The Interaction Of PHB1 With STAT3 And Its Effect On TH17 Cells

Objective: In the previous study,the research team found that PHB12,another PHB1 family member,interacts with the key transcription factor ROR?t of Th17 cells,and also reported that PHB12 interacts with another important transcription factor,STAT3,so we would like to explore Whether PHB1 interacts with STAT3 and whether this interaction regulates Th17 cell differentiation and function.The study of the differentiation and regulation of Th17 cells can find a new therapeutic target and treatment idea for the treatment of Th17 cell-related inflammation and autoimmune diseases.Methods: Firstly,Th17 cells at different time points were induced to differentiate and the expression of PHB1 and STAT3 was detected by fluorescence quantitative PCR.Then the HA-PHB1 and Flag-STAT3 expression plasmids were constructed by gene recombination technology and transferred into HEK293 T cells.PHB1 and STAT3 were co-localized in HEK293 T cells by immunofluorescence.Then HA-PHB1 and Flag-STAT3 expression plasmids were co-transfected into HEK293 T cells,and the interaction between PHB1 and STAT3 was detected by co-immunoprecipitation;then Na?ve CD4+ T cells were sorted from human umbilical cord blood and differentiated in vitro.Th17 cells were obtained and their interaction in Th17 cells was verified by co-immunoprecipitation.Finally,the PHB1 interference plasmid was constructed and packaged into lentivirus to achieve the expression of the effector gene that reduced PHB1 in Th17 cells.The expression of IL-17 at the transcriptional level was detected by fluorescence quantitative PCR.Results:In the process of Th17 induction of differentiation,PHB1 and STAT3 gene expression is consistent,and the two in the early phase of induction of differentiation,that is,within 72 hours of the two more close.Both PHB1 and STAT3 can be localized in the nucleus and cytoplasm,and their fluorescence intensity in the cytoplasm is stronger than in the nucleus.PHK1 interacts with STAT3 in HEK293 T cells and interacts with human Th17 cells.Reduction of PHB1 in human Th17 cells down-regulated the expression of IL-17 secreted by Th17 cells.In addition,the expression of STAT3 was also reduced.