Renjifo X, Letellier C, Keil G M, Ismaili J, Vanderplasschen A, Michel P, Godfroid J, Walravens K, Charlier G, Pastoret P P, Urbain J, Denis M, Moser M, Kerkhofs P
Département de Biologie Moléculaire, Université Libre de Bruxelles, 1640 Rhode-Saint-Genèse, Belgium.
J Virol. 1999 Jun;73(6):4840-6. doi: 10.1128/JVI.73.6.4840-4846.1999.
The aim of the present study was to develop an in vitro system for presentation of bovine herpesvirus 1 (BHV-1) antigens to bovine T lymphocytes and to characterize the antigen-presenting cells (APC) which efficiently activate CD4(+) T cells. Two approaches were used to monitor the infection of APC by BHV-1 as follows: (i) detection of viral glycoproteins at the cell surface by immunofluorescence staining and (ii) detection of UL26 transcripts by reverse transcription-PCR. The monocytes were infected, while dendritic cells (DC) did not demonstrate any detectable viral expression. These data suggest that monocytes are one site of replication, while DC are not. The capacities of monocytes and DC to present BHV-1 viral antigens in vitro were compared. T lymphocytes (CD2(+) or CD4(+)) from BHV-1 immune cattle were stimulated in the presence of APC previously incubated with live or inactivated wild-type BHV-1. DC stimulated strong proliferation of Ag-specific T cells, while monocytes were poor stimulators of T-cell proliferation. When viral attachment to the surface of the APC was inhibited by virus pretreatment with soluble heparin, T-cell proliferation was dramatically decreased. Unexpectedly, incubation of DC and monocytes with the deletion mutant BHV-1 gD-/-, which displays impaired fusion capacity, resulted in strong activation of T lymphocytes by both APC types. Collectively, these results indicate that presentation of BHV-1 antigens to immune T cells is effective in the absence of productive infection and suggest that BHV-1 gD-/- mutant virus could be used to induce virus-specific immune responses in cattle.
本研究的目的是开发一种体外系统,用于将牛疱疹病毒1型(BHV-1)抗原呈递给牛T淋巴细胞,并鉴定能有效激活CD4(+) T细胞的抗原呈递细胞(APC)。采用两种方法监测BHV-1对APC的感染,具体如下:(i)通过免疫荧光染色检测细胞表面的病毒糖蛋白;(ii)通过逆转录PCR检测UL26转录本。单核细胞被感染,而树突状细胞(DC)未表现出任何可检测到的病毒表达。这些数据表明,单核细胞是一个复制位点,而DC不是。比较了单核细胞和DC在体外呈递BHV-1病毒抗原的能力。在预先用活的或灭活的野生型BHV-1孵育的APC存在下,刺激来自BHV-1免疫牛的T淋巴细胞(CD2(+)或CD4(+))。DC刺激Ag特异性T细胞强烈增殖,而单核细胞对T细胞增殖的刺激作用较差。当用可溶性肝素对病毒进行预处理以抑制病毒附着于APC表面时,T细胞增殖显著降低。出乎意料的是,用融合能力受损的缺失突变体BHV-1 gD-/-孵育DC和单核细胞,导致两种APC类型均强烈激活T淋巴细胞。总体而言,这些结果表明,在没有生产性感染的情况下,将BHV-1抗原呈递给免疫T细胞是有效的,并表明BHV-1 gD-/-突变病毒可用于诱导牛的病毒特异性免疫反应。