皮下注射鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中IgA、IgM和IgG的发生规律

为探讨鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中抗体发生的规律,将鸭瘟病毒Cha株弱毒苗皮下注射免疫20日龄樱桃谷鸭后60 d内定时随机剖杀5只鸭,采集血清、胆汁、气管和消化道(食道、十二指肠、空肠、回肠、盲肠和直肠)分泌液,应用间接ELISA检测抗DPV的IgA、IgM和IgG滴度(以Log10表示).结果表明:①血清:抗体滴度由高到低为IgG、IgM和IgA,相应的检测到的时间段分别为免疫后6～60,3～15,12～36 d.②胆汁:抗体滴度由高到低为IgA、IgG和IgM,相应的检测到的时间段分别为免疫后9～21,15～27,3～12 d.③分泌液:气管和消化道分泌液中抗体滴度由高到低均为IgA、IgM和IgG,其中IgA抗体滴度由高到低为十二指肠、食道、气管、空肠、盲肠、回肠和直肠,相应的检测到IgA的时间段分别为免疫后3～60,9～60,3～60,9～60,12～60,12～27,6～36d;IgM由高到低为气管、食道、十二指肠、空肠、盲肠、直肠和回肠,相应的检测到IgM的时间段分别为免疫后3～12,6～15,3～12,6～12,9～12,6～9,6～9 d;IgG由高到低为食道、十二指肠、气管、空肠、盲肠、直肠和回肠,相应的检测到IgG的时间段分别为免疫后9～36,12～27,6～36,9～36,12～36,9～21,15～21 d.综上,鸭瘟弱毒疫苗皮下免疫鸭后,IgM和IgG分别是系统免疫中体液免疫的先锋抗体和主要抗体;IgA是气管、消化道和胆汁中的主要抗体.     

Analysis of the IgG antibody response against Paramphistomidae trematoda in naturally infected cattle. Application to serological surveys

The IgG antibody response to Calicophoron daubneyi (Digenea: Paramphistomidae) excretory/secretory antigens was evaluated in naturally infected cattle from Lugo (Galicia, NW Spain) by using an ELISA procedure. Two studies were conducted, first a survey in 524 cattle separated into three groups according to age, G-1 (0–2 years old), G-2 (3–5 years old) and G-3 (>6 years old). In the second study, three groups of cattle were employed: G-I, naturally infected; G-T, naturally infected and treated with oxyclozanide plus levamisole (Nilzan Plus^®); G-C, cattle maintained in a farm where C. daubneyi has never diagnosed. Variations on egg-output and haematic parameters (erythrocytes, haematocrite, leukocytes and lymphocytes) were also analyzed.

The ELISA procedure showed that 61.2% of the cattle in the first study had been exposed to the trematode, but only 10.1% passed eggs in the feces. Age-association with egg-output was shown but not with the IgG values. In the second experiment, the administration of the anthelmintic reduced significantly the IgG kinetic levels and the C. daubneyi-egg-output was suppressed during 12 weeks in the G-T group. The values of red cells, haematocrite, leukocytes and lymphocytes increased significantly in the treated cattle 5 weeks after chemotherapy; however, new reduction after week 5 was recorded, as results of the challenge of these cattle.

This is the first investigation in which evaluation of the IgG humoral response against C. daubneyi in cattle has been carried out. We proved that a notable IgG response in naturally infected cattle is induced, and can be detected by using an ELISA procedure. The IgG antibodies did not increase after challenge infection. Our results proved an important percentage of cattle were exposed to this trematode in the area of study and suitable measures for preventing this relationship must be considered.     

兔抗猪瘟高免血清的制备及纯度鉴定

利用猪瘟弱毒疫苗(C株)免疫新西兰大白兔,制备抗猪瘟高免血清IgG。运用间接ELISA方法测血清效价,再经饱和硫酸铵盐析沉淀法提纯血清IgG,SDS-PAGE电泳法鉴定提纯IgG的纯度,结合无菌检查试验和仔猪接种试验,检测血清IgG的安全性。结果表明,获得了蛋白含量为6 mg/mL,效价为1∶6400的高效价、安全性高、成本低的抗猪瘟高免血清IgG。为猪瘟病毒的检测和猪瘟的防制提供了理想的生物制剂。     

初乳清粉的货架期预测和贮藏稳定性

为了掌握初乳清粉的贮藏特性,采用货架期预测方程来估计初乳清粉在室温(25℃)、50％相对湿度(RH)条件下PET包装的货架期,并对90d贮藏期过程中其理化指标以及生化指标的变化进行研究.研究结果表明:在PET包装90d贮藏时间内,初乳清粉的水分含量、硫代巴比妥酸(TBA)值和羟甲基糠醛(HMF)含量均随贮藏时间的延长而显著增加(P＜0.05),色差随贮藏时间的延长有所波动,但总体呈现上升趋势,而免疫球蛋白(IgG)的含量在60d内逐渐降低,60d后其下降不显著;在25℃、50％ RH条件下PET包装的初乳清粉的货架期为99.40d.     

载板蓝根多糖MPEG-PLA聚合物微球对小鼠免疫力的影响

为研究载板蓝根多糖微球的免疫增强作用。利用实验室制备的载板蓝根多糖MPEG-PLA聚合物微球免疫小鼠,并测定小鼠血液的常规指标,即白细胞和淋巴细胞总数以及总IgG水平和腹腔巨噬细胞吞噬能力等免疫指标。结果表明：在给药后第7天和第14天,载板蓝根多糖微球40、20、10 mg组和板蓝根多糖组小鼠的各项免疫指标均高于空白微球组和生理盐水组,并且载板蓝根多糖微球各组在第14天时的免疫效果与第7天时的免疫效果相比较差异不显著,其中,微球组中以载板蓝根多糖微球20 mg组的免疫效果为最好,在第7天和第14天时小鼠的白细胞和淋巴细胞总数分别为(0.96±0.46)和(11.03±0.35)以及(8.73±0.51)和(9.44±0.42),总IgG水平为(33.541±0.458)和(32.713±0.481),巨噬细胞吞噬指数和吞噬百分率则分别达到(56.00±2.16)和(1.61±0.04)。载板蓝根多糖微球具有一定的免疫增强作用,虽然微球的短期免疫效果均低于纯板蓝根多糖,但是可达到长效免疫。     

Efficacy of an oral live vaccine for veterinary use against pseudotuberculosis

Pseudotuberculosis, an infection caused by the ubiquitous enteropathogenic bacterium Yersinia pseudotuberculosis, is a recurrent veterinary problem in livestock and zoo animals. The only vaccine currently available in zoos is Pseudovac (a mixture of killed strains of various serotypes), but its efficacy is not well established. We show here that Pseudovac does not protect guinea pigs against a severe Y. pseudotuberculosis infection. We thus evaluated the possibility of using a live attenuated Y. pseudotuberculosis strain (IP32680) as an oral vaccine against animal pseudotuberculosis. We report that IP32680 is avirulent for guinea pigs and induces a strong IgG response against various serotypes of Y. pseudotuberculosis. One and two oral inoculations of IP32680 provided 50% and 83% protection, respectively against a severe infection with a highly pathogenic strain. The avirulent Y. pseudotuberculosis IP32680 is therefore much more protective than Pseudovac and may represent a valuable oral vaccine against pseudotuberculosis in zoo animals.     

Lipase-ISCOM免疫对小鼠乳中IgG的影响

选择100只健康昆明小白鼠,研究小鼠不同生理阶段以及免疫条件下,乳中总IgG浓度和抗LipaseIgG类抗体效价变化。分娩后4 d分别注射灭菌生理盐水、脂肪酶(Lipase)+灭菌生理盐水、空免疫刺激复合物(ISCOM)、Lipase-ISCOM。分娩后8和12 d(即注射后4和8 d),用ELISA方法测定血清及乳中总IgG浓度和抗Lipase IgG类抗体。结果表明,Lipase-ISCOM免疫小鼠后,血清和乳中抗Lipase IgG类抗体效价(log2)分别达到4.03和3.38,显著高于对照组(P0.05)。ISCOM为免疫乳生产候选疫苗的研究奠定了基础。     

马抗猫细小病毒特异性免疫球蛋白的研制

由猫细小病毒(feline parvovirus,FPV)引起的猫泛白细胞减少症目前仍呈上升势头,发病率和死亡率较高。迄今国内外尚未见到较理想的治疗药物。本试验选用马作为免疫动物,收集血清。通过对传统正辛酸—硫酸铵沉淀法进行优化,用Protein A亲和层析、胃蛋白酶消化,获得精制抗猫瘟免疫球蛋白(IgG)生物制剂。对该生物制剂进行物理性状、热原性、安全性、无菌检验、SDS-PAGE纯度鉴定、血凝抑制试验、双向琼脂扩散试验、细胞中和试验和动物治疗试验。结果表明,该生物制剂为略带乳光的澄明液体,溶液稳定无分层,无可见颗粒,无微生物污染,保存期长;对家兔、小白鼠、豚鼠等试验动物安全稳定、无毒副作用;其热原性符合规定,F(ab′)2分子质量约为110 ku,中和效价在128倍以上,对5只人工感染FPV猫进行治疗,治愈率达100%,证实该制品对猫泛白细胞减少症有很好的治疗效果。     

藏药川西獐牙菜抗小鼠免疫性肝损伤的研究

【目的】探讨川西獐牙菜对卡介苗(BCG)/脂多糖(LPS)所致的免疫性肝损伤小鼠免疫器官脏器指数和血清免疫球蛋白含量的影响。【方法】采用尾静脉注射BCG和LPS构建小鼠(体质量20g)免疫性肝损伤模型。制备川西獐牙菜提取物,按高剂量(5.380mg/只)、中剂量(2.690mg/只)、低剂量(0.538mg/只)灌胃免疫性肝损伤小鼠,连续10d后,取小鼠肝脏、脾脏和胸腺,计算其脏器指数,并取血清,测定其中IgG、IgA、IgM的含量。【结果】川西獐牙菜的各给药组能有效防止BCG/LPS对小鼠各脏器的免疫性损伤,并能影响IgG、IgA、IgM的含量,调节B淋巴细胞网络的紊乱,使异常的免疫状态恢复正常。【结论】川西獐牙菜预防性给药能减轻免疫性肝损伤对小鼠肝脏的破坏,减弱体液免疫功能的亢进,从而起到保肝、抗病、抗损伤的作用。