Role Of Toll-like Receptors,Co-stimulators And Checkpoints In The Pathogenesis Of Takayasu’s Arteritis

Takayasu’s Arteritis(TAK)is an autoimmune disease in which autoreactive T-cells attack the medium and large arteries,destroying the structure of the blood vessel wall,leading to stenosis,closure or dilation of the blood vessel lumen,causing ischemia.Toll-like receptors(TLR)directly or indirectly regulate the activation and differentiation of T-cells.The presentation of antigen to T-cell by dendritic cells is a representative of indirect regulation,in which the co-stimulators regulate the differentiation direction of T-cells and the intensity of the immune response.Co-stimulating molecules include positive co-stimulation molecules(including CD28,CD40L,etc.)and negative co-stimulation molecules(i.e.immune checkpoints,including CTLA4,PD-1,etc.).In our study,the expression level of genes struck by real-time fluorescence quantitative polymerase chain reaction was compared between subgroups of TAK and healthy controls,and explore the association among TLR molecules,T-cell-related co-stimulation molecules and the differentiation activation of T-cell subgroups in TAK by experimental data mining,establish a TAK molecular mechanism model to validate immunoregulation targets.Autoimmune disease may influence the gene expression stability,so in the first part,we analyzed the expression stability of 9 commonly used reference genes in human PBMCs for the subsequent relative gene quantification.In the second part of the study,16 differential expression genes of TAK were analyzed by RT-qPCR,including:TLR2,TLR4,p50,p65,/κBα,CD28,CD40,CTLA4,PD-1,LAG3,TCR,CD3,GATA3,RORC,FOXP3 and BCL6.Among them,p50,CD28,TCR,GATA3,RORC and FOXP3are related to TAK activity.Then,we performed correlation analysis and cluster analysis in gene expression level,and structured a dynamic network of gene expression based on TAK,therapeutic and disease activity.We compared experiemental network with the network based public databases.The analysis indicates that the NFκB,CD28,Th2 cell,Th17 cell,Treg cell are closely related to TAK activity,p65is more closely related to the activation and differentiation of T-cell compared with p50,and in the nonactive TAK(treated),several TLRs are co-expressed with several T-cell activation or differentiation-related genes,and the comparison with healthy comtrol suggesting that it is TAK’s pathological manifestation;in active TAK(treated),PD-1/PD-L1,LAG3,TIGIT,CD40/CD40L,BCL6network,mediated TLR and T-cell activation or differentiation related gene communication;This molecular mechanism model provides experimental evidence and directions for the follow-up exploration of the mechanism of regulations between TLR and T-cell in TAK.