Pub Date : 2020-07-31DOI: 10.3760/CMA.J.CN112309-20200320-00130
Wei-jin Huang, Youchun Wang
Antibody dependent enhancement (ADE) is a common phenomenon in virology. It is involved in the mechanisms of infections caused by Dengue virus (DV), severe acute respiratory syndrome coronavirus (SARS-CoV), influenza virus, HIV and other viruses and affects the research and development of vaccines against them. Because the pre-existing specific antibodies or antibodies at sub-neutralizing titer can enhance the infectivity of viruses, leading to disease aggravation, vaccination may promote infection instead of preventing it. This article focused on the impact of ADE on the research and development of vaccines and the assessment of ADE.
{"title":"Influence of antibody dependent enhancement (ADE) on vaccine development/ 抗体依赖性增强作用对疫苗研发的影响","authors":"Wei-jin Huang, Youchun Wang","doi":"10.3760/CMA.J.CN112309-20200320-00130","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200320-00130","url":null,"abstract":"Antibody dependent enhancement (ADE) is a common phenomenon in virology. It is involved in the mechanisms of infections caused by Dengue virus (DV), severe acute respiratory syndrome coronavirus (SARS-CoV), influenza virus, HIV and other viruses and affects the research and development of vaccines against them. Because the pre-existing specific antibodies or antibodies at sub-neutralizing titer can enhance the infectivity of viruses, leading to disease aggravation, vaccination may promote infection instead of preventing it. This article focused on the impact of ADE on the research and development of vaccines and the assessment of ADE.","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"7 1","pages":"558-562"},"PeriodicalIF":0.0,"publicationDate":"2020-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44209459","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}
Pub Date : 2020-05-31DOI: 10.3760/CMA.J.CN112309-20200303-00096
Xingui Tian, R. Zhou
Safe and effective vaccines are urgently needed to prevent and control the outbreak of COVID-19. SARS-CoV-2 belongs to the genus Betacoronavirus like severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV). Here we summarized the current progress and problems in the development of vaccines against SARS-CoV and MERS-CoV in order to provide reference for COVID-19 vaccine development. Key words: SARS-CoV-2; COVID-19; SARS; MERS; Vaccine
{"title":"Progress in SARS and MERS vaccines: lessons for the development of COVID-19 vaccine","authors":"Xingui Tian, R. Zhou","doi":"10.3760/CMA.J.CN112309-20200303-00096","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200303-00096","url":null,"abstract":"Safe and effective vaccines are urgently needed to prevent and control the outbreak of COVID-19. SARS-CoV-2 belongs to the genus Betacoronavirus like severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV). Here we summarized the current progress and problems in the development of vaccines against SARS-CoV and MERS-CoV in order to provide reference for COVID-19 vaccine development. \u0000 \u0000Key words: \u0000SARS-CoV-2; COVID-19; SARS; MERS; Vaccine","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49371168","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}
Pub Date : 2020-04-15DOI: 10.3760/CMA.J.CN112309-20200303-00097
M. Tan, Jing-Rong Qu, Ying Huang, Yan Li, Jiewen Mai, Chenghui Ma, Yaling Shi
Objective To evaluate the performance of three antibody kits for novel coronavirus (SARS-CoV-2) and to investigate the feasibility and advantages of them in clinical application. Methods A total of 104 patients who were admitted to Guangzhou Eighth People's Hospital with COVID-19 from January to February 2020 were selected as research group. Fifty-one healthy subjects were selected during the same period as negative control group. Serum antibodies (IgM/IgG) against SARS-CoV-2 were detected using two kinds of colloidal gold kits (A and B kits) and one chemiluminescence kit (C kit). The positive rates of SARS-CoV-2 nucleic acid in different samples from patients with COVID-19 were retrospectively analyzed. Results The clinical sensitivity of A kit to detect SARS-CoV-2-specific IgM and IgG was 77.88% (81/104) and 65.38% (68/104), respectively, and the clinical specificity was 70.59% (36/51) and 100.00% (51/51). However, the false positive rate in IgM detection was as high as 29.41% (15/51). The sensitivity of B kit to test total antibodies to SARS-CoV-2 was 63.46% (66/104), and the clinical specificity was 94.12% (48/51). The clinical sensitivity of C kit to detect SARS-CoV-2-specific IgM and IgG were respectively 31.73% (33/104) and 64.42% (67/104), and the clinical specificity were both 98.04% (50/51). There was a moderate correlation between the detection results of two colloidal gold kits and the chemiluminescence kit with the Kappa values of 0.462 and 0.587 (Z=6.157, P<0.01; Z=7.345, P<0.01). C kit had the highest positive detection rate for IgG, and would be more reliable to be used for IgG detection in COVID-19 patients 14 d after onset. The total positive detection rate of nucleic acid in all types of samples was 63.46% (66/104). The highest positive detection rate was in throat swabs or sputum samples, followed by those in blood samples and anal swabs. No viral nucleic acid was detected in urine samples for the time being. Conclusions SARS-CoV-2-specific antibodies could be detected in the early or late stage of COVID-19. The method of antibody detection has the advantages of shorter detection time, simple operation and high biological safety, indicating that it could be used as a supplementary or auxiliary detection for the diagnosis of suspected COVID-19 cases with negative nucleic acid test results. The chemiluminescence kit has good sensitivity and specificity, and is well recommended for clinical laboratories. Key words: SARS-CoV-2; Chemiluminescence immunoassay; Colloidal gold immunoaasay; Nucleic acid detection; Performance evaluation; Clinical application
{"title":"Performance evaluation and clinical application of three antibody test kits for novel coronavirus (SARS-CoV-2)","authors":"M. Tan, Jing-Rong Qu, Ying Huang, Yan Li, Jiewen Mai, Chenghui Ma, Yaling Shi","doi":"10.3760/CMA.J.CN112309-20200303-00097","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200303-00097","url":null,"abstract":"Objective \u0000To evaluate the performance of three antibody kits for novel coronavirus (SARS-CoV-2) and to investigate the feasibility and advantages of them in clinical application. \u0000 \u0000 \u0000Methods \u0000A total of 104 patients who were admitted to Guangzhou Eighth People's Hospital with COVID-19 from January to February 2020 were selected as research group. Fifty-one healthy subjects were selected during the same period as negative control group. Serum antibodies (IgM/IgG) against SARS-CoV-2 were detected using two kinds of colloidal gold kits (A and B kits) and one chemiluminescence kit (C kit). The positive rates of SARS-CoV-2 nucleic acid in different samples from patients with COVID-19 were retrospectively analyzed. \u0000 \u0000 \u0000Results \u0000The clinical sensitivity of A kit to detect SARS-CoV-2-specific IgM and IgG was 77.88% (81/104) and 65.38% (68/104), respectively, and the clinical specificity was 70.59% (36/51) and 100.00% (51/51). However, the false positive rate in IgM detection was as high as 29.41% (15/51). The sensitivity of B kit to test total antibodies to SARS-CoV-2 was 63.46% (66/104), and the clinical specificity was 94.12% (48/51). The clinical sensitivity of C kit to detect SARS-CoV-2-specific IgM and IgG were respectively 31.73% (33/104) and 64.42% (67/104), and the clinical specificity were both 98.04% (50/51). There was a moderate correlation between the detection results of two colloidal gold kits and the chemiluminescence kit with the Kappa values of 0.462 and 0.587 (Z=6.157, P<0.01; Z=7.345, P<0.01). C kit had the highest positive detection rate for IgG, and would be more reliable to be used for IgG detection in COVID-19 patients 14 d after onset. The total positive detection rate of nucleic acid in all types of samples was 63.46% (66/104). The highest positive detection rate was in throat swabs or sputum samples, followed by those in blood samples and anal swabs. No viral nucleic acid was detected in urine samples for the time being. \u0000 \u0000 \u0000Conclusions \u0000SARS-CoV-2-specific antibodies could be detected in the early or late stage of COVID-19. The method of antibody detection has the advantages of shorter detection time, simple operation and high biological safety, indicating that it could be used as a supplementary or auxiliary detection for the diagnosis of suspected COVID-19 cases with negative nucleic acid test results. The chemiluminescence kit has good sensitivity and specificity, and is well recommended for clinical laboratories. \u0000 \u0000 \u0000Key words: \u0000SARS-CoV-2; Chemiluminescence immunoassay; Colloidal gold immunoaasay; Nucleic acid detection; Performance evaluation; Clinical application","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43173603","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}
Pub Date : 2020-04-15DOI: 10.3760/CMA.J.CN112309-20200318-00124
Hong-xia Ma, Jing-jing Pan, Yi Li, Y. Ye, Baifan Zhang, Hai-feng Wang, Jia-yong Zhao, A. You, Jin Xu, Xueyong Huang
Objective To investigate the performance of real-time RT-PCR for the detection of SARS-CoV-2 nucleic acid in clinical diagnosis of COVID-2019. Methods Laboratory test data and basic case information of Henan COVID-19 cases were collected from the China's Infectious Disease Information System as of March 5, 2020. All information was entered by local hospitals and Center for Disease Control and Prevention (CDC). Local hospitals or country CDC were responsible for sampling and municipal CDC was responsible for nucleic acid testing. Results A total of 6 714 specimens were detected and the positive rate of SARS-CoV-2 nucleic acid was 23.82%. The specimens were collected from 1 200 confirmed cases, 2 178 suspected cases and 77 asymptomatic cases. The nucleic acid diagnosis rate of COVID-19 was 36.96% (1 277/3 455). In all cases, the positive rates of SARS-CoV-2 nucleic acid in nasal swabs, sputum samples and throat swabs were 19.38%, 28.59% and 23.53%, respectively (χ2=15.896, P<0.01). The positive rate of SARS-CoV-2 nucleic acid in confirmed COVID-19 cases was 63.10%. The positive rates in nasal swabs, sputum samples and throat swabs were 50.80%, 58.71% and 65.21 (χ2=18.612, P<0.01). The positive rates of SARS-CoV-2 nucleic acid were 43.51%, 23.98%, 22.82%, 12.17%, 14.46% and 13.21% in samples collected on the day of symptom onset and one week, two weeks, three weeks, four weeks, five weeks and above five weeks after the onset, respectively. The positive rates in confirmed cases were respectively 89.03%, 86.57%, 52.30%, 17.53%, 17.69% and 24.14% at those time points. Conclusions Real-time RT-PCR is the most effective method for early pathogenic diagnosis of COVID-19. The highest detection rate of nucleic acid is achieved within one week after the onset of COVID-19, and the latest time for nucleic acid detection is 38 d after the onset. Key words: SARS-CoV-2; COVID-19; Real-time RT-PCR
{"title":"Real-time RT-PCR for the detection of SARS-CoV-2 nucleic acid","authors":"Hong-xia Ma, Jing-jing Pan, Yi Li, Y. Ye, Baifan Zhang, Hai-feng Wang, Jia-yong Zhao, A. You, Jin Xu, Xueyong Huang","doi":"10.3760/CMA.J.CN112309-20200318-00124","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200318-00124","url":null,"abstract":"Objective \u0000To investigate the performance of real-time RT-PCR for the detection of SARS-CoV-2 nucleic acid in clinical diagnosis of COVID-2019. \u0000 \u0000 \u0000Methods \u0000Laboratory test data and basic case information of Henan COVID-19 cases were collected from the China's Infectious Disease Information System as of March 5, 2020. All information was entered by local hospitals and Center for Disease Control and Prevention (CDC). Local hospitals or country CDC were responsible for sampling and municipal CDC was responsible for nucleic acid testing. \u0000 \u0000 \u0000Results \u0000A total of 6 714 specimens were detected and the positive rate of SARS-CoV-2 nucleic acid was 23.82%. The specimens were collected from 1 200 confirmed cases, 2 178 suspected cases and 77 asymptomatic cases. The nucleic acid diagnosis rate of COVID-19 was 36.96% (1 277/3 455). In all cases, the positive rates of SARS-CoV-2 nucleic acid in nasal swabs, sputum samples and throat swabs were 19.38%, 28.59% and 23.53%, respectively (χ2=15.896, P<0.01). The positive rate of SARS-CoV-2 nucleic acid in confirmed COVID-19 cases was 63.10%. The positive rates in nasal swabs, sputum samples and throat swabs were 50.80%, 58.71% and 65.21 (χ2=18.612, P<0.01). The positive rates of SARS-CoV-2 nucleic acid were 43.51%, 23.98%, 22.82%, 12.17%, 14.46% and 13.21% in samples collected on the day of symptom onset and one week, two weeks, three weeks, four weeks, five weeks and above five weeks after the onset, respectively. The positive rates in confirmed cases were respectively 89.03%, 86.57%, 52.30%, 17.53%, 17.69% and 24.14% at those time points. \u0000 \u0000 \u0000Conclusions \u0000Real-time RT-PCR is the most effective method for early pathogenic diagnosis of COVID-19. The highest detection rate of nucleic acid is achieved within one week after the onset of COVID-19, and the latest time for nucleic acid detection is 38 d after the onset. \u0000 \u0000 \u0000Key words: \u0000SARS-CoV-2; COVID-19; Real-time RT-PCR","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47693187","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}
Pub Date : 2020-04-15DOI: 10.3760/CMA.J.CN112309-20200312-00115
Kai Chen, Suwen Jiang, A. Hu
In December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the outbreak of COVID-19 in Wuhan. It is a kind of enveloped positive-strand RNA viruses, belonging to the subgenus Sarbecovirus of the genus Betacoronavirus. It is similar to other viruses of the subgenus Sarbecovirus in genomic structure and closely related to the bat coronavirus (RaTG13), indicating that its natural host is likely to be bat, but the intermediate host is still controversial. SARS-CoV-2 infects host cells with angiotensin-converting enzyme 2 (ACE2) as a receptor. At present, SARS-CoV-2 has undergone some mutations. This article reviewed the current research situation and progress in the etiology of COVID-19. Key words: COVID-19; Severe acute respiratory syndrome coronavirus 2; Etiology
{"title":"Advances in the etiology of COVID-19","authors":"Kai Chen, Suwen Jiang, A. Hu","doi":"10.3760/CMA.J.CN112309-20200312-00115","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200312-00115","url":null,"abstract":"In December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the outbreak of COVID-19 in Wuhan. It is a kind of enveloped positive-strand RNA viruses, belonging to the subgenus Sarbecovirus of the genus Betacoronavirus. It is similar to other viruses of the subgenus Sarbecovirus in genomic structure and closely related to the bat coronavirus (RaTG13), indicating that its natural host is likely to be bat, but the intermediate host is still controversial. SARS-CoV-2 infects host cells with angiotensin-converting enzyme 2 (ACE2) as a receptor. At present, SARS-CoV-2 has undergone some mutations. This article reviewed the current research situation and progress in the etiology of COVID-19. \u0000 \u0000Key words: \u0000COVID-19; Severe acute respiratory syndrome coronavirus 2; Etiology","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42543417","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}
Objective To investigate the therapeutic effects of leflunomide on salivary gland secretion hypofunction in NOD mice with Sjogren′s syndrome. Methods NOD mice were randomly divided into four groups: prevention group, prevention control group, treatment group and treatment control group. Salivary flow rate was measured after pilocarpine stimulation. Changes in the average number and area of infiltrating lesions were compared after hematoxylin and eosin (HE) staining. The percentages of CD3+ T, CD4+ T, CD8+ T, CD44+ CD62L-CD4+ T, CD19+ B and CD138+ B cells in submandibular gland and spleen were detected by flow cytometry. The levels of TNF-α, IL-17A and IL-6 in serum were detected by CBA method. Results The salivary flow rate (t=-5.81, P<0.001; t=-3.61, P<0.05), the number of infiltrating lesions(t=3.95, P<0.01; t=4.94, P<0.001)and the average area of infiltrating lesions(t=3.18, P<0.05; t=2.35, P<0.05)were significantly ameliorated in the prevention and treatment groups. Moreover, CD3+ CD4+ T cells(t=2.39, P<0.05; t=3.82, P<0.01)and CD44+ CD62L-CD4+ T cells(t=3.53, P<0.05; t=3.36, P<0.05)in the submandibular gland were significantly decreased. CD3+ T(t=6.08, P<0.001; t=2.76, P<0.05), CD3+ CD4+ T (t=3.73, P<0.05; t=2.39, P<0.05), CD19+ B (t=5.88, P<0.001; t=4.23, P<0.01) and CD138+ B cells (t=4.30, P<0.001; t=4.46, P<0.01) in the spleen were also significantly reduced. Serum IL-17A (t=4.15, P<0.01; t=3.36, P<0.01) in the two groups and TNF-α (t=4.56, P<0.001) in the prevention group were down-regulated, but no significant difference was observed in IL-6 level. Conclusions This study suggested that leflunomide could prevent and improve salivary gland hypofunction and inhibit immune activation in NOD mice, providing reference for evaluating leflunomide in the treatment of Sjogren′s syndrome. Key words: Sjogren′s syndrome; Leflunomide; NOD mice; Salivary gland function; Lymphocyte; Inflammatory cytokine
{"title":"Protective effects of leflunomide on salivary function of submandibular gland in NOD mice","authors":"Chao Yang, Xu Zheng, Xiaoxiao Yang, Guo-sheng Wang, Xiang-Pei Li, Xiao-mei Li","doi":"10.3760/CMA.J.CN112309-20191016-00330","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191016-00330","url":null,"abstract":"Objective \u0000To investigate the therapeutic effects of leflunomide on salivary gland secretion hypofunction in NOD mice with Sjogren′s syndrome. \u0000 \u0000 \u0000Methods \u0000NOD mice were randomly divided into four groups: prevention group, prevention control group, treatment group and treatment control group. Salivary flow rate was measured after pilocarpine stimulation. Changes in the average number and area of infiltrating lesions were compared after hematoxylin and eosin (HE) staining. The percentages of CD3+ T, CD4+ T, CD8+ T, CD44+ CD62L-CD4+ T, CD19+ B and CD138+ B cells in submandibular gland and spleen were detected by flow cytometry. The levels of TNF-α, IL-17A and IL-6 in serum were detected by CBA method. \u0000 \u0000 \u0000Results \u0000The salivary flow rate (t=-5.81, P<0.001; t=-3.61, P<0.05), the number of infiltrating lesions(t=3.95, P<0.01; t=4.94, P<0.001)and the average area of infiltrating lesions(t=3.18, P<0.05; t=2.35, P<0.05)were significantly ameliorated in the prevention and treatment groups. Moreover, CD3+ CD4+ T cells(t=2.39, P<0.05; t=3.82, P<0.01)and CD44+ CD62L-CD4+ T cells(t=3.53, P<0.05; t=3.36, P<0.05)in the submandibular gland were significantly decreased. CD3+ T(t=6.08, P<0.001; t=2.76, P<0.05), CD3+ CD4+ T (t=3.73, P<0.05; t=2.39, P<0.05), CD19+ B (t=5.88, P<0.001; t=4.23, P<0.01) and CD138+ B cells (t=4.30, P<0.001; t=4.46, P<0.01) in the spleen were also significantly reduced. Serum IL-17A (t=4.15, P<0.01; t=3.36, P<0.01) in the two groups and TNF-α (t=4.56, P<0.001) in the prevention group were down-regulated, but no significant difference was observed in IL-6 level. \u0000 \u0000 \u0000Conclusions \u0000This study suggested that leflunomide could prevent and improve salivary gland hypofunction and inhibit immune activation in NOD mice, providing reference for evaluating leflunomide in the treatment of Sjogren′s syndrome. \u0000 \u0000 \u0000Key words: \u0000Sjogren′s syndrome; Leflunomide; NOD mice; Salivary gland function; Lymphocyte; Inflammatory cytokine","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"211-217"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45894973","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}
Pub Date : 2020-03-31DOI: 10.3760/CMA.J.CN112309-20190917-00301
Xiaojuan Wang, Ziyu Xiao, Y. Liu, M. Wang, Junfei Huang, Q. Ma, Yue Wang, Xu Chen, H. Yong, F. Hong, Shijun Li
Objective To investigate the effects of caspase-11 non-canonical inflammasome on the Leptospira interrogans (L.interrogans)-induced secretion of inflammatory cytokines in J774A.1 cells. Methods Murine mononuclear macrophage cells (J774A.1) were infected with L. interrogans strain 56601. Expression of caspase-11, IL-1β, IL-1α and IL-18 at mRNA level in J774A.1 cells were detected by real-time RT-PCR. The levels of caspase-11, IL-1β, IL-1α and IL-18 in the culture supernatants of J774A.1 cells were detected by ELISA. Results Real-time RT-PCR showed that caspase-11 expression at mRNA level was 5.12, 14.21, 8.94, 14.06, 18.58 and 0.93 times of that in uninfected cells after 1, 2, 4, 8, 12 and 24 h of L. interrogans infection, and respectively decreased to 0.10, 0.07, 0.10, 0.09, 0.07 and 0.45 times after caspase-11 inhibitor intervention (P<0.05). Expression of IL-1β, IL-1α and IL-18 at mRNA level was significantly increased after infection (P<0.05). After the intervention with caspase-11 inhibitor, IL-1β mRNA decreased to 0.05, 0.03, 0.02, 0.05, 0.06 and 0.02 times (P<0.05); IL-1α mRNA decreased to 0.14, 0.07, 0.15, 0.10, 0.03 and 0.06 times (P<0.05); IL-18 mRNA decreased to 0.08, 0.10, 0.16, 0.18, 0.10 and 0.07 times (P<0.05). ELISA results showed that the expression of caspase-11, IL-1β, IL-1α and IL-18 at protein level was significantly increased. After the intervention with caspase-11 inhibitor, caspase-11 level decreased to 43.07, 41.64, 51.96, 86.56, 105.36, and 129.95 pg/ml (P<0.05); IL-1β level decreased to 15.01, 14.19, 68.02, 31.20, 173.13 and 104.98 pg/ml (P<0.05); IL-1α level decreased to 12.14, 15.40, 38.01, 21.97, 24.48 and 27.09 pg/ml (P<0.05); IL-18 level decreased to 96.27, 102.21, 85.34, 116.28, 155.36 and 114.03 pg/ml (P<0.05). Conclusions Caspase-11 non-canonical inflammasome was involved in the mediation of IL-1β, IL-1α and IL-18 secretion in mouse mononuclear macrophages after L. interrogans infection. Key words: Leptospira interrogans; Caspase-11 non-canonical inflammasome; Inflammatory cytokine; Macrophage; Gene silencing
{"title":"Effects of caspase-11 non-canonical inflammasome on Leptospira interrogans-induced secretion of inflammatory cytokines in J774A.1 cells","authors":"Xiaojuan Wang, Ziyu Xiao, Y. Liu, M. Wang, Junfei Huang, Q. Ma, Yue Wang, Xu Chen, H. Yong, F. Hong, Shijun Li","doi":"10.3760/CMA.J.CN112309-20190917-00301","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190917-00301","url":null,"abstract":"Objective \u0000To investigate the effects of caspase-11 non-canonical inflammasome on the Leptospira interrogans (L.interrogans)-induced secretion of inflammatory cytokines in J774A.1 cells. \u0000 \u0000 \u0000Methods \u0000Murine mononuclear macrophage cells (J774A.1) were infected with L. interrogans strain 56601. Expression of caspase-11, IL-1β, IL-1α and IL-18 at mRNA level in J774A.1 cells were detected by real-time RT-PCR. The levels of caspase-11, IL-1β, IL-1α and IL-18 in the culture supernatants of J774A.1 cells were detected by ELISA. \u0000 \u0000 \u0000Results \u0000Real-time RT-PCR showed that caspase-11 expression at mRNA level was 5.12, 14.21, 8.94, 14.06, 18.58 and 0.93 times of that in uninfected cells after 1, 2, 4, 8, 12 and 24 h of L. interrogans infection, and respectively decreased to 0.10, 0.07, 0.10, 0.09, 0.07 and 0.45 times after caspase-11 inhibitor intervention (P<0.05). Expression of IL-1β, IL-1α and IL-18 at mRNA level was significantly increased after infection (P<0.05). After the intervention with caspase-11 inhibitor, IL-1β mRNA decreased to 0.05, 0.03, 0.02, 0.05, 0.06 and 0.02 times (P<0.05); IL-1α mRNA decreased to 0.14, 0.07, 0.15, 0.10, 0.03 and 0.06 times (P<0.05); IL-18 mRNA decreased to 0.08, 0.10, 0.16, 0.18, 0.10 and 0.07 times (P<0.05). ELISA results showed that the expression of caspase-11, IL-1β, IL-1α and IL-18 at protein level was significantly increased. After the intervention with caspase-11 inhibitor, caspase-11 level decreased to 43.07, 41.64, 51.96, 86.56, 105.36, and 129.95 pg/ml (P<0.05); IL-1β level decreased to 15.01, 14.19, 68.02, 31.20, 173.13 and 104.98 pg/ml (P<0.05); IL-1α level decreased to 12.14, 15.40, 38.01, 21.97, 24.48 and 27.09 pg/ml (P<0.05); IL-18 level decreased to 96.27, 102.21, 85.34, 116.28, 155.36 and 114.03 pg/ml (P<0.05). \u0000 \u0000 \u0000Conclusions \u0000Caspase-11 non-canonical inflammasome was involved in the mediation of IL-1β, IL-1α and IL-18 secretion in mouse mononuclear macrophages after L. interrogans infection. \u0000 \u0000 \u0000Key words: \u0000Leptospira interrogans; Caspase-11 non-canonical inflammasome; Inflammatory cytokine; Macrophage; Gene silencing","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"225-230"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49203601","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}
Pub Date : 2020-03-31DOI: 10.3760/CMA.J.CN112309-20190830-00280
G. Miao, Lin Zhang, X. Hou, Q. Hao
Objective To clone and express the 126-274 aa OspA peptide (OspA-pep) of Chinese Borrelia garinii (B.garinii) strain PD91 and to preliminarily study its immune protectivity. Methods The gene encoding the 126-274 aa OspA-pep of B. garinii PD91 was amplified by polymerase chain reaction (PCR) and then cloned into the prokaryotic expression vector pET-30a to construct the recombinant plasmid pET-30a-OspA-pep. Escherichia coli BL21 (DE3) competent cells transfected with the recombinant plasmid were induced by IPTG to express the target protein. The recombinant OspA-pep (rOspA-pep) was purified with Ni-IDA resin chromatography and its immunogenicity was analyzed by Western blot. New Zealand white rabbits were immunized with different doses of rOspA-pep (20, 30, 40, 50, 60, 80 and 100 μg). The titers of specific IgG antibodies in rabbit serum samples before and after immunization were detected by indirect immunofluorescence assay (IFA). The optimal immune dose was determined according to the antibody titer after immunization. In vitro neutralization test was performed to detect the immune protection of rOspA-pep using serum samples of the optimal immunization group. The optimal dose of rOspA-pep was used to immunize New Zealand white rabbits to observe the changes in antibody titer. Results The recombinant plasmid pET-30a-OspA-pep was successfully constructed and highly expressed in host bacteria. Western blot showed that rOspA-pep had obvious antigen-antibody reaction with polyclonal antibody against B. garinii PD91 strain. IFA results showed the titers of IgG antibody in serum samples of rabbits immunized with rOspA-pep increased significantly (up to 1∶2 480) and 40 μg was the optimal dose. The neutralization rates of antibodies induced by 40 μg of rOspA-pep were 100% against 106 strain/ml of representative B. garinii PD91 and Borrelia afzelii (B.afzelii) FP1 strains, 100% against 107 strain/ml of FP1 strain, and 60% against 107 strain/ml of PD91 strain. After immunization with 40 μg rOspA-pep on 1 d and 30 d, the titers of specific IgG antibody in rabbit serum samples reached the peak within two months, and maintained at that level for about 3-4 months before a gradual decline. Conclusions The 126-274 aa OspA peptide fragment of Chinese B. garinii PD91 strain possessed good immunogenicity and induced antibodies with better in vitro neutralizing activity, which suggested that it could be used as a candidate component of the second generation subunit vaccine in China. Key words: Borrelia garinii; OspA peptide; Cloning and expression; Immunoprotectivity
目的克隆并表达中国加里伯氏疏螺旋体(B.garinii)菌株PD91的126-274 aa OspA肽(OspA-pep)，并初步研究其免疫保护作用。方法采用聚合酶链反应(PCR)扩增出加里尼布氏杆菌PD91 126-274 aa OspA-pep基因，并将其克隆到原核表达载体pET-30a中，构建重组质粒pET-30a-OspA-pep。用IPTG诱导转染重组质粒的大肠杆菌BL21 (DE3)感受态细胞表达目的蛋白。重组OspA-pep (rOspA-pep)采用Ni-IDA树脂层析纯化，Western blot检测其免疫原性。用不同剂量的rOspA-pep(20、30、40、50、60、80、100 μg)免疫新西兰大白兔。采用间接免疫荧光法(IFA)检测免疫前后兔血清中特异性IgG抗体的滴度。根据免疫后抗体滴度确定最佳免疫剂量。采用体外中和试验检测最佳免疫组血清样品的免疫保护作用。采用最佳剂量的rOspA-pep免疫新西兰大白兔，观察抗体效价的变化。结果成功构建了重组质粒pET-30a-OspA-pep，并在宿主菌中高表达。Western blot结果显示，rOspA-pep与加里尼布氏杆菌PD91多克隆抗体存在明显的抗原抗体反应。IFA结果显示，经rOspA-pep免疫的家兔血清中IgG抗体滴度显著升高(达1∶2 480)，最佳剂量为40 μg。40 μg rOspA-pep对代表性布氏加里尼布氏杆菌PD91和阿夫泽利伯氏疏螺旋体FP1菌株106株/ml的抗体中和率为100%，对FP1菌株107株/ml的抗体中和率为100%，对PD91菌株107株/ml的抗体中和率为60%。40 μg rOspA-pep免疫1 d和30 d后，兔血清样品中特异性IgG抗体滴度在2个月内达到峰值，并在3-4个月左右维持该水平，随后逐渐下降。结论中国加里尼布氏杆菌PD91菌株126-274 aa OspA肽片段具有良好的免疫原性，诱导的抗体具有较好的体外中和活性，可作为中国第二代亚单位疫苗的候选组分。关键词:加里氏疏螺旋体;OspA肽;克隆与表达;Immunoprotectivity
{"title":"Molecular cloning and expression of OspA peptide from a Chinese Borrelia garinii strain PD91 and preliminary study on its immunoprotectivity","authors":"G. Miao, Lin Zhang, X. Hou, Q. Hao","doi":"10.3760/CMA.J.CN112309-20190830-00280","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190830-00280","url":null,"abstract":"Objective \u0000To clone and express the 126-274 aa OspA peptide (OspA-pep) of Chinese Borrelia garinii (B.garinii) strain PD91 and to preliminarily study its immune protectivity. \u0000 \u0000 \u0000Methods \u0000The gene encoding the 126-274 aa OspA-pep of B. garinii PD91 was amplified by polymerase chain reaction (PCR) and then cloned into the prokaryotic expression vector pET-30a to construct the recombinant plasmid pET-30a-OspA-pep. Escherichia coli BL21 (DE3) competent cells transfected with the recombinant plasmid were induced by IPTG to express the target protein. The recombinant OspA-pep (rOspA-pep) was purified with Ni-IDA resin chromatography and its immunogenicity was analyzed by Western blot. New Zealand white rabbits were immunized with different doses of rOspA-pep (20, 30, 40, 50, 60, 80 and 100 μg). The titers of specific IgG antibodies in rabbit serum samples before and after immunization were detected by indirect immunofluorescence assay (IFA). The optimal immune dose was determined according to the antibody titer after immunization. In vitro neutralization test was performed to detect the immune protection of rOspA-pep using serum samples of the optimal immunization group. The optimal dose of rOspA-pep was used to immunize New Zealand white rabbits to observe the changes in antibody titer. \u0000 \u0000 \u0000Results \u0000The recombinant plasmid pET-30a-OspA-pep was successfully constructed and highly expressed in host bacteria. Western blot showed that rOspA-pep had obvious antigen-antibody reaction with polyclonal antibody against B. garinii PD91 strain. IFA results showed the titers of IgG antibody in serum samples of rabbits immunized with rOspA-pep increased significantly (up to 1∶2 480) and 40 μg was the optimal dose. The neutralization rates of antibodies induced by 40 μg of rOspA-pep were 100% against 106 strain/ml of representative B. garinii PD91 and Borrelia afzelii (B.afzelii) FP1 strains, 100% against 107 strain/ml of FP1 strain, and 60% against 107 strain/ml of PD91 strain. After immunization with 40 μg rOspA-pep on 1 d and 30 d, the titers of specific IgG antibody in rabbit serum samples reached the peak within two months, and maintained at that level for about 3-4 months before a gradual decline. \u0000 \u0000 \u0000Conclusions \u0000The 126-274 aa OspA peptide fragment of Chinese B. garinii PD91 strain possessed good immunogenicity and induced antibodies with better in vitro neutralizing activity, which suggested that it could be used as a candidate component of the second generation subunit vaccine in China. \u0000 \u0000 \u0000Key words: \u0000Borrelia garinii; OspA peptide; Cloning and expression; Immunoprotectivity","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"218-224"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47156905","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}
Pub Date : 2020-03-31DOI: 10.3760/CMA.J.CN112309-20191114-00376
Yaya Pian, Zhenxiang Gao, Jingjing Nie, Jihong Hu
Objective To study the molecular mechanism of β-1, 4-galactosyltransferase 6 (β4galt6) in regulating lymphocyte migration under inflammatory conditions. Methods CRISPR/Cas9 system was used to knock out the β4galt6 gene of mouse islet vascular endothelial cells (MS1). Adhesion assay was performed to compare the adhesion ability of lymphocytes to wild-type cells and gene knockout cells. Expression of adhesion molecules on the surface of wild-type cells and gene knockout cells were compared using RT-PCR and flow cytometry. Transwell model was used to compare the transmigration ability of lymphocytes across wild-type cells and gene knockout cells. Results The β4galt6 gene knockout cell line, β4galt6 KO, was successfully constructed. The percentage of lymphocytes adhereing to wild-type MS1 cells was significantly higher than that to β4galt6 KO cells under inflammatory conditions. The expression of adhesion molecules including intercellular adhesion molecule 1 (ICAM1), vascular cell adhesion molecule 1 (VCAM1) and P-selectin on the surface of wild-type MS1 cells was significantly higher than that on β4galt6 KO cells. Moreover, the percentage of lymphocytes passing through wild-type MS1 cells was significantly higher than that through β4galt6 KO cells. Conclusion Under inflammatory conditions, β4galt6 could promote the migration of lymphocytes. Key words: β-1, 4-galactosyltransferase 6; Lymphocyte; Migration; Inflammation
{"title":"Molecular mechanism of β-1, 4-galactosyltransferase 6 in regulating lymphocyte migration","authors":"Yaya Pian, Zhenxiang Gao, Jingjing Nie, Jihong Hu","doi":"10.3760/CMA.J.CN112309-20191114-00376","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191114-00376","url":null,"abstract":"Objective \u0000To study the molecular mechanism of β-1, 4-galactosyltransferase 6 (β4galt6) in regulating lymphocyte migration under inflammatory conditions. \u0000 \u0000 \u0000Methods \u0000CRISPR/Cas9 system was used to knock out the β4galt6 gene of mouse islet vascular endothelial cells (MS1). Adhesion assay was performed to compare the adhesion ability of lymphocytes to wild-type cells and gene knockout cells. Expression of adhesion molecules on the surface of wild-type cells and gene knockout cells were compared using RT-PCR and flow cytometry. Transwell model was used to compare the transmigration ability of lymphocytes across wild-type cells and gene knockout cells. \u0000 \u0000 \u0000Results \u0000The β4galt6 gene knockout cell line, β4galt6 KO, was successfully constructed. The percentage of lymphocytes adhereing to wild-type MS1 cells was significantly higher than that to β4galt6 KO cells under inflammatory conditions. The expression of adhesion molecules including intercellular adhesion molecule 1 (ICAM1), vascular cell adhesion molecule 1 (VCAM1) and P-selectin on the surface of wild-type MS1 cells was significantly higher than that on β4galt6 KO cells. Moreover, the percentage of lymphocytes passing through wild-type MS1 cells was significantly higher than that through β4galt6 KO cells. \u0000 \u0000 \u0000Conclusion \u0000Under inflammatory conditions, β4galt6 could promote the migration of lymphocytes. \u0000 \u0000 \u0000Key words: \u0000β-1, 4-galactosyltransferase 6; Lymphocyte; Migration; Inflammation","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"206-210"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49309675","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}
Pub Date : 2020-03-31DOI: 10.3760/CMA.J.CN112309-20190605-00176
Lixiang Chen, Shun Li, Xiaohui Zhou
Cell death is an important event in the life cycle. Physical injury can cause cell death in eukaryotes. Besides, specific signaling pathway-mediated programmed cell death has attracted increasing attention. Currently, programmed cell death mainly includes apoptosis, programmed necrosis (necroptosis) and pyroptosis. Necroptosis and pyroptosis, as two new ways of programmed cell death, have been found to play a key role in the process of pathogen infection. Both necroptosis and pyroptosis have the characteristics of programmed lytic cell death, but the signaling pathways involved in them have significant differences. This review focused on the morphological characteristics, signal transduction pathways and the role played in the process of pathogen infection of necroptosis and pyroptosis. Key words: Necroptosis; Pyroptosis; Signaling pathway
{"title":"Overview of necroptosis and pyroptosis signaling pathways","authors":"Lixiang Chen, Shun Li, Xiaohui Zhou","doi":"10.3760/CMA.J.CN112309-20190605-00176","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190605-00176","url":null,"abstract":"Cell death is an important event in the life cycle. Physical injury can cause cell death in eukaryotes. Besides, specific signaling pathway-mediated programmed cell death has attracted increasing attention. Currently, programmed cell death mainly includes apoptosis, programmed necrosis (necroptosis) and pyroptosis. Necroptosis and pyroptosis, as two new ways of programmed cell death, have been found to play a key role in the process of pathogen infection. Both necroptosis and pyroptosis have the characteristics of programmed lytic cell death, but the signaling pathways involved in them have significant differences. This review focused on the morphological characteristics, signal transduction pathways and the role played in the process of pathogen infection of necroptosis and pyroptosis. \u0000 \u0000 \u0000Key words: \u0000Necroptosis; Pyroptosis; Signaling pathway","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"231-237"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43226179","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}
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