接种疫苗和用 CRFK 细胞系超强感染的猫体内针对猫肾细胞（CRFK）系抗原、α-烯醇化酶和膜联蛋白 A2 的抗体。
Antibodies against Crandell Rees feline kidney (CRFK) cell line antigens, alpha-enolase, and annexin A2 in vaccinated and CRFK hyperinoculated cats.
机构信息
Department of Clinical Sciences, Veterinary Medical Center, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
出版信息
J Vet Intern Med. 2010 Mar-Apr;24(2):306-13. doi: 10.1111/j.1939-1676.2010.0476.x.
BACKGROUND
Cats inoculated with feline herpesvirus 1, calicivirus, and panleukopenia (FVRCP) vaccines grown on the Crandell Rees feline kidney (CRFK) cell line have been shown to develop anti-CRFK antibodies. The identities of common CRFK antigens are unknown.
HYPOTHESIS
Cats inoculated with CRFK lysates and FVRCP vaccines will develop autoantibodies measurable by Western blot immunoassay. Antigens associated with these antibodies can be isolated for further study.
ANIMALS
One CRFK hyperinoculated rabbit, 44 age-matched unvaccinated kittens purchased from a commercial vendor.
METHODS
Commonly recognized CRFK antigens were identified by comparison of Western blot immunoassays using sera from a hyperinoculated rabbit and kittens inoculated with CRFK lysate or 1 of 4 commercially available FVRCP vaccines. Antigens were purified from CRFK lysates and sequenced. Antigen recognition was confirmed by Western blot immunoassay and indirect ELISA for 2 proteins using sera from CRFK and FVRCP inoculated kittens.
RESULTS
CRFK antigens 47, 40, and 38 kD in size were identified. Protein isolation and sequencing identified 3 CRFK proteins as alpha-enolase, annexin A2, and macrophage capping protein (MCP). Sera from FVRCP and CRFK inoculated cats were confirmed to recognize annexin A2 and alpha-enolase by Western blot immunoassay and indirect ELISA.
CONCLUSIONS AND CLINICAL RELEVANCE
This study validated the use of Western blot immunoassay for detection of antibodies against CRFK proteins and identified 3 CRFK antigens. In humans, alpha-enolase antibodies are nephritogenic; alpha-enolase and annexin A2 antibodies have been associated with autoimmune diseases. Further research will be necessary to determine the clinical relevance of these findings.
背景
接种了在猫肾细胞（CRFK）上生长的猫疱疹病毒 1、杯状病毒和细小病毒（FVRCP）疫苗的猫已被证明会产生抗 CRFK 抗体。然而，CRFK 常见抗原的身份仍不清楚。
假说
接种了 CRFK 裂解物和 FVRCP 疫苗的猫会产生可通过 Western blot 免疫印迹测定的自身抗体。与这些抗体相关的抗原可被分离出来进行进一步研究。
动物
一只 CRFK 高度接种的兔子，以及从商业供应商处购买的 44 只年龄匹配的未接种小猫。
方法
通过比较用高度接种的兔子血清和接种了 CRFK 裂解物或 4 种市售 FVRCP 疫苗之一的小猫血清进行 Western blot 免疫印迹，确定了常见的 CRFK 抗原。从 CRFK 裂解物中分离和测序了抗原。使用来自接种了 CRFK 和 FVRCP 的小猫的血清，通过 Western blot 免疫印迹和间接 ELISA 对 2 种蛋白质进行确认，证实了抗原识别。
结果
鉴定出大小为 47、40 和 38 kD 的 CRFK 抗原。蛋白质分离和测序鉴定出 3 种 CRFK 蛋白，分别为α-烯醇酶、膜联蛋白 A2 和巨噬细胞封顶蛋白（MCP）。使用来自接种了 FVRCP 和 CRFK 的猫的血清，通过 Western blot 免疫印迹和间接 ELISA 证实了抗膜联蛋白 A2 和α-烯醇酶的识别。
结论和临床相关性
本研究验证了 Western blot 免疫印迹用于检测针对 CRFK 蛋白的抗体的用途，并鉴定了 3 种 CRFK 抗原。在人类中，α-烯醇酶抗体具有肾毒性；α-烯醇酶和膜联蛋白 A2 抗体与自身免疫性疾病有关。需要进一步的研究来确定这些发现的临床相关性。
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