Luce R, Shepherd M, Paillot R, Blacklawst B, Wood J L N, Kydd J H
Animal Health Trust, Centre for Preventive Medicine, Lanwades Park, Kentford, Newmarket, Suffolk CB8 7UU, UK.
Equine Vet J. 2007 May;39(3):202-9. doi: 10.2746/042516407x174216.
An assay has been developed that measures EHV-1 specific interferon gamma synthesis (IFNgamma), a cytokine produced following the activation of memory T lymphocytes and therefore a measure of cell mediated immunity. The method requires validation in the field.
To measure the frequency of EHV-1 specific, IFNgamma synthesising peripheral blood mononuclear cells (PBMC) in a population of Thoroughbred horses, and examine its relationship with age, gender, premises and history of vaccination or field infection with EHV-1.
Lymphocytes from 200 Thoroughbred horses were stimulated with EHV-1 in vitro, and IFNgamma detected using a monoclonal antibody and indirect immunofluorescence. Percent positive cells were enumerated by flow cytometric analysis and the results described and compared statistically between groups.
The frequency of IFNgamma+ PBMC was significantly higher in animals age >5 years compared with 2-4 years, in females vs. males, on stud farms vs. training yards and following vaccination of 2-year-olds with inactivated virus compared with nonvaccinates. Age strongly confounded all these associations and care must therefore be taken interpreting these results. Mares exposed to a field infection with EHV-1 also had higher frequencies of IFNgamma+ PBMC than other vaccinated horses.
The frequency of EHV-1 specific, IFNgama+ PBMC among the sample Thoroughbred population was diverse but lowest in young, unvaccinated horses-in-training.
The frequency of EHV-1 specific lymphocytes synthesising IFNgamma in this population may be associated with its susceptibility to infection with this virus. This easy technique may be applied to monitor the antigenicity of vaccines and their effectiveness at stimulating cellular immunity.
已开发出一种检测方法，可测量EHV-1特异性γ干扰素合成（IFNγ），γ干扰素是记忆T淋巴细胞激活后产生的一种细胞因子，因此可作为细胞介导免疫的一项指标。该方法需要在实际应用中进行验证。
测定纯种马群体中EHV-1特异性、合成IFNγ的外周血单核细胞（PBMC）的频率，并研究其与年龄、性别、饲养场所及EHV-1疫苗接种或野外感染史的关系。
用EHV-1体外刺激200匹纯种马的淋巴细胞，使用单克隆抗体和间接免疫荧光法检测IFNγ。通过流式细胞术分析计数阳性细胞百分比，并对结果进行描述及组间统计学比较。
年龄大于5岁的动物中IFNγ+ PBMC的频率显著高于2-4岁的动物；雌性高于雄性；种马场的高于训练场的；2岁马匹接种灭活病毒疫苗后高于未接种疫苗的。年龄强烈混淆了所有这些关联，因此在解释这些结果时必须谨慎。暴露于EHV-1野外感染的母马中IFNγ+ PBMC的频率也高于其他接种疫苗的马匹。
在抽样的纯种马群体中，EHV-1特异性、IFNγ+ PBMC的频率各不相同，但在未接种疫苗的年轻训练马匹中最低。
该群体中合成IFNγ的EHV-1特异性淋巴细胞的频率可能与其对该病毒感染的易感性有关。这种简便技术可用于监测疫苗的抗原性及其刺激细胞免疫的效果。
