Vaccination with purified microgamete antigens prevents transmission of rodent malaria

Malaria vaccination with preparations of microgametes has been shown to inhibit transmission of Plasmodium spp. to the mosquito vectors of avian, rodent and simian parasites. This transmission-blocking immunity results from the induction of microgamete-agglutinating antibodies in the vaccinated host which, when ingested in a mosquito blood meal, react with exflagellating microgametes in the midgut to prevent fertilization and oocyst development. Here we have passively transferred the immunity with gamete-specific monoclonal antibodies raised against the rodent malaria parasite Plasmodium yoelii nigeriensis, and an IgG2a isotype monoclonal antibody from a hybridoma cell line, PYG-1, has been used to identify the target antigens on the gametes and to affinity-purify sufficient quantities of specific gamete antigen to facilitate vaccination studies. This transmission-blocking monoclonal antibody immunoprecipitated microgamete antigens of apparent relative molecular mass (Mr), 67K (67,000), 59K, 57K, 42K and 35K. Immunization of mice with these proteins before infection and mosquito feeding led to a 85-99.7% reduction in transmission to the mosquito vector; vaccination via intravenous or intramuscular routes was equally effective and did not require an adjuvant.     

Rapid cloning of high-affinity human monoclonal antibodies against influenza virus

Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14-21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.     

Adjuvant-free IgG responses induced with antigen coupled to antibodies against class II MHC

The generation of strong serological responses to protein antigens in experimental animals usually requires the use of potent adjuvants, most of which cannot be used in human or veterinary vaccines because of deleterious side effects. Attempting to circumvent this problem, we have assessed an adjuvant-free antigen-delivery system based on the hypothesis that antigen coupled to monoclonal antibodies (mAbs) specific for class II major histocompatibility complex (MHC) determinants should be 'targeted' onto antigen-presenting cells, thus facilitating recognition by helper T cells. We found that the biotin-binding protein avidin could generate a serological response in mice, without adjuvant, when injected coupled to a biotinylated anti-class II MHC mAb. Equivalent amounts of avidin mixed with the non-biotinylated form of the same mAb failed to elicit a response. A targeting effect was demonstrated at low levels of injected conjugate because only mice bearing the appropriate class II antigens responded. Responses were also seen with a protein antigen other than avidin, offering a new, adjuvant-free approach to subunit vaccine construction.     

HBV包膜大蛋白、中蛋白和HBsAg疫苗免疫原性的比较

目的:检测HBV包膜大蛋白(L蛋白)、中蛋白(M蛋白)和HBsAg疫苗的免疫原性.方法:采用本实验室表达纯化得到的HBV包膜大蛋白、中蛋白与市售疫苗(主要成分为HBsAg),以2、10g剂量免疫家兔.ELISA检测抗HBsAg抗体滴度水平变化.结果:注射了L蛋白、M蛋白和HBsAg疫苗的家兔实验组产生的抗HBsAg抗体滴度水平有着非常明显的差别.注射了L蛋白、M蛋白的家兔在14d即可检测出抗HBsAg抗体,而注射HBsAg的家兔分别在21d和28d才能检测到抗HBsAg抗体.同时注射L蛋白实验组产生的抗HBsAg抗体滴度水平要比注射同剂量HBsAg实验组高出50%,并且注射2gL蛋白的实验组比注射10gHBsAg蛋白的实验组更快出现了抗HBsAg抗体,且抗体滴度水平更高.结论:L蛋白比M蛋白及市售疫苗的免疫原性强,不但产生抗体时间提前,而且滴度高,是新一代高效乙肝疫苗的优选蛋白.     

IgM-autoantibodies against isologous erythrocytes also react with isologous IgG(Fc)

The mechanisms by which the adaptive immune system of animals discriminate between foreign and self components are not known. It is generally believed that antibodies are produced only in response to foreign agents. However, against this axiom there is mounting evidence that normal animals produce autoantibodies against a variety of self components. In two murine model systems, a high proportion of IgM-producing cells secrete autoantibodies. In one model, the autoantibodies are specific for isologous mouse IgG(Fc) and in the other, the autoantibodies are specific for antigens revealed on the surface of mouse erythrocytes (RBC) by proteolytic enzymes. The function of these two autoimmune responses is a mystery. Here we show that the two responses are related. The lysis of mouse RBC, modified with bromelain (brom) by IgM autoantibodies was completely inhibited by isologous IgG(Fc), and antibodies against mouse IgG(Fc) formed precipitin bands with solubilized membranes from mouse RBC. We conclude that mouse RBC membranes and IgG(Fc) have at least one antigenic determinant in common and that this determinant is the target for autoimmune responses in normal mice.     

An anti-idiotype vaccine against experimental schistosomiasis

Schistosomiasis is a parasitic infection of man which is widespread in tropical countries, and which so far has resisted attempts at control. We have been approaching the problem from an immunological angle. We have previously reported the production of a rat monoclonal IgG2a antibody against Schistosoma mansoni which exhibits marked cytoxicity for schistosomula in the presence of eosinophils and a high degree of protection by passive transfer in naive rats. This antibody, IPLSm1, was shown to bind specifically to a schistosomulum membrane target antigen defined as a glycoprotein of relative molecular mass 38,000 (38K), which is strongly immunogenic in schistosome infection of various animal species including man. Although theoretically the 38K protein represents an excellent candidate for a potential vaccine against schistosomiasis, the glycanic nature of the epitope recognized by IPLSm1 limits its production by DNA recombinant technology. It was, moreover, shown that, together with protective antibodies, the 38K molecule was able to induce the production of blocking IgG2c antibodies that inhibit the functional properties of IPLSm1 both in vitro and in vivo. Therefore, following Jerne's network theory, we considered an alternative approach, the possibility of immunization using anti-idiotype antibodies. In the present study, rat monoclonal anti-idiotype antibodies were produced against IPLSm1 (AB1). Anti-idiotype antibodies (AB2) were selected by their capacity to inhibit the binding of radioiodinated AB1 to its 38K target antigen. Sera from naive LOU rats immunized with a purified AB2 preparation contained specific anti-schistosome antibodies (AB3) which bound to 38K. AB3 antibodies were strongly cytotoxic for schistosomula in the presence of rat eosinophils and conferred highly significant protection by passive transfer. Most importantly, rats immunized with AB2 demonstrated marked protection (50-80%) to a challenge infection.     

HIV感染的治疗性DNA疫苗（综述）

DNA疫苗被称为继完速病原体疫苗和基因工程重组蛋白疫苗后的第3代疫苗,其兼有重组亚单位疫苗的安全性和减毒活疫苗的有效性。HIV感染的防治则以激发细胞免疫为主,以激发细胞免疫为主的DNA疫苗给HIV感染防治带来曙光。目前DNA疫苗在HIV感染防治研究方面已取得显著进展,其临床研究已证明具有明显的刺激机体细胞免疫的效果,尤其是一些细胞因子、趋化因子可明显增强HIV DNA疫苗的免疫反应。就DNA免疫佐剂、HIV病毒基因DNA疫苗作用机制以及加强HIV DNA疫苗的各种调节因素方面的研究进展进行综述,以促进HIV DNA疫苗防治的研究。     

Prevention of HIV-2 and SIVsm infection by passive immunization in cynomolgus monkeys.

Infection of macaques with simian immunodeficiency virus (SIV) and human immunodeficiency virus type 2 (HIV-2) are useful models for studies of immunotherapy and vaccination against HIV as well as for testing of antiviral drugs. Vaccine research showing protective immunity in immunized monkeys has indicated that it will be possible to develop a vaccine for prevention of human HIV infection, although many hurdles remain. The design of an HIV vaccine would be helped if the basis of the protective immunity could be elucidated. Passive immune prophylaxis offers a means to determine the relative role of antibodies in protection against infection. We have studied whether a transfer of antibodies can prevent HIV-2 and SIVsm (SIV of sooty mangabey origin) infection in cynomolgus monkeys. Sera with high antibody titres were collected, heat-treated and injected into naive animals 6 h before challenge with 10-100 monkey-infectious doses of live homologous virus. All control animals treated with normal monkey serum (n = 6) or no serum (n = 39) became infected by the challenge virus, whereas five out of seven animals pretreated with antibody-containing serum at a dose of 9 ml kg-1 resisted infection. Thus passively transferred antibodies can protect against a low-dose lentivirus challenge in a nonhuman primate.     

鸭病毒性肝炎综合防制措施的研究

肉鸭养殖场采取严格消毒、隔离饲养、加强种鸭免疫和雏鸭免疫、利用血清或抗体对雏鸭被动预防、药物预防等综合防制措施,对选定养殖户的养殖跟踪表明,在20日龄的鸭病毒性肝炎发生率从60%降低到20%。对来自非免疫种鸭和免疫种鸭的雏鸭群分别采用雏鸭疫苗接种、免疫血清注射和喂中药“鸭肝灵”,结果表明,疫苗的免疫接种是防制DVH所必须的,且与中草药“鸭肝灵”配合应用防制效果明显;注射血清能起到一定的防制作用,但其效价维持的时间较短,至少要经过两次注射才可能起到有效的保护作用。而对来自规则免疫种鸭的健康雏鸭其母源抗体有一定的保护作用,对预防DHV的早期感染起关键的作用,但保护不到15日龄,最早需在5日龄进行首免,15日龄才能达到有效的保护作用,同时饲喂中药“鸭肝灵”保护效果明显。无论雏鸭的母源抗体如何,隔离饲养、严格消毒,疫苗免疫(首免时间依种鸭免疫情况而定),并同时饲喂中药“鸭肝灵”保护效果明显。     

2010—2011年宾川县麻疹IgM抗体检测结果分析

目的：通过IgM抗体检测结果,分析宾川县麻疹有效接种率状况,为预防和消除麻疹提供科学依据。方法：采用描述流行病学方法对宾川县2010-2011年麻疹IgM抗体检测资料进行统计分析。结果：宾川县2010-2011年共检测麻疹血液标本781人份,其中麻疹IgM抗体阳生率95.51%,7～10岁组阳性率最高达96.36%,0～1岁组阳性率最低为94.56%。结论：宾川县近年来麻疹疫苗免疫接种工作成效明显,人群麻疹IgM抗体处于一个较高水平,下一步要继续抓好麻疹基础免疫接种工作,注重接种质量和效果,消除人群免疫空白,提高人群麻疹抗体水平。     

甘油-3-磷酸对甲肝疫苗诱导的体液免疫影响*

目的 研究甘油-3-磷酸对甲肝疫苗诱导的小鼠体液免疫应答的影响。方法 选取49只雌性ICR小鼠,分7组,每组7只,这7组小鼠分别为阴性对照、单纯抗原组、铝佐剂组和4组甘油-3-磷酸组,分别在免疫之后的第4、8、12、16、20周用酶联免疫法(ELISA)测定小鼠血清中的特异性的anti-HAV IgG水平。结果 在小鼠免疫之后的第4、8、12、16、20周单纯抗原组、铝佐剂组和4组不同剂量甘油-3-磷酸组均能检测到特异性抗HAV抗体,并且趋势为先升高后降低,且在免疫后的第8周最高;第4周和第8周的甘油-3-磷酸实验组的结果均高于单纯抗原组,且差异均具有统计学意义(P<0.05),并且第4周甘油-3-磷酸组抗体水平均超过铝佐剂组,并具有统计学意义(P<0.05),其中甘油-3-磷酸2 mg为最佳剂量组,其抗体水平在免疫后的第4、8、12、16、20周均高于单纯抗原组,且在第8周抗体水平达到峰值,抗HAV IgG水平为lg(3.064±0.07851),铝佐剂的也是在第8周抗体水平达到峰值,抗HAV IgG水平为lg(3.150 ±0.1382)。结论 甘油-3-磷酸有免疫佐剂的作用,能够有效、迅速地增强HAV诱导的体液免疫应答,是一种潜在、安全有效的新型疫苗佐剂。     

ELISA定量测定单克隆抗体方法改进

本文介绍了以亲和层析纯化的免抗鼠IgG类抗体及其酶标物作为捕获及检测抗体进行鼠杂交瘤培养上清及腹水中IgG类单克隆抗体的ELISA定量测定方法。结果表明:小牛血清(BS)、免血清(RS)、马血滑(HS)、人血清(HuS)及非抗体产生细胞的培养上清对本项εLISA实验无干扰,使测定的结果较可靠,测定的鼠IgG各亚类的标准曲线的线性范围均在5～100ng/ml之间,因此这个快速、可靠的方法可为鼠IgG类单克隆抗体的定量测定提供一种实用的手段。     

pH-responsive vaccine delivery systems for improving cellular immunity

Immune vaccination is a critically important strategy in disease prevention and treatment. In vaccination, efficient vaccine carriers are necessary to augment the immune response initiated by vaccines generally with weak immunogenecity. Nowadays, commercially available vaccine/carrier formulations induce effective humoral immunity but weak or none cellular immunity. However, in practice, cellular rather than humoral immunity plays the key role in treating some intractable diseases such as AIDS and cancers, in which the elicited cytotoxic T lymphocytes can directly kill HIV-infected or cancer cells. To trigger potent cellular immunity, the carriers should help antigens escape from endosomal/lysosomal degradation and release them directly into the cytosol of antigen presenting cells. To this aim, such kind of vaccine carriers should be rationally designed and prepared in terms of their structure and physiochemical properties. Here, we summarized the recent advances in pH-responsive vaccine carriers exclusively developed for improving cellular immunity.     

57例不规则抗体干扰ABO血型鉴定分析

通过研究干扰血型鉴定的不规则抗体的特异性及类型,为正确的进行血型鉴定提供实验参考依据。2013年7月至2014年7月对22 558例患者进行血型及不规则抗体筛查检测,对因不规则抗体干扰血型鉴定患者进行抗体鉴定,明确抗体特异性及类型,并选择与抗体对应抗原阴性的红细胞制备反定型细胞进行血型鉴定,以排除干扰明确血型。结果:发现57名患者因MNS、Rh、Lewis、Duffy、P血型系统产生的不规则抗体干扰血型鉴定,其中抗-M最多,共37例,占64.91%,其次为抗-E和抗-e相关抗体,均为5例,分别占8.77%;抗-Lea3例,占5.26%;Ig G与Ig M类混合抗体37例,单纯Ig G类抗体14例,单纯Ig M类抗体6例。研究结果表明,Ig M和Ig G类不规则抗体均可干扰血型鉴定,以Ig M类为主。     

Cell-mediated immunity in the liver of mice vaccinated against malaria.

Mice can be protected against several species of lethal malaria infection by vaccination, and their recovery correlates well with increased anti-malarial antibody levels, particularly IgG (ref.2). However, there is also a good correlation between protection by vaccines and priming for delayed-type hypersensitivity in the skin, although there is no obvious explanation for this effect. We now report an apparent relationship between protection and a cell-mediated immune response involving the migration of various types of cell capable of killing malaria parasites in vitro to the liver. We suggest that the effect of vaccination is to bring together parasites, specific antibody and nonspecific cytotoxic cells, and that the liver may be a major site for their interaction.     

1,25(OH)_2VD_3在过敏原特异性免疫治疗花粉过敏哮喘中的应用

以1,25(OH)₂VD₃为佐剂制备软叶针葵花粉变应原疫苗,以小鼠致敏哮喘模型为研究对象进行特异性免疫治疗.通过检测小鼠气道高反应性、血清中特异性抗体、细胞因子以及肺组织病理学切片等指标对治疗模型的构建进行评价,探讨1,25(OH)₂VD₃在过敏原特异性免疫治疗花粉过敏哮喘免疫机制中的作用.结果表明:以1,25(OH)₂VD₃为变应原疫苗能有效抑制花粉特异性IgE的产生和Th2细胞因子IL-4分泌,促进封闭性抗体IgG2a产生和Th1细胞因子IFN-γ的分泌,增加耐受性细胞因子IL-10生产,使Th2反应向Th1反应转变.1,25(OH)₂VD₃在花粉过敏性哮喘治疗中能大幅提高过敏原特异性免疫治疗的疗效.     

分子佐剂PHA增强小鼠特异性体液免疫的效应

目的:探讨PHA对小鼠特异性体液免疫的作用和应用价值.方法:经SRBC腹腔免疫昆明种小鼠30只,并随机等量分两组,实验组每只腹腔注射PHA 0.1ug/g,对照组注入等量N.S.5d后,两组均眼球取血,并颈椎离断处死取脾脏制备脾细胞悬液.用体外抗体形成细胞实验.(Plaque forming cell,PFC)和定量溶血分光光度测定法(Quantitativehemolysisspectrophotomentry,QHS),分别测取两组的抗体形成细胞率和QHS(495nm)OD值,经X±SD组间比较的t检验,观察有无差异性.结果:实验组的PFC形成率和QHS的OD值均显著高于对照组(p<0.01).结论:PHA有增强小鼠PFC的形成率和特异性抗体的表达.提示:PHA为分子佐剂在提高体液免疫应答的应用有较高参考价值.     

BOG初次免疫联合结核分枝杆菌Ag85A DNA疫苗加强免疫效果的研究

为探讨BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强免疫的序贯免疫策略对小鼠的免疫效果。采用BCG及结核分枝杆菌Ag85A DNA疫苗依次免疫小鼠,在末次免疫后的4、6、8周通过检测小鼠血清总IgG抗体、特异性淋巴细胞增殖．细胞因子的水平,观测BCG初次免疫,Ag85A DNA疫苗加强免疫的策略对小鼠的免疫效果。结果显示,采用BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强免疫的策略组的小鼠与其它免疫方式组相比,IgG明显升高（P〈0．05）、特异性淋巴细胞明显增殖、IFN7、IL-2、IL-4水平明显增高（P〈0．05）。由此可知,在采用BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强的免疫策略后,能增强对小鼠的免疫效应,尤其是Thl型细胞免疫反应增强明显,为进一步在动物体内进行保护性效应试验的研究提供了实验依据。     

纳米铝佐剂诱导鸡提前产生抗AIVH9体液免疫应答

接种灭活油佐剂疫苗是目前国内防控禽流感主要的措施,为有效防控禽流感的流行发挥了重要的作用,但灭活苗首次接种后需要较长时间抗体才能达到有效保护水平.纳米颗粒具有独特的生物学特性,作为疫苗佐剂能引起机体强烈的免疫应答.本研究用纳米铝颗粒作为AIV H9的佐剂,结果显示,小鸡产生有效免疫保护抗体时间较常规油佐剂疫苗提前4天且无副反应,但抗体峰值和持续时间均低于油佐剂疫苗.氢氧化铝纳米颗粒作为佐剂可能对禽流感等重大传染病的紧急预防接种具有潜在的应用价值.     

Chemical basis of antigenic variation in foot-and-mouth disease virus

One of the difficulties in controlling foot and mouth disease by vaccination is the occurrence of the virus as seven distinct serotypes because immunity conferred by vaccination against one serotype leaves the animals susceptible to infection by the other six. Moreover, the antigenic variation, even within a serotype, can be so great that immunity against the homologous strain of virus need not necessarily ensure protection against infection by other viruses within that serotype. Here we report the separation of three natural antigenic variants, distinguishable in cross-neutralization tests from an isolate of foot-and-mouth disease virus (FMDV). The serological differences could also be demonstrated by antisera elicited by synthetic peptides corresponding to residues 141-160 of the capsid polypeptide VP1, showing that this region contains a major immunogenic site of the virus. The results have practical implications for the choice of viruses for vaccine production.