Objective:To detect IgG antibody to Chlamydophila pneumoniae(CP) in sera of HIV/AIDS patients and provide rationale for inclusion of routine screening for anti-CP antibodies and anti-chlamydial agents in the Nigerian ...Objective:To detect IgG antibody to Chlamydophila pneumoniae(CP) in sera of HIV/AIDS patients and provide rationale for inclusion of routine screening for anti-CP antibodies and anti-chlamydial agents in the Nigerian National H1V/AIDS Management Plan.Methods:Serum samples from 34 consenting HIV/AIDS patients attended a Government-approved Antiretroviral Treatment Facility in Abuja were screened by enzyme-linked immunosorbent assay for anti-CP IgG antibody using ImmunoComb? Chlamydia Bivalent IgG Test kit(Orgenics,Israel).Results: Anti-CP IgG antibody was detected in 20(58.8%) of 34 patients tested.The detection rale was higher among the males(8/13:61.5%) than the females(12/21:57.1%).Patients of the age group 16-30 years had the highest(7/10:70%) detection of anti-CP IgG antibody.Conclusions:The result of the present study suggests the presence of anti-CP antibodies in sera of the HIV/AIDS patients,and reinforces the need for routine screening for anti-CP antibodies as a necessary intervention to reduce the burden of Chlamydophila pneumoniae(C.pneumoniae) infections and to reduce HIV-positive morbidity in Nigeria.The outcome of this study also provides justification for the possible inclusion of anti-chlamydial agents in the National HIV/AIDS Management Plan to provide prophylaxis against or treat active C.pneumoniae infections.展开更多
The role of Chlamydophila pneumoniae in pathogenesis of atherosclerosis is one of the most important discussions in coronary artery diseases. In this study, the relationship between C. pneumoniae seropositivity and at...The role of Chlamydophila pneumoniae in pathogenesis of atherosclerosis is one of the most important discussions in coronary artery diseases. In this study, the relationship between C. pneumoniae seropositivity and atherosclerotic plaque was evaluated among two groups: one group with significant coronary stenosis and one group with normal coronary angiography. Serum C. pneumonia IgM and IgG were evaluated and compared in case and control groups. The seropositivity rates of IgM and IgG antibodies were not statistically differ between case and control groups, although the incidences of positivity were more in case group. The results of our study did not show a correlation between C. pneumonia infection and atherosclerotic plaque or coronary artery stenosis neither in acute nor in chronic infection. More precise studies are needed to clarify the probable inflammatory cascade that starts with C. pneumonia infection and lead to development of atherosclerotic plaque.展开更多
To clone the gene coding the immunodominant region in the chlamydial protease-like activity factor （CPAF） from Chlaroydophila pneumoniae, to analyze immunoeoropetenee of the expressed protein, and to evaluate its va...To clone the gene coding the immunodominant region in the chlamydial protease-like activity factor （CPAF） from Chlaroydophila pneumoniae, to analyze immunoeoropetenee of the expressed protein, and to evaluate its value in serodiagnosis, the CPAF immunodominant region gene was amplified, ligated into a pGEX6p-2 vector, and then the expressed recombinant protein was purified with glutathione Stransferase （GST） agarose gel FF after renaturation, then identified by SDS-PAGE and Western blot. A new indirect ELISA was developed with the purified protein as coating antigen. The immunogenicity of the recombinant protein was evaluated by immunization to New Zealand rabbits, and its immunoreactivity was analyzed by reacting with anti-C, pneumoniae antibody. 300 clinical sera samples were respectively detected by mieroimmunofluorescenee （MIF） as reference method and the indirect ELISA, and the difference between the two methods was analyzed. Cross-reactivity against Chlamydia trachomatis was investigated with the indirect ELISA to detect anti-C, trachomatis positive antisera. The results indicated that a 51.3 kDa recombinant protein was obtained. Western blot assay proved that the recombinant protein could merely specifically react with human anti- C. pneurnoniae antisera. The titers of the specific IgG antibodies in the immunized New Zealand rabbits were above 1 ： 16 000. Anti- C. pneumoniae IgG positive and negative reference sera were detected with the indirect ELISA, and the concordance rate of negative and positive results were both 100% （40/40）. The sensitivity and specificity of the indirect ELISA in comparison with MIF were 93.8% （45/48） and 100% （252/252） separately by detecting 300 clinical sera samples, and the concordance rate between the two methods was 99.0%. No cross reaction against C. trachomatis was found with the indirect ELISA to detect anti-C, trachomatis positive antisera. In conclusion, the prepared recombinant protein of the CPAF immunodominant region shows excellent immunocompetence and can be used to develop a new indirect ELISA as a method to detect anti-C, pneumoniae antibody for diagnosis of C. pneumoniae infection.展开更多
Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection is suggested to have a role. Co-infection with Chlamydophila pneumoniae (Cp) and Mycoplasma pneumoniae (Mp) was linked ...Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection is suggested to have a role. Co-infection with Chlamydophila pneumoniae (Cp) and Mycoplasma pneumoniae (Mp) was linked with coronary plaque rupture, in association with vessel dilatation and adventitial inflammation. Pathogens are recognized by Toll-like receptors (TLRs) development of the inflammatory process. Objective: Here, we studied whether co-infection by Cp and Mp was involved in the increased inflammation present in AAA and if it could be associated with deficient expression of TLRs. We compared human samples of AAA with non-dilated human aortic atherosclerotic lesions, regarding the amount of Cp and Mp antigens, and expression of TLR2 and TLR4. Methods: Two groups of aorta fragments were analyzed: G1 (n = 13) moderate atherosclerosis and G2 (n = 14) AAA samples, through immunohisto-chemistry and in situ hybridization methods. Results: Mp and Cp antigens in intima/medial layer were greater in G2 than G1, with no difference in adventitia. TLR2 and TLR4 were higher in G2 than G1 adventitia fat. There was a correlation between Mp versus TLR2 and of TLR4 in intima/medial layer and in adventitia of G1, but there was a lack of correlation in G2. In Cp adventitia, the correlation in G1 was high with TLR2 but not with TLR4, and in G2 the correlation was positive for both TLRs. Conclusion: This study favors the concept that symbiotic co-infection by Cp and Mp participates in the pathogenesis of AAA. It also emphasizes that adventitial fat is the initial site for colonization of these bacteria that probably reach the tissue through vasa vasorum. An exacerbated immune reaction is not efficient to control the infection that reaches and proliferates in high levels at the medial and intimal layer, contributing to the development of vessel dilatation.展开更多
Objective This paper aims to develop a monoclonal antibodies （MAbs）- based ELISA for detecting Chlamydophila pneumoniae （C. pneumonioe） antigens in humans with the variable domains （VD） 2 and 3 of the major oute...Objective This paper aims to develop a monoclonal antibodies （MAbs）- based ELISA for detecting Chlamydophila pneumoniae （C. pneumonioe） antigens in humans with the variable domains （VD） 2 and 3 of the major outer membrane protein （MOMPvD2-VD~） and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae. Methods MOMPvo2-vo3were overexpressed in Escherichia coil and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I （156 patients with typical respiratory illness clinically confirmed before） and Group II （57 healthy donors）. Results In Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II ＂healthy donors＂, 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively. Conclusion The novel MOMPvD2.VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.展开更多
目的分析经宏基因组二代测序技术(metagenopmic next generation sequencing,mNGS)确诊的28例鹦鹉热衣原体肺炎病人的临床特点。方法回顾性分析2019年10月1日至2022年4月1日长沙市第一医院采用mNGS诊断的28例鹦鹉热衣原体肺炎病人的诊...目的分析经宏基因组二代测序技术(metagenopmic next generation sequencing,mNGS)确诊的28例鹦鹉热衣原体肺炎病人的临床特点。方法回顾性分析2019年10月1日至2022年4月1日长沙市第一医院采用mNGS诊断的28例鹦鹉热衣原体肺炎病人的诊治情况。结果28例病人中10例为女性,18例为男性,中位年龄60岁;27例病人有明确禽类接触史,1例无。重症病例占32.1%,多为有慢性阻塞性肺疾病(COPD)、糖尿病、高血压、冠心病或尿毒症等基础疾病病人。临床表现包括发热(100%)、畏寒(28.6%)、咳嗽、咳痰(71.4%)、气促(53.6%)。外周血中白细胞总数多在正常范围,96.4%的病人中性粒细胞比例增高、淋巴细胞比例降低;96.4%病人PCT升高,所检测病人中ESR和CRP升高者达100%;分别有71.4%和50%病人乳酸脱氢酶和肌酸激酶增高;有82.1%的病人谷草转氨酶增高,有78.6%的病人白蛋白降低。胸部CT表现单侧病变25例、其中右肺病变19例,常见影像改变是斑片状阴影(78.6%)、大片实变(21.4%),26例伴有胸腔积液。治疗情况:单用氟喹诺酮类(莫西沙星或者左氧氟沙星)治疗9例,单用多西环素治疗2例;联合用药13例,其中青霉素类(哌拉西林他唑巴坦或头孢哌酮舒巴坦钠)联用喹诺酮类有10例,青霉素类联合多西环素3例。有2例病人初始选择青霉素类加喹诺酮类药物治疗无效,改用多西环素或联用多西环素后症状缓解。28例病人均预后良好,无死亡病例。结论鹦鹉热衣原体肺炎病人的职业史、临床表现、实验室检查结果及肺部CT具有一定的特点;临床对可疑病例应尽早行mNGS检测,快速过渡到精准治疗,能明显改善预后。展开更多
基金funded by NIPRD through a Research Grant awarded in 2005 to Ngwai YB and Izebe KS
文摘Objective:To detect IgG antibody to Chlamydophila pneumoniae(CP) in sera of HIV/AIDS patients and provide rationale for inclusion of routine screening for anti-CP antibodies and anti-chlamydial agents in the Nigerian National H1V/AIDS Management Plan.Methods:Serum samples from 34 consenting HIV/AIDS patients attended a Government-approved Antiretroviral Treatment Facility in Abuja were screened by enzyme-linked immunosorbent assay for anti-CP IgG antibody using ImmunoComb? Chlamydia Bivalent IgG Test kit(Orgenics,Israel).Results: Anti-CP IgG antibody was detected in 20(58.8%) of 34 patients tested.The detection rale was higher among the males(8/13:61.5%) than the females(12/21:57.1%).Patients of the age group 16-30 years had the highest(7/10:70%) detection of anti-CP IgG antibody.Conclusions:The result of the present study suggests the presence of anti-CP antibodies in sera of the HIV/AIDS patients,and reinforces the need for routine screening for anti-CP antibodies as a necessary intervention to reduce the burden of Chlamydophila pneumoniae(C.pneumoniae) infections and to reduce HIV-positive morbidity in Nigeria.The outcome of this study also provides justification for the possible inclusion of anti-chlamydial agents in the National HIV/AIDS Management Plan to provide prophylaxis against or treat active C.pneumoniae infections.
文摘The role of Chlamydophila pneumoniae in pathogenesis of atherosclerosis is one of the most important discussions in coronary artery diseases. In this study, the relationship between C. pneumoniae seropositivity and atherosclerotic plaque was evaluated among two groups: one group with significant coronary stenosis and one group with normal coronary angiography. Serum C. pneumonia IgM and IgG were evaluated and compared in case and control groups. The seropositivity rates of IgM and IgG antibodies were not statistically differ between case and control groups, although the incidences of positivity were more in case group. The results of our study did not show a correlation between C. pneumonia infection and atherosclerotic plaque or coronary artery stenosis neither in acute nor in chronic infection. More precise studies are needed to clarify the probable inflammatory cascade that starts with C. pneumonia infection and lead to development of atherosclerotic plaque.
文摘To clone the gene coding the immunodominant region in the chlamydial protease-like activity factor （CPAF） from Chlaroydophila pneumoniae, to analyze immunoeoropetenee of the expressed protein, and to evaluate its value in serodiagnosis, the CPAF immunodominant region gene was amplified, ligated into a pGEX6p-2 vector, and then the expressed recombinant protein was purified with glutathione Stransferase （GST） agarose gel FF after renaturation, then identified by SDS-PAGE and Western blot. A new indirect ELISA was developed with the purified protein as coating antigen. The immunogenicity of the recombinant protein was evaluated by immunization to New Zealand rabbits, and its immunoreactivity was analyzed by reacting with anti-C, pneumoniae antibody. 300 clinical sera samples were respectively detected by mieroimmunofluorescenee （MIF） as reference method and the indirect ELISA, and the difference between the two methods was analyzed. Cross-reactivity against Chlamydia trachomatis was investigated with the indirect ELISA to detect anti-C, trachomatis positive antisera. The results indicated that a 51.3 kDa recombinant protein was obtained. Western blot assay proved that the recombinant protein could merely specifically react with human anti- C. pneurnoniae antisera. The titers of the specific IgG antibodies in the immunized New Zealand rabbits were above 1 ： 16 000. Anti- C. pneumoniae IgG positive and negative reference sera were detected with the indirect ELISA, and the concordance rate of negative and positive results were both 100% （40/40）. The sensitivity and specificity of the indirect ELISA in comparison with MIF were 93.8% （45/48） and 100% （252/252） separately by detecting 300 clinical sera samples, and the concordance rate between the two methods was 99.0%. No cross reaction against C. trachomatis was found with the indirect ELISA to detect anti-C, trachomatis positive antisera. In conclusion, the prepared recombinant protein of the CPAF immunodominant region shows excellent immunocompetence and can be used to develop a new indirect ELISA as a method to detect anti-C, pneumoniae antibody for diagnosis of C. pneumoniae infection.
基金CNPQ (National Counsel of Technological and Scientific Development of Federal Government grant 132905/ 2006-0.) FAPESP (Foundation that supports research in the State of São Paulo, grant 562444/2007).
文摘Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection is suggested to have a role. Co-infection with Chlamydophila pneumoniae (Cp) and Mycoplasma pneumoniae (Mp) was linked with coronary plaque rupture, in association with vessel dilatation and adventitial inflammation. Pathogens are recognized by Toll-like receptors (TLRs) development of the inflammatory process. Objective: Here, we studied whether co-infection by Cp and Mp was involved in the increased inflammation present in AAA and if it could be associated with deficient expression of TLRs. We compared human samples of AAA with non-dilated human aortic atherosclerotic lesions, regarding the amount of Cp and Mp antigens, and expression of TLR2 and TLR4. Methods: Two groups of aorta fragments were analyzed: G1 (n = 13) moderate atherosclerosis and G2 (n = 14) AAA samples, through immunohisto-chemistry and in situ hybridization methods. Results: Mp and Cp antigens in intima/medial layer were greater in G2 than G1, with no difference in adventitia. TLR2 and TLR4 were higher in G2 than G1 adventitia fat. There was a correlation between Mp versus TLR2 and of TLR4 in intima/medial layer and in adventitia of G1, but there was a lack of correlation in G2. In Cp adventitia, the correlation in G1 was high with TLR2 but not with TLR4, and in G2 the correlation was positive for both TLRs. Conclusion: This study favors the concept that symbiotic co-infection by Cp and Mp participates in the pathogenesis of AAA. It also emphasizes that adventitial fat is the initial site for colonization of these bacteria that probably reach the tissue through vasa vasorum. An exacerbated immune reaction is not efficient to control the infection that reaches and proliferates in high levels at the medial and intimal layer, contributing to the development of vessel dilatation.
基金supported by grants from the National Natural Science Foundation of China (Grant No. 30901352)Innovative Research Team in University of Hunan Province (Number: [2008] 51)Hunan Provincial Innovation Foundation for Postgraduate and Hunan Provincial Training and Innovation Base for Post-graduate
文摘Objective This paper aims to develop a monoclonal antibodies （MAbs）- based ELISA for detecting Chlamydophila pneumoniae （C. pneumonioe） antigens in humans with the variable domains （VD） 2 and 3 of the major outer membrane protein （MOMPvD2-VD~） and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae. Methods MOMPvo2-vo3were overexpressed in Escherichia coil and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I （156 patients with typical respiratory illness clinically confirmed before） and Group II （57 healthy donors）. Results In Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II ＂healthy donors＂, 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively. Conclusion The novel MOMPvD2.VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.
文摘目的分析经宏基因组二代测序技术(metagenopmic next generation sequencing,mNGS)确诊的28例鹦鹉热衣原体肺炎病人的临床特点。方法回顾性分析2019年10月1日至2022年4月1日长沙市第一医院采用mNGS诊断的28例鹦鹉热衣原体肺炎病人的诊治情况。结果28例病人中10例为女性,18例为男性,中位年龄60岁;27例病人有明确禽类接触史,1例无。重症病例占32.1%,多为有慢性阻塞性肺疾病(COPD)、糖尿病、高血压、冠心病或尿毒症等基础疾病病人。临床表现包括发热(100%)、畏寒(28.6%)、咳嗽、咳痰(71.4%)、气促(53.6%)。外周血中白细胞总数多在正常范围,96.4%的病人中性粒细胞比例增高、淋巴细胞比例降低;96.4%病人PCT升高,所检测病人中ESR和CRP升高者达100%;分别有71.4%和50%病人乳酸脱氢酶和肌酸激酶增高;有82.1%的病人谷草转氨酶增高,有78.6%的病人白蛋白降低。胸部CT表现单侧病变25例、其中右肺病变19例,常见影像改变是斑片状阴影(78.6%)、大片实变(21.4%),26例伴有胸腔积液。治疗情况:单用氟喹诺酮类(莫西沙星或者左氧氟沙星)治疗9例,单用多西环素治疗2例;联合用药13例,其中青霉素类(哌拉西林他唑巴坦或头孢哌酮舒巴坦钠)联用喹诺酮类有10例,青霉素类联合多西环素3例。有2例病人初始选择青霉素类加喹诺酮类药物治疗无效,改用多西环素或联用多西环素后症状缓解。28例病人均预后良好,无死亡病例。结论鹦鹉热衣原体肺炎病人的职业史、临床表现、实验室检查结果及肺部CT具有一定的特点;临床对可疑病例应尽早行mNGS检测,快速过渡到精准治疗,能明显改善预后。
