狂犬病免疫球蛋白应用研究进展

中国是全球第二大狂犬病流行国家,每年有数以百万计的狂犬病Ⅲ级暴露案例需要联合应用狂犬病免疫球蛋白和狂犬病疫苗。狂犬病免疫球蛋白价格较昂贵,使用方法较复杂,在狂犬病Ⅲ级暴露后处置中使用率长期以来偏低。降低狂犬病免疫球蛋白的使用剂量可以减少狂犬病Ⅲ级暴露后处置费用。简化狂犬病免疫球蛋白的使用方法可以使狂犬病Ⅲ级暴露后处置更加便于实施。降低狂犬病免疫球蛋白的使用剂量和简化狂犬病免疫球蛋白的使用方法有助于提高狂犬病Ⅲ级暴露后处置中狂犬病免疫球蛋白的使用率。狂犬病免疫球蛋白对狂犬病疫苗免疫效果的影响研究中存在矛盾的结论,探索狂犬病免疫球蛋白影响狂犬病疫苗免疫效应的机制有助于解释狂犬病免疫球蛋白对狂犬病疫苗免疫效果影响的复杂性。本文系统回顾关于狂犬病免疫球蛋白应用研究的进展,以期为制订新的具备实际操作可行性的狂犬病免疫球蛋白应用准则提供参考。     

马狂犬病免疫球蛋白副反应

<正>新近一项使用法国巴斯德疫苗研究所生产的马狂犬病免疫球蛋白(ERIG)的485人的前瞻性研究表明,副反应率较低(疫苗批号:5212,蛋白含量为0.87%)。未发生过敏反应,仅有0.8%的暴露者发生血清病。我们于1987年共给3812例狂犬病暴露者注射ERIG;所用制品的84%来自巴斯德疫苗研究所,而16%是由瑞土血清及疫苗研究所(伯尔尼)生产的。上述前瞻性研究中有8例(1.64%)使用伯尔尼ERIG。在发展中国家,不能给大多数暴露者提供人狂犬病免疫球蛋白(HRIG),在泰国亦很少使用。不过,我们研究所对ERIG皮试阳性的暴露者,每当有可能时都使用价格高为ERIG 10倍的HRIG。在1987年及1988年     

人狂犬病免疫球蛋白使用效果观察

为了解人狂犬病免疫球蛋白的作用效果,我们将观察对象随机分成了A、B、C三组,分别采用三种措施进行狂犬病的预防治疗,即：A组联合使用狂犬病疫苗与人狂犬病免疫球蛋白;B组联合使用狂犬病疫苗与抗狂犬病血清(马源);C组仅注射狂犬病疫苗,并采用小鼠中和试验对这三组成员在免疫后3、7、14、45天及1年时的中和抗体水平进行检测。结果表明：狂犬病疫苗与人狂犬病免疫球蛋白或抗狂犬病血清联合使用,可使体内更早出现抗狂犬病的中和抗体。注射人狂犬病免疫球蛋白后未发生临床副反应。     

抗狂犬病毒中和抗体研究进展

狂犬病是一种人兽共患传染病,人和动物一旦发病后死亡率几乎百分之百,而有效的暴露后预防措施可以将死亡风险降至零。根据WHO推荐的狂犬病暴露后预防方案,一般狂犬病暴露者需要进行疫苗注射,严重者则需在进行疫苗注射的同时注射抗狂犬病毒中和抗体。常用的中和抗体有马抗狂犬病毒免疫球蛋白和人抗狂犬病毒免疫球蛋白,然而两者都存在引起过敏反应或血液疾病的风险。人源抗狂犬病毒中和抗体则因为具有安全性高、成本低、可量产等优点有望代替免疫球蛋白用于暴露后预防。基因工程抗体技术的发展加速了抗体人源化的进程。就抗狂犬病毒中和抗体的发展历程,不同类型中和抗体的优缺点以及中和抗体的未来研究方向作了综述及展望,以期为新一代狂犬疫苗的研发提供参考。     

世界卫生组织狂犬病专家会议报告

<正>一、前言 自从1983年第七次狂犬病专家委员会会议以来,对狂犬病研究和控制有许多新进展,因此有必要召开一系列专家国际会议来形成一些新建议。 由于过去三年发生一些注射过现代细胞培养疫苗而仍然得狂犬病的报告,因此有必要对这些病例进行认真的分析,并对现有WHO狂犬病暴露后治疗建议进行审查,有关暴露后处理的进一步改进的建议在本报告的第一部分介绍。另一重要问题是关     

用人体细胞制备狂犬病疫苗的进展和远景(续完)

<正>同时使用二倍体细胞疫苗和人狂犬病免疫球蛋白 狂犬病感染后,同时以被动抗体处理。被动抗体的辅助效果,首先在动物模型中被显示出来,然后动物血清被应用于人类。近来为了避免血清病,动物血清由HRIG(人狂犬病免疫球蛋白)所代替,早为人们所知,这种被动抗体与狂犬疫苗同时使用可以抑制免疫反应的效果,所以对HRIG和HDCV联合使用研究特别重要。     

健康教育在狂犬病预防控制中的价值分析

目的 分析健康教育在狂犬病预防控制中的应用价值.方法 选取2020年1～11月,因动物致伤至我院犬伤处置重点门诊就诊的200例患者作为研究对象,对其开展狂犬病预防相关知识的健康教育,比较健康教育开展前后患者对狂犬病相关知识的知晓情况.结果 经健康教育后,患者对狂犬病相关知识知晓率明显高于健康教育前,差异有统计学意义(P...     

Vero细胞衍生的新纯化疫苗的随机非劣性临床试验:评估暴露后肌内和皮内接种的狂犬疫苗Rabivax-S的免疫效果

<正>狂犬病是100%致死的疾病,但可用疫苗和免疫球蛋白进行预防。作者开发了Vero细胞狂犬病疫苗(Rabivax-S)并通过肌内(IM)和皮内(ID)途径接种评价了它的安全性和免疫原性。这是180名(5岁及以上)疑似狂犬病暴露者(WHO II类和III类各90人暴露)参与的随机非劣性对照研究。参与者或者接种Rabivax-S(1 mL IM;5剂),Rabivax-S(ID为0.1 mL;8剂量)或接种纯化     

狂犬病免疫球蛋白及疫苗对感染狂犬病绵羊的治疗效果

<正>抗狂犬马免疫球蛋白(ERIG)或人狂犬免疫球蛋白(HRIG)用于感染后的治疗已成功的建立。这是以动物实验和在人类中不断积累的临床研究及回顾分析为基础的。但是在人类,我们尚不清楚有任何可监控的预期研究。如何处理感染了狂犬病的家养动物仍处于争论中。最新研究表明,感染了狂犬病的狗只注射疫苗不能保护。WHO的一个工作组1988     

高效价冻干人用狂犬病疫苗暴露后免疫程序的研究

初步确定高效价冻干人用狂犬病疫苗（6．0IU／剂）暴露后免疫程序。制备高效价的冻干人用狂犬病疫苗（6．0IU／剂）,以狂犬病街毒CNX8601和BD06分别攻击小鼠和比格犬的咬肌,接种不同效价的狂犬病疫苗,以RFFIT法检测中和抗体,根据动物死亡情况,计算暴露后疫苗保护率,对不同效价的疫苗进行中和抗体测定和保护率统计分析。在以小鼠为实验动物的疫苗保护率研究中,冻干人用狂犬病疫苗（3．1IU／剂）0／3／7／14／28免疫程序的保护率为40．6％,高效价的冻干人用狂犬病疫苗（6．0IU／剂）0／3／14免疫程序的保护率为56．2％,中和抗体比较,P〈0．05,2组间有显著性差异;在以比格犬为实验动物的保护效果研究中,冻干人用狂犬病疫苗的保护率（3．1IU／剂）为70％;高效价的冻干人用狂犬病疫苗（6．0IU／剂）的保护率为80％,中和抗体的比较,P〉0．05,没有显著性差异。高效价冻干人用狂犬病疫苗暴露后免疫程序可初步确定为0、3、14d免疫。     

Novel, chimpanzee serotype 68-based adenoviral vaccine carrier for induction of antibodies to a transgene product

An E1-deletion-containing adenoviral recombinant based on the chimpanzee serotype 68 (AdC68) was developed to express the rabies virus glycoprotein. Mice immunized with this construct (AdC68rab.gp) developed antibodies to rabies virus and remained resistant to challenge with an otherwise lethal dose of rabies virus. In na?ve mice immunized intranasally, the rabies virus-specific antibody responses elicited by AdC68rab.gp were comparable with regard to both titers and isotype profiles to those induced by an adenoviral recombinant based on human serotype 5 (Adhu5) expressing the same transgene product. In contrast, subcutaneous immunization with the AdC68rab.gp vaccine resulted in markedly lower antibody responses to the rabies virus glycoprotein than the corresponding Adhu5 vaccine. Antibodies from AdC68rab.gp-immunized mice were strongly biased towards the immunoglobulin G2a isotype. The antibody response to the rabies virus glycoprotein presented by Adhu5rab.gp was severely compromised in animals preexposed to the homologous adenovirus. In contrast, the rabies virus-specific antibody response to the AdC68rab.gp vaccine was at most marginally affected by preexisting immunity to common human adenovirus serotypes, such as 2, 4, 5, 7, and 12. This novel vaccine carrier thus offers a distinct advantage over adenoviral vaccines based on common human serotypes.     

用Vero细胞制备马抗狂犬病血清抗原

以Vero细胞为基质制备马抗狂犬病血清用抗原,以期建立有效、经济、简便的抗原制备方法。用含10％马血清营养液对Vero细胞作适应性培养,接种狂犬病毒,以含1％～3％马血清营养液作维持液培养病毒,于第5,8天收获病毒液,经灭活、浓缩、离心等制成抗原。第5,8天收获病毒滴度可稳定在7．010gLD50／mL以上,灭活抗原具良好的抗原性,用NIH法测定效价达6．0IU／mL以上,可用作抗原生产抗狂犬病血清。     

Intravenous human rabies immunoglobulin for post-exposure prophylaxis: serum rabies neutralizing antibody concentrations and side-effects

The beneficial effect of passive immunization for post-exposure rabies prophylaxis is associated with the appearance of serum neutralizing antibody (SNA) earlier than occurs with vaccine alone. We compared the SNA response and the side-effects in 30 previously unimmunized healthy volunteers given a commercially available human rabies immunoglobulin (HRIG) intramuscularly (i.m.) or an experimental HRIG prepared by DEAE Sephadex column chromatography, intravenously (i.v.) with or without human diploid-cell culture rabies vaccine (HDCS). The subjects were divided into five equal groups: HDCS alone, HDCS + i.m. HRIG 20 IU/kg (currently recommended), i.v. HRIG alone 15 IU/kg, HDCS + i.v. HRIG 15 IU/kg or HDCS + HRIG 5 IU/kg i.v. plus 10 IU/kg i.m. to simulate local bite wound infiltration. HDCS, 1.0 ml, was injected subcutaneously (s.c.) on days 0, 3, 7, 14 and 28. Only local discomfort at injection sites was observed without differences between groups. SNA was demonstrated in all HRIG recipients at day 1, but the concentrations were higher in those receiving it intravenously. No difference in the SNA response to vaccine was observed between the i.v. and i.m. HRIG groups given the same vaccine lot. It would appear that i.v. HRIG 15 IU/kg can be substituted for i.m. HRIG 20 IU/kg for post-exposure prophylaxis. Since the current regimen is almost 100% protective, there is no way of proving that i.v. HRIG 15 IU/kg is more efficacious. The immediate SNA level and economy are the chief advantages of i.v. HRIG 15 IU/kg.     

Novel Vaccines to Human Rabies

Rabies, the most fatal of all infectious diseases, remains a major public health problem in developing countries, claiming the lives of an estimated 55,000 people each year. Most fatal rabies cases, with more than half of them in children, result from dog bites and occur among low-income families in Southeast Asia and Africa. Safe and efficacious vaccines are available to prevent rabies. However, they have to be given repeatedly, three times for pre-exposure vaccination and four to five times for post-exposure prophylaxis (PEP). In cases of severe exposure, a regimen of vaccine combined with a rabies immunoglobulin (RIG) preparation is required. The high incidence of fatal rabies is linked to a lack of knowledge on the appropriate treatment of bite wounds, lack of access to costly PEP, and failure to follow up with repeat immunizations. New, more immunogenic but less costly rabies virus vaccines are needed to reduce the toll of rabies on human lives. A preventative vaccine used for the immunization of children, especially those in high incidence countries, would be expected to lower fatality rates. Such a vaccine would have to be inexpensive, safe, and provide sustained protection, preferably after a single dose. Novel regimens are also needed for PEP to reduce the need for the already scarce and costly RIG and to reduce the number of vaccine doses to one or two. In this review, the pipeline of new rabies vaccines that are in pre-clinical testing is provided and an opinion on those that might be best suited as potential replacements for the currently used vaccines is offered.     

A Novel Rabies Vaccine Based on a Recombinant Parainfluenza Virus 5 Expressing Rabies Virus Glycoprotein

Untreated rabies virus (RABV) infection leads to death. Vaccine and postexposure treatment have been effective in preventing RABV infection. However, due to cost, rabies vaccination and treatment have not been widely used in developing countries. There are 55,000 human death caused by rabies annually. An efficacious and cost-effective rabies vaccine is needed. Parainfluenza virus 5 (PIV5) is thought to contribute to kennel cough, and kennel cough vaccines containing live PIV5 have been used in dogs for many years. In this work, a PIV5-vectored rabies vaccine was tested in mice. A recombinant PIV5 encoding RABV glycoprotein (G) (rPIV5-RV-G) was administered to mice via intranasal (i.n.), intramuscular (i.m.), and oral inoculation. The vaccinated mice were challenged with a 50% lethal challenge dose (LD₅₀) of RABV challenge virus standard 24 (CVS-24) intracerebrally. A single dose of 10⁶ PFU of rPIV5-RV-G was sufficient for 100% protection when administered via the i.n. route. The mice vaccinated with a single dose of 10⁸ PFU of rPIV5-RV-G via the i.m. route showed very robust protection (90% to 100%). Intriguingly, the mice vaccinated orally with a single dose of 10⁸ PFU of rPIV5-RV-G showed a 50% survival rate, which is comparable to the 60% survival rate among mice inoculated with an attenuated rabies vaccine strain, recombinant LBNSE. This is first report of an orally effective rabies vaccine candidate in animals based on PIV5 as a vector. These results indicate that rPIV5-RV-G is an excellent candidate for a new generation of recombinant rabies vaccine for humans and animals and PIV5 is a potential vector for oral vaccines.     

Human rabies immunoglobulin assayed by the rapid fluorescent focus inhibition test suppresses active rabies immunization

The rabies antibody content of each of ten lots of human rabies immunoglobulin was titrated by both the mouse neutralization test and the rapid fluorescent focus inhibition test. The two tests did not give comparable results, the antibody titres obtained by the mouse neutralization test being 1·4–9·6 times higher than those obtained by the rapid fluorescent focus inhibition test. This titre difference was associated with a consistently lower antibody response in human volunteers who had received post-exposure rabies vaccine treatment which included the administration of RIG assayed by the RFFIT.     

狂犬病病毒CTN-1V株在人二倍体细胞Walvax-2株的适应传代

目的建立狂犬病病毒固定毒CTN-1V株在人二倍体细胞Walvax-2株上的传代适应株。方法用狂犬病病毒固定毒CTN-1V株经昆明小鼠鼠脑回传后的病毒接种Walvax-2细胞,连续传代,检测各代次病毒的滴度及免疫原性。结果 CTN-1V株能较好地适应Walvax-2细胞,通过连续带毒传代至第7代,病毒滴度可达6.78 lg LD50/mL,并在第10~15代内滴度维持在7.0 lg LD50/mL以上,15~30代滴度稳定在7.0 lg LD50/mL左右。以15代适应毒株CTN-1V-HDC P15制备的疫苗原液,各项指标均符合《中华人民共和国药典》三部(2010版)的要求,疫苗效力在6.0IU/剂以上。结论所获人胚肺二倍体细胞Walvax-2株传代适应狂犬病毒固定毒株CTN-1V-HDC可用于人用狂犬病疫苗的生产开发。     

Rabies healthcare-seeking behaviors of urban and peri-urban residents: Results from a rabies knowledge,attitudes, and practices survey,Bangladesh, 2018

Rabies is one of the most lethal infectious diseases, with those living in Asia and Africa having the highest risk of dying from rabies. We conducted a knowledge, attitudes and practices survey in urban and peri-urban areas of Bangladesh to describe canine bite rates, rabies knowledge, and healthcare seeking behaviors and barriers to human and dog vaccination. A bite risk assessment score (BRAS) and healthcare-seeking behavior score (HSBS) was calculated for each bite victim. Respondents were given two hypothetical situations to assess potential behaviors after a bite and willingness to pay for rabies vaccine and immunoglobulin. In total, 2,447 households participated in the survey and 85 bite victims were identified. The BRAS identified that 31% of bites posed no risk of rabies transmission. Multivariate analyses showed that living in Chittagong (β = 1.4; 95% CI: 0.1, 2.7) was associated with a higher HSBS. Findings presented here provide useful information regarding bite occurrences, healthcare-seeking behaviors, and a need for strategies to increase rabies awareness.     

Antibody response to purified chick embryo cell vaccine in equines for production of equine rabies immune globulin

Rabies is endemic in India. The post exposure treatment of class three bite cases, as recommended by the World Health Organization, must involve the use of rabies immunoglobulin as soon as possible and up to the seventh day of start of anti rabies vaccination. The annual requirement in India as projected by the Ministry of Health, Government of India is about 1500 liters of purified anti rabies serum (ERIG). Central Research Institute (CRI), Kasauli, being the sole producer of ERIG in India, an effort was made to increase the production of ERIG by the use of tissue culture vaccine (Human) for primary immunization of equines. It also involved changing the vaccine from horse brain suspension to tissue culture vaccine by eliminating horse sacrifice for antigen preparation. A better immune response was obtained.     

狂犬病毒糖蛋白DNA疫苗的研制及其免疫效果的观察

构建了含有狂犬病毒（RV）CVS株糖蛋白（GP）基因的重组质粒pCMVCVSRG,将其转染至鼠NIH3T3细胞中,用间接免疫荧光法和APAAP法均证实RVGP能在真核细胞中表达。分别将合RV不同毒株的GP基因的质粒（DNA疫苗）及空白载体质粒（对照组）免疫小鼠,仅DNA疫苗免疫的小鼠产生了中和抗体。以RV攻击后,DNA疫苗免疫组小鼠的存活率与对照组相比,差异有极显著性意义（P＜0．01）;不同的启动子（CMV或SV40）与不同GP基因（来源于CVS株或ERA株）对DNA疫苗的免疫效果无明显影响。在注射120d后．用PCR方法仍可检测出RVGP基因。结果表明：狂犬病DNA疫苗能够诱生低水平的中和抗体和记忆性B淋巴细胞,并能保护小鼠抵抗RV的攻击。该疫苗能在体内稳定存在。狂犬病DNA疫苗的研制为狂犬病免疫开辟了一条新途径,并可为防治其他疾病的DNA疫苗的研制奠定基础。