Double‐negative T cells mediate M1 polarization of microglial cells via TNF‐α‐NLRP3 to aggravate neuroinflammation and cognitive impairment in Alzheimer's disease mice

• Published in: Journal of cellular physiology 2022-10, Vol.237 (10), p.3860-3871
• Main Authors: Han, Chenyang, Sheng, Yongjia, Wang, Jin, Zhou, Xiaohong, Li, Wenyan, Zhang, Caiqun, Guo, Li, Yang, Yi
• Format: Article
• Language: English
• Subjects: Alzheimer's disease; Antibodies; CD11b antigen; Cell activation; Cognitive ability; cognitive impairment; double‐negative T cells; Flow cytometry; Immunoassay; Impairment; Inflammasomes; Inflammation; Inflammatory response; Lymphocytes; Lymphocytes T; M1 polarization; Microglia; microglial cells; Neurodegenerative diseases; Polarization; Presenilin 1; Regulatory mechanisms (biology); siRNA; Tumor necrosis factor receptors; Tumor necrosis factor-TNF; Tumor necrosis factor-α
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.30839

We mainly study the role and regulatory mechanism of double‐negative T cells (DNTs) in Alzheimer's disease (AD). The mice splenic DNTs were separated and amplified by Rosettesep antibody adsorption method and Easysep magnetic activated cell sorting. DNTs were intraperitoneally injected into the APP/PS1‐AD mice model, which was found to aggravate cognitive impairment in mice. DNTs secreted tumor necrosis factor α (TNF‐α) to promote the activation of NLRP3 and the M1 polarization of microglial cells, and silencing NLRP3 with small interfering RNA (siRNA) suppressed the effect of DNTs. DNTs were later cocultured with mice microglial cell line BV2, then fluorescence staining was conducted to detect NLRP3 expression, and enzyme‐linked immunoassay was performed to measure the expression of inflammatory factors. Moreover, the levels of NLRP3, ASC, and TNFR1 proteins were detected by western‐blot assay, and the proportion of F4/80 + CD11b + M1 cells was detected by flow cytometry. DNTs promoted the M1 polarization of BV2 cells and the activation of NLRP3 inflammasome. After treatment of BV2 cells with NLRP3 inhibitor, the effect of DNTs was weakened. Later, TNF‐α siRNA was transfected into DNTs, and it was found that DNTs with TNF‐α silencing had markedly weakened polarization effect on BV2 cells. We discovered that the proportion of DNTs increased in AD patients. DNTs secreted TNF‐α to regulate the activation of NLRP3 inflammasome and the M1 polarization of microglial cells, thus promoting the central inflammatory response and aggravating the cognitive impairment in AD mice.