Laboratorio de Inmunofisiología y Proteómica, FMVZ, Univerisdad Nacional Autónoma de México, 04510, México.
Laboratorio de Virología, FMVZ, Univerisdad Nacional Autónoma de México, 04510, Mexico.
Res Vet Sci. 2020 Aug;131:131-136. doi: 10.1016/j.rvsc.2020.04.014. Epub 2020 Apr 15.
Bovine viral diarrhea virus (BVDV) infects different cell types including antigen-presenting cells such as macrophages. The infection induces pro-inflammatory cytokines like interleukin 1 beta (IL-1β), which is necessary to trigger a successful inflammatory response against infections. Several authors have reported differences between IL-1β gene expression and protein detection in BVDV-infected macrophages. These patterns may be related to inflammasome assembly, which promote the formation of active caspase 1 in order to produce mature IL-1β molecules. Our goal was to assess BVDV ability to induce the release of IL-β through a caspase 1 dependent pathway in bovine macrophages. We infected peripheral blood monocyte-derived macrophages using BVDV NADL strain at 0.001, 0.1, 2 and 10 multiplicities of infection (MOI) and we measured IL-1β at different times 2, 6, 12, 24, 48, 72 h. We found an increase of 1140-2154 pg for a MOI of 10:1 and 2:1 respectively. To inhibit caspase 1, we used either carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (Z-VAD) or carbobenzoxy-tyr-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (Y-VAD). We found decreased IL-1β secretion 2154 pg/ml to 854 pg/ml IL-1β secretion using Y-VAD and we observed decrease from 2154 pg/ml to 22.33 pg/ml with Z-VAD, and this inhibition was followed by diminished viral replication from 2.25 × 10 to 2.1 × 10 CCID, which suggests that caspase 1-dependent secretion of the IL-1β active molecule is important for viral replication. This is the first report showing that BVDV infected-bovine macrophages trigger the caspase 1 dependent pathway for IL-1β activation and that activation increases viral replication.
牛病毒性腹泻病毒（BVDV）感染不同的细胞类型，包括抗原呈递细胞如巨噬细胞。该感染诱导白细胞介素 1β（IL-1β）等促炎细胞因子的产生，这对于引发针对感染的成功炎症反应是必要的。一些作者已经报道了 BVDV 感染的巨噬细胞中 IL-1β 基因表达和蛋白检测之间的差异。这些模式可能与炎症小体的组装有关，炎症小体可促进活性半胱天冬酶 1 的形成，以产生成熟的 IL-1β 分子。我们的目标是评估 BVDV 通过牛巨噬细胞中的半胱天冬酶 1 依赖途径诱导 IL-β释放的能力。我们使用 BVDV NADL 株以 0.001、0.1、2 和 10 感染复数（MOI）感染外周血单核细胞衍生的巨噬细胞，并在不同时间 2、6、12、24、48 和 72 小时测量 IL-1β。我们发现 MOI 为 10:1 和 2:1 时，IL-1β 的增加分别为 1140-2154pg。为了抑制半胱天冬酶 1，我们使用了 carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone（Z-VAD）或 carbobenzoxy-tyr-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone（Y-VAD）。我们发现使用 Y-VAD 时，IL-1β 分泌从 2154pg/ml 降低到 854pg/ml，使用 Z-VAD 时从 2154pg/ml 降低到 22.33pg/ml，并且这种抑制伴随着病毒复制从 2.25×10 到 2.1×10 CCID 的减少，这表明半胱天冬酶 1 依赖性的 IL-1β 活性分子的分泌对于病毒复制很重要。这是第一项表明 BVDV 感染的牛巨噬细胞触发 IL-1β 激活的半胱天冬酶 1 依赖性途径的报告，并且激活增加了病毒复制。
