禽流感灭活抗原与佐剂配合鼻腔免疫对鸽呼吸道抗体分泌细胞的影响

通过禽流感灭活抗原配合黏膜免疫佐剂鼻腔免疫乳鸽,研究鸽呼吸道各段抗体分泌细胞的分布和数量。结果显示,首免后第3、5周,应用CpG免疫后肺IgG分泌细胞面积显著高于对照组（P〈0.05）,首免后第5,7周,应用CpG免疫后肺IgA分泌细胞面积显著高于对照组（P〈0.05）;首免后第3、5、7周,应用灭活禽流感抗原免疫后,鸽呼吸道各部位IgG分泌细胞和IgA分泌细胞面积与对照组无显著差异;应用禽流感抗原配合CpG和胆酸钠免疫后鸽呼吸道各部位单位面积中IgA分泌细胞和IgG分泌细胞面积均显著或极显著高于对照组（P〈0.01,P〈0.05）。结果表明,灭活禽流感抗原配合黏膜免疫佐剂通过鼻腔免疫能够提高呼吸道中IgA分泌细胞和IgG分泌细胞的面积,增强局部呼吸道体液免疫应答水平。     

Serum neutralizing antibody titers in dairy cattle administered an inactivated vesicular stomatitis virus vaccine

Two doses of a formalin-killed, cell culture-derived vesicular stomatitis virus (vsv)-New Jersey serotype vaccine were administered intramuscularly, 30 days apart, to all lactating and nonlactating cows in a 350-cow dairy herd. Serum specimens were obtained serially from 96 cows before vaccination and at 30, 52 and 80 days after vaccination and from 24 of these cows 175 days after vaccination. Serum neutralizing antibody titers to vsv-New Jersey serotype were determined from serum-dilution, plaque-reduction tests. Serum neutralizing antibody titers also were determined during the same period for 67 nonvaccinated heifers in the herd. Peak group geometric mean serum neutralizing antibody titers of 1:530.46 +/- 1.14 (group geometric mean titer log10, 2.725 +/- 0.055) developed 21 days after the second vaccination, but decreased to a low value of 1:65.36 +/- 1.38 (group geometric mean titer log10, 1.815 +/- 0.142) by 175 days after vaccination. The nonvaccinated group had no detectable antibody titer to vsv-New Jersey serotype throughout the study. All serum specimens from the vaccinates and controls were negative for heterologous reactivity to vsv-Indiana serotype.     

A lack of antibody formation against inactivated influenza virus after aerosol vaccination in presence or absence of adjuvantia

In the poultry industry, infections with avian influenza virus (AIV) can result in significant economic losses. The risk and the size of an outbreak might be restricted by vaccination of poultry. A vaccine that would be used for rapid intervention during an outbreak should be safe to use, highly effective after a single administration and be suitable for mass application. A vaccine that could be applied by spray or aerosol would be suitable for mass application, but respiratory applied inactivated influenza is poorly immunogenic and needs to be adjuvanted. We chose aluminum OH, chitosan, cholera toxin B subunit (CT-B), and Stimune as adjuvant for an aerosolized vaccine with inactivated H9N2. Each adjuvant was tested in two doses. None of the adjuvanted vaccines induced AIV-specific antibodies after single vaccination, measured 1 and 3 weeks after vaccination by aerosol, in contrast to the intramuscularly applied vaccine. The aerosolized vaccine did enter the chickens' respiratory tract as CT-B-specific serum antibodies were detected after 1 week in chickens vaccinated with the CT-B-adjuvanted vaccine. Chickens showed no adverse effects after the aerosol vaccination based on weight gain and clinical signs. The failure to detect AIV-specific antibodies might be due to the concentration of the inactivated virus.     

Intraocular vaccination with an inactivated highly pathogenic avian influenza virus induces protective antibody responses in chickens

Because it is expected to induce cross-reactive serum and mucosal antibody responses, mucosal vaccination against highly pathogenic avian influenza (HPAI) is potentially superior to conventional parenteral vaccination. Here, we tested whether intraocular vaccination with an inactivated AI virus induced protective antibody responses in chickens. Chickens were inoculated intraocularly twice with 10⁴ hemagglutination units of an inactivated H5N1 HPAI virus. Four weeks after the second vaccination, the chickens were challenged with a lethal dose of the homologous H5N1 HPAI virus. Results showed that most of the vaccinated chickens mounted positive antibody responses. The median serum hemagglutination inhibition titer was 1:80. Addition of CpG oligodeoxynucleotide 2006 or cholera toxin to the vaccine did not enhance serum antibody titers. Cross-reactive anti-hemagglutinin IgG, but not IgA, was detected in oropharyngeal secretions. In accordance with these antibody results, most vaccinated chickens survived a lethal challenge with the H5N1 HPAI virus and did not shed the challenge virus in respiratory or digestive tract secretions. Our results show that intraocular vaccination with an inactivated AI virus induces not only systemic but also mucosal antibody responses and confers protection against HPAI in chickens.     

Duration of circulating antibody and immunity following infection with equine influenza virus

The duration of immunity as measured by virological, serological and clinical responses following infection with influenza A/equine/Newmarket/79 (H3N8) was assessed in repeated challenge experiments in which ponies were infected by exposure to aerosols of infectious virus. Previous infection stimulated complete clinical protection which persisted for at least 32 weeks as demonstrated by the absence of febrile responses and coughing in two groups of ponies infected 16 weeks or 32 weeks after the first infection. Partial clinical protection persisted for over a year as demonstrated by the absence of coughing and a reduction in the number of febrile responses in a group of ponies infected 62 weeks after their first infection. These results contrasted with those observed in immunologically naive control ponies which developed pyrexia, dyspnoea and nasal discharge and coughing. The kinetics of virus specific antibody production in primary and secondary infections with equine influenza were studied by the single radial haemolysis test and a radioisotopic antiglobulin binding assay which measured virus specific IgGab antibody isotype. Antibody to the haemagglutinin, as measured by the single radial haemolysis test, declined rapidly after primary infection whereas the IgGab responses to whole virus antigens persisted for longer. The single radial haemolysis test was therefore particularly useful for the detection of antibody responses in multiple infections or exposures to influenza antigens. The radioisotopic antiglobulin binding assay was more sensitive for identifying infections which had occurred more than six months previously, as evidenced by anamnestic IgGab responses in ponies with low levels of antibody before rechallenge.     

鸵鸟禽流感H5N1灭活疫苗免疫抗体监测

选择50日龄健康鸵鸟40只,随机均分为4组,于50日龄首免、75日龄强化免疫不同剂量的高致病性禽流感H5N1灭活疫苗,结果只有首免3mL、强化免疫5mL组,在125日龄时平均抗体水平在1：32以上,可以产生免疫保护反应。因此认为生产中可以于50日龄对鸵鸟首免、75日龄强化免疫高致病性禽流感H5N。灭活疫苗,剂量应在3mL以上。     

Comparison of humoral and cellular immune responses to inactivated swine influenza virus vaccine in weaned pigs

Humoral and cellular immune responses to inactivated swine influenza virus (SIV) vaccine were evaluated and compared. Fifty 3-week-old weaned pigs were randomly divided into the non-vaccinated control group and vaccinated group containing 25 pigs each. Pigs were vaccinated intramuscularly twice with adjuvanted UV-inactivated A/SW/MN/02011/08 (MN/08) H1N2 SIV vaccine at 6 and 9 weeks of age. Whole blood samples for multi-parameter flow cytometry (MP-FCM) and serum samples for hemagglutination inhibition (HI) assay were collected at 23 and 28 days after the second vaccination, respectively. A standard HI assay and MP-FCM were performed against UV-inactivated homologous MN/08 and heterologous pandemic A/CA/04/2009 (CA/09) H1N1 viruses. While the HI assay detected humoral responses only to the MN/08 virus, the MP-FCM detected strong cellular responses against the MN/08 virus and significant heterologous responses to the CA/09 virus, especially in the CD4+CD8+ T cell subset. The cellular heterologous responses to UV-inactivated virus by MP-FCM suggested that the assay was sensitive and potentially detected a wider range of antigens than what was detected by the HI assay. Overall, the adjuvanted UV-inactivated A/SW/MN/02011/08 H1N2 SIV vaccine stimulated both humoral and cellular immune responses including the CD4-CD8+ T cell subset.     

新-流二联灭活疫苗不同方法免疫后抗体水平检测

为探讨鸡新城疫-禽流感(H9N2 HP株)二联灭活疫苗与禽流感抗体不同组合方法免疫商品蛋鸡和商品肉鸡后抗体水平变化规律,试验分别采用A、B、C三种方法免疫商品蛋鸡与商品肉鸡各40羽,A免疫方法为只接种新-流二联灭活苗,B免疫方法为接种新-流二联灭活苗的同时注射禽流感抗体,C免疫方法为接种新-流二联灭活苗7 d后注射禽流感抗体,D组为空白对照组,各组均于免前及免后2d、6d、10d、14d、21d、28d、35d采血检测新城疫与禽流感抗体水平.结果发现,商品蛋鸡与商品肉鸡各免疫组,免后新城疫抗体均先开始下降,免后第10 d抗体开始上升,第14 d开始快速上升,第35 d达到最高,蛋鸡组最高达8log2,而肉鸡组最高达7log2.商品蛋鸡组中配合注射禽流感抗体免疫组,禽流感抗体先上升,然后下降,再继续上升,禽流感抗体保持较高的水平,不低于3log2.而未注射抗体组,禽流感抗体先下降,至第10 d下降至2log2以下,然后开始快速上升.而商品肉鸡各组禽流感抗体免后均开始下降,未注射抗体组,下降至第10 d后开始上升,注射抗体组免后第10 d时下降的幅度比未注射抗体组小,且下降至第15 d后才开始上升.说明商品蛋鸡免疫新-流二联灭活疫苗时配合注射禽流感抗体,短期内对机体禽流感抗体水平提升明显,而在商品肉鸡上配合注射流感抗体对机体禽流感抗体水平提升不明显.     

一种免疫增强剂对新-流二联灭活苗抗体消长规律的影响

免疫增强剂是一种能增加疫苗免疫效果的生物活性物质。试验A组为添加了免疫增强剂的新城疫、禽流感(H9亚型)二联灭活疫苗(La Sota株+HP株),试验B组为未添加免疫增强剂的新城疫、禽流感(H9亚型)二联灭活疫苗(La Sota株+HP株),分别比较A组与B组的抗体水平。试验结果表明:7日龄SPF鸡A组抗体与B组抗体无差异;在28日龄SPF鸡上:免后10、15、21、28、35 d,ND部分A组抗体比B组分别高0.2log2、0.6log2、1.5log2、0.9log2、0.5log2,AI部分抗体高出0.1log2、0.3log2、1.2log2、1.0log2、0.6log2;在7日龄商品蛋鸡上A组抗体比B组抗体在免后15、21、28、35 d,ND部分抗体高出1.6log2、2.0log2、1.9log2、3.7log2,AI部分分别高1.3log2、2.7log2、1.8log2、2.0log2。     

Immunostimulatory properties of a novel adjuvant administered with inactivated influenza virus vaccine

The immunopotentiating activity of a new delivery system was investigated comparatively to Alhydrogel adjuvant, as an antiviral inactivated vaccine after one injection. The efficiency of the new formulation (BioMed) was evaluated with an inactivated porcine strain of influenza (A/Sw/IN/1726/88 H1N1) in the pig model. The first assessment criteria was the follow-up of selected immunological parameters such as, antibody levels, lymphoproliferation, double positive CD4+CD8+ T lymphocytes and cytokine production (IL-2, IL-4, IFN-gamma). The second criteria was the estimate of the protection level of animals exposed to a homologous challenge of 50 PID50 one month after a single immunizing or control injection. In the BioMed group of animals, 4 pigs (out of 6) were free of macroscopic lesion, while lesions could be seen in all individuals of other groups and virus was isolated in only one animal, whereas all other animals of other groups had virus in their lungs. This better protection of BioMed animals seems to be correlated mainly with higher levels of antibodies and to a lesser extent with a slightly better CMI response and probably with the production of memory CD4+CD8+ T cells.     

Comparison of antibody responses after vaccination with two inactivated rabies vaccines

Thirty laboratory dogs were randomly assigned to two groups (A and B) of 15 dogs and subcutaneously vaccinated with a single dose of one of two commercially available monovalent inactivated rabies vaccines: RABISIN (Merial, France) (group A) and NOBIVAC Rabies (Intervet International) (group B). Rabies antibodies were measured over a period of 4 months using the fluorescent antibody virus neutralization (FAVN) test. The two vaccines performed differently in terms of magnitude and persistence of rabies antibodies titers in dogs. Two weeks after vaccination, average rabies antibody titers peaked at 2.53 IU/mL (range, 0.17-13.77 IU/mL) and 1.26 IU/mL (range, 0.50-4.56 IU/mL) in groups A and B dogs, respectively. The average FAVN antibody titres against rabies on D28, D56, D84, D112 and D120 were significantly higher in group A than in group B. Although all dogs from group B serologically responded to vaccination, the proportion of dogs with antibody titres >or=0.5 IU/mL dropped significantly after D28 and was statistically significantly lower on D56, D84 and D112 compared to group A dogs. In conclusion, in the context of international trade, the choice of the vaccine and the timing of blood tests are critical factors in achieving successful serological test results after rabies vaccination. RABISIN induces high and sustained antibody titres against rabies, increasing the flexibility for the time of blood sampling after primo-vaccination.     

不同剂型新城疫-禽流感二联灭活疫苗免疫SPF鸡后抗体水平对比试验

不同剂型的佐剂对疫苗的免疫效果有较大的影响,本试验旨在对比不同剂型佐剂的新城疫-禽流感(简称"新-流")二联灭活疫苗的免疫效果,从而为生产上选择最佳剂型佐剂应用于新城疫-禽流流感二联灭活疫苗提供依据。分别将油包水型、水包油型、水包油包水型佐剂与新城疫及禽流感灭活抗原乳化成不同剂型的新城疫-禽流感(H9亚型,HP株)二联灭活疫苗,将这三种不同剂型的新城疫-禽流感(H9亚型,HP株)二联灭活疫苗分别免疫一组7日龄SPF鸡。每羽颈部皮下注射0.2 m L,同时设一组未免7日龄SPF鸡作为对照组,各免疫组与对照组SPF鸡于免疫组免后6、10、15、21、28、35 d进行采血,检测新城疫与禽流感抗体水平。结果表明:免疫油包水型疫苗组SPF鸡免疫后新城疫与禽流感抗体上升最快,在免后10、15、21、28、35 d的新城疫与禽流感抗体也最高,其次是免疫水包油包水型的,而免疫水包油型疫苗组的SPF鸡新城疫与禽流感抗体上升最慢,而且在免后10、15、21、28、35 d的新城疫与禽流感抗体也最低。此外,从各免疫组可以看出,在免后6 d时新城疫抗体已开始产生,在1log2以下,而禽流感抗体仍未产生。可见免疫新-流二联苗后,新城疫抗体比禽流感抗体更早产生,油包水型新-流二联苗的免后抗体高于水包油包水及水包油型,而且能持续刺激机体产生高水平抗体,更具优势。     

Needle-free delivery of an inactivated avian influenza H5N3 virus vaccine elicits potent antibody responses in chickens

A needle-free delivery system was assessed as a route for providing quick, safe, and effective vaccination against avian influenza (AI). Two groups of chickens were vaccinated with a commercially available inactivated H5N3 virus vaccine delivered either with a needle-free device or with the conventional syringe-and-needle method recommended by the vaccine manufacturer. The kinetic aspects of seroconversion, peak antibody levels, and antibody titers were measured by a combination of an indirect enzyme-linked immunosorbent assay and the hemagglutination-inhibition test and were all found to be similar in the 2 groups of chickens. We conclude that the needle-free delivery system could result in effective immunization against H5N1 AI epidemics and pandemics in chickens.     

重组禽流感病毒H5亚型二价灭活疫苗(H5N1,Re-5+Re-4)免疫鸡和鸭后抗体消长规律

禽流感(AI)是由A型流感病毒引起的一种禽类传染病.根据致病性的不同将禽流感分为高致病性禽流感(HPAI)和低致病性禽流感(IPAI),HPAI被世界动物卫生组织(OIE)列为A类传染病,我国将其列为一类动物疫病 [1].近几年来,禽流感在世界各地反复出现,2004年亚洲发生了高致病性禽流感,2006年底和2007年初东南亚又出现疫情,并发现了人感染禽流感.我国养禽业也一直受到禽流感的威胁.因此,它不仅给畜牧业造成了巨大的损失,而且给人们的公共卫生安全带来了严重的威胁.     

猪流感病毒多表位抗原的小鼠黏膜免疫试验

为探讨经黏膜途径免疫接种猪流感病毒(SIV)多肽疫苗的可行性,选择STV主要结构蛋白血凝素(HA)的T、B抗原表位,将多个表位片段串联,定向克隆到原核表达载体pET-30a(+)中,对串联表达的重组猪流感病毒蛋白SIV-ep1、SW-ep2、SIV-el2进行了小鼠免疫试验。重组蛋白以包涵体形式表达,经洗涤定量后,采用灌胃途径免疫6周龄KM小鼠,检测免疫小鼠抗体及细胞免疫水平。结果发现三免后各免疫组血清IgG抗体与PBS对照组差异极显著(P<0.01);黏膜分泌型IgA抗体,所有免疫蛋白组与PBS组差异均极显著(P<0.01);细胞免疫检测,所有免疫组小鼠外周血T淋巴细胞增殖明显;猪流感病毒血凝抗体检测,SIV-ep1免疫组、SIV-ep2免疫组、SIV-ep1与LT蛋白免疫组抗SIVH1N1亚型HI效价达1:1024,SIV-el2蛋白免疫组HI效价达1:512。结果表明,猪流感病毒重组多表位抗原灌胃免疫小鼠,能产生理想的黏膜免疫抗体和血清抗体。     

Functional antibody responses in pigs vaccinated with live and inactivated Aujeszky's disease virus

Functional antibody tests, including virus neutralising activity of serum, antibody dependent cellular cytotoxicity and complement mediated lysis, were used to measure the response of pigs given either live or inactivated Aujeszky's disease virus vaccines. Pigs were then challenged with virulent Aujeszky's disease virus and antibody responses were analysed and found not to correlate with protection. Reasons for this lack of correlation are discussed and it is suggested that these results indicate that more emphasis should be placed on measuring the local immune response.     

Comparison of intramuscular and footpad subcutaneous immunization with DNA vaccine encoding HSV-gD2 in mice

Herpes simplex virus type 2 is the most common infectious agent in humans that causes genital herpes disease and vaccination is a desirable method to prevent herpes infections. An effective therapeutic vaccine will need to elicit virus-specific immune responses. The route of immunization has important role in immune responses. In this study, DNA vaccine encoding glycoprotein D of herpes simplex virus type 2 (HSV-gD2) was prepared and injected via intramuscular and footpad routes to determine the optimal method of delivery for immune stimulation. The control manipulation of immune response by concerning route of administration is highly appreciated issue by researches. Although DNA vaccine containing HSV-gD2 is effective in both intramuscular and footpad injection routes, the latter could induce significantly higher cellular responses against HSV-2.     

Immunoglobulins and anti-Marek's disease virus antibody synthesis in chickens after passive immunization with immunoglobulin Y anti-Marek's disease virus antibody.

The effect of passive immunization with immunoglobulin Y (IgY) antibody against Marek's disease virus (MDV) was examined in MDV-susceptible chickens. The production of IgY, immunoglobulin M, and probably also immunoglobulin A was depressed in passively immunized chickens when compared with that in MDV-exposed chickens which had not been given IgY anti-MDV antibody. In passively immunized chickens, the synthesis of immunoglobulin M and IgY anti-MDV antibodies in response to MDV infection also was delayed as determined by agar gel precipitin and indirect fluorescence antibody tests.