狂犬病疫苗接种人群中狂犬病病毒中和抗体水平分析

目的 通过分析狂犬病疫苗接种人群中狂犬病病毒RABV中和抗体特征,为狂犬病防控提供依据。方法 收集2019-2020年杭州市职业病防治院犬伤后暴露预防接种门诊抗体检测者信息,经筛选后确定157例RABV中和抗体检测者作为研究对象,采用快速荧光灶抑制试验检测狂犬病毒中和抗体,利用横断面研究方法对狂犬病毒中和抗体水平进行人群特征分析。结果 研究对象中,初种者98人,复种者59人,复种者中2-1-1程序比5针法产生的抗体水平更高,其余特征均无统计学差异(P>0.05)。除1例初种者中和抗体效价低于0.5 IU/mL,其余检测者中和抗体效价均高于0.5 IU/mL。狂犬病毒中和抗体水平时间趋势分析显示,狂犬病毒中和抗体水平与时间呈负相关,回归方程为(F=4.974,P=0.031),标准化回归系数为-0.322(t=-2.230,P=0.031)。结论 复种者狂犬病毒中和抗体水平与免疫程序有关,2-1-1程序对人体再次免疫应答的效果优于5针法。接种疫苗2年后部分个体出现失保护状态,再次暴露有必要全程接种狂犬病疫苗。     

BCG-CpG-DNA佐剂人用狂犬病疫苗(MRC-5细胞)在食蟹猴体内的免疫原性

目的 评价BCG-CpG-DNA佐剂人用狂犬病疫苗(MRC-5细胞)在食蟹猴体内的免疫原性。方法 将食蟹猴随机分为对照组、3针组及4针组,经后肢肌肉免疫,1.0 mL/剂。3针组及4针组均接种BCG-CpG-DNA佐剂人用狂犬病疫苗,3针组于0 d、7 d、21 d各接种1剂,4针组于0 d、3 d、7 d、14 d各接种1剂;对照组接种市售国产同类疫苗,于0 d注射2剂、7 d及21 d各1剂。初免前(0 d)及初免后7 d、14 d、28 d、35 d、49 d、63 d后肢隐静脉采血,分离血清及外周血淋巴细胞。快速荧光灶抑制实验(Rapid Fluorescent focus Inhibition Test, RFFIT)及酶联免疫斑点检测技术(Enzyme-linked Immunospot Assay, ELISPOT)分别检测血清中抗狂犬病病毒中和抗体水平和外周血淋巴细胞中细胞因子IFN-γ、IL-2的表达水平。结果 食蟹猴血清中和抗体水平于免疫前均呈阴性(抗体效价<0.5 IU/mL),3针组和对照组在初免后7～28 d呈上升趋势,于28 d达最高,4针组在14 d...     

HIV抗体阴性结核性脑膜炎患者治疗前后外周血CD细胞水平的变化情况

目的: 分析HIV抗体阴性结核性脑膜炎(tuberculous meningitis,TBM)患者抗结核治疗前后外周血CD细胞水平的变化及临床意义。方法: 采用前瞻性研究的方法,选取2016年3月至2020年2月于新疆医科大学第八附属医院就诊且符合入组标准的102例TBM患者作为研究对象,其中,临床分期Ⅰ期22例、Ⅱ期37例、Ⅲ期43例。于研究对象抗结核治疗前及治疗2周后,检测其外周血CD3⁺、CD4⁺、CD8⁺T细胞计数及CD4⁺/CD8⁺T细胞比值。分析研究对象治疗前后外周血CD细胞水平变化情况,及其与TBM临床分期和患者治疗转归的相关性。结果: 抗结核治疗2周后,研究对象CD3⁺、CD4⁺ T细胞计数和CD4⁺/CD8⁺ T细胞比值[分别为(808.27±183.25)×10⁶个/μl、(413.09±134.53)×10⁶个/μl和1.23±0.29]均明显高于治疗前[分别为(631.38±150.14)×10⁶个/μl、(366.78±98.03)×10⁶个/μl和0.99±0.23],差异均有统计学意义(t=7.541,P=0.000;t=2.809,P=0.005;t=6.548,P=0.000)。抗结核治疗2周后,Ⅲ期TBM患者的CD3⁺、CD4⁺T细胞计数与CD4⁺/CD8⁺T细胞比值[分别为(613.23±140.29)×10⁶个/μl、(342.53±98.36)×10⁶个/μl、0.95±0.18]明显低于Ⅱ期患者[分别为(753.33±153.47)×10⁶个/μl、(399.57±112.26)×10⁶个/μl、1.22±0.21]和Ⅰ期患者[分别为(989.23±194.35)×10⁶个/μl、(501.11±139.25)×10⁶个/μl、1.42±0.31],差异均有统计学意义(F值分别为40.875、13.372、32.151,P值均为0.000)。抗结核治疗2周后,病情好转患者的CD3⁺、CD4⁺ T细胞计数与CD4⁺/CD8⁺T细胞比值[分别为(941.25±204.17)×10⁶个/μl、(487.35±134.25)×10⁶个/μl、1.36±0.31]明显高于病情恶化者[分别为(683.43±155.76)×10⁶个/μl、(389.64±120.38)×10⁶个/μl、1.02±0.19],差异均有统计学意义(t=4.206,P=0.000;t=2.394,P=0.018;t=3.698,P=0.000)。结论: TBM患者抗结核治疗2周后,免疫功能明显改善,且病情好转患者免疫功能改善程度好于病情恶化者。TBM患者临床分期越高,免疫功能受损越严重,对调整治疗方案的需求越高。     

替诺福韦治疗慢性乙型肝炎患者疗效及安全性分析*

目的 观察比较应用替诺福韦（TDF）和恩替卡韦（ETV）治疗慢性乙型肝炎（CHB）患者的疗效和安全性。方法 2015年1月~2016年12月我中心收治的初始治疗的CHB患者60例,采用随机数字表法将患者分为两组,每组30例,分别给予TDF或ETV治疗,观察96 w,比较两组血清HBV DNA和丙氨酸氨基转移酶（ALT）水平和阴转率或复常率及不良反应发生情况。结果 在治疗前、治疗第12 w、24 w、48 w、72 w和96 w,TDF治疗组患者血清HBV DNA水平分别为（7.5±0.9）lg IU/mL、（1.7±1.4）lg IU/mL、（1.1±1.0）lg IU/mL、（1.0±0.8）lg IU/mL、（0.7±0.6）lg IU/mL和（0.6±0.5）lg IU/mL,ETV治疗组则分别为（6.8±1.5）lg IU/mL、（2.1±1.1）lg IU/mL、（1.8±1.1）lg IU/mL、（1.0±0.7）lg IU/mL、（0.8±0.8）lg IU/mL和（0.9±0.8）lg IU/mL,在治疗24 w前,TDF治疗组血清HBV DNA水平显著低于ETV治疗组,差异具有统计学意义（P<0.05）; TDF治疗组血清ALT水平分别为（205.8±23.8） U/L、（80.7±8.6） U/L、（49.7±12.5）U/L、（46.2±6.7）U/L、（42.5±6.5）U/L和（41.8±2.4）U/L,ETV治疗组则分别为（199.0±25.9） U/L、（99.8±11.0）U/L、（59.3±9.4）U/L、（53.9±8.8）U/L、（44.62±7.4）U/L和（42.7±4.5）U/L,在治疗24 w前,TDF组血清ALT水平显著低于ETV组（P<0.05）;在治疗第12 w、24 w、48 w、72 w和96 w,TDF组病毒学应答率分别为50.0%、70.0%、73.3%、80.0%和96.7%,而ETV组则分别为23.3%、56.7%、60.0%、70.0%和93.3%,在治疗12 w时,TDF组显著高于ETV组（P<0.05）;TDF组血清ALT复常率分别为63.3%、96.7%、96.7%、100.0%和100.0%,ETV组则分别为50.0%、86.7%、96.7%、96.7%和100.0%,两组无统计学差异（P>0.05）。结论 在当前情况下,应用TDF或ETV任一药物治疗CHB初治患者,均有很好的近期疗效,TDF显示出更早的病毒学应答率,值得进一步观察。     

早期肠内免疫微生态营养对原发性肝癌患者术后临床疗效的影响

目的 观察原发性肝癌患者行肝癌切除术后早期给予肠内免疫微生态营养支持的临床疗效。方法 120例行原发性肝癌切除术患者被随机分为观察组60例和对照组60例,对照组术后常规肠内营养,观察组在对照组的基础上在肠内营养物中加入L-谷氨酰胺、L-精氨酸和双歧三联活菌,比较两组患者术后肝功能、凝血功能、血清免疫球蛋白、内毒素、肿瘤坏死因子α（TNF-α）、白介素6 （IL-6）、外周血CD4⁺和CD8⁺淋巴细胞、一般状况和胃肠道功能。结果 观察组患者术后第7天血清白蛋白、前白蛋白、血清IgA、IgG、IgM、血CD4⁺和CD8⁺淋巴细胞相对计数分别为（41.65±4.58） g/L、（196.65±25.64） mg/L、（2.12±0.56） g/L、（9.65±0.86） g/L、（0.98±0.25） g/L、（0.42±0.14）和（0.32±0.05）,均显著高于对照组的[（35.52±5.38） g/L、（178.38±30.54） mg/L、（1.83±0.48） g/L、（8.22±0.68） g/L、（0.80±0.25） g/L、（0.32±0.06）和（0.24±0.08）,P均<0.05];国际标准化比值、总胆红素、直接胆红素、内毒素、TNF-α和IL-6分别为（1.05±0.16）、（20.34±8.65）μmol、（11.35±2.38）μmol/L、（1.82±0.25）ng/L、（258.62±28.65） pg/mL和（156.24±9.38） pg/mL,均显著低于对照组的[（1.46±0.18）、（38.65±9.46）μmol、（19.35±4.28）μmol/L、（2.14±0.15） ng/L、（314.65±41.28） pg/mL和（171.35±20.38） pg/mL,P均<0.05];观察组患者术后排气、排便、下床活动、进食半流质、住院时间分别为（44.52±8.36）h、（74.35±14.68）h、（30.17±9.65）h、（85.34±16.25）h和（10.25±1.88） d,均显著短于对照组的[（68.65±10.48） h、（112.58±14.95） h、（41.35±10.38）h、（115.36±25.68）h、（14.58±2.36） d,P均<0.01]。结论 在原发性肝癌肝切除术后早期给予肠内免疫微生态营养支持有助于改善患者营养状态,提高免疫功能,减轻炎症反应和肝功能损害,促进患者早期康复。     

减毒沙门氏菌投递的TGEV S基因疫苗口服免疫猪的动态规律

目的 研究TGEV S基因疫苗SL7207(PVAXD-TS)口服免疫仔猪后的动态免疫诱导规律。方法 将口服疫苗SL7207(PVAXD-TS)、空载体菌株SL7207(PVAXD)以1.6×10¹¹CFU/头的剂量口服接种20日龄仔猪,以PBS为空白对照,2周后以2.0×10¹¹CFU的剂量加强免疫。分别测定0、2、4、6和8周的血清IgG、IgA、TGEV中和抗体、细胞免疫反应 IL-4、IFN-γ、外周血淋巴细胞增殖情况。结果 仔猪口服免疫后第4周即可检测抗TGEV的特异性IgG和粪黏液IgA抗体;在免疫后第6周IgA、IgG、IL-4、IFN-γ抗体和外周血T淋巴细胞增殖与SL7207(PVAXD)、PBS间均存在统计学差异(P<0.01),并在第6周达到最高值;中和实验显示该疫苗诱导的血清IgG具有中和活性。结论 证实以减毒沙门菌为载体的TGEV S基因疫苗口服免疫仔猪有较好的免疫原性。     

双黄连对MHV-3诱导的暴发型肝衰竭小鼠的保护作用研究*

目的 探讨双黄连对病毒感染诱导的暴发性肝衰竭小鼠的保护作用。方法 取BABL/cJ小鼠36只,随机均分为对照组、模型组和双黄连处理组。取滴度为1×10⁶ PFU/ml的3型鼠肝炎病毒（MHV-3）,采用高压水注射法经小鼠尾静脉注入小鼠体内,诱导暴发性肝衰竭模型。在造模后给予10%双黄连或生理盐水灌胃治疗7 d。采用RT-PCR法检测小鼠肝组织IP-10 mRNA和血清MHV-3 DNA水平,并检测血清转化生长因子-βl（TGF-βl）、白细胞介素-10（IL-10）、层黏连蛋白（LN）、透明质酸（HA）和肝功能指标。结果 在治疗7 d,模型组和双黄连组小鼠血清谷丙转氨酶（ALT）分别为（229.03±30.56） IU/L和（22.83±6.02） IU/L,谷草转氨酶（AST）为（139.01±11.75） IU/L和（32.45±4.59） IU/L,LN分别为（99.86±10.57） nmol/L和（57.02±7.24） nmol/L,HA为117.05±16.72） nmol/L和（53.01±11.35） nmol/L,TGF-βl分别为（319.46±39.12） pg/ml和（93.87±11.20）pg/ml,IL-10为25.70±3.87） pg/ml和（1.07±0.09） pg/ml, MHV-3 DNA分别为12.47±3.05） lg copies/mL和（1.24±0.10）lg copies/mL,肝组织IP-10 mRNA分别为【（25.70±3.87）和（12.79±3.08）,P<0.05】。结论 双黄连有一定的抗MHV-3作用,可能是通过抑制肝内IP-10 mRNA水平,减少IL-10对肝细胞的炎性损伤,阻止了肝组织纤维化而促进了肝功能的恢复。     

慢性丙型肝炎患者外周血滤泡辅助性T淋巴细胞表面PD-1受体表达及意义研究*

目的 探讨不同血清病毒载量的慢性丙型肝炎（CHC）患者外周血滤泡辅助性T淋巴细胞（Tfh）表面程序性死亡受体-1（PD-1）表达情况。方法 根据血清HCV RNA水平不同,将180例CHC患者分为低病毒载量组76例,3 lg copies/ml≤血清HCV RNA<6 lg copies/ml和高病毒载量组104例,血清HCV RNA≥6 lg copies/ml。比较两组外周血Tfh细胞百分比、Tfh细胞表面PD-1阳性率、外周血T、B淋巴细胞亚群、外周血CD4⁺T淋巴细胞和CD8⁺T淋巴细胞表面PD-1表达和血清白细胞介素21（IL-21）水平的差异。结果 低病毒载量组和高病毒载量组血清HCV RNA水平分别为（4.5±1.2）lg copies/ml和（6.4±0.7）lg copies/ml,Tfh细胞表面PD-1阳性百分比分别为（26.2±2.2）%和（37.2±1.1）%,Tfh细胞百分比分别为（7.9±0.7）%和（5.1±0.4）%,血清IL-21水平分别为（46.8±1.3） ng/L和（21.7±1.1） ng/L,差异均有统计学意义（P<0.01）;低病毒载量组和高病毒载量组外周血CD4⁺T细胞百分比分别为（51.1±4.6）%和（37.6±4.4）%,CD8⁺T细胞百分比分别为（24.0±3.1）%和（31.7±3.9）%, CD4⁺T/CD8⁺T细胞比值分别为（3.3±0.2）和（2.3±0.1）,CD19⁺B细胞百分比分别为（16.7±3.9）%和（11.8±3.2）%,差异具有统计学意义（P<0.01）;低病毒载量组和高病毒载量组外周血CD4⁺T细胞表面PD-1阳性率分别为（10.1±2.3）%和（2.4±0.6）%],CD8⁺T细胞表面PD-1阳性率分别为（6.3±2.2）%和（1.0±0.3）%,差异也具有统计学意义（P<0.01）。结论 不同血清病毒载量的CHC患者外周血Tfh和T淋巴细胞亚群以及血清白细胞介素21水平存在显著差异,进一步探讨它们对病情、抗病毒治疗应答和预后的关系,将具有十分重要的临床意义。     

围手术期应用乌司他丁治疗对肝细胞癌患者肝切除术后外周血Th22细胞百分比和血浆白细胞介素-22的影响*

目的 观察围手术期应用乌司他丁对肝细胞癌（HCC）肝切除手术患者术后外周血Th22细胞百分比和血浆白细胞介素（IL）-22水平的影响。方法 2015年7月~2017年4月解放军昆明总医院普通外科行肝切除手术的HCC患者88例,术后51例接受常规治疗,37例另加乌司他丁治疗7 d。另选30例健康人作为对照。分离外周血单个核细胞（PBMC）,使用流式细胞术检测CD3⁺CD4⁺IL-22⁺的Th22细胞百分比,采用ELISA法检测血浆IL-22水平。对符合正态分布的计量资料比较采用独立样本t检验,对于非正态分布的计量资料比较采用Mann-Whitney检验。结果 HCC患者外周血Th22细胞百分比为（1.4±0.3） %,显著高于健康人【（0.8±0.1） %,P<0.0001】;HCC患者血浆IL-22水平为（116.9±32.6） pg/ml,亦显著高于健康人【（42.1±18.2） pg/ml,P<0.0001】;在术后3 d,乌司他丁治疗组和常规治疗组血清ALT分别为【219.4（40.1,510.9） IU/L和450.6（56.1,820.7） IU/L,P<0.05】,血清AST分别为【108.8（82.5,439.1） IU/L和257.3（115.3,7265） IU/L,P<0.01】;在术后7 d,乌司他丁治疗组和常规治疗组血清ALT分别为【72.4（25.6,471.5） IU/L和115.4（35.7,625.2） IU/L,P<0.05】,血清AST分别为【61.4（29.4,351.4） IU/L和90.5（45.5,293.8） IU/L,P<0.05】;术后3 d和术后7 d,乌司他丁治疗组外周血Th22细胞百分比分别为（1.2±0.4）%和（1.1±0.3）%,较常规治疗组显著降低【分别为（1.3±0.3）%,P<0.05和（1.3±0.2） %,P<0.05】;血浆IL-22水平分别为（98.1±20.1） pg/ml和（94.1±24.8） pg/ml,亦较常规治疗组显著降低 【分别为（109.7±29.8） pg/ml,P<0.05和（110.7±37.1）%,P<0.05】。结论 肝切除术后应用乌司他丁治疗可降低外周血Th22细胞比例和血浆IL-22水平,对肝功能具有一定的保护作用     

射频消融联合TACE治疗原发性肝癌患者疗效及对外周血T细胞亚群的影响

目的 探讨射频消融联合TACE治疗原发性肝癌患者的疗效及其对外周血T细胞亚群的影响。方法 2014年1月~2016年2月间于我院接受TACE治疗的原发性肝癌患者97例,按照TACE术后是否接受射频消融治疗,将患者分为联合组52例和TACE组45例。采用ELISA法检测血清细胞因子。对两组患者的治疗效果、治疗前后T细胞亚群变化情况以及治疗后生存质量进行比较。结果 联合组总有效率为67.31%,显著高于TACE组的40.00% （P<0.05）;联合组疾病控制率为90.38%,显著高于TACE组的73.33%（P<0.05）;联合组治疗后3个月外周血CD3⁺T细胞、CD4⁺T细胞、CD4⁺/CD8⁺比值分别为（68.40±10.20）%、（45.76±6.83）%和（1.96±0.31）,均明显高于TACE组【（56.14±6.75）%、（33.27±5.16）%和（1.21±0.22）,P<0.05】,联合组CD8⁺T细胞百分比为（22.08±2.55）%,显著低于TACE组【（28.42±3.97）%,P<0.05】;联合组患者治疗后3个月外周血IL-6、IL-8和TNF-α水平分别为（129.45±67.14） pg/mL、（0.49±0.13） ng/mL和（134.78±88.20） pg/mL,均明显低于TACE组【（165.30±65.91）pg/mL、（0.72±0.20） ng/mL和（200.43±84.02）pg/mL,P<0.05】;联合组患者生活质量改善率为55.77%,显著高于TACE组的40.00%（P<0.05）,联合组患者并发症发生率为73.08%,与TACE组的71.11%相比无统计学差异（P>0.05）。结论 射频消融联合TACE治疗原发性肝癌患者近期疗效较好,可改善患者免疫功能,提高生存质量。     

Vero细胞口服狂犬病疫苗的研究

本试验采用ＣＴＮ－１株经Ｖｅｒｏ细胞传１５代，病毒滴度均在７．０～８．０ｌｏｇＬＤ５０／ｍｌ之间。给８只犬分别口服８．１ｌｏｇＬＤ５０的疫苗３０天的中和抗体几何平均效价为１：１１０，中和抗体的平均国际单位为２．５９ＩＵ／ｍｌ，阳转率为１００％；给４只犬分别口服７．４３ｌｏｇＬＤ５０的疫苗，阳转率为５０％，中和抗体的平均国际单位为２．４８ＩＵ／ｍｌ。选用３５－６０日龄的乳犬１６只口服１０个剂量的疫苗，并在口服后５、１０、２０天、３个月各杀３只犬取脑及唾液腺分别在ＢＨＫ２１细胞上盲传２代，同时在昆明种小白鼠乳鼠脑内盲传２代，进行病毒分离，均为阴性，余下犬观察１２个月均正常。总之该口服狂犬病疫苗对犬具有较好的安全性及口服免疫原性。     

对乙酰氨基酚诱导的急性肝损伤小鼠肝组织糖基转移酶Colgalt2表达的变化*

目的 探讨对乙酰氨基酚（APAP）诱导的急性肝损伤小鼠肝组织糖基转移酶Colgalt2表达的变化。方法 取C57BL/6小鼠100只,采用腹腔注射APAP溶液法建立小鼠急性肝损伤模型。首先建立不同浓度干预组,每组10只,分别给予【0 mg·kg^-1 APAP（对照组）、100 mg·kg^-1 APAP处理组、250 mg.kg^-1 APAP处理组、500 mg·kg^-1 APAP处理组和1000 mg·kg^-1 APAP处理组】,再建立不同时间干预组,每组10只,均给予500 mg·kg^-1 APAP腹腔注射,分别观察【0 h APAP（对照组）、2 h APAP处理组、4 h APAP处理组、8 h APAP处理组和12 h APAP处理组】。采用Real-time quantitative PCR 和Western blot法检测肝组织对糖基转移酶Colgalt2 基因及其蛋白表达情况。结果 在250 mg·kg^-1、500 mg·kg^-1和1000 mg·kg^-1 APAP处理组,血清ALT分别为（1360.5±189.8）IU/L、（3191.2±118.6）IU/L和（5022.7±234.6）IU/L,AST分别为（2147.1±133.4）IU/L、（3628.8±107.9）IU/L和（5854.5±295.1）IU/L,较对照组显著升高（P<0.05）;在4 h、8 h和12 h APAP处理组,血清ALT水平有类似的变化;在100 mg·kg^-1、250 mg·kg^-1、500 mg·kg^-1、1000 mg·kg^-1 APAP处理组,肝组织Colgalt2基因水平较对照组显著升高;在2 h、4 h、8 h和12 h APAP处理组,肝组织Colgalt2基因水平较对照组逐渐显著升高,肝组织Colgalt2蛋白表达水平也随APAP干预剂量的增加和干预时间的延长也呈逐渐增强趋势。结论 肝组织糖基转移酶Colgalt2在对乙酰氨基酚诱导的急性肝损伤小鼠发病过程中发挥着重要的作用。     

Calculating rabies virus neutralizing antibodies titres by flow cytometry

The determination of the rabies neutralizing antibody (VNA) response after immunization against rabies is an acceptable index of the efficacy of a vaccine and a successful treatment. Several tests have been developed in attempt to improve the assessment of VNA, from mice inoculation to cell-culture fluorescence inhibition tests. All of them, however, present special difficulties in terms of reading or accuracy. The present study describes a neutralization test performed in cell-culture appraised by flow cytometry (FC). Serial dilutions of the serum samples were mixed in vitro with rabies virus before the addition of BHK-21 cells. After 24h-incubation, cells were released by trypsin treatment, fixed and permeabilized with a p-formaldehyde solution and stained with a rabies virus nucleocapsid protein-specific antibody conjugate. The percentage of virus infection inhibition caused by specific antibodies present in the serum were evaluated in a Beckton Dickinson FACSCalibur flow cytometer. A correlation curve between the IU/ml content and the percentage of infective inhibition was built with a reference serum and the VNA titers of serum samples were obtained by extrapolation. Titers obtained by FC and standard test showed an effective pairing results (p < 0.01), with a correlation coefficient (r) = 0.7. These results permit to envisage the FC as a suitable technique to evaluate VNA in sera from immunized animals and likely in human serum samples. Nevertheless, new studies comparing FC to gold-standard techniques are required for determining the FC values of Sensibility and Specificity.     

Evaluation of the safety, immunogenicity, and pharmacokinetic profile of a new, highly purified, heat-treated equine rabies immunoglobulin, administered either alone or in association with a purified, Vero-cell rabies vaccine

A clinical evaluation of a new, purified, heat-treated equine rabies immunoglobulin (PHT-Erig), F(ab′)₂ preparation, was carried out in Thailand and in the Philippines—two countries where rabies is endemic. An initial prospective, randomised, controlled trial (Study 1), compared the safety and pharmacokinetics (serum concentrations of rabies antibodies) after administration either of PHT-Erig or of a commercially-available, equine rabies immune globulin (Erig PMC). A second trial (Study 2) simulated post-exposure rabies prophylaxis by using a reference cell culture vaccine, the purified Vero-cell rabies vaccine (PVRV), administered in association with either Erig PMC or PHT-Erig. In Study 1, 27 healthy, Thai adults received a 40 IU kg⁻¹ dose of either Erig PMC (n=12) or PHT-Erig (n=15) via the intramuscular (IM) route; half of the dose was injected into the deltoid area and the other half into the buttocks. Serum for rabies antibody determination and F(ab′)₂ concentration was collected at hours (H) 0, 6 and 12, and on day (D) 2, 3, 4, 6, 8, 10, 12 and 15. Both products were safe, with no serious adverse events, and in particular, no anaphylactic reactions or serum sickness was reported. A statistical comparison of the pharmacokinetic parameters did not demonstrate bioequivalence of the two products. Nonetheless, the relative bioavaibility of 93% and the similar absorption rates suggest the pharmacokinetic profiles of Erig and PHT-Erig are similar. The antibody level in either group were low throughout the 15-day study period. The geometric mean titer (GMT) values ranged from group 0.027–0.117 IU ml⁻¹ in the Erig group and from 0.029 to 0.072 IU ml⁻¹ in the PHT-Erig. There was no significant difference between the evolution of GMT values for the two groups. In Study 2, 71 healthy volunteers received 40 IU kg⁻¹ via the intramuscular route of either Erig PMC (n=36) or PHT-Erig (n=35) on D0, in association with five doses of PVRV on D0, D3, D7, D14 and D28. The safety evaluation was performed during the 28-day follow-up and serum samples for anti-rabies antibody titration were collected on D0 (before injection) D3, D7, D14 and D28. No serious reactions were reported in either group. In particular, no immediate (anaphylactic type) or delayed (serum sickness) allergic reactions were observed. Over the 28-day follow-up period, GMT profiles of the two groups were statistically equivalent. On D14, 100% of the subjects had protective antibody titers (anti-rabies antibodies ≥0.5 IU ml⁻¹, which is the WHO-recommended level of seroconversion), and Erig PMC and PHT-Erig were indistinguishable according to the clinical definition chosen. On D28, the GMT values were 33.2 IU ml⁻¹ (95% CI, 23.8–46.1 IU ml⁻¹) in the Erig PMC/PVRV group and 31.4 IU ml⁻¹ (95% confidence interval, CI, 23.4–42.2 IU ml⁻¹) in the PHT-Erig/PVRV group, showing evidence of adequate vaccine-induced antibody responses in both groups. The increased purity, the heat-treatment step introduced in the manufacturing process of PHT-Erig, and the good clinical results substantiate the use of this new generation, purified equine F(ab′)₂ preparation in the post-exposure prophylaxis of rabies.     

Failure of pre- and postexposure rabies vaccinations in a child infected with HIV.

We report the case of a 6-y-old HIV-infected girl with severe immune deficiency who failed to respond to intramuscular pre-exposure rabies vaccination using human diploid cell rabies vaccine on days 0, 7 and 28. She also failed to respond to an intradermal postexposure rabies regimen using purified verocell rabies vaccine at 4 sites on days 0, 3 and 7 and at 2 sites on days 30 and 90 (double the usual regimen). Sequentially monitored rabies neutralizing antibody titers were below the WHO minimum acceptable level (> 0.15 IU/ml) in all specimens. Rabies prevention in HIV-infected persons with severe immune suppression requires further study.     

Immune responses of humans to a human diploid cell strain of rabies virus vaccine: lymphocyte transformation, production of virus-neutralizing antibody, and induction of interferon.

Lymphocyte transformation, production of neutralizing antibody, and interferon activity of 20 healthy volunteers in response to a human diploid cell strain (HDCS) of rabies virus vaccine were studied. Ten vaccines received 1.0 ml of HDCS vaccine on days 0, 3, 7, and 14, and five of these volunteers received, in addition, 20 international units of human rabies immune globulin/kg of body weight on day 0. All 10 volunteers developed high titers of neutralizing antibody, and eight of the 10 had lymphocytes that were immunologically stimulated by HDCS rabies virus antigen. The interferon responses of eight vaccines to 1.0 ml of HDCS vaccine given intramuscularly on days 0 and 28 and of two vaccines to 1.0 ml of vaccine given intradermally in eight body sites on the same days were measured; low levels of interferon-like activity were found in eight of nine volunteers after the first vaccination, and no activity was found upon revaccination. The high titers of neutralizing antibody that developed did not correlate with lymphocyte stimulation or interferon-like activity, but it is true that all 20 volunteers did develop high titers of neutralizing antibody after 1.0 ml of HDCS vaccine was given intradermally.     

Isolation and Characterization of Cross-Reactive Human Monoclonal Antibodies That Potently Neutralize Australian Bat Lyssavirus Variants and Other Phylogroup 1 Lyssaviruses

Australian bat lyssavirus (ABLV) is a rhabdovirus that circulates in four species of pteropid bats (ABLVp) and the yellow-bellied sheath-tailed bat (ABLVs) in mainland Australia. In the three confirmed human cases of ABLV, rabies illness preceded fatality. As with rabies virus (RABV), post-exposure prophylaxis (PEP) for potential ABLV infections consists of wound cleansing, administration of the rabies vaccine and injection of rabies immunoglobulin (RIG) proximal to the wound. Despite the efficacy of PEP, the inaccessibility of human RIG (HRIG) in the developing world and the high immunogenicity of equine RIG (ERIG) has led to consideration of human monoclonal antibodies (hmAbs) as a passive immunization option that offers enhanced safety and specificity. Using a recombinant vesicular stomatitis virus (rVSV) expressing the glycoprotein (G) protein of ABLVs and phage display, we identified two hmAbs, A6 and F11, which completely neutralize ABLVs/ABLVp, and RABV at concentrations ranging from 0.39 and 6.25 µg/mL and 0.19 and 0.39 µg/mL respectively. A6 and F11 recognize overlapping epitopes in the lyssavirus G protein, effectively neutralizing phylogroup 1 lyssaviruses, while having little effect on phylogroup 2 and non-grouped diverse lyssaviruses. These results suggest that A6 and F11 could be effective therapeutic and diagnostic tools for phylogroup 1 lyssavirus infections.     

Efficacy of Hepatitis B vaccine in those who lost Hepatitis B surface antigen during follow-up: Efficacy of HBV vaccine in those who lost HBsAg

Background

The level of HBsAg in some chronic hepatitis B virus (HBV)-infected individuals may decline over time so that it is not detectable in serum.

Objective

To assess the efficacy of HBV vaccine in those who lost their HBsAg without seroconverssion to anti-HBs antibody.

Patients and Methods

From April 1993 to December 2008, of 1603 chronic HBV-infected individuals, 34 (22 men and 12 women) became HBsAg-negative in follow-up visits, with no detectable anti-HBs antibody and HBV DNA in their sera. They received HBV vaccination at 0, 1 and 6 months (case group). Fifty-two subjects (30 men and 22 women) who were negative for HBsAg, anti-HBs and anti-HBc antibody, received HBV vaccination according to the said schedule (control group). Anti-HBs antibody was assessed one month after the last dose of vaccination in the both groups.

Results

The mean±SD age of the case and control groups was 38±12.7 and 33.4 ± 8.6 years, respectively (p = 0.07). The sex distribution between these two groups were similar (p = 0.652). The mean ± SD years of follow-up for the case group was 7.6 ± 4.5 years. Anti-HBs antibody level ≥ 10 IU/L was found in 8 (24%) subjects in the case group and in 45 (87%) in the control group (p < 0.001). The mean±SD anti-HBs antibody level in the case group was 68 ± 32.66 and in the control group 344.6 ± 38.9 IU/L (p < 0.001).

Conclusions

We found that nearly 24% of chronic HBsAg-positive subjects who lost their HBsAg responded to HBV and the remaining cases need to be followed for occult HBV infection.