白细胞滤器中的NK及T细胞回收数量及其相关因素研究

Number of NK and T Cells Recovered from Leukocyte Reduction Filters and Its Correlation Factors

目的为了探索白细胞滤器中NK及T细胞回收数量及其相关因素,本项目将分析来源于白细胞滤器及全血的NK细胞及T细胞的数量,并进一步分析白细胞滤器来源的与全血来源的细胞数量是否与年龄、性别等因素相关。方法2023年7月—2024年6月于深圳市血液中心共收集51名重复献血者的全血样本及白细胞滤器(400 mL)。通过PBS缓冲液冲洗白细胞滤器得到白细胞,通过血细胞分析仪分别对来源全血及白细胞滤器来源的细胞进行计数,密度梯度离心法分离得到单个核细胞,流式细胞术分别检测两组来源的CD3-CD56+NK、CD3+CD4+T、CD3+CD8+T细胞的比例,GraphPad Prism 9.0软件进行统计分析。结果细胞计数结果显示,在一定量的冲洗液范围内,白细胞滤器来源的总淋巴细胞数量随着冲洗液体积的增多而增多,但细胞密度随着冲洗液的增多而降低。且白细胞滤器中的总淋巴细胞数量在不同年龄和不同性别的献血者中并未体现出明显的统计学差异(P>0.05)。细胞数量与细胞比例的结果通过计算可知,来源于白细胞滤器的CD3-CD56+NK细胞数量在不同年龄组间存在差异(P<0.05),而在性别分组中未见明显差异(P>0.05)。而来源全血的男性CD3-CD56+NK细胞数量要明显高于女性(P>0.05)。无论是来源白细胞滤器还是全血的CD3+CD4+T细胞、CD3+CD8+T细胞数量在不同年龄组和性别组中都未见明显统计学差异(P>0.05)。结论在一定量的冲洗液范围内,来源白细胞滤器的总淋巴细胞数量与冲洗液的体积相关,CD3+CD4+T细胞、CD3+CD8+T细胞的数量与献血者年龄、性别无明显相关,而CD3-CD56+NK细胞数量与献血者年龄相关。

Objective To assess the recover cells from Leukocyte reduction filters(LRFs)and their related factors,this study analyzed the number of natural killer(NK)cells and T cells from LRFs and whole blood(WB).The correlation between the number of these cells and age,sex and other factors was further analyzed.Methods WB and LRFs(400 mL)were collected from 51 repeat blood donors in Shenzhen Blood Center from July 2023 to June 2024.White blood cells(WBCs)were obtained by washing LRFs with PBS buffer.The cells from WB and LRFs were counted by blood cell analyzer.Mononuclear cells were isolated by density gradient centrifugation.The proportions of CD3^(-)CD56^(+)NK,CD3^(+)CD4^(+)T and CD3^(+)CD8^(+)T cells were detected by flow cytometry.Statistical analysis was performed using GraphPad Prism v.9.0 software.Results In a certain flush volume,the number of total lymphocytes from LRFs increased with the increase of flush volume,but the cell density decreased with the increase of flush solution.There was no significant difference in the total number of lymphocytes from LRFs among donors of different ages and genders(P>0.05).According to the results of cell number and cell ratio,the number of CD3^(-)CD56^(+)NK cells from LRFs was different in age groups(P<0.05),but no significant difference was found in gender groups(P>0.05).The number of CD3^(-)CD56^(+)NK cells from WB in male donor were significantly higher than that in female donor(P>0.05).Whether from LRFs or WB,there was no significant difference in the number of CD3^(+)CD4^(+)T cells and CD3^(+)CD8^(+)T cells in different age groups and gender groups(P>0.05).Conclusion In a certain flush volume,the total number of lymphocytes from LRFs was correlated with the volume of the flush solution.The number of CD3^(+)CD4^(+)T cells and CD3^(+)CD8^(+)T cells were not significantly correlated with the age and gender of blood donor,while the number of CD3^(-)CD56^(+)NK cells was correlated with the age of blood donor.