红细胞调控CD4／CD8的血细胞快速培养实验研究

目的利用系统免疫学血细胞培养系统模式，在体外研究红细胞调控CD4／CD8规律及其意义。方法22例人枸橼酸抗凝新鲜血（11例正常人和11例原发性肝癌患者）。全血细胞培养组（自然组）：0．2ml全血细胞悬液加0．3ml血浆和0．2ml生理盐水，白细胞培养组（分离组）：0．2ml白细胞悬液加0．3ml血浆和0．2m1生理盐水，充分混匀，37℃温育1小时，2000转／分离心5分钟，两管的白细胞CD4和CD8表达水平用PC-5酶标抗CD4单抗和FITC-酶标抗CD8单抗测定．红细胞调节率=（全血细胞培养组CD4或CD8或CD4／CD8水平-白细胞培养组CD4或CD8或CD4／CD8水平）÷白细胞培养组CD4或CD8或CD4／CD8水平。结果22例全血细胞培养组CD4／CD8水平（1．40±0．59）明显高于白细胞培养组（0．48±0．26），t=6．6918，P〈0．01，正常人的白细胞培养组CD4／CD8水平（0．58±0．27）明显高于肝癌的白细胞培养组CD4／CD8水平（0．35±0．19，P〈0．01），正常人CD4／CD8调节率（2．42±2．39）高于肝癌患者CD4／CD8调节率（2．07±1．27）。结论本实验研究属于系统免疫学和计算免疫学领域，正常人红细胞能高调节CD4／CD8水平，在原发性肝癌中，细胞免疫功能低下，表现CD4／CD8表达低水平与红细胞对CD4／CD8低调节率密切相关。

Red Blood Cells Can Regulate CD4/CD8 in the Blood Cells Culture

Objective Using systems immunology method of rapid blood culture system,we have studied red blood cells to regulate the levels of CD4/CD8 in vitro.Method Whole blood cells culture group：0.2 ml of whole blood cells suspension and 0.3 ml of auto-plasma and 0.2 ml of saline;white blood cells culture group：0.2 ml of white blood cells suspension and 0.3 ml of auto-plasma and 0.2 ml of saline,mixed fully,and incubated at 37 ℃ for 60 minutes,then were centrifuged at 2 000 rounds per minute for 5 minutes.Expression level of CD4 and CD8 on white blood cells in two tube bottom mixture was determined by FITC-Conjugated Abs（CD8） and PC-5-conjugated Abs（CD4）.Regulation rate of red blood cells =（level of CD4 or CD8 or CD4/CD8 in whole blood cells culture-level of CD4 or CD4/CD8 in white blood cells culture） ÷level of CD4 or CD8 or CCD4/CD88 in white blood cells culture.Result The level（1.40 ± 0.19） of CD4/CD8 in whole blood cells culture was significantly higher than that（0.48 ± 0.26） of CD4/CD8 in white blood cells culture（n=22,t=6.6918,P 0.01）.The level（0.58 ± 0.27） of CD4/CD8 in white blood cells culture of normal people was significantly higher than that（0.35 ± 0.19） of CD4/CD8 in white blood cells culture of patient whit primary hepatic cancer（n=11,t=4.4003,P 0.01）,CD4/CD8 regulation rate（2.42 ± 2.39） in normal people was higher than that（2.07 ± 1.27） CD4/CD8 regulation rate in patient with P.H.C.Conclusion The experimental study is a systems immunology and computational immunology.Red blood cells can regulate level of CD4/CD8.In patients with primary hepatic cancer,there is a close relationship between low levels of CD4/CD8 and low CD4 /CD8 regulate rate of red blood cells.