In:
PeerJ, PeerJ, Vol. 6 ( 2018-09-21), p. e5638-
Abstract:
The benefits of IL-35 treatment have been verified in multiple animal models of diseases, while its influence on T cells immunity under normal condition still needs to be elucidated. The present study was designed to investigate the effects modulating IL-35 levels in vivo and in vitro on T cells, response and also the effects on T cells subsets in normal mice. Methods A plasmid pMSCV-IL-35-GFP carrying mouse linear IL-35 fragment with two subunits joint together was constructed and the heterodimer expression was confirmed. Normal mice were randomly divided into three groups and received an intravenous injection of PBS, pMSCV-GFP and pMSCV-IL-35-GFP respectively. After 72 h, spleen tissues and peripheral blood were harvested for following analysis. Meanwhile, splenic T cells were isolated and incubated with 10, 30, or 50 ng/mL recombinant IL-35 factor for 24 h with the addition of anti-CD3/CD28 in vitro . T-cell subsets were assessed by Fluorescence activated cell sorting (FACS) and related cytokines together with effector molecules were determined by real time PCR. Results Western blotting confirmed a 52 kDa band in the cell lysate of HEK 293T transducted with pMSCV-IL-35-GFP plasmid, indicating a successful expression of IL-35. Ebi3 and IL-12A, two subunits of IL-35, could be identified 72 h post DNA injection. IL-35 upregulation in vivo effectively inhibit CD4 + and CD8 + T cell proliferation and Th1 cytokine secretion. Effector molecules of CD8 + T cells were also remarkably suppressed. On the contrary, high level of IL-35 significantly induced CD4 + CD25 + Tregs and Th2 enhancement. The in vitro study provided similar results. Conclusion The results indicated Th1 and CD8 + T cell inhibition and Th2 and Tregs bias in the presence of IL-35 under a normal state which partly contributed to its therapeutic potential.
Type of Medium:
Online Resource
ISSN:
2167-8359
DOI:
10.7717/peerj.5638/fig-1
DOI:
10.7717/peerj.5638/fig-2
DOI:
10.7717/peerj.5638/fig-3
DOI:
10.7717/peerj.5638/fig-4
DOI:
10.7717/peerj.5638/fig-5
DOI:
10.7717/peerj.5638/fig-6
DOI:
10.7717/peerj.5638/table-1
DOI:
10.7717/peerj.5638/supp-1
DOI:
10.7717/peerj.5638/supp-2
DOI:
10.7717/peerj.5638/supp-3
DOI:
10.7717/peerj.5638/supp-4
Language:
English
Publisher:
PeerJ
Publication Date:
2018
detail.hit.zdb_id:
2703241-3
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