Detection of Immunodominant Proteins of Felis catus Gammaherpesvirus 1

Stutzman-Rodriguez K , Troyer R , VandeWoude S
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Abstract

We recently identified and sequenced a novel herpesvirus of domestic cats, Felis catus gammaherpesvirus 1 (FcaGHV1). FcaGHV1 is a member of the gammaherpesvirus subfamily which also includes the human cancer-associated herpesviruses, Epstein-Barr virus (EBV) and Kaposi’s sarcoma-associated herpesvirus (KSHV). As a first step toward developing a serologic assay to detect exposure to FcaGHV1, we are seeking to determine which viral proteins elicit an antibody response in naturally occurring domestic cat infections. We cloned selected FcaGHV1 genes into a mammalian expression vector and performed transfections of a feline cell line for expression of recombinant FcaGHV1 proteins. We fixed cells with paraformaldehyde and methanol-acetone and tested reactivity to serum from cats naturally infected with FcaGHV1 using immunofluorescent antibody staining. An FIV immunoflouresence test was developed as a positive control for transfection and assay function. Serum from specific pathogen-free laboratory cats served as negative controls. Preliminary data from 9 cats with FcaGHV1 infection indicates that capsid protein ORF 65 and tegument protein ORF38 may elicit antibodies during naturally occurring FcaGHV1 infection. Results of this study will suggest which FcaGHV1 proteins are immunodominant during natural infection. With this information we plan to develop a serologic assay and further evaluate FcaGHV1 as a model for EBV and KSHV.

猫γ疱疹病毒1型免疫优势蛋白的检测
我们最近鉴定并测序了家猫的一种新型疱疹病毒——猫γ疱疹病毒1 (Felis catus gammaherpesvirus 1, FcaGHV1)。FcaGHV1是伽玛疱疹病毒亚家族的成员,该亚家族还包括人类癌症相关疱疹病毒、eb病毒和卡波西肉瘤相关疱疹病毒。作为开发检测FcaGHV1暴露的血清学分析的第一步,我们正在寻求确定在自然发生的家猫感染中哪些病毒蛋白会引起抗体反应。我们将选定的FcaGHV1基因克隆到哺乳动物表达载体中,并转染猫细胞系表达重组FcaGHV1蛋白。我们用多聚甲醛和甲醇-丙酮固定细胞,并使用免疫荧光抗体染色检测对自然感染FcaGHV1的猫血清的反应性。建立了FIV免疫荧光试验作为转染和检测功能的阳性对照。特异性无病原体实验猫血清作为阴性对照。来自9只感染FcaGHV1的猫的初步数据表明,在自然发生的FcaGHV1感染过程中,衣壳蛋白orf65和被膜蛋白ORF38可能引发抗体。本研究的结果将提示在自然感染过程中,哪些FcaGHV1蛋白具有免疫优势。有了这些信息,我们计划开发一种血清学检测方法,并进一步评估FcaGHV1作为EBV和KSHV的模型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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