Decourcey Michelle Athena, Davis William Charles, de Souza Cleverson
Veterinarian, Veterinary Medical Diagnostic Laboratory, University of Missouri College of Veterinary Medicine, Columbia, CO, USA.
Veterinarian, PhD, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA.
Braz J Vet Med. 2024 Jan 16;46:e002723. doi: 10.29374/2527-2179.bjvm002723. eCollection 2024.
Analysis of the primary and recall responses to a membrane molecule (MMP), encoded by MAP2121c demonstrated that tri-directional signaling between the antigen-presenting cell (APC), CD4 and CD8 is essential for eliciting a CD8 cytotoxic T cell (CTL) response against Mycobacterium avium subsp. paratuberculosis. As reported here, RNA-sequencing was used to initiate the characterization of the signaling pathways involved in eliciting the development of CD8 CTL, starting with the characterization of the activation status of genes in monocyte-derived macrophages (MoMΦ) following uptake and processing MMP for the presentation of antigenic epitopes to CD4 and CD8 T cells. Activation status was compared with the uptake and processing of LPS, a nonspecific stimulator of macrophages. 1609 genes were identified that were upregulated, and 1277 were downregulated three hours after uptake and processing MMP. No significant difference was observed in the cytokine genes selected for analysis of the signaling that must occur between APC, CD4, and CD8 for the development of CTL. The initial observations indicate screening of the transcriptome should include genes involved in signaling between APC and CD4, and CD8 regardless of their activation status. Four genes of interest in this study, IL12A, IL12B, IL15, and IL23A, were not significantly different from control values. The initial studies also indicate MoMΦ can be included with dendritic cells and monocyte-derived dendritic cells for further analysis of the tri-directional signaling required for the development of CTL.
对由MAP2121c编码的一种膜分子（MMP）的初次和回忆反应分析表明，抗原呈递细胞（APC）、CD4和CD8之间的三向信号传导对于引发针对副结核分枝杆菌的CD8细胞毒性T细胞（CTL）反应至关重要。如本文所报道，RNA测序被用于启动对参与引发CD8 CTL发育的信号通路的表征，首先是对单核细胞衍生的巨噬细胞（MoMΦ）在摄取和处理MMP以将抗原表位呈递给CD4和CD8 T细胞后基因的激活状态进行表征。将激活状态与巨噬细胞的非特异性刺激物LPS的摄取和处理进行比较。在摄取和处理MMP三小时后，鉴定出1609个基因上调，1277个基因下调。在为分析CTL发育过程中APC、CD4和CD8之间必须发生的信号传导而选择的细胞因子基因中未观察到显著差异。初步观察表明，转录组筛选应包括参与APC与CD4以及CD8之间信号传导的基因，无论它们的激活状态如何。本研究中感兴趣的四个基因IL12A、IL12B、IL15和IL23A与对照值无显著差异。初步研究还表明，MoMΦ可与树突状细胞和单核细胞衍生的树突状细胞一起用于进一步分析CTL发育所需的三向信号传导。
