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目的为了探讨质粒DNA体外刺激淋巴细胞的增殖状况,建立了一种方便可靠的评价豚鼠细胞免疫水平的试验方法。方法用羧基荧光素乙酰乙酸琥珀酰亚胺酯(CFSE)染色豚鼠全血,经植物血凝素(PHA)和质粒DNA刺激培养,利用流式细胞术分析细胞的增殖状况。结果豚鼠全血经PHA和DNA刺激,淋巴细胞增殖能力不同;未免疫组经DNA和PHA刺激后增殖的差异显著,免疫组差异不显著。DNA质粒在体内外均可作为刺激源刺激淋巴细胞增殖。结论建立了一种基于活细胞染料CFSE染色的豚鼠全血淋巴细胞增殖试验方法,可方便、快速、有效地评价细胞免疫水平。
Objective To investigate the proliferation of lymphocytes stimulated by plasmid DNA in vitro and to establish a convenient and reliable method for the evaluation of cellular immunity in guinea pigs. Methods Whole blood of guinea pigs was stained with carboxyfluorescein acetoacetate succinimidyl ester (CFSE). The cells were stimulated with PHA and plasmid DNA. The proliferation of cells was analyzed by flow cytometry. Results The whole blood of guinea pigs were stimulated by PHA and DNA, and the ability of proliferation of lymphocytes was different. There was significant difference in proliferation between untreated group and control group after DNA and PHA stimulation. There was no significant difference in immune group. DNA plasmids can stimulate lymphocyte proliferation both in vitro and in vivo. Conclusions A method for testing whole blood lymphocyte proliferation of guinea pigs based on the CFSE staining of living cells was established, which can evaluate the cellular immunity conveniently, rapidly and effectively.