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目的 构建ＩＦＮ－γ基因真核表达重组质粒作为基因佐剂，观察其与ｐｃＤＮＡ３－ＲＯＰ１（ｐｃ－ＲＯＰ１） ＤＮＡ共同免疫小鼠所诱导的免疫应答。方法 将ＩＦＮ－γ基因片段定向插入真核表达载体ｐｃＤＮＡ３，双酶切鉴定，获得ｐｃＩＦＮ 重组子；碱裂解法大量制备，经肌肉注射免疫ＢＡＬＢ／ｃ小鼠，每只鼠注射ｐｃＩＦＮ、ｐｃ－ＲＯＰ１ 各１００μｇ，两周后同量加强免疫一次，以ｐｃＤＮＡ３空质粒及生理盐水组为对照。分别于免疫后第３０ 天、５０ 天、７０天共三次用ＭＴＴ法测定小鼠脾脏Ｔ淋巴细胞增殖活性及ＮＫ细胞活性；双夹心ＥＬＩＳＡ 测定血清细胞因子ＩＦＮ－γ、ＩＬ－２及酶法测定ＮＯ含量；ＥＬＩＳＡ法测定ＩｇＧ抗体滴度。结果 构建成功的作用下，该两项指标均明显提高，且ＩＦＮ－γ、ＩＬ－２ 及ＮＯ水平均较不加佐剂组显著提高（Ｐ＜ ０．０１）；而对ＩｇＧ抗体滴度无显著影响（Ｐ＞ ０．０５。结论 ＩＦＮ－γ基因佐剂具有协同ｐｃ－ＲＯＰ１ＤＮＡ免疫的作用，可增强免疫鼠细胞免疫应答，ＩＦＮ－γ、ＩＬ－２细胞因子及ＮＯ的产生
OBJECTIVE: To construct an eukaryotic expression recombinant plasmid of IFN-γ gene as a gene adjuvant and observe its immune response induced by co-immunization with pcDNA3-ROP1 (pc-ROP1) DNA. Methods The IFN-γ gene fragment was inserted into the eukaryotic expression vector pcDNA3 by double enzyme digestion to obtain pcIFN recombinant. After alkaline lysis, a large amount of IFN-γ gene fragment was prepared and immunized BALB / c mice by intramuscular injection. Each mouse was injected with pcIFN and pc-ROP1 Each 100μg, two weeks after the same amount of booster immunization to pcDNA3 empty plasmid and saline group as a control. The activity of T lymphocyte proliferation and the activity of NK cells in spleens were measured by MTT method three times after immunization on the 30th, 50th and 70th days respectively. Serum cytokines IFN-γ and IL-2 were determined by enzyme-linked immunosorbent assay NO content; ELISA method for the determination of IgG antibody titers. Results The successful construction of these two indicators were significantly increased, and the levels of IFN-γ, IL-2 and NO were significantly increased compared with those without adjuvant (P <0.01), while the titer of IgG antibody was not significant (P> 0.05) .Conclusion IFN-γ gene adjuvant has the effect of cooperating pc-ROP1 DNA immunization, and can enhance the immune response of immunized mice, the production of IFN-γ, IL-2 cytokines and NO