High IF Citations

【Cited FineTest Elisa Kit】 Current Research on Autophagy and Autoimmune Disorder

FineTest Elisa kit contributes to the research on autophagy and autoimmune disorder. The immunoassay is designed to measure IL-8 and IL-1β concentration in THP-1 and RAW 264.7 cells.

Publication Details
Article Title: Vitamin D3 and cArbamazepine Protect against Clostridioides Difficile Infection in Mice by Restoring Macrophage Lysosome Acidification
Journal Title: Autophagy
DOI: 10.1080/15548627.2021.2016004
IF: 16.016

Abstract:        Clostridioides difficile infection (CDI) is a common cause of nosocomial diarrhea. TcdB is a major C. difficile exotoxin that activates macrophages to promote inflammation and epithelial damage. Lysosome impairment is a known trigger for inflammation. Herein, we hypothesize that TcdB could impair macrophage lysosomal function to mediate inflammation during CDI. Effects of TcdB on lysosomal function and the downstream pro-inflammatory SQSTM1/p62-NFKB (nuclear factor kappa B) signaling were assessed in cultured macrophages and in a murine CDI model. Protective effects of two lysosome activators (i.e., vitamin D3 and carbamazepine) were assessed. Results showed that TcdB inhibited CTNNB1/β-catenin activity to downregulate MITF (melanocyte inducing transcription factor) and its direct target genes encoding components of lysosomal membrane vacuolar-type ATPase, thereby suppressing lysosome acidification in macrophages. The resulting lysosomal dysfunction then impaired autophagic flux and activated SQSTM1-NFKB signaling to drive the expression of IL1B/IL-1β (interleukin 1 beta), IL8 and CXCL2 (chemokine (C-X-C motif) ligand 2). Restoring MITF function by enforced MITF expression or restoring lysosome acidification with 1α,25-dihydroxyvitamin D3 or carbamazepine suppressed pro-inflammatory cytokine expression in vitro. In mice, gavage with TcdB-hyperproducing C. difficile or injection of TcdB into ligated colon segments caused prominent MITF downregulation in macrophages. Vitamin D3 and carbamazepine lessened TcdB-induced lysosomal dysfunction, inflammation and histological damage. In conclusion, TcdB inhibits the CTNNB1-MITF axis to suppress lysosome acidification and activates the downstream SQSTM1-NFKB signaling in macrophages during CDI. Vitamin D3 and carbamazepine protect against CDI by restoring MITF expression and lysosomal function in mice.
Keywords:      Autophagic Flux, Clostridium Difficile, MITF, Macrophages, Toxin B

Immunoassay

FineTest Product Sample Detection Target Species
Mouse IL-8/cxcl15(Interleukin 8) ELISA Kit (EM1592) RAW 264.7 cell IL-8 Mouse
Human IL-1β(Interleukin 1 Beta) ELISA Kit (EH0185) THP-1 cell IL-1β Human

Validated Image

Autophagy

Figure Source: Autophagy, 2022 Jan 6;1-18. doi: 10.1080/15548627.2021.2016004.

Figure 1. Pro-inflammatory cytokine induction, autophagic flux impairment and NFKB activation by TcdB in human PMA-differentiated THP-1 macrophages: (E-F)Knockdown of SQSTM1 with siRNA abolished TcdB (10 ng/ml; 6 h)-induced (E) NFKB RELA/p65 phosphorylation at serine 536 and (F) mRNA expression of IL1B, IL8, and CXCL2.