整合素α6单克隆抗体对人乳头瘤病毒6/11病毒颗粒体外感染HaCaT细胞的影响

目的 探讨整合素α6单克隆抗体(GoH3)对人乳头瘤病毒(HPv)6/11病毒颗粒(VP)体外感染HaCaT细胞的影响.方法 ①采用4组不同浓度的HPV6/11 VP体外感染HaCaT细胞;②加入6组不同浓度的GoH3后,以106拷贝/ml HPV6/11 VP体外感染HaCaT细胞;③加入稀释倍数为1∶100的GoH...     

Influence of an anti-integrin α6 monoclonal antibody on the in vitro infection of human HaCaT keratinocytes with human papillomavirus (HPV) 6/11 virus particles

Objective To investigate the influence of an anti-integrin α6 monoclonal antibody (GoH3) on the in vitro infection of a human keratinocyte cell line HaCaT with HPV6/11 virus particles (VP). Methods HaCaT cells were infected in vitro with 4 different concentrations of HPV6/11 VP alone, HPV6/11 VP of 106 copies/ml after incubation with 6 different dilutions of GoH3, or 8 clinical isolates of HPV6/11 VP of 106 copies/ml before or after the incubation with 1∶ 100 dilution of GoH3. After additional culture, the infected HaCaT cells were collected and fluorescence quantitative (FQ)-PCR was performed to detect the HPV DNA load in cells. The inhibition rate of CoH3 on the infection was calculated. Results The viral load was different among the HaCaT cells infected with different concentrations of HPV6/11 VP (P < 0.01). The inhibition rate on the infection positively correlated with the concentration of CoH3 when the dilution was more than 1∶ 100; however, when the dilution was less than 1∶ 100, the increase in CoH3 concentration had no influence on the inhibition rate. The average viral load in HaCaT cells infected with clinical isolates of HPV6/11 VP was (5.81 ± 2.51) × 104 copies/ml in the absence of GoH3, (3.02 ± 1.21) × 104 copies/ml with the presence of CoH3, and the average inhibition rate of GoH3 was (46.9 ± 4.7)%. Conclusions GoH3 could partially suppress the adhesion of HPV6/11 VP to HaCaT cells, hinting that integrin a6 is an important HPV6/11 VP receptor on host cells.     

Influence of an anti-integrin α6 monoclonal antibody on the in vitro infection of human HaCaT keratinocytes with human papillomavirus (HPV) 6/11 virus particles

Objective To investigate the influence of an anti-integrin α6 monoclonal antibody (GoH3) on the in vitro infection of a human keratinocyte cell line HaCaT with HPV6/11 virus particles (VP). Methods HaCaT cells were infected in vitro with 4 different concentrations of HPV6/11 VP alone, HPV6/11 VP of 106 copies/ml after incubation with 6 different dilutions of GoH3, or 8 clinical isolates of HPV6/11 VP of 106 copies/ml before or after the incubation with 1∶ 100 dilution of GoH3. After additional culture, the infected HaCaT cells were collected and fluorescence quantitative (FQ)-PCR was performed to detect the HPV DNA load in cells. The inhibition rate of CoH3 on the infection was calculated. Results The viral load was different among the HaCaT cells infected with different concentrations of HPV6/11 VP (P < 0.01). The inhibition rate on the infection positively correlated with the concentration of CoH3 when the dilution was more than 1∶ 100; however, when the dilution was less than 1∶ 100, the increase in CoH3 concentration had no influence on the inhibition rate. The average viral load in HaCaT cells infected with clinical isolates of HPV6/11 VP was (5.81 ± 2.51) × 104 copies/ml in the absence of GoH3, (3.02 ± 1.21) × 104 copies/ml with the presence of CoH3, and the average inhibition rate of GoH3 was (46.9 ± 4.7)%. Conclusions GoH3 could partially suppress the adhesion of HPV6/11 VP to HaCaT cells, hinting that integrin a6 is an important HPV6/11 VP receptor on host cells.     

E型沙眼衣原体重组主要外膜蛋白诱导出的小鼠免疫效应

目的 检验自行研制的E型沙眼衣原体重组主要外膜蛋白(rMOMP)诱导小鼠产生的特异性免疫应答效应.方法 3～4周清洁级BALB/C雌鼠36只,分为佐剂组、单独组和对照组,每组12只.佐剂组采用纯化的rMOMP 50 μg和等体积的弗氏佐剂、单独组单用纯化的rMOMP 50μg、对照组单用PBS200μL,于第0、2、4周分别双侧股四头肌进行免疫.ELISA法检测小鼠血清中沙眼衣原体特异性IgG抗体以及阴道冲洗液中沙眼衣原体特异性sIgA抗体,ELISA法检测细胞因子IFN-γ,MTT法检测小鼠脾淋巴细胞特异性增殖反应;同源攻击后阴道宫颈脱落细胞沙眼衣原体培养,观察小鼠的迟发型超敏反应以及小鼠的血清抗体中和试验.结果 佐剂组小鼠血清沙眼衣原体特异性IgG抗体A_(405)值为0.641±0.059,淋巴细胞增殖指数5.085±1.291,迟发型超敏反应右足足垫增厚0.324 ±0.054 mm.单独组小鼠血清特异性IgG抗体A_(405)值为0.424±0.015,淋巴细胞增殖指数3.123±0.840,迟发型超敏反应右足足垫增厚0.272±0.064mm.佐剂组各项指标的检测结果都高于单独组(P<0.05)和对照组(P<0.01).结论 沙眼衣原体rMOMP能刺激机体产生有效的特异性体液和细胞免疫.     

性病门诊人乳头瘤病毒感染的流行病学调查

目的：了解性病门诊就诊者中人乳头瘤病毒（human papillomavirus,HPV）及其它性传播疾病（sexuallytransmitted disease,STD）病原体混合感染的流行情况。方法：对90例临床样本采用聚合酶链反应（polymerase chain reac-tion,PCR）技术检测尿道或宫颈拭子HPV DNA,同时检测其它STD病原体。结果：在90例就诊者中,HPV DNA阳性13例（14.4%）,女性、混合感染数多者及教育程度低者HPV感染率高（P〈0.05）,HPV感染者至少同时伴有1种其它STD病原体感染。结论：HPV在性病门诊就诊者中存在一定的感染率,HPV感染与性别、混合感染数、教育程度等相关,HPV与其它STD病原体的混合感染多见。     

N-乙酰基转移酶2基因多态性与染发皮炎易感性关系

目的 探讨N-乙酰基转移酶2（NAT2）基因多态性与中国人染发皮炎遗传易感性的关系.方法 应用聚合酶链反应（PCR）及限制性片段长度多态性（RFLP）技术检测和分析天津地区60例染发皮炎患者及73例正常对照者的NAT2野生型等位基因（NAT2＊4）和3个常见突变位点（NAT2＊5A,NAT2＊6B和NAT2＊7A）的基因多态性,比较NAT2各等位基因及基因型频率在染发皮炎组与对照组间分布差异.结果 NAT2等位基因NAT2＊4,NAT2＊5A,NAT2＊6B和NAT2＊7A在染发皮炎组中的分布频率分别是52.5%,5.0%,26.7%,15.8%,在对照组中为55.5%,3.4%,27.4%,13.7%,两组相比差异均无统计学意义（P〉0.05）;快型、中间型、慢型基因型在染发皮炎组中频率分别是26.7%,51.7%,21.7%,在对照组中为30.1%,50.7%,19.2%,与对照组相比差异无统计学意义（P〉0.05）.结论 NAT2基因多态性与中国汉族人染发皮炎遗传易感性无关.     

衣原体噬菌体Chp3衣壳蛋白Vp1二级结构及B细胞抗原表位预测

目的：预测衣原体噬菌体Chp3Vp1蛋白的二级结构和B细胞表位。方法：以Chp3Vp1氨基酸序列为基础,采用Gamier-Robson法、Chou—Fasman法和Karplus-Schulz法预测蛋白二级结构;按Kyte—Doolittle法、Hopp—Woods法、Emini法和Jameson-Wolf法预测蛋白的抗原表位。结果：Chp3Vp1蛋白含多个抗原位点,预测其N端1—10,101—112,159—166,174—184,189—195,288—299,323—333,419-435,477-490为优势表位。结论：Chp3Vp1蛋白有较强的免疫原性,预测的抗原位点为之后蛋白相互作用研究和单抗制备提供了理论依据。     

E型沙眼衣原体重组主要外膜蛋白诱导恒河猴的交叉免疫效应

目的 探讨E型重组主要外膜蛋白(rMOMP)对恒河猴诱导产生的衣原体交叉免疫应答效应.方法 恒河猴分3组,每组2只,分别为佐剂蛋白组、佐剂组、对照组.于第0、2、4周双侧肱三头肌注射.末次免疫后两周,ELISA检测恒河猴血清中沙眼衣原体特异性IgG抗体和细胞因子IFN-γ,MTT法检测恒河猴淋巴细胞特异性增殖反应,观察恒河猴的迟发型超敏反应,以及恒河猴的血清抗体中和试验.结果 佐剂蛋白组产生了较强的抗rMOMP反应和高水平细胞因子.淋巴细胞特异性增殖反应、迟发型超敏反应明显强于对照组,抗体中和试验蛋白佐剂组血清能抑制D/E/H/L2型沙眼衣原体生长.结论 沙眼衣原体rMOMP能刺激恒河猴产生有效的交叉免疫.     

衣原体GPIC噬菌体衣壳蛋白Vp1的克隆、表达和鉴定

目的获得衣原体GPIC噬菌体衣壳蛋白Vp1基因及其蛋白。方法提取噬菌体φCPG1及其核酸，并对Vp1克隆、表达，鉴定。结果所获得的Vp1基因片段经测序，长度为1661bp，检索确认其序列与Genebank一致。对重组质粒进行诱导表达，SDS-PAGE和蛋白质印迹均显示获得相对分子质量约62000的衣原体GPIC噬菌体衣壳蛋白Vp1。结论成功获得了噬菌体衣壳蛋白Vp1。为进一步研究打下基础。     

急性心肌梗死患者溶栓治疗对患者血浆脑钠肽水平的影响

目的观察急性心肌梗死（AMI）患者应用瑞替普酶溶栓再通患者与未通患者血浆脑钠肽（BNP）、肌钙蛋白I（cTnI）浓度及对左心室功能的影响。方法AMI患者95例,溶栓治疗再通组65例,溶栓治疗未再通组30例。检测入院即刻、入院后24h、48h、7d、14d、28d血浆BNP及cTnI浓度并进行比较,同时比较入院后第3～4d、第28d左室射血分数（LVEF）并计算它的变化量（ΔLVEF）。结果入院时2组患者BNP浓度差异无统计学意义（P〉0.05）,入院后24h、48h、7d、14d、28d溶栓再通组BNP浓度显著低于溶栓治疗未再通组（P〈0.01）;入院后24h、48h、7d溶栓再通组cTnI浓度显著低于溶栓治疗未再通组（P〈0.01）,入院第3～4d两组患者LVEF差异无显著性（P〉0.05）,入院第28d再灌注组LVEF和ΔLVEF显著高于未再灌注组（P〈0.01）。结论溶栓治疗再通能显著降低AMI患者血浆BNP及cTnI水平,同时改善左室功能。血浆BNP、cTnI浓度可作为溶栓治疗再通的观察指标。