title: Evaluation of humoral and cellular immune responses to a DNA vaccine encoding chicken type II collagen for rheumatoid arthritis in normal rats.
abstract: A major challenge in the development of effective therapies for rheumatoid arthritis (RA) is finding a method for the specific inhibition of the inflammatory disease processes without the induction of generalized immunosuppression. Of note, the development of therapeutic DNA vaccines and boosters that may restore immunological tolerance remains a high priority. pcDNA-CCOL2A1 is a therapeutic DNA vaccine encoding chicken type II collagen(CCII). This vaccine was developed by our laboratory and has been shown to exhibit efficacy comparable to that of the current "gold standard" treatment, methotrexate (MTX). Here, we used enzyme-linked immunosorbent assays with anti-CII IgG antibodies, quantified the expression levels of Th1, Th2, and Th3 cytokines, and performed flow cytometric analyses of different T-cell subsets, including Th1, Th2, Th17, Tc, Ts, Treg, and CD4(+)CD29(+)T cells to systemically evaluate humoral and cellular immune responses to pcDNA-CCOL2A1 vaccine in normal rats. Similar to our observations at maximum dosage of 3 mg/kg, vaccination of normal rats with 300 μg/kg pcDNA-CCOL2A1 vaccine did not induce the production of anti-CII IgG. Furthermore, no significant changes were observed in the expression levels of pro-inflammatory cytokines interleukin (IL)-1α, IL-5, IL-6, IL-12(IL-23p40), monocyte chemotactic protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, regulated on activation in normal T-cell expressed and secreted (RANTES), receptor activator for nuclear factor-κB ligand (RANKL), and granulocyte colony-stimulating factor (G-CSF) or anti-inflammatory cytokines IL-4 and IL-10 in vaccinated normal rats relative to that in controls(P > 0.05). However, transforming growth factor (TGF)-β levels were significantly increased on days 10 and 14, while interferon (IFN)-γ and tumor necrosis factor (TNF)-α levels were significantly decreased on days 28 and 35 after vaccination(P < 0.05). Similarly, there were no significant differences in the percentages of Tc, Ts, Th1/Th2, and Th17 cells between the 2 groups(P > 0.05), with the exception of Treg cells, which were significantly reduced on days 14 and 21 after vaccination (P < 0.05), and CD4(+)CD29(+)T cells, which were significantly increased on days 7 and 14 after vaccination(P < 0.05).Taken together, these results suggested that pcDNA-CCOL2A1 vaccine did not markedly affect the balance of immune system components in vaccinated normal rats, indicating that this DNA vaccine may have clinical applications in the treatment of RA.
authors.affiliation: a Department of Immunology and National Center for Biomedicine Analysis ; Beijing 307 Hospital Affiliated to Academy of Military Medical Sciences ; Beijing , China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Xiao
authors.forename: Long
authors.initials: L
authors.lastname: Juan
authors.forename: Yun
authors.initials: Y
authors.lastname: Song
authors.forename: Zhang
authors.initials: Z
authors.lastname: Zhijian
authors.forename: Jin
authors.initials: J
authors.lastname: Jing
authors.forename: Yu
authors.initials: Y
authors.lastname: Kun
authors.forename: Hao
authors.initials: H
authors.lastname: Yuna
authors.forename: Dai
authors.initials: D
authors.lastname: Dongfa
authors.forename: Ding
authors.initials: D
authors.lastname: Lili
authors.forename: Tan
authors.initials: T
authors.lastname: Liuxin
authors.forename: Liang
authors.initials: L
authors.lastname: Fei
authors.forename: Liu
authors.initials: L
authors.lastname: Nan
authors.forename: Yuan
authors.initials: Y
authors.lastname: Fang
authors.forename: Sun
authors.initials: S
authors.lastname: Yuying
authors.forename: Xi
authors.initials: X
authors.lastname: Yongzhi
mesh_terms: D000818:Animals; D001172:Arthritis, Rheumatoid / Q000188:drug therapy*; D002645:Chickens; D024043:Collagen Type II / Q000276:immunology*; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D007111:Immunity, Cellular / Q000276:immunology*; D056724:Immunity, Humoral / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008297:Male; D051381:Rats; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D019444:Vaccines, DNA / Q000276:immunology* / Q000627:therapeutic use*
chemical_list: D024043:Collagen Type II; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D019444:Vaccines, DNA; D016753:Interleukin-10; D016179:Granulocyte Colony-Stimulating Factor; D015847:Interleukin-4; D007371:Interferon-gamma
keywords: CCOL2A1; cellular immunity; chicken type II collagen; RA; chicken type II procollagen; chicken type II procollagen gene; CCII; collagen induced arthritis; MTX; disease modifying antirheumatic drug; experimental autoimmune encephalomyelitis; NOD; granulocyte colony-stimulating factor; EAE; humoral immunity; interleukin; MCP-1; macrophage inflammatory protein-1 α; RANTES; methotrexate; IL; monocyte chemotactic protein-1; MIP-1α; multiple sclerosis; DMARD; receptor activator for nuclear factor-κB ligand; G-CSF; regulated on activation in normal T-cell expressed and secreted; RANKL; rheumatoid arthritis; rheumatoid arthritis; CIA; spontaneous nonobese diabetic; MS; therapeutic DNA vaccine