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牛新生粒细胞减少症——两种 BVD 疫苗和生产细胞表面蛋白质组(MDBK)的比较蛋白质组学特征。

Bovine neonatal pancytopenia--comparative proteomic characterization of two BVD vaccines and the producer cell surface proteome (MDBK).

机构信息

Institute of Animal Physiology, Department of Veterinary Sciences, LMU Munich, Veterinärstr 13, München D-80539, Germany.

出版信息

BMC Vet Res. 2013 Jan 23;9:18. doi: 10.1186/1746-6148-9-18.

Abstract

BACKGROUND

Bovine neonatal pancytopenia (BNP) is a disease syndrome in newborn calves of up to four weeks of age, first observed in southern Germany in 2006. By now, cases have been reported in several countries around the globe. Many affected calves die within days due to multiple haemorrhages, thrombocytopenia, leukocytopenia and bone marrow depletion. A certain vaccine directed against Bovine Virus Diarrhoea Virus (BVDV) was recently shown to be associated with BNP pathogenesis. Immunized cows develop alloantibodies that are transferred to newborn calves via colostrum intake. In order to further elucidate BNP pathogenesis, the purpose of this study was to characterize and compare the protein composition of the associated vaccine to another vaccine directed against BVDV not related to BNP and the cell surface proteome of MDBK (Madin-Darby Bovine Kidney) cells, the cell line used for production of the associated vaccine.

RESULTS

By SDS-PAGE and mass spectrometry, we were able to detect several coagulation-related and immune modulatory proteins, as well as cellular and serum derived molecules being shared between the associated vaccine and MDBK cells. Furthermore, the number of proteins identified in the BNP related vaccine was almost as high as the number of surface proteins detected on MDBK cells and exceeded the amount of proteins identified in the non-BNP related vaccine over 3.5 fold. The great amount of shared cellular and serum derived proteins confirm that the BNP associated vaccine contained many molecules originating from MDBK cells and vaccine production.

CONCLUSIONS

The respective vaccine was not purified enough to prevent the development of alloantibodies. To narrow down possible candidate proteins, those most likely to represent a trigger for BNP pathogenesis are presented in this study, giving a fundament for further analysis in future research.

摘要

背景

牛新生全血细胞减少症(BNP)是一种发生于 4 周龄以内新生犊牛的疾病综合征,于 2006 年在德国南部首次观察到。目前,在全球多个国家都有病例报告。由于多发性出血、血小板减少症、白细胞减少症和骨髓耗竭,许多受影响的犊牛在数天内死亡。最近发现,一种针对牛病毒性腹泻病毒(BVDV)的特定疫苗与 BNP 的发病机制有关。免疫母牛会产生同种异体抗体,通过初乳摄入转移到新生犊牛体内。为了进一步阐明 BNP 的发病机制,本研究的目的是对与 BNP 相关的疫苗与另一种不相关的 BVDV 疫苗的蛋白质组成进行特征分析和比较,以及比较与生产相关疫苗的 MDBK(Madin-Darby 牛肾)细胞的细胞表面蛋白质组。

结果

通过 SDS-PAGE 和质谱分析,我们能够检测到几种与凝血相关和免疫调节的蛋白质,以及与相关疫苗和 MDBK 细胞共享的细胞和血清来源的分子。此外,在与 BNP 相关的疫苗中鉴定出的蛋白质数量几乎与在 MDBK 细胞上检测到的表面蛋白质数量一样多,超过了非 BNP 相关疫苗中鉴定出的蛋白质数量的 3.5 倍以上。大量共享的细胞和血清来源的蛋白质证实,BNP 相关疫苗包含了许多源自 MDBK 细胞和疫苗生产的分子。

结论

相应的疫苗没有被充分纯化以防止同种异体抗体的产生。为了缩小可能的候选蛋白范围,本研究提出了最有可能代表 BNP 发病机制触发因素的那些蛋白,为未来研究中的进一步分析奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f5f/3560244/de48f1423dca/1746-6148-9-18-1.jpg

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