Study of microfilament cytoskeleton-binding proteins phagocytosis antigen in immature dendritic cells for 3 hours

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    Objective Analysis of key filament cytoskeletal binding proteins that affect the performance of antigenic phagocytosis by immature dendritic cells Methods Monocytes (MOs) were isolated from healthy human peripheral blood and imDCs and mature dendritic cells (mDCs) were obtained by cytokine-induced culture; ImDCs were cultured with low molecular weight (40 kD) and high molecular weight (150 kD) dextrans for 1, 3 and 6 hours, respectively, and the percentage of imDCs phagocytosed by flow cytometry was detected; imDCs were incubated with 40 kD dextrose for 3 h, immunophenotypic molecules of imDCs were detected by flow cytometry; localization of microfilaments and PFN1, WASP, α-actinin in cells were observed by immunofluorescence technology; mRNA and protein expression differences of MCBPs were detected by qPCR and WB; The stepwise regression and principal component analysis methods in the systems biology algorithm were used to screen the MCBPs with the largest component coefficients. Results The antigen phagocytosis of small molecules was faster, and the engulfment time lasted about 3 hours; 3 hours after imDCs phagocytosis antigen, cell phenotype and morphology gradually differentiated to mDCs, and the microfilament cytoskeleton underwent obvious remodeling; the expression of binding proteins including PFN1, CDM, WASP, CAPZB, Filamin A and α-actinin were down-regulated, the expression of WAVE1, Arp2/3 complex and Fascin were upregulated; The mRNA expression of signaling protein Rac1 was up-regulated, and the mRNA expression of CDC42 and RhoA was down-regulated; The immunofluorescence results showed that PFN1, WASP, and α-actinin transposed during the antigen engulfment of imDCs; PFN1 had the highest component coefficient. Conclusion PFN1 may be the key MCBPs in the process of antigenic phagocytosis of imDCs, which is of great significance for understanding the immunological function of imDCs.

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  • Received:June 07,2023
  • Revised:June 02,2024
  • Adopted:June 05,2024
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