Lim Y S, Kang B Y, Kim E J, Kim S H, Hwang S Y, Kim K M, Kim T S
College of Pharmacy, Institute of Biotechnology, Chonnam National University, Kwangju, Korea.
Arch Pharm Res. 1998 Oct;21(5):537-42. doi: 10.1007/BF02975371.
To more effectively drive immune responses toward antigen-specific T helper type 2 (Th2) cell-mediated responses, we constructed a mammalian expression vector (pOVA/IL4) carrying a fused gene in which the ovalbumin (OVA) cDNA was covalently linked to murine interleukin-4 (IL-4) cDNA. A biologically active OVA/IL4 protein was expressed by the transfected COS cells with the pOVA/IL4 DNA, as demonstrated by Western blotting and cytokine bioassay. Intramuscular injection of BALB/c mice with the pOVA/IL4 DNA increased both the production of OVA-specific IL-4 by CD4+ T cells and the ratio of anti-OVA IgG1 to anti-OVA IgG2a isotypes, while the injection with the pOVA DNA alone, or with the mixture of the pOVA and pIL4 DNA did no or little increase. Furthermore, the OVA-specific, Th2 cell-mediated immune responses were significantly enhanced by multiple injections with the pOVA/IL4 DNA. These studies indicate that the direct linkage of an OVA gene to an IL-4 gene in the expression plasmid confines the effects of IL-4 to the OVA-specific cells, efficiently driving the immune response toward OVA-specific, Th2 cell-mediated responses.
为了更有效地驱动免疫反应向抗原特异性2型辅助性T细胞（Th2）介导的反应方向发展，我们构建了一个哺乳动物表达载体（pOVA/IL4），其携带一个融合基因，其中卵清蛋白（OVA）cDNA与小鼠白细胞介素-4（IL-4）cDNA共价连接。转染了pOVA/IL4 DNA的COS细胞表达了具有生物活性的OVA/IL4蛋白，蛋白质免疫印迹和细胞因子生物测定证明了这一点。用pOVA/IL4 DNA对BALB/c小鼠进行肌肉注射，增加了CD4+ T细胞产生的OVA特异性IL-4以及抗OVA IgG1与抗OVA IgG2a同种型的比例，而单独注射pOVA DNA或注射pOVA与pIL4 DNA的混合物则没有增加或增加很少。此外，多次注射pOVA/IL4 DNA显著增强了OVA特异性、Th2细胞介导的免疫反应。这些研究表明，表达质粒中OVA基因与IL-4基因的直接连接将IL-4的作用局限于OVA特异性细胞，有效地驱动免疫反应向OVA特异性、Th2细胞介导的反应方向发展。
