Hu Xuqiao, Li Fei, Zeng Jieying, Zhou Zhenru, Wang Zhaoyang, Chen Jing, Cao Dongyan, Hong Yifan, Huang Laixin, Chen Yongsheng, Xu Jinfeng, Dong Fajin, Yu Rongmin, Zheng Hairong
Department of Ultrasound, First Affiliated Hospital of Southern University of Science and Technology, Second Clinical Medical College of Jinan University (Shenzhen People's Hospital), Shenzhen 518020, China.
Integrated Chinese and Western Medicine Postdoctoral Research Station, Jinan University, Guangzhou 510632, China.
Research (Wash D C). 2022 Dec 19;2022:0013. doi: 10.34133/research.0013. eCollection 2022.
Rheumatoid arthritis (RA) is a common autoimmune disease characterized by chronic and progressive inflammation of the synovium. Focused ultrasound therapy is an increasingly attractive alternative for treating RA owing to its noninvasiveness; however, it remains unclear which immune subsets respond to ultrasound stimulation. In this study, we showed that spleen-targeted low-frequency pulsed focused ultrasound (LFPFU) effectively improved the severity of arthritis in an arthritis mouse model established in DBA/1J mice. Additionally, we performed in-depth immune profiling of spleen samples from RA mice, RA mice that underwent ultrasound therapy, and healthy controls using mass cytometry along with extensive antibody panels and identified the immune composition of 14 cell populations, including CD4/CD8 T cells, B cells, natural killer cells, and dendritic cells. Moreover, multidimensional analysis according to cell-surface markers and phenotypes helped in identifying 4 and 5 cell subpopulations among T and myeloid cells, respectively, with 6 T cell subsets and 3 myeloid cell subsets responsive to ultrasound therapy among the 3 groups. Of these cell subsets, CD8 T cell subsets showed a unique response to ultrasound stimulation in RA mice. Specifically, CD8 T cells show a noticeable correlation with the degree of arthritis progression and could serve as an indicator for spleen-focused ultrasound-based therapy. Furthermore, single-cell RNA sequencing of spleen cells revealed the importance of T, B, and myeloid cell populations in the anti-inflammatory pathway. These results elucidated the unique cell subsets and transcriptome of splenic cells responsive to LFPFU and demonstrated the potential of spleen-focused ultrasound stimulation in the treatment of inflammatory diseases.
类风湿性关节炎（RA）是一种常见的自身免疫性疾病，其特征是滑膜的慢性进行性炎症。聚焦超声疗法因其无创性而成为治疗RA越来越有吸引力的替代方法；然而，尚不清楚哪些免疫亚群对超声刺激有反应。在本研究中，我们表明，在DBA/1J小鼠建立的关节炎小鼠模型中，脾脏靶向低频脉冲聚焦超声（LFPFU）有效地改善了关节炎的严重程度。此外，我们使用质谱流式细胞术以及广泛的抗体组合，对来自RA小鼠、接受超声治疗的RA小鼠和健康对照的脾脏样本进行了深入的免疫分析，并确定了14个细胞群体的免疫组成，包括CD4/CD8 T细胞、B细胞、自然杀伤细胞和树突状细胞。此外，根据细胞表面标志物和表型进行的多维度分析分别有助于在T细胞和髓系细胞中鉴定出4个和5个细胞亚群，在这3组中，有6个T细胞亚群和3个髓系细胞亚群对超声治疗有反应。在这些细胞亚群中，CD8 T细胞亚群在RA小鼠中对超声刺激表现出独特的反应。具体而言，CD8 T细胞与关节炎进展程度呈现出显著的相关性，并且可以作为基于脾脏聚焦超声治疗的一个指标。此外，脾脏细胞的单细胞RNA测序揭示了T细胞、B细胞和髓系细胞群体在抗炎途径中的重要性。这些结果阐明了对LFPFU有反应的脾脏细胞的独特细胞亚群和转录组，并证明了脾脏聚焦超声刺激在治疗炎症性疾病中的潜力。
