| 刘露丝,张乐,邵安良,徐丽明.基于Gal抗原缺失小鼠的腹腔巨噬细胞吞噬鸡血红细胞试验敏感性初探[J].中国药事,2021,35(3):300-306 |
| 基于Gal抗原缺失小鼠的腹腔巨噬细胞吞噬鸡血红细胞试验敏感性初探 |
| On Sensitivities of Phagocytosis of Mouse Peritoneal Macrophages on Chicken Erythrocytes Based on Gal Antigen-deficient Mice |
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| DOI：10.16153/j.1002-7777.2021.03.009 |
| 中文关键词: Gal抗原缺失小鼠 巨噬细胞 鸡血红细胞 吞噬作用 免疫原性 固有免疫 |
| 英文关键词: Gal antigen-deficient mice macrophages chicken erythrocytes phagocytosis immunogenicity innate immunity |
| 基金项目:国家重点研发计划(2016YFC1103200，2016YFC1103203) |
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| 中文摘要: |
| 目的：采用腹腔巨噬细胞吞噬鸡红细胞试验考察Gal抗原缺失小鼠评价动物源材料对固有免疫影响的敏感性。方法：Gal抗原缺失小鼠(GGTA1 KO)经兔血预免后，随机分为空白对照组(生理盐水)、地塞米松抑制性阳性对照组、植物血凝素刺激性阳性对照组、牛跟腱样品植入组。地塞米松阳性对照组与植物血凝素阳性对照组参照YY/T 1465.4-2017进行给药。牛跟腱植入组皮下植入牛跟腱(2.4 cm2 /只)，周期为4周。最后所有实验组小鼠均腹腔注射鸡血红细胞，参照YY/T 1465.4-2017计数巨噬细胞和被吞噬的鸡血红细胞个数，并计算吞噬百分数和吞噬指数。结果：Gal抗原缺失小鼠的植物血凝素阳性对照组的细胞吞噬百分数和吞噬指数为39.05%和2.02，略高于空白对照组的32.15%和1.87，但无统计学差异；牛跟腱植入组的细胞吞噬百分数和吞噬指数分别为55%和2.59，均高于空白对照组，存在显著性差异(P＜0.01)；地塞米松抑制性阳性对照组吞噬百分数和吞噬指数分别为20.60%和1.43，低于空白对照组(P＜0.05，P＜0.01)；研究结果表明，Gal抗原缺失小鼠对植物血凝素有一定程度的刺激反应， 而地塞米松可明显降低吞噬百分数和吞噬指数，说明试验体系基本成立。牛跟腱植入物引起了吞噬细胞吞噬百分数和吞噬指数的显著升高，提示牛跟腱具有免疫原性风险。结论：在腹腔巨噬细胞吞噬鸡血红细胞试验中，Gal抗原缺失小鼠对地塞米松抑制性阳性对照表现出显著的抑制效应，而牛跟腱植入物的刺激引起了明显的刺激效应，表明Gal抗原缺失小鼠对于评价动物源性生物材料对固有免疫的影响具有一定应用价值。 |
| 英文摘要: |
| Objective: By using the test of phagocytosis of mouse peritoneal macrophages on chicken erythrocytes to evaluate the sensitivities of Gal antigen-deficient mice on the effect of innate immunity caused by biomaterial derived from animal tissues. Methods: Gal antigen-deficient mice (pre-immunized with rabbit redblood cells) were randomly divided into blank control group (saline solution administration), the positive control group inhibited with dexamethasone, the positive control group stimulated with phytohemagglutinin (PHA), and the bovine-tendon-implanted group. The mice were administrated with dexamethasone and PHA according to the phagocytosis of mouse peritoneal macrophages on chicken erythrocytes ex-vivo method (YY/T 1465.4-2017), respectively. And in the bovine-tendon-implanted group, bovine tendon (2.4 cm2 ) was subcutaneously implanted to the backside of the mice for 4 weeks. Finally, chicken erythrocytes were intraperitoneally injected into all the experimental mice, the numbers of macrophages and chicken erythrocytes ingested by macrophages were counted, and the percentage and index of phagocytosis were calculated. Results: There was a slight increase in the percentage and index of phagocytosis in the positive control group stimulated with PHA (39.05%, 2.02) compared with those in the blank control group (32.15%, 1.87) with no significance. The percentage and index of phagocytosis in the bovine bovine-tendon-implanted group were 55% and 2.59, which were significantly higher than those in the blank control group (P＜0.01). And in the positive control group inhibited with dexamethasone, the percentage and index of phagocytosis were 20.60% and 1.43, which were lower than those in the blank control group (P＜0.05, P＜0.01). These results suggested that PHA has increased the percentage and index of phagocytosis slightly, but the dexamethasone has decreased the percentage and index of phagocytosis significantly, so the test system was basically established. Furthermore, bovine tendon has increased the percentage and index of phagocytosis obviously, suggesting that bovine tendon has immunogenicity risk. Conclusion: According to phagocytosis of mouse peritoneal macrophages on chicken erythrocytes ex-vivo method, dexamethasone could inhibit the phagocytosis function of macrophages. In contrast, stimulation of the bovine tendon implant elicited a significant stimulatory effect, suggesting that Gal antigen-deficient mice could be used to evaluate the effect of innate immunity caused by biomaterials derived from animal tissues. |
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