猪伪狂犬病油乳剂灭活疫苗的制备及安全性与免疫性试验

用本室分离鉴定的猪伪狂犬病毒鄂A经毒株接种仓鼠肾细胞（BHK－21）,制备病毒抗原液，毒价不低于10^-6TCID50/0.1ml。经一定浓度的甲醛溶液灭活后与油相佐剂乳化研制成油乳剂灭活油疫苗4批。本研究对该制品的安全性、免疫性进行了测定。对18g左右小白鼠接种0.3ml，初生仔猪、断奶仔猪及妊娠母猪加倍剂量注射，均未出现不良反应，安全性良好。对母猪的繁殖性能不产生影响。后备母猪及妊娠母清中和抗体指数于免疫后21d达到316以上，间隔35d加强免疫一次后，中和抗体指数可达1000以上。断奶仔猪及初生仔猪免疫后对强毒的攻击，保护率分别为100％及90.62％。     

牛病毒性腹泻／粘膜病O系弱毒冻干苗的试验生产和检验报告

１９９２￣１９９４年共试生产牛病毒性腹泻／粘膜病（ＢＶＤ／ＭＤ）Ｏ系细胞培养弱毒冻干苗６０万头份，通过实验室内安全检验、效力检验、无菌检验、水份测定、真空度检验、物理性状检验及在玉树、称多和泽库三个疫病发生严重的县试用，证明《ＢＶＤ／ＭＤＯ系细胞培养弱毒冻干苗制造与检验试行办法》可行，并为制定该苗的制造与检验规程提供了依据。     

Vaccination experiments on pregnant mice with E. coli vaccines prepared from septicaemic and enteropathogenic strains of E. coli

Experiments have been carried out with vaccination of pregnant mice against E. coli, followed by i.p. challenge of the offspring at one week of age.With a septicaemic strain the results were highly significant, and the method is therefore recommendable for testing of vaccines against such strains of E. coli.Results were less clear-cut with enteropathogenic strains of E. coli. However, with mortality rates of 40 to 45 % in baby mice born by non-vaccinated mothers and less than 15 % in baby mice born by vaccinated mothers, the difference in percentage mortality seems sufficient to warrant the use of the method also in the control of vaccines against enteropathogenic E. coli strains.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518     

Poxviruses as vaccine vectors

The discovery of Jenner in 1798 founded the science of immunology and eventually led to smallpox eradication from the earth in 1980 after a world-wide vaccination campaign with vaccinia virus (another poxvirus) and paradoxically, despite the eradication of smallpox, there has been an explosion of interest in vaccinia virus in the eighties. This interest has stemmed in part from the application of molecular genetics to clone and express foreign genes from recombinant vaccinia viruses. Vaccinia is also gaining renewed interest due to bioterrorism.

These recombinant viruses have multiple applications in research and vaccinology and led to the development of vectored vaccines, such as the recombinant vaccinia rabies vaccine used to eliminate rabies in Western Europe and, more recently, in the United States. Secondly, alternative poxvirus vectors, such as avipox viruses, were proved to be even safer and efficacious non-replicating vectors (suiciole vectors) when used in non-avian species.     

New gene-based approaches for an AIDS vaccine

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.     

Comparison of antibody values in sera of pigs vaccinated with a subunit or an attenuated vaccine against classical swine fever

Ten pigs, aged 85 days, were vaccinated with a subunit vaccine containing 32 g of classical swine fever virus glycoprotein E2 (gp E2) (group 1), and a further 10 pigs were vaccinated with a C strain vaccine (10^4±0.15 TCID₅₀/ml), produced by amplification in minipig kidney (MPK) cell culture (group 2). Nine non-vaccinated pigs served as a control group (group 3). Serum samples were collected before (day 0) and at 4, 10, 21 and 28 days after vaccination and were analysed by two commercially available enzyme immunoassays and by a neutralizing peroxidase-linked assay (NPLA). At the same times, peripheral blood was taken for determining the total leukocyte count and the body temperature was taken daily. Antibodies were not detected in serum samples collected before vaccination (day 0), and no side-effects that could be connected with vaccination were observed during the trial. Ten days after vaccination 6/10 pigs vaccinated with the subunit vaccine were seropositive. On days 21 and 28, the ratios of serologically positive to vaccinated pigs were 9/10 and 10/10, respectively. Four of the ten pigs that were vaccinated with the C strain vaccine were positive on day 21 and 9/10 on day 28. However, the results of the NPLA showed that only 4/10 pigs had an antibody titre >1:32 at the end of the trial in both the vaccinated groups, even though the subunit vaccine initiated an earlier and higher level of neutralizing antibodies than the vaccine produced from the C strain. Challenge was performed 28 days after vaccination on four randomly selected pigs from both vaccinated groups. The pigs survived the challenge without showing any clinical signs of classical swine fever (CSF), while two nonvaccinated control pigs died on the 10th and 12th days after infection.     

Seasonal atopic dermatitis in dogs sensitive to a major allergen of Japanese cedar (Cryptomeria japonica) pollen

Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme-linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs.     

Immune Responses to Intermediate Strain IBD Vaccine at Different Levels of Maternal Antibody in Broiler Chickens

Tropical Animal Health and Production -     

犬冠状病毒基因疫苗表达载体的构建及其免疫原性研究

以pVAX1为载体首次构建了犬冠状病毒病基因的3种真核表达质粒。首先将犬冠状病毒大熊猫株（CCVDXMV）纤突蛋白（S）、膜蛋白（M）和核蛋白（N）基因进行了克隆和序列测定。将测序后的质粒pTS、pTM和pTN分别双酶切，回收目的基因片段并将其定向克隆到真核表达质粒pVAX1中得到重组质粒pVAXS、pVAXM和pVAXN。将这3种真核表达质粒通过脂质体介导法转染MDCK细胞，通过RT—PCR法进行转录水平的检测，并用间接ELISA法检测目的蛋白的表达情况。结果在pVAXS、pVAXM、pVAXN转染MDCK细胞36h后就可检测到目的基因的转录；在转染72h后可检测到3种目的蛋白的表达。动物免疫试验表明，3种真核表达载体能有效地诱导机体产生细胞免疫与体液免疫应答，这为CCV基因疫苗的研究奠定了良好的基础。