牙周炎基因疫苗佐剂IL⁃15对SD大鼠肠系膜淋巴结B细胞增殖的影响
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国家自然科学基金(81560186);贵州省科技计划项目(黔科合基础[2019]1333号);遵义市科技计划课题[遵市科合HZ字(2019)21号];贵州省高等学校口腔疾病研究特色重点实验室开放课题(SKLODR?2016?04)


Effects of periodontitis gene vaccine adjuvant IL⁃15 on the proliferation of mesenteric lymph node B cells in SD rats
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    目的:观察牙周炎疫苗佐剂白细胞介素-15(interleukin-15,IL-15)对SD大鼠肠系膜淋巴结B淋巴细胞增殖的影响,初步探讨用IL-15提高基因疫苗免疫效应的可能性。方法:将牙周炎基因疫苗pVAX1-HA2-FimA/IL-15(A组)、pVAX1-HA2-FimA(B组)和生理盐水(C组)鼻滴免疫SD大鼠,采用磁珠分选法分离纯化肠系膜淋巴结B细胞,CCK-8法检测B细胞的数量,RT-qPCR法检测CKs2基因相对表达量变化,ELISA法检测培养液中特异性IgA的表达水平。结果:A组B细胞增殖水平高于B组和C组(P < 0.05);A组B淋巴细胞CKs2 mRNA相对表达量稍高于B组,但差异无统计学意义(P>0.05)。A组B淋巴细胞分泌的特异性IgA水平均高于B、C组(P<0.05)。结论:IL-15是一种有效的基因疫苗佐剂,使用本课题组构建的牙周炎基因疫苗能够促进B细胞增殖,进而提高特异性IgA抗体的表达水平。

    Abstract:

    Objective:To observe the effect of periodontitis vaccine adjuvant interleukin-15(IL-15)on the proliferation of B lymphocytes in the mesenteric lymph nodes of SD rats,and explore the possibility of using IL-15 to improve the immune effect of gene vaccine. Methods:SD rats were immunized with nasal spray of the periodontitis gene vaccine pVAX1-HA2-FimA/IL-15(group A),pVAX1-HA2-FimA(group B),and normal saline(group C). Then mesenteric lymph node B cells were isolated and purified by magnetic bead sorting method,the number of B cells was detected by CCK8 method,the relative expression level of CKs2 gene was detected by RT-qPCR,and the expression level of specific IgA in culture medium was detected by ELISA. Results:The proliferation level of B lymphocytes in group A was higher than those in group B and group C(P < 0.05). The relative expression level of CKs2 mRNA of B lymphocytes in group A was higher than that in group B,but the difference was not statistically significant(P > 0.05). The level of specific IgA secreted by B lymphocytes in group A was higher than those in groups B and C(P < 0.05). Conclusion:IL-15 is an effective gene vaccine adjuvant to promote the proliferation of B cells and increase the expression level of specific IgA antibodies in the periodontitis gene vaccine constructed by our team.

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田 源,高晓佩,曾凤娇,于 航,刘建国,白国辉.牙周炎基因疫苗佐剂IL⁃15对SD大鼠肠系膜淋巴结B细胞增殖的影响[J].南京医科大学学报(自然科学版),2020,(10):1456-1459

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  • 收稿日期:2019-05-10
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  • 在线发布日期: 2020-11-10
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