Molecular mechanism of β-1, 4-galactosyltransferase 6 in regulating lymphocyte migration / 中华微生物学和免疫学杂志

ABSTRACT

Objective:To study the molecular mechanism of β-1, 4-galactosyltransferase 6 (β4galt6) in regulating lymphocyte migration under inflammatory conditions.Methods:CRISPR/Cas9 system was used to knock out the β4galt6 gene of mouse islet vascular endothelial cells (MS1). Adhesion assay was performed to compare the adhesion ability of lymphocytes to wild-type cells and gene knockout cells. Expression of adhesion molecules on the surface of wild-type cells and gene knockout cells were compared using RT-PCR and flow cytometry. Transwell model was used to compare the transmigration ability of lymphocytes across wild-type cells and gene knockout cells.Results:The β4galt6 gene knockout cell line, β4galt6 KO, was successfully constructed. The percentage of lymphocytes adhereing to wild-type MS1 cells was significantly higher than that to β4galt6 KO cells under inflammatory conditions. The expression of adhesion molecules including intercellular adhesion molecule 1 (ICAM1), vascular cell adhesion molecule 1 (VCAM1) and P-selectin on the surface of wild-type MS1 cells was significantly higher than that on β4galt6 KO cells. Moreover, the percentage of lymphocytes passing through wild-type MS1 cells was significantly higher than that through β4galt6 KO cells.Conclusion:Under inflammatory conditions, β4galt6 could promote the migration of lymphocytes.