Regulation of bovine Th2 differentiation in Vitro

ABSTRACT: Naive CD4+ T cells, sorted from PBMCs of grass-fed beef cattle, were stimulated with anti-bovine CD3 under Th2 differentiation conditions with or without Ostertagia ostertagi (OO) protein extract. Th1 differentiation conditions were included for comparison. The effect of recombinant bovine IL-4 (rbIL-4) and weakening TCR signal strength on Th2 differentiation were also tested. Flow cytometry, qPCR and proteomic assay were performed to analyze the differentiated cells. The majority of differentiated cells expressed IFNgamma (a hallmark cytokine for Th1) and a small percentage of cells expressed both IFNgamma and IL-4 (a hallmark cytokine for Th2), indicating the presence of Th0 cells, as previously reported in cattle. While weakening TCR signal strength reduced Th2 cell expansion, adding rbIL-4 or OO protein extract enhanced Th2 cell expansion but without significant changes in IFNgamma and IL-4 expression. Proteomic data predicted that, as in mice and humans, bovine Th2 differentiation results from three upstream stimulation with CD3, CD28, and IL-4, which were inhibited when OO protein extract was added into the Th2 differentiation media. Importantly, protein profiling indicated that the addition of rbIL-4 enhanced IL-4 signaling but inhibited IL-12 signaling. Soon, we are planning to explore if other cytokines like IL-10 can be applied to optimize this Th2 differentiation in vitro. Naive bovine CD4+ T cells differentiate into a mixed population containing a high percentage of IFNgamma producing cells and a small percentage of Th0 cells. Furthermore, bovine Th2 differentiation is sensitive to regulation such as by OO or rbIL-4.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Bos Taurus (ncbitaxon:9913)

SUBMITTER: Yan Wang  

PROVIDER: MSV000091527 | MassIVE | Wed Mar 22 12:13:00 GMT 2023

SECONDARY ACCESSION(S): PXD041034

REPOSITORIES: MassIVE