Insights gained from single-cell analysis of immune cells in tofacitinib treatment of Vogt-Koyanagi-Harada disease

Abstract

Vogt-Koyanagi-Harada (VKH) disease is an important refractory uveitis mediated by pathological T cells (TCs). Tofacitinib (TOFA) is a Janus kinases (JAKs) targeted therapy for several autoimmune diseases. However, the specific pathogenesis and targeted therapeutics for VKH remain largely unknown. Based on single-cell RNA sequencing and mass cytometry, we present the first multimodal high-dimensional analysis to determine a comprehensive human immune atlas of VKH patients undergoing TOFA therapy in the context of subset composition, gene signatures, enriched pathways, and intercellular interactions. VKH patients are characterized by TCs polarization from naive to effector and memory subsets, altogether with accrued monocytes, upregulated cytokines and JAK-STAT signaling pathways. In vitro, TOFA reversed Th17/ regulatory T-cell (Treg) imbalance and inhibited IL-2-induced STAT1/3 phosphorylation. TOFA alleviated VKH symptoms by restoring pathological TCs polarization and functional marker expression and downregulating cytokine signaling and lymphocyte function. Remarkably, inflammation-related responses and intercellular interactions decreased after TOFA treatment, particularly in monocytes. Notably, we identified two inflammation-and JAK-associated monocyte subpopulations that were strongly implicated in VKH pathogenesis and mechanisms involved in TOFA treatment. Here, we provide a novel JAK-targeted therapy for VKH and elaborate on the possible therapeutic mechanisms of TOFA, expanding our knowledge of VKH pathological patterns.

Sleep loss potentiates Th17‐cell pathogenicity and promotes autoimmune uveitis

“…Clinical trials are on-going for JAK inhibitors as autoimmune treatments including AU. 27,66 More importantly, we discovered that SL enhanced inter-cellular communication among myeloid cells, TCs, and BCs, contributing to the activation and differentiation of TCs (CD28-CD80/86), BCs (CD40-CD40LG) and myeloid cells (CSF2-CSF2RA). The interactions of CSF2 and its receptors between Th17 cells and myeloid cell subsets, were specifically presented in IRBP 1−20 -immunized EAU mice, but not in blank and control mice, and were enhanced following SL.…”

“…We performed CyTOF experiments and the subsequent bioinformatics analysis to map the immune cells based on surface markers and intracellular cytokines with single‐cell resolution 25 (Supporting Information Table S3). We annotated the FlowSOM‐defined nodes into classical immune cell types, including CD4 + naïve (CD4NA), central memory, regulatory, effector memory (CD4TEM) and cytotoxic T cells (CD4CTL), CD8 + naïve (CD8NA), effector memory, and cytotoxic T cells (TCs), natural killer cells, B cells (BCs), classical monocytes (CMCs), non‐classical monocytes, conventional dendrite cells (CDCs) and plasmacytoid dendrite cells (Figure 1A, Supporting Information Figure S1A,B) based on previous studies 26,27 …”

“…As downstream effectors of pro‐inflammatory cytokines, the JAK‐STAT pathway is involved in inflammatory processes. Clinical trials are on‐going for JAK inhibitors as autoimmune treatments including AU 27,66 . More importantly, we discovered that SL enhanced inter‐cellular communication among myeloid cells, TCs, and BCs, contributing to the activation and differentiation of TCs (CD28‐CD80/86), BCs (CD40‐CD40LG) and myeloid cells (CSF2‐CSF2RA).…”

“…We annotated the FlowSOM-defined nodes into classical immune cell types, including CD4 + naïve (CD4NA), central memory, regulatory, effector memory (CD4TEM) and cytotoxic T cells (CD4CTL), CD8 + naïve (CD8NA), effector memory, and cytotoxic T cells (TCs), natural killer cells, B cells (BCs), classical monocytes (CMCs), non-classical monocytes, conventional dendrite cells (CDCs) and plasmacytoid dendrite cells (Figure 1A, Supporting Information Figure S1A,B) based on previous studies. 26,27 We explored the effects of SL on cell composition in the unstimulated state because stimulation affected myeloid cell signals. 26 After SL, we validated that monocyte decreased, which was the main component of myeloid cells (Figure 1B).…”

Progesterone attenuates Th17-cell pathogenicity in autoimmune uveitis via Id2/Pim1 axis

“…Differential expression analysis for each cell type between different groups was performed by using the Wilcoxon rank-sum test implement in the ''Find-Markers'' function of the Seurat package (version 4.0.5). Similar with previous studies [28,29], DEGs between the EAU and naïve groups were identified to generate an EAU-related DEG dataset (EAU-DEGs) (|LogFC|> 0.25, P value < 0.05). DEGs between the PRG and EAU groups were identified to establish a PRG-related DEG dataset (PRG-DEGs) (|LogFC|> 0.25, P value < 0.05).…”