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.     

Fundamentals of host immune response against Brucella abortus: what the mouse model has revealed about control of infection

The studies reviewed here evaluated the role cellular immune system components play in control of brucellosis by conducting comparative studies with brucella-resistant C57BL/10 or C57BL/6 mice and susceptible BALB/c mice. We have shown by both in vitro and in vivo studies that activation of macrophages with interferon-gamma (IFN-γ) is an important factor for control of infection with B. abortus in the mouse model and that the mechanism of anti-brucella activity largely involved reactive oxygen intermediates. Differences in control of the organism by resistant and susceptible mice was not related to inherent differences in the ability of their macrophages to control infection either with or without IFN-γ activation nor was it attributable to NK cells since we found no role for them in control of brucellosis in either mouse strain. However, relative resistance to brucellosis did correlate with increased production of IFN-γ by CD4 T cells during the first weeks after infection while IL-10 contributed to susceptibility in BALB/c mice. Moreover, by 3 weeks post-infection splenocytes from the susceptible BALB/c mice failed to produce IFN-γ and relied on TNF- as well as CD8 T cells to control infection until the end of the plateau phase around 6 weeks post-infection when IFN-γ production resumed and clearance began. In contrast, IFN-γ was crucial for control throughout the infection in the more resistant C57BL/6 mice and the mice died in its absence by 6 weeks post-infection compared to 12 weeks for the more susceptible mice that relied on additional mechanisms of control. In contrast to the IFN-γ knock-out mice, both β2 microglobulin knock-out C57BL/6 mice, which do not express conventional MHC class I molecules and thus cannot present antigen to CD8 T cells, or perforin knock-out C57BL/6 mice, which have no T cell cytotoxic activity, controlled and cleared the infection as well as normal C57BL/6 mice. The hiatus of IFN-γ production in BALB/c mice correlated with very high levels of total IL-12 and it was postulated that the lack of IFN-γ was a consequence of p40 homodimer blocking activity. However, reduction of p40 IL-12 in vivo through administration of indomethacin reduced the infection without a concomitant measurable increase in IFN-γ. Current studies are aimed at elucidating the mechanism of the IFN-γ hiatus.     

二乙基己烯雌酚对成年仓鼠睾丸一氧化氮生成和生精细胞凋亡的影响

探讨了在二乙基己烯雌酚（DES）诱发成年动物生精细胞凋亡过程中睾丸一氧化氮（NO）生成和生精细胞一氧化氮合酶（eNOS和iNOs）表达的变化，以期为阐明DES诱发生精细胞凋亡机理的研究提供基础资料。成年雄性仓鼠皮下注射不同剂量DES（分别为0．01、0．1和1mg／kg体重），连续注射7d后取其睾丸，进行NO含量的测定和eNOS、iNOS免疫组化染色。电镜观察生精细胞超微结构的变化，并用TUNEL法检测睾丸中生精细胞凋亡的变化，苏丹Ⅲ染色法检测睾丸生精小管内脂滴分布的变化。结果显示：NO的生成与DES呈剂量依赖性。DES处理后，在1mg／kg体重剂量组，大量生精细胞表达eNOS和iNOS，并出现大量凋亡，退化的生精细胞胞浆内有大量髓样结构，并有大量脂滴分布于生精细胞内和细胞间。eNOS和iNOS阳性生精细胞与凋亡的生精细胞数量和类型基本一致，主要为精母细胞和圆形精子细胞。     

造血和神经相关标志物在人胎盘的表达

为探讨人胎盘组织在造血中的作用以及研究造血和神经标志物在胎盘中的表达和在胚胎发育中的相关性，用免疫组织化学法对人胎盘组织和体外培养的胎盘组织中的贴壁细胞进行染色，观察其造血因子和神经标志物的表达。结果发现人7月龄胎盘组织血管内血细胞表达多种造血细胞因子，如SCF、VEGF、BMP-4和FGF-1和KDR，血管内皮细胞上也有表达；胎盘组织同时表达造血干细胞标记CD34、CD133和神经细胞标记物Nestin和MAP2等。而足月龄胎盘CD34、CD133、KDR和造血相关因子SCF、VEGF、BMP-4、FGF-1等弱表达，不表达Nestin和MAP2。两组胎盘切片都不表达GFAP和MBP。胎盘贴壁细胞（hPDACs）碱性磷酸酶、波形蛋白、CD133、Nestin和MAP2染色呈阳性，CD34、GFAP和MBP阴性表达。人胎盘组织和胎盘贴壁细胞表达多种造血相关因子和神经细胞标志物，提示胎盘具有造血功能，造血形成和神经发生之间可能存在基因表达叠加现象。     

毒害艾美耳球虫初次感染雏鸡免疫器官IL-2和淋巴细胞增殖功能变化

应用组织匀浆涂片和酸性α-醋酸萘酯酶（ANAE）染色及细胞培养技术和四甲基偶氮唑盐（MTT）测定法对毒害艾美耳球虫（eimeria necatrix，E．necatrix）初次感染雏鸡免疫器官的T细胞比例、白细胞介素-2（interleukin-2，IL-2）诱生活性、T细胞和B细胞对ConA或PMA的增殖功能的动态变化进行了较全面系统的研究。结果发现，E．necatrix初次感染雏鸡，其胸腺和脾脏T细胞比例分别于感染后7～21d和7～24d明显高于对照雏鸡；IL-2诱生活性分别于感染后16～18d和18～21d较对照雏鸡显著升高；T细胞对ConA的增殖反应分别在感染后14～16d和10～18d明显增加。法氏囊和脾脏B细胞对PMA的增殖反应分别于感染后14～24d和14～21d显著高于对照雏鸡。表明E．necatrix初次感染雏鸡免疫器官的IL-2调节及细胞免疫和体液免疫功能均明显提高。     

鲁西黄牛体细胞克隆保种技术的研究

以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体，研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛（1♂，4♀）成纤维细胞系，作为核移植供体细胞，利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植，试验结果表明：重构胚的融合率为62．5％（242／387），分裂率为63．6％（154／242），体外培养第7天囊胚发育率为42．9％（66／154）。体外培养第7天的囊胚的内细胞团细胞（ICM）与滋养层细胞数平均为37和47，ICM占44．2％。体外发育到7d的囊胚新鲜胚胎的移植妊娠率（新鲜胚胎）移植受体10头，60d妊娠率为20％（2／10）。     

不同体外培养条件对家兔早期囊胚贴壁行为的影响

采集家兔自然交配后96h的早期囊胚，以低糖DMEM＋150mL／L胎牛血清＋0．1mmol／L非必需氨基酸＋100IU／mL青霉素＋100IU／mL链霉素为基础培养基，比较了不同胚胎处理方法、不同饲养层以及培养液中添加不同成分对兔早期囊胚贴壁和增殖的影响，以完善兔胚胎干细胞的建系方法。结果表明，以胚胎分割法和链霉蛋白酶-E（proteinaseE）处理掉黏蛋白及部分透明带的胚胎容易贴壁和增殖；在小鼠成纤维细胞饲养层和兔胎儿成纤维细胞饲养层上，胚胎的脱带率差别不大，但在小鼠成纤维细胞饲养层上贴壁率明显提高，且贴壁后内细胞团增殖较快；添加胰岛素、白血病抑制因子和伊巯基乙醇均利于抑制ES的分化和促进内细胞团的增殖。     

饲养层细胞对胚胎干细胞的影响及其分离培养的影响因素

胚胎干细胞及其相关研究是生命科学领域中最有活力、最有影响力、最有应用前景的研究方向之一，它与遗传工程，转基因技术等相结合具有重大的理论意义与实践价值。饲养层细胞在胚胎干细胞分离培养的过程中起着异常重要的作用。作者结合实践经验就饲养层细胞对胚胎干细胞的影响进行了概述，介绍了饲养层细胞的分类，并分析了饲养层细胞分离培养的影响因素。     

黄芩及其成分对RU486诱导小鼠流产的保胎作用及脾脏CD80+、CD86+细胞数量的影响

为了研究共刺激分子CD80(B7-1)、CD86(B7-2)在流产发生机制中的意义,探讨中药黄芩及其成分的安胎作用及机理,本试验用米非司酮(RU486)皮下注射(每鼠90μg)诱导BALB/c小鼠流产,流式细胞仪测定脾脏中CD80 、CD86 细胞的数量。发现RU486诱导流产的小鼠脾脏中CD80 细胞显著升高,CD86 细胞显著减少。预先经口给予保胎中药黄芩及其单体成分黄芩苷和黄芩素,则能显著抑制RU486的流产作用,使母鼠胚胎吸收率降低,脾脏中CD80 细胞数量显著降低,CD86 细胞数量显著升高。这些结果表明,RU486诱导流产与机体共刺激分子CD80 、CD86 有关。保胎中药黄芩及单体成分有调节共刺激分子CD80 、CD86 细胞数量的作用。