector encoding EGFP","authors":"Yong Chen, Xingui Tian","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.09.001","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.09.001","url":null,"abstract":"Objective \u0000To rapidly and efficiently construct a replication-competent human recombinant adenovirus type 14 vector expressing enhanced green fluorescence protein (rAd14-EGFP) using in vitro homologous recombination. \u0000 \u0000 \u0000Methods \u0000The skeleton plasmid pBRAd14 was constructed using homologous recombination in Escherichia coli (E.coli) BJ5183 competent cells. The plamid was linearized and transfected into AD293 cells to rescue Ad14. Exnase, a recombinase, was used to construct the shuttle plasmid pSK14-EGFP in vitro using homologous recombination among four fragments. The overlapping sequence was 15-20 bp. Three exogenous fragments generated with PCR including Ad14 E3L fragment, EGFP gene and Ad14 E3R fragment were cloned into the plasmid pBluescript Ⅱ SK(-) simultaneously. Recombinant plasmid pBRAd14-EGFP was constructed by in vitro homologous recombination between 27 kb fragment of plasmid pBRAd14 obtained through double digestion and Ad14 E3L-EGFP-Ad14 E3R fragment amplified by PCR using the shuttle plasmid pSK14-EGFP as template. The plasmid pBRAd14-EGFP was linearized and transfected into cells to obtain the viral vector rAd14-EGFP, which was then used to immunize mice to detect the induced immune responses. \u0000 \u0000 \u0000Results \u0000A replication-competent E3-deleted adenovirus vector rAd14-EGFP expressing EGFP was successfully constructed. Intracellular proliferation properties and immunogenicity of the vector were no significantly differences compared with those of Ad14. \u0000 \u0000 \u0000Conclusions \u0000In vitro homologous recombination using the commercial recombinase Exnase can be a rapid, efficient and accurate method to construct adenoviral vector. \u0000 \u0000 \u0000Key words: \u0000Human adenovirus type 14; Recombinase Exnase; In vitro homologous recombination; Adenovirus vector","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"645-651"},"PeriodicalIF":0.0,"publicationDate":"2019-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46804636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}