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抗革兰氏阴性外膜菌重组共同抗原猪超免疫血清的制备与特性分析

Production and characterization of swine hyperimmune serum against recombinant, common antigens of Gram-negative outer membrane bacteria.

作者信息

Rząsa A, Pietrasina O, Czerniecki M, Bajzert J, Stefaniak T

机构信息

Department of Immunology, Pathophysiology and Veterinary Preventive Medicine, Wroclaw University of Environmental and Life Sciences, C.K. Norwida 31, 50-375 Wroclaw, Poland.

出版信息

Pol J Vet Sci. 2019 Mar;22(1):117-125. doi: 10.24425/pjvs.2019.127078.

DOI:10.24425/pjvs.2019.127078
PMID:30997780
Abstract

The application of immune serum is one of the most efficient method used formerly in the protection of raised piglets'/weaners' health . The objective of the study was to determine specific antibody response during hyperimmunization of fatteners with a self-prepared subunit vaccine, and to propose production method of immune serum against Gram-negative bacteria antigens. The vaccine was administered every two weeks, 4 times. Individual and pooled serum samples were assayed for IgM, IgG and IgA antibodies against Histophilus somni recombinant Hsp60, H.somni rOMP40 and Pasteurella multocida LPS. Additionally total serum IgG and haptoglobin concentrations were measured. Two weeks after the first vaccination IgM antibody raised significantly against H.s. rOMP40 and LPS, whereas after 4 weeks it increased against rHsp60 antigens. Anti-LPS IgM antibody raised up stepwise till the end of the observation, but IgM antibody against H.s. rHsp60 and H.s. rOMP40 decreased in further samplings. A significant raise in IgG class H.s. rHsp60- -antibody was found 4 weeks after the first immunization and a similar raise against two remain- ing antigens after 6 weeks. The intensity of the reaction increased till the end of the experiment. The raise in IgA antibody level was observed only for H.s. rHsp60 antigen. Clinically observed, proper animal health and welfare were confirmed by haptoglobin concentration, which remained in physiological range. At least 4 booster doses were necessary to obtain hyperimmune serum containing a high level of antibodies against examined antigens. The number of immunizations influenced response profiles for specific IgM, IgG, IgA antibodies.

摘要

免疫血清的应用是过去用于保护断奶仔猪健康的最有效方法之一。本研究的目的是确定育肥猪用自制亚单位疫苗进行超免疫期间的特异性抗体反应,并提出抗革兰氏阴性菌抗原免疫血清的生产方法。疫苗每两周接种一次,共接种4次。对个体和混合血清样本检测针对睡眠嗜组织菌重组热休克蛋白60(Hsp60)、睡眠嗜组织菌重组外膜蛋白40(rOMP40)和多杀性巴氏杆菌脂多糖(LPS)的IgM、IgG和IgA抗体。此外,还测量了血清总IgG和触珠蛋白浓度。首次接种疫苗两周后,针对睡眠嗜组织菌rOMP40和LPS的IgM抗体显著升高,而4周后针对重组热休克蛋白60抗原的IgM抗体升高。抗LPS IgM抗体在观察期结束前逐步升高,但在后续采样中针对睡眠嗜组织菌重组热休克蛋白60和睡眠嗜组织菌重组外膜蛋白40的IgM抗体下降。首次免疫4周后发现IgG类睡眠嗜组织菌重组热休克蛋白60抗体显著升高,6周后针对其余两种抗原的IgG抗体也有类似升高。反应强度在实验结束前一直增加。仅观察到针对睡眠嗜组织菌重组热休克蛋白60抗原的IgA抗体水平升高。临床观察发现,触珠蛋白浓度保持在生理范围内,证实动物健康和福利状况良好。至少需要4次加强剂量才能获得含有高水平针对检测抗原抗体的超免疫血清。免疫次数影响特异性IgM、IgG、IgA抗体的反应谱。

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