论文部分内容阅读
目的从成份血浆除白细胞过滤板中分离T淋巴细胞,并检测其免疫原性。方法以白细胞过滤板为原料,从白细胞滤板中分离培养T淋巴细胞,计数单核细胞数量,并检测细胞活力。去除细胞中的红细胞和血小板,纯化后,对T淋巴细胞进行诱导、分化和培养,并检测其对猪的免疫原性。结果从每个白细胞滤板中平均可分离3.94×107个单核细胞,细胞平均存活率达93.18%。纯化后T淋巴细胞在体外增殖培养8~10 d处于增殖高峰期,14 d后达平衡期;增殖倍数最高可达100倍,平均为38.5倍。用分离的淋巴细胞免疫猪后,具有较强的免疫反应,致1头猪死亡;E玫瑰花抑制试验显示,有1头猪合格;淋巴细胞毒试验结果显示,有1头猪合格。结论成份血浆除白细胞过滤板中能分离培养T淋巴细胞,初步分离培养的淋巴细胞可产生免疫原性,但结果并不理想,其免疫条件和检测方法有待进一步优化。本研究为今后用分离培养的淋巴细胞制备猪抗人淋巴细胞免疫血浆提供了参考。
Objective To isolate T lymphocytes from the plasma leukocyte filter and to test their immunogenicity. Methods Using leukocyte filter as raw material, T lymphocytes were isolated and cultured from leucocyte filter. The number of monocytes was counted and cell viability was measured. After removal of erythrocytes and platelets in the cells, T lymphocytes were induced, differentiated and cultured after purification, and their immunogenicity to the pigs was examined. Results On average, 3.94 × 107 mononuclear cells were isolated from each leukocyte filter, and the average cell viability was 93.18%. The purified T lymphocytes proliferated in vitro for 8-10 days and reached the proliferative peak at 14 days and reached the equilibrium stage after 14 days. The highest proliferation rate was 100 times and the average was 38.5 times. After immunized pigs with isolated lymphocytes, it had a strong immune response, resulting in 1 pig died; E-rosette inhibition test showed that 1 pig passed the test. Lymphocyte cytotoxicity test showed that 1 pig passed the test. CONCLUSION: T lymphocytes can be isolated and cultured in the plasma of leucocyte filter plate. The initial isolation and culture of lymphocytes can produce immunogenicity, but the results are not satisfactory. The immunological conditions and detection methods need to be further optimized. This study provides a reference for the preparation of anti-human lymphocyte immune plasma from lymphocytes cultured in the future.