Inflammasome plays a major role in innate immune responses by activating caspase-1, resulting in secretion of interleukin-1β (IL-1β) and inflammatory pathologic responses. IL-1β release is widely used as an indirect readout to study inflammasome activation. Here we report an iGLuc reporter (pro-IL-1β-Gluc) of pig origin to monitor cytosolic pro-IL-1β cleavage and mature IL-1β release. Based on the iGLuc reporter, we reconstructed the inflammasome system in vitro and screened PRRSV- and ASFV-encoded proteins involved in regulating inflammasome activation. We found that three non-structural proteins (nsps) of PRRSV, nsp1β, nsp2 and nsp5, activate the NLRP3 inflammasome, and four nsps of PRRSV, nsp1ɑ, nsp7, nsp10 and nsp11, inhibit NLRP3 inflammasome activation, of which nsp10 and nsp11 have a highly significant inhibitory effect. In addition, we also found that four ASFV-encoded proteins, S183L, E199L, O61R and I7L activate the inflammatory response and four ASFV-encoded proteins, I226L, A151R, NP419L and QP383R, inhibit the inflammatory response. Our results provide a highly sensitive and high-throughput tool to screen for proteins that regulate inflammasome activation in vitro.

炎症小体通过激活caspase-1在先天性免疫反应中起主要作用，导致白介素1β（IL-1β）的分泌和炎症病理反应。IL-1β释放被广泛用作研究炎症小体激活的间接读数。在这里，我们报告了一个来自猪的iGLuc报告基因（pro-IL-1β-Gluc），以监测胞浆中的pro-IL-1β裂解和成熟的IL-1β释放。基于iGLuc记者，我们在体外重建了炎性体系统并筛选了涉及调节炎症小体活化的PRRSV和ASFV编码蛋白。我们发现PRRSV的三个非结构蛋白（nsps），nsp1β，nsp2和nsp5激活了NLRP3炎性小体，而PRRSV的四个nsps，nsp1ɑ，nsp7，nsp10和nsp11抑制了NLRP3炎性体的激活，其中nsp10和nsp11具有高度显着的抑制作用。此外，我们还发现四种ASFV编码蛋白S183L，E199L，O61R和I7L激活炎症反应，而四种ASFV编码蛋白I226L，A151R，NP419L和QP383R抑制炎症反应。我们的结果提供了一种高灵敏度和高通量的工具，可筛选出可调节体外炎症小体活化的蛋白质。