鸡干扰素γ与白介素2对外周血中Th1细胞分化相关细胞因子的影响

Effects of chicken interferon-γ and interleukin-2 on cytokines related to Th1 cell differentiation in peripheral blood

摘要:

为了探讨鸡干扰素γ(chicken interferon-γ,ChIFN-γ)与白介素2(chicken interleukin-2,ChIL-2)对1型辅助性T细胞(type 1 helper T lymphocytes,Th1)分化的影响，本研究原核表达、纯化了ChIFN-γ与ChIL-2，并以不同浓度刺激刀豆球蛋白A (concanavalin A,Con A)活化的鸡外周血淋巴细胞，检测其对Th1细胞分化相关细胞因子表达的影响。结果表明，不同浓度的ChIFN-γ与ChIL-2均可上调Th1细胞分化相关细胞因子的转录水平，最佳刺激浓度分别为12.5μg/mL和25.0 μg/mL。然后将ChIFN-γ和ChIL-2分别与H9N2灭活疫苗联用，以口服或肌肉注射方式免疫无特定病原体(specific pathogen free,SPF)鸡，检测其对Th1细胞分化相关细胞因子表达的影响。结果表明，与单独使用H9N2疫苗相比，ChIFN-γ和ChIL-2均能显著上调H9N2疫苗诱导的Th1细胞分化相关细胞因子的转录水平，且肌肉注射的效果优于口服。本研究通过对Th1细胞分化相关细胞因子进行检测，从体外和体内水平证实了ChIFN-γ与ChIL-2能够增强ConA或H9N2灭活疫苗诱导的Th1细胞分化，为其作为疫苗佐剂使用提供了理论依据。

Abstract:

This paper aims to explore the effects of chicken interferon-γ (ChIFN-γ) and interleukin-2 (ChIL-2) on type 1 helper (Th1) T lymphocyte differentiation. To be specific, ChIFN-γ and ChIL-2 were first expressed in Escherichia coli competent cells and then purified by Ni-NTA affinity chromatography. Different concentration of ChIFN-γ and ChIL-2 were employed to stimulate the lymphocytes in chicken peripheral blood which had been activated by concanavalin A (Con A), and the mRNA levels of cytokines related to Th1 cell differentiation were detected by real-time quantitative PCR (RT-qPCR). The results showed that both ChIFN-γ and ChIL-2 can significantly up-regulate mRNA levels of cytokines related to Th1 cell differentiation and the optimal concentration was 12.5 μg/mL and 25.0 μg/mL, respectively. In addition, specific-pathogen-free (SPF) chickens were immunized with ChIL-2 or ChIFN-γ together with H9N2 vaccine, or H9N2 vaccine alone by oral administration or intramuscular injection, respectively. The mRNA levels of cytokines related to Th1 cell differentiation were detected after immunization. The results showed that ChIFN-γ and ChIL-2 significantly up-regulated the mRNA levels of cytokines related to Th1 cell differentiation induced by H9N2 vaccine compared with H9N2 vaccine alone, and that the intramuscular injection was better than oral administration. In this study, we verified that ChIFN-γ and ChIL-2 can significantly enhance mRNA levels of cytokines related to Th1 cell differentiation induced by ConA or H9N2 vaccine in vitro and in vivo. The results of this study can lay a theoretical basis for using ChIFN-γ and ChIL-2 as vaccine adjuvants.