title: Surface-Displayed IL-10 by Recombinant Lactobacillus plantarum Reduces Th1 Responses of RAW264.7 Cells Stimulated with Poly(I:C) or LPS.
abstract: Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surfacedisplayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.|College of Veterinary Medicine, Jilin University, Changchun 130062, P.R. China.
authors.forename: Ruopeng
authors.initials: R
authors.lastname: Cai
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Yanlong
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Wentao
authors.initials: W
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Shi
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Chunwei
authors.initials: C
authors.lastname: Shi
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Jingtao
authors.initials: J
authors.lastname: Hu
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.|Shandong Baolai-Leelai Bio-Tech Co., LTD, Taian, Shandong Province 171000, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gu
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Liping
authors.initials: L
authors.lastname: Ye
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Fangyu
authors.initials: F
authors.lastname: Zhou
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Qinglong
authors.initials: Q
authors.lastname: Gong
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun 130062, P.R. China.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Han
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Guilian
authors.initials: G
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun 130118, P.R. China.
authors.forename: Chunfeng
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D015153:Blotting, Western; D018920:Coculture Techniques; D016753:Interleukin-10 / Q000235:genetics*; D015850:Interleukin-6 / Q000378:metabolism; D007780:Lacticaseibacillus casei / Q000235:genetics; D048191:Lactiplantibacillus plantarum / Q000187:drug effects / Q000235:genetics*; D008070:Lipopolysaccharides / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D016328:NF-kappa B / Q000235:genetics; D011070:Poly I-C / Q000276:immunology; D000067996:RAW 264.7 Cells; D011994:Recombinant Proteins / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D001426:Bacterial Proteins; C556062:IL10 protein, mouse; D015850:Interleukin-6; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D016328:NF-kappa B; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D011070:Poly I-C
keywords: IL-10; Lactobacillus plantarum; RAW264.7; pgsA; surface display component