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目的探讨tuftsin衍生物T肽(TP)对T细胞存活、分化的影响。方法无菌分离正常小鼠和荷瘤(B16-F10)小鼠胸腺细胞,分为正常T细胞组、正常T细胞+TP组、荷瘤T细胞组和荷瘤T细胞+TP组,TP终浓度为4.18μmol·L~(-1),处理72 h后收集培养的胸腺细胞,Guava ViaCount流式细胞术检测细胞存活率,Annexin V-FITC和PI检测细胞凋亡,CD4-FITC和CD8a-Per CP双标记检测细胞的分化成熟。结果正常T细胞组细胞存活率为(26.70±2.40)%,而加入TP后细胞存活率为(36.50±2.00)%,两组比较有显著差异(P<0.05);荷瘤T细胞+TP组细胞存活率为(20.70±2.20)%,与荷瘤T细胞组细胞存活率(17.80±1.30)%相比无显著差异(P>0.05)。正常T细胞组早期凋亡和晚期凋亡细胞百分比分别是(28.07±1.43)%和(8.45±0.48)%,加入TP后,早期凋亡和晚期凋亡细胞百分比分别是(15.90±0.45)%和(5.10±0.21)%,加药前后比较均有显著差异(P<0.01)。正常T细胞组CD4~+和CD8~+双阳细胞的比例为(70.50±1.91)%,经TP处理后CD4~+和CD8~+双阳细胞的比例降低为(55.30±3.81)%,比较有显著差异(P<0.01)。正常T细胞组CD4~+或CD8~+单阳细胞的比例分别为(22.13±1.12)%和(6.74±0.36)%,而经TP处理后CD4~+或CD8~+单阳细胞的比例分别为(34.87±2.77)%和(9.65±0.61)%,均显著增加(均P<0.01)。结论 TP能促进正常胸腺T细胞存活,抑制凋亡,并诱导其分化成熟。
Objective To investigate the effect of tuftsin derivative T peptide (TP) on the survival and differentiation of T cells. Methods The thymocytes of normal mice and tumor-bearing mice (B16-F10) were isolated aseptically and divided into normal T cell group, normal T cell + TP group, tumor-bearing T cell group and tumor-bearing T cell + TP group, Thymocytes were collected after 72 h of treatment. Cell viability was detected by Guava ViaCount flow cytometry. Apoptosis was detected by Annexin V-FITC and PI. CD4-FITC and CD8a- Per CP double labeling was used to detect the differentiation and maturation of cells. Results The survival rate of T cell group was (26.70 ± 2.40)%, while the survival rate of T cell group was (36.50 ± 2.00)%, there was significant difference between the two groups (P <0.05) The cell survival rate was (20.70 ± 2.20)%, which was not significantly different from that of tumor-bearing T cells (17.80 ± 1.30)% (P> 0.05). The percentages of early apoptosis and late apoptotic cells in normal T cell group were (28.07 ± 1.43)% and (8.45 ± 0.48)%, respectively. The percentages of early apoptosis and late apoptotic cells were (15.90 ± 0.45)% And (5.10 ± 0.21)%, respectively, before and after treatment were significantly different (P <0.01). The proportion of CD4 ~ + and CD8 ~ + double positive cells in normal T cells was (70.50 ± 1.91)%, and the percentage of CD4 ~ + and CD8 ~ + double positive cells decreased to (55.30 ± 3.81)% after TP treatment Difference (P <0.01). The percentage of CD4 ~ + or CD8 ~ + single-positive cells in normal T cells were (22.13 ± 1.12)% and (6.74 ± 0.36)% respectively, while the proportion of CD4 ~ + or CD8 ~ + single positive cells (34.87 ± 2.77)% and (9.65 ± 0.61)%, both significantly increased (all P <0.01). Conclusion TP can promote normal thymic T cell survival, inhibit apoptosis and induce its differentiation and maturation.