三种荧光色素标记的CD4抗体在流式细胞术研究T辅助细胞中的有效性评价

Evaluating the Efficacy of Three Different Fluorochrome-labeled CD4 Antibodies in Studying T Helper Cells by Flow Cytometry

探讨三种荧光色素标记的CD4抗体在流式细胞术检测培养刺激前后的T辅助(Th)细胞中的有效性,从而确定Th细胞研究时优选且有效的CD4抗体。将佛波脂(PMA)和离子霉素(Ion)培养刺激前后的正常人外周血单个核细胞(PBMC)分别用CD3-APC和CD8-PerCP确定Th细胞(CD3+CD8-T细胞)百分率,用CD4-PerCP、CD4-FITC和CD4-APC确定Th细胞(CD4+T细胞)百分率。并用免疫磁珠分选后的CD4+T细胞验证CD4-APC抗体在Th细胞研究中的有效性。结果显示:Th细胞(CD3+CD8-T细胞)百分率在培养刺激前后基本无差别(P>0.05),而Th细胞(CD4+T细胞)百分率有所下降,其中CD4-PerCP下降的最明显(P<0.05),CD4-FITC和CD4-APC轻微下降(P>0.05)。就平均荧光强度(MFI)而言,CD4-APC在三种荧光色素的CD4抗体中最强,且与培养刺激前相比,培养刺激后的CD4-APC下降不明显(P>0.05),而CD4-PerCP和CD4-FITC平均荧光强度明显下降(P<0.05)。免疫磁珠分选后的细胞亦证实培养刺激对于CD4-APC表达的影响很小。结论:三种荧光色素抗体中,优选的CD4-APC抗体在流式细胞术中用于Th细胞研究是有效的。

To determine the preferred and effective CD4 antibody used for T helper（Th） cell study,Th cells before and after stimulation were investigated by flow cytometry with three different fluorochrome-labeled CD4 antibodies.Before and after stimulation with PMA and Ion,the percentage of Th cells（CD3＋CD8-T cells） was determined by CD3-APC and CD8-PerCP,while the percentage of Th cells（CD4＋ T cells） was detected by CD4-PerCP,CD4-FITC and CD4-APC.Furthermore,CD4＋ T cells sorted by immunomagnetic beads were used to confirm the efficacy of CD4-APC antibody in Th cell research.The results showed that there was no significant difference in the percentages of Th cells（CD3＋CD8-T cells） before and after stimulation（P0.05）.The percentages of Th cells（CD4＋CD8-T cells） before and after stimulation detected by three fluorochrome-labeled CD4 antibodies were all declined.Among them,CD4-PerCP declined significantly（P0.05）,while CD4-FITC and CD4-APC declined slightly（P0.05）.The MFI of CD4-APC was the strongest among three fluorochrome-labeled CD4 antibodies and did not decrease significantly after stimulation（P0.05）,while the MFI of CD4-PerCP and CD4-FITC decreased sharply（P0.05）.Cells sorted by immunomagnetic beads also confirmed that the stimulation had little effect on the expression of CD4-APC.It is thus showed that the preferred CD4-APC antibody was effective in Th cell research among the three fluorochrome-labeled CD4 antibodies.