结核病（TB）由结核分枝杆菌（Mtb）引起，仍然是全球性的健康负担之一。各种病例的发生和多药耐药性的存在证实了结核病尚未完全被征服。基于这些原因，本研究旨在利用Mtb分泌的蛋白质探索结核病疫苗和药物候选物的可能性。在这些蛋白质中，已知MPT32具有抗原性和免疫原性。目前还没有关于宿主免疫反应和巨噬细胞中的调节的报告。本研究使用RAW 264.7小鼠巨噬细胞进行了MPT32试验，这些细胞通过感知病原体入侵和环境变化来调控免疫反应。我们发现MPT32能够激活脂多糖（LPS）诱导的MMP-9基因表达和RAW 264.7细胞的炎症反应。在用MPT32处理细胞后，观察到类风湿关节炎因子-α（TNF-α）、白细胞介素（IL）-1β（IL-1β）和IL-6等促炎细胞因子的增加。此外，活化的巨噬细胞会表达诱导型一氧化氮合酶（iNOS）和环氧合酶-2（COX-2），生成各种炎症介质分子，如一氧化氮（NO）。增加的iNOS和COX-2水平，作为MMP-9表达的上调调节因子，也得到了确认。这些生化事件与激活的MAPK信号传导和NF-κ B转录因子的转位有关。本研究结果证实了MPT32对RAW 264.7小鼠巨噬细胞的免疫调节效应，提出了使用MPT32进行结核病疫苗和药物候选物的可能性，并有助于预防结核病。© 2023 Wiley Periodicals LLC.

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb) and is still one of the global health burdens. The occurrence of various cases and multidrug resistance confirm that TB has not been completely conquered. For these reasons, the present research has been conducted to explore TB vaccine and drug candidate possibility using Mtb-secreted proteins. Among these proteins, MPT32 is known to have antigenicity and immunogenicity. There has not been a report on the host immune responses and regulation in macrophage cells. The present study was conducted with MPT32 in RAW 264.7 murine macrophage cells that control immune responses by sensing pathogen invasion and environmental change. We have found that MPT32 could activate lipopolysaccharide (LPS)-induced gene expression of metalloproteinase-9 (MMP-9) and inflammation in RAW 264.7 cells. After treating cells with MPT32, the increase in pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β (IL-1β) and IL-6, was observed. In addition, activated macrophages expressed inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) to generate various inflammatory mediator molecules, such as nitric oxide (NO). The increase in iNOS and COX-2 levels, which are up-regulators of MMP-9 expression, was also confirmed. The biochemical events are involved in the downstream of activated MAPK signaling and translocation of NF-κ B transcription factor. The present results prove the immunomodulatory effect of MPT32 in the RAW 264.7 murine macrophage cells. it claims the possibility of a TB vaccination and drug candidate using MPT32, contributing to the prevention of TB.© 2023 Wiley Periodicals LLC.