禽流感特异性转移因子的制备及其免疫作用

目的制备禽流感病毒特异性转移因子并探讨其对禽流感灭活疫苗的免疫增效作用。方法用禽流感病毒H5N1血清亚型灭活疫苗免疫鸡,用国标血凝抑制方法检测病毒特异性血凝抑制抗体效价。当抗体效价达到高峰时,翅静脉采取外周血,分离淋巴细胞并制备细胞单层、传代后获得禽流感病毒H5N1血清亚型特异性转移因子。用所获得的特异性转移因子进行疫苗免疫增效试验。结果采用本法可获得禽流感病毒特异性转移因子。免疫增效试验表明,在进行禽流感病毒灭活疫苗免疫的同时使用禽流感病毒特异性转移因子,可在一定幅度内提高禽流感病毒抗体水平并能延长抗体维持时间。不同给药途径比较试验表明,口服途径给药的疫苗增效作用优于注射途径给药。结论通过淋巴细胞体外培养可以制备禽流感病毒特异性转移因子。禽流感病毒H5N1血清亚型特异性转移因子对禽流感病毒灭活疫苗具有明显的增效作用,且口服途径给药的疫苗免疫增效作用优于注射途径给药。     

地衣芽孢杆菌对实验性家兔阴道炎金葡菌的消除作用

本文以地衣芽孢杆菌菌液对实验性家兔金葡萄感染阴道炎进行治疗,观察把衣芽孢杆菌对阴道内葡菌的消除作用。结果表明,地衣芽孢杆菌对金黄色葡萄球菌有明确的消除作用,可逆转由金葡菌引起的阴道感染性炎症。     

用对流电泳提纯兔出血症病毒

兔出血症病毒（ＲＨＤＶ）在ｐＨ８．６的琼脂糖凝胶上电泳,在负极侧有一血凝峰,免疫电镜观察有大硅ＲＨＤＶ颗粒,证实ＲＨＤＶ病毒粒子带正电荷。利用这一特性可用对流电泳提纯病毒。将抗体与病毒颗粒形成的沉淀带切下,作ＳＤＳ－ＰＡＧＥ,经免疫转印出现６条区带,其中６０ｋＤ的ＶＰ_１为主要结构多肽。用免疫复合物提取核酸,以狄高辛标记制成探针,能与病毒核酸和克隆的ＲＨＤＶｃＤＮＡ２．２ｋｂ片段和４．８ｋｂ片段杂交,探针灵敏度达ｐｇ水平。能用病毒核酸作模板制备探针,证实ＲＨＤＶ的核酸为ＤＮＡ。     

不同抑制剂对地衣芽孢杆菌生长影响的研究

报道了地衣芽孢杆菌对不同抑制剂的反应。结果表明,当ｐＨ在７．０时,链霉素浓度为５００μｇ／ｍＬ,青霉素浓度１０００μｇ／ｍＬ,头孢唑啉钠浓度为５００μｇ／ｍＬ时对地衣芽孢杆菌有明显抑制作用。这对正确使用地衣芽孢杆菌制剂（整肠生）提供了可靠的理论依据。     

新生幼犬的被动免疫研究

新生幼犬通过初乳获得的被动免疫,对它的生存非常重要,本研究用犬的多价免疫血清代替初乳作为幼犬保护性免疫球蛋白,对生后2 d内不能获得初乳的幼犬,以及虽然获得初乳但初乳中特异性免疫球蛋白含量较低的幼犬进行了被动免疫试验。对5窝35只幼犬在生后,随机分为吃初乳的对照组、口服血清组、皮下注射血清组、以及口服和皮下注射血清又吃初乳5个组,在幼犬出生时、及出生后48 h和第5天采血,测定犬细小病毒、犬传染性肝炎、和犬副流感的血凝抑制（HI）抗体效价。结果表明：对没有吃到初乳的仔犬,通过早期口服或皮下注射途径给予成年犬的血清,均能代替初乳供给幼犬免疫球蛋白。对吃到初乳的仔犬,能进一步提高幼犬体内特异性抗体水平,但以皮下注射途径最佳。     

地衣芽胞杆菌对实验性家兔阴道炎影响的研究

目的通过探讨地衣芽胞杆菌活菌制剂对实验性家兔细菌性阴道病的影响,恢复家兔阴道微生态平衡和正常菌群环境。方法（1）采用注射用氨苄西林和甲硝唑生理盐水溶液注入家兔阴道进行冲洗,建立家兔阴道脱污染动物模型。（2）取40只阴道脱污染家兔,其中20只接种大肠埃希菌,20只接种金黄色葡萄球菌,建立家兔阴道感染模型。（3）地衣芽胞杆菌对家兔阴道菌群失调的调整作用：采用不同浓度的地衣芽胞杆菌菌液（10^6CFU／ml、10^8CFU／ml、10^9CFU／ml）对感染家免阴道进行接种,分析和考察地衣芽胞杆菌对家兔阴道菌群失调的影响以及对家兔阴道黏膜的影响。结果（1）动物经过金黄色葡萄球菌感染,通过地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、肠杆菌、乳杆菌数量明显上升,葡萄球菌数量明显下降,自细胞数量减少,黏膜红肿减轻、分泌物减少,治疗作用明显。（2）大肠埃希菌感染动物经地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、乳杆菌数量明显上升,肠杆菌数量明显降低,白细胞数量减少,黏膜红肿消失、分泌物减少。结论地衣芽胞杆菌对实验性家兔金黄色葡萄球菌和大肠埃希菌阴道炎的治疗有效。地衣芽胞杆菌与乳杆菌的作用相似,具有维持阴道菌群平衡的作用。     

H7N9灭活疫苗联合MF59佐剂在小鼠体内诱导的长效体液免疫应答

以BALB/c小鼠为模型,探讨H7N9流感病毒灭活疫苗免疫小鼠后所诱导的长效体液免疫应答的动态变化。不同剂量的流感H7N9全病毒灭活疫苗单独或辅以MF59佐剂肌肉注射免疫小鼠一次。连续采集免疫后小鼠15个月的血清,用ELISA方法检测特异性IgG抗体水平,血凝抑制(hemagglutination inhibition,HI)试验和微量中和(microneutralization,MN)试验检测第15个月时的HI抗体和中和抗体效价。实验结果发现,小鼠血清中的特异性IgG抗体水平随时间变化持续缓慢上升,第5个月时达到顶峰,随后略有下降但一直持续平稳状态;IgG抗体滴度与疫苗剂量成正相关,且添加佐剂能提高抗体滴度。HI及MN抗体检测表明,免疫后第15个月产生的抗体能有效中和病毒,且抗体跟疫苗剂量成正比。以上研究表明,H7N9流感病毒灭活疫苗免疫小鼠一次诱导产生的特异性抗体能在较长期内保持比较平稳的抗体滴度,为小鼠提供免疫保护;增加抗原剂量和添加MF59佐剂能增加疫苗特异性抗体水平。该研究为H7N9流感疫苗产生的长期保护效应提供了一定的数据积累和参考。     

副地衣芽孢杆菌FA6全基因组测序及比较基因组学分析

副地衣芽孢杆菌(Bacillus paralicheniformis)FA6是一株从草鱼肠道内分离出来的细菌, 其具有淀粉酶和纤维素酶等多种碳水化合物酶活性。为深入研究副地衣芽孢杆菌FA6可能的益生机制, 研究通过三代测序技术测定了副地衣芽孢杆菌FA6的全基因组序列, 运用生物信息学方法进行基因组组装、基因预测和功能注释。同时通过比较基因组学方法, 比较分析了副地衣芽孢杆菌FA6与4株基因组序列已经发表的芽孢杆菌基因组结构和功能的差异。结果发现副地衣芽孢杆菌FA6全基因组由1条环状染色体组成, 大小为4450579 bp, GC含量为45.9%。副地衣芽孢杆菌FA6基因组中含有128个蛋白酶基因, 32个脂肪酶基因和72个糖苷水解酶基因, 这些基因与食物降解相关; 此外, 细菌基因组中还含有7个编码羊毛硫抗生素相关的基因。比较基因组结果显示, 副地衣芽孢杆菌FA6与其他4株芽孢杆菌的基因组共线性关系较好, 但是与地衣芽孢杆菌菌株相比, 菌株FA6基因组特征更接近于副地衣芽孢杆菌菌株。副地衣芽孢杆菌FA6基因组中编码纤维素酶、半纤维素酶和淀粉酶的基因数量分别为5、7和5个, 多于其他菌株, 能够更好地降解植物多糖。研究结果表明副地衣芽孢杆菌FA6高度适应植物性成分, 反映了该菌株在草鱼肠道中的适应性进化, 该菌株可能可以作为益生菌用于水产养殖。     

地衣芽孢杆菌活菌制剂对成人急性肠炎的疗效观察

1986年我院检验科从一名正常待产妇女阴道分离出一株革兰氏阳性需氧芽孢杆菌,经中科院微生物研究所鉴定为地衣芽孢杆菌。(Baccillue licheniformis)、两年来研究表明该菌生物毒性极低。我们应用地衣芽孢杆菌活菌制剂治疗成人急性肠炎59例,七天治愈率达93.22％,这种疗效的机     

流行性出血热病毒沙鼠肾细胞培养灭活疫苗免疫家兔的抗体反应及保护试验

利用自流行性出血热(EHF)病人血液中分离的EHF病毒浙10株,感染长爪沙鼠肾单层细胞,经培养后,按流行性乙型脑炎的生物制品法规要求研制成灭活疫苗。取1ml经肌肉免疫家兔,6天后即可在血液中检出荧光抗体,至第14～21天为最高峰,荧光抗体效价可达320～2560。经活EHF病毒攻击,有明显的保护作用,保护率达90％以上。疫苗经1:10或1:30稀释后免疫家兔,荧光抗体阳转率仍达100％,但抗体滴度明显降低。肌肉接种优于皮下接种。本研究证明,甲醛灭活EHF病毒可破坏血凝素活性,从而影响疫苗血凝抑制抗体的产生,可能也会影响中和病毒的能力。     

Nachweis humoraler und lokaler Antikörper nach Einsatz einer bordetella Bronchiseptica-Lebendvakzine am Schwein

The aim of this work was to prove if a Bordetella bronchiseptica living vaccine is able to induce humoral and local antibodies in pigs. The living vaccine was compared with a killed Bordetella bronchiseptica adjuvans vaccine. The results show that piglets of sows vaccinated parenterally with living vaccine and killed adjuvans vaccine develop a stronger humoral immune reaction than piglets vaccinated locally with living vaccine. Sows immunized parenterally with living vaccine permint their piglets in the first three weeks colostral antibodies like sows immunized with killed adjuvans vaccine. The living vaccine induces especially local antibodies.     

Synergies between vaccination and dietary arginine and glutamine supplementation improve the immune response of channel catfish against Edwardsiella ictaluri

Channel catfish was used to investigate the enhancement of vaccine efficacy following dietary supplementation with arginine (ARG, 4% of diet), glutamine (GLN, 2% of diet), or a combination of both. After vaccination against Edwardsiella ictaluri, humoral and cellular immune responses, along with lymphoid organ responses were evaluated. E.?ictaluri-specific antibody titers in plasma were higher (P?相似文献   

Immunization with surface antigen vaccine alone and after treatment with 1-(2-fluoro-5-methyl-beta-L-arabinofuranosyl)-uracil (L-FMAU) breaks humoral and cell-mediated immune tolerance in chronic woodchuck hepatitis virus infection

Woodchucks chronically infected with the woodchuck hepatitis virus (WHV) were treated with the antiviral drug 1-(2-fluoro-5-methyl-beta-L-arabinofuranosyl)-uracil (L-FMAU) or placebo for 32 weeks. Half the woodchucks in each group then received four injections of surface antigen vaccine during the next 16 weeks. Vaccination alone elicited a low-level antibody response to surface antigen in most carriers but did not affect serum WHV DNA and surface antigen. Carriers treated first with L-FMAU to reduce serum WHV DNA and surface antigen and then vaccinated had a similar low-level antibody response to surface antigen. Following vaccinations, cell-mediated immunity to surface antigen was demonstrated in both groups, independent of serum viral and antigen load, but was significantly enhanced in woodchucks treated with L-FMAU and was broadened to include other viral antigens (core, e, and x antigens and selected core peptides). Cell-mediated immunity and antibody responses to surface antigen were observed after drug discontinuation in half of the carriers that received L-FMAU alone. Surface antigen vaccine alone or in combination with drug broke humoral and cell-mediated immune tolerance in chronic WHV infection, but the combination with drug was more effective. This suggested that a high viral and antigen load in carriers is important in maintaining immunologic tolerance during chronicity. The humoral and cellular immunity associated with the combination of L-FMAU and vaccine resembled that observed in self-limited WHV infection. Such combination therapy represents a potentially useful approach to the control of chronic hepatitis B virus infection in humans.     

Cutting Edge: IFN-gamma-producing CD4 T lymphocytes mediate spore-induced immunity to capsulated Bacillus anthracis

Virulent strains of Bacillus anthracis produce immunomodulating toxins and an antiphagocytic capsule. The toxin component-protective Ag is a key target of the antianthrax immune response that induces production of toxin-neutralizing Abs. Coimmunization with spores enhances the antitoxin vaccine, and inactivated spores alone confer measurable protection. We aimed to identify the mechanisms of protection induced in inactivated-spore immunized mice that function independently of the toxin/antitoxin vaccine system. This goal was addressed with humoral and CD4 T lymphocyte transfer, in vivo depletion of CD4 T lymphocytes and IFN-gamma, and Ab-deficient (muMT(-/-)) or IFN-gamma-insensitive (IFN-gammaR(-/-)) mice. We found that humoral immunity did not protect from nontoxinogenic capsulated bacteria, whereas a cellular immune response by IFN-gamma-producing CD4 T lymphocytes protected mice. These results are the first evidence of protective cellular immunity against capsulated B. anthracis and suggest that future antianthrax vaccines should strive to augment cellular adaptive immunity.     

Tailoring Subunit Vaccine Immunity with Adjuvant Combinations and Delivery Routes Using the Middle East Respiratory Coronavirus (MERS-CoV) Receptor-Binding Domain as an Antigen

The development of an effective vaccine is critical for prevention of a Middle East respiratory syndrome coronavirus (MERS-CoV) pandemic. Some studies have indicated the receptor-binding domain (RBD) protein of MERS-CoV spike (S) is a good candidate antigen for a MERS-CoV subunit vaccine. However, highly purified proteins are typically not inherently immunogenic. We hypothesised that humoral and cell-mediated immunity would be improved with a modification of the vaccination regimen. Therefore, the immunogenicity of a novel MERS-CoV RBD-based subunit vaccine was tested in mice using different adjuvant formulations and delivery routes. Different vaccination regimens were compared in BALB/c mice immunized 3 times intramuscularly (i.m.) with a vaccine containing 10 µg of recombinant MERS-CoV RBD in combination with either aluminium hydroxide (alum) alone, alum and polyriboinosinic acid (poly I:C) or alum and cysteine-phosphate-guanine (CpG) oligodeoxynucleotides (ODN). The immune responses of mice vaccinated with RBD, incomplete Freund’s adjuvant (IFA) and CpG ODN by a subcutaneous (s.c.) route were also investigated. We evaluated the induction of RBD-specific humoral immunity (total IgG and neutralizing antibodies) and cellular immunity (ELISpot assay for IFN-γ spot-forming cells and splenocyte cytokine production). Our findings indicated that the combination of alum and CpG ODN optimized the development of RBD-specific humoral and cellular immunity following subunit vaccination. Interestingly, robust RBD-specific antibody and T-cell responses were induced in mice immunized with the rRBD protein in combination with IFA and CpG ODN, but low level of neutralizing antibodies were elicited. Our data suggest that murine immunity following subunit vaccination can be tailored using adjuvant combinations and delivery routes. The vaccination regimen used in this study is promising and could improve the protection offered by the MERS-CoV subunit vaccine by eliciting effective humoral and cellular immune responses.     

MHC-restricted protection of cats against FIV infection by adoptive transfer of immune cells from FIV-vaccinated donors

The role of cellular immunity in vaccine protection against FIV infection was evaluated using adoptive cell transfer studies. Specific-pathogen-free cats received two adoptive transfers of washed blood cells from either vaccinated or unvaccinated donors with varying MHC compatibility at 1-week intervals, and a homologous FIV(Pet) challenge 1 day after the first adoptive transfer. FIV-specific CTL, IFN-gamma production, and proliferation responses were detected in the PBMC from the vaccinated donors. Seven of eleven (64%) recipients of cells from half-matched/vaccinated donors remained negative for FIV-antibodies after FIV challenge and four of those were completely protected. Two of two recipients of cells from MHC-identical/vaccinated donors were completely protected. All recipients of cells from unrelated/vaccinated, half-matched/unvaccinated, or unrelated/unvaccinated donors were unprotected. Thus, protection mediated by adoptive transfer of immunocytes from vaccinated cats was MHC-restricted, occurred in the absence of antiviral humoral immunity, and correlated with the transfer of cells with FIV-specific CTL and T-helper activities.     

迟缓爱德华菌菌蜕疫苗对罗非鱼注射和口服免疫的效果分析

通过腹腔注射、口服两种免疫途径探讨迟缓爱德华菌菌蜕疫苗对罗非鱼Oreochromis niloticus的免疫保护效果。将制备的迟缓爱德华菌菌蜕疫苗（ETG）和福尔马林灭活疫苗（FKC）采用腹腔注射、口服两种免疫途径免疫罗非鱼,分别于免疫后14d、21d和28d采集罗非鱼血清、头肾、脾脏,测定血清中抗体IgM水平,血清中酸性磷酸酶（ACP酶）、超氧化物歧化酶（SOD酶）活性及罗非鱼头肾和脾脏中白介素（IL-1）、肿瘤坏死因子（TNF）、干扰素（IFN）、Caspase3等细胞因子的相对表达量,并通过攻毒试验得到菌蜕疫苗、福尔马林灭活疫苗两种疫苗的相对免疫保护率。免疫组罗非鱼的血清抗体水平均极显著高于（P<0.01）对照组,ETG注射组抗体效价极显著高于（P<0.01）FKC口服组。免疫28d,免疫组SOD、ACP酶活力显著高于（P<0.05）对照组（Group E、F）;在头肾中,免疫组（Group A、B）TNF、IL-1和IFN的相对表达量显著高于（P<0.05）对照组（Group E、F）。在免疫保护试验中,所有免疫组的免疫保护率均显著高于（P<0.05）对照组,注射、口服菌蜕疫苗的相对保护率分别为79%、77%,注射、口服灭活疫苗组的的相对保护率分别为62%、60%,但两种疫苗免疫保护率无显著差异。试验研制的菌蜕疫苗得到更高的免疫保护率,菌蜕疫苗在预防罗非鱼爱德华菌病中有良好的研究价值和应用前景。     

Passive antibody protection of cats against feline immunodeficiency virus infection.

All six cats passively immunized with sera from either feline immunodeficiency virus (FIV)-vaccinated cats or cats infected with FIV (Petaluma strain) were protected from homologous FIV infection at a challenge dose that infected all six control cats. Passive immunization with sera from cats vaccinated with uninfected allogeneic T cells used to grow the vaccine virus did not protect either of two cats against the same FIV challenge. These results suggest that antiviral humoral immunity, perhaps in synergy with anticellular antibodies, may be responsible for previously reported vaccine protection.     

The protective role of postvaccinal immunity in mumps in children

The immunological study of children with infectious parotitis (IP) without complications and with such complications as pancreatitis, meningitis or orchitis in the glandular form was carried out. In accordance with the previously proposed principle, 4 types of immune response (IR) were established on the basis of differences in initial resistance and the IR profile: cell-mediated immunity (types I and III) and humoral immunity (types II and IV). The patients included nonvaccinated children, as well as children vaccinated on epidemic indications, 3-6, 7-9, 10 and more years before infection. The comparative analysis of the number of IP cases with and without complications in the groups of children, divided according to their immunization history and the type of IR, revealed that postvaccinal immunity in children vaccinated on epidemic indications (less than a month ago) or 3-6 years before infection had protective potential, sufficient for the prevention of complicated forms of IP. Immunity obtained 7-9 years ago was effective for the protection from IP complications only in cell-mediated, but not humoral IR. Postvaccinal immunity obtained more than 10 years ago did not ensure the decrease in the occurrence of complicated forms of IP (in comparison with that in nonvaccinated patients) in children with any type of IR.     

IgA-driven T cell-mediated anti-bacterial immunity in man after live oral Ty 21a vaccine

Cellular immunity against Salmonella typhi was observed by using a direct anti-bacterial in vitro assay in volunteers orally vaccinated with the live S. typhi mutant strain Ty 21a. With this experimental approach, it was demonstrated that Ty 21a vaccine also induces cellular immunity against S. paratyphi A and B. Interestingly, the mechanism involved in cellular immunity against bacteria seems to be of an antibody-dependent cellular cytotoxicity (ADCC) type, with IgA acting as the humoral arm and CD4+ T lymphocytes as the cellular one. In accordance with the increase in IgA-driven ADCC against S. typhi, a major rise in IgA against O and H antigens was observed in the serum of vaccinees in parallel to an increase in IgG of identical specificity. Furthermore, a Ty 21 vaccine induced cellular activity against flagellar antigens. These results indicate that IgA-ADCC by T lymphocytes against bacteria can originate from local stimulation of the gut mucosal immune system. This cellular defense mechanism might be at the origin of the protection induced by Ty 21a vaccine.