PMID- 32194180 OWN - NLM STAT- MEDLINE DCOM- 20210329 LR - 20210329 IS - 1096-1208 (Electronic) IS - 0882-4010 (Linking) VI - 143 DP - 2020 Jun TI - Recombinant ubiquitin-conjugating enzyme of Eimeria maxima induces immunogenic maturation in chicken splenic-derived dendritic cells and drives Th1 polarization in-vitro. PG - 104162 LID - S0882-4010(20)30449-6 [pii] LID - 10.1016/j.micpath.2020.104162 [doi] AB - Dendritic cells (DCs) are key linkages between innate immunity and acquired immunity. The antigens that promote the functions of DCs might be the effective candidates of novel vaccine. In this research, the ability of ubiquitin-conjugating enzyme (UCE), a recognized common antigens among chicken Eimeria species, to stimulate DCs of chickens were evaluated. We cloned UCE gene from Eimeria maxima (EmUCE), and its protein expression was confirmed by SDS-PAGE and western-blot. Immunofluorescence assay confirmed the binding of rEmUCE on the surface of chicken splenic-derived DCs (ChSP-DCs). Flow cytometric analysis showed that rEmUCE-treated ChSP-DCs increased MHCII, CD1.1, CD11c, CD80, and CD86 phenotypes. qRT-PCR indicated that transcript levels of maturation markers CCL5, CCR7, and CD83 in ChSP-DCs were upregulated in response to rEmUCE. Following rEmUCE treatment, chSP-DCs activated TLR signaling and inhibited Wnt signaling. Moreover, rEmUCE promoted DC-mediated T-cell proliferation in DC/T-cell co-incubation. Interestingly, CD3(+)/CD4(+) T-cells were significantly enhanced when rEmUCE-treated chSP-DCs were co-incubated with T-cells. Cytokine secretion pattern of rEmUCE-stimulated ChSP-DCs revealed that the production of IL-12 and IFN-gamma was increased whereas IL-10 and TGF-beta were unchanged. Likewise, the co-incubation of ChSP-DCs with T-cells indicated increased production of IFN-gamma but not IL-4. Collectively, rEmUCE could polarize DCs to immunogenic phenotype and shift the immune cells towards Th1 response. Our observations provide valuable insight for future research aimed at vaccine development against avian coccidiosis. CI - Copyright (c) 2020 The Authors. Published by Elsevier Ltd.. All rights reserved. FAU - Lakho, Shakeel Ahmed AU - Lakho SA AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2017207046@njau.edu.cn. FAU - Haseeb, Muhammad AU - Haseeb M AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2016207041@njau.edu.cn. FAU - Huang, Jianmei AU - Huang J AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2016207025@njau.edu.cn. FAU - Hasan, Muhammad Waqqas AU - Hasan MW AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2015207037@njau.edu.cn. FAU - Naqvi, Muhammad Ali-Ul-Husnain AU - Naqvi MA AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2017207047@njau.edu.cn. FAU - Zhou, Zhouyang AU - Zhou Z AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: 2016107051@njau.edu.cn. FAU - Song, XiaoKai AU - Song X AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: songxiaokai@njau.edu.cn. FAU - Yan, RuoFeng AU - Yan R AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: yanruofeng@njau.edu.cn. FAU - Xu, Lixin AU - Xu L AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: xulixin@njau.edu.cn. FAU - Li, XiangRui AU - Li X AD - MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China. Electronic address: lixiangrui@njau.edu.cn. LA - eng PT - Journal Article DEP - 20200317 PL - England TA - Microb Pathog JT - Microbial pathogenesis JID - 8606191 RN - 0 (Protozoan Proteins) RN - 0 (Recombinant Proteins) RN - EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes) SB - IM MH - Animals MH - Cell Differentiation MH - Chickens MH - Cloning, Molecular MH - Dendritic Cells/*metabolism/physiology MH - Eimeria/*enzymology/genetics MH - Flow Cytometry MH - Fluorescent Antibody Technique MH - Protozoan Proteins/genetics/*metabolism MH - Recombinant Proteins MH - Sequence Analysis, DNA MH - Th1 Cells/*metabolism/physiology MH - Ubiquitin-Conjugating Enzymes/genetics/*metabolism OTO - NOTNLM OT - Cytokines OT - Dendritic cells OT - Eimeria species OT - T-cell mediated immunity OT - Toll-like receptors OT - Ubiquitin-conjugating enzyme COIS- Declaration of competing interest The authors declare that they have no conflict with competing interests. EDAT- 2020/03/21 06:00 MHDA- 2021/03/30 06:00 CRDT- 2020/03/21 06:00 PHST- 2019/11/19 00:00 [received] PHST- 2020/03/12 00:00 [revised] PHST- 2020/03/13 00:00 [accepted] PHST- 2020/03/21 06:00 [pubmed] PHST- 2021/03/30 06:00 [medline] PHST- 2020/03/21 06:00 [entrez] AID - S0882-4010(20)30449-6 [pii] AID - 10.1016/j.micpath.2020.104162 [doi] PST - ppublish SO - Microb Pathog. 2020 Jun;143:104162. doi: 10.1016/j.micpath.2020.104162. Epub 2020 Mar 17.