神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用

梅传忠; 沈继龙; 陈勇; 李柏青

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神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用

梅传忠 , 沈继龙 , 陈勇 , 李柏青

文章导航 > 蚌埠医学院学报 > 2004, 29 > 3: (193-196)

引用本文:

Citation:

神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用

1. 安徽医科大学病原生物学教研室, 安徽合肥 230032;
2. 蚌埠医学院生化与分子生物学教研室, 安徽蚌埠 233003;
3. 蚌埠医学院免疫学教研室, 安徽蚌埠 233003

作者简介: 梅传忠(1972-),男,安徽宿州人,硕士研究生,讲师,研究方向:细胞免疫与细胞凋亡。

通讯作者: 李柏青, 教授(bb_bqli@yahoo.com)

基金项目:
国家自然科学基金(NO30070721)资助项目
中图分类号: R329.25

Apoptosis of γδT cells and αβT cells induced by ceramide

Department of Microbiology and Parasitology, Anhui Medical University, Hefei, Anhui 230032, China

Corresponding author: LI Bai-qing, 教授(bb_bqli@yahoo.com)

CLC number: R329.25

摘要: 目的: 观察神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用。方法: 用结核杆菌低分子多肽抗原(Mtb-Ag)和CD3单抗(CD3mAb)分别刺激正常人外周血单个核细胞,获得Mtb-Ag激活的T细胞(γδT细胞为主)及CD3mAb激活的T细胞(αβT细胞为主);用不同浓度的神经酰胺(C2-Cer)分别作用于γδT细胞和αβT细胞,培养3 h后收获细胞,应用Annexin-V-FITC/PI染色,流式细胞术检测不同亚群T细胞的凋亡,应用四甲基噻唑氮蓝(MTT)比色法检测T细胞的活性。结果: Mtb-Ag激活的T细胞中γδT细胞比例可达70%~90%,CD3mAb激活的T细胞中αβT细胞比例>90%;与对照组比较,低浓度的C2-Cer (≤ 10 μmol/L)对人γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用均不明显(P>0。05);而高浓度的C2-Cer对γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用存在显著差异:100 μmol/L的C2-Cer诱导γδT细胞、αβT细胞凋亡比例分别增加10。4%(P>0。05)及73。5%(P<0。01),活细胞区(R1区)的细胞比例分别为对照组的88。1%(P>0。05)和7。8%(P<0。01);而500 μmol/L时诱导γδT细胞和αβT细胞凋亡比例分别增加71。9%和73。8%(P<0。01),R1区的细胞比例分别为对照组的36。5%和2。7%(P<0。01)。结论: 神经酰胺能诱导活化的人γδT细胞和αβT细胞发生凋亡,诱导γδT细胞发生凋亡所需的C2-Cer浓度明显高于诱导αβT细胞所需浓度。
- 细胞凋亡 /
- 神经酰胺 /
- γδT细胞 /
- αβT细胞 /
- 结核杆菌抗原
Abstract: Objective: To investigate the role of apoptosis of γδT cells and αβT cells induced by ceramide.Methods: The peripheral blood mononuclear lymphocytes of healthy donors were stimulated respectively with the low molecular peptide antigen derived from Mycobacterium tuberculosis antigen(Mtb Ag) and CD3mAb to generate Mtb Ag activated T cells(γδT cells were predominent) and CD3mAb activated T cells(αβT cells were predominent).These two T cell subsets were treated with Ceramide(C2 Cer) at different concentration;after 3hours of cultivation,the apoptosis of different subsets of T cells was measured by flowcytometry(FCM),using Annexin V FITC/PI staining;and the availability of T cells was detected by methl thiazolyl tetrazolium(MTT) colorimetric assay.Results: The percentage of γδT cells in Mtb Ag activated T cells and αβT cells in CD3mAb activated T cells were 70% to 90% and >90%,respectively.The apoptosis and the inhibition of the availability of γδT cells and αβT cells induced by lower concentration of C2 Cer(≤ 10 μmol/L)appeared weak in contrast to their control.There were significant differences in the role of apoptosis of γδT cells and αβT cells induced by higher concentration of C2 Cer;the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(100 μmol/L) increased by 10.4%(P>0.05) and 73.5%(P<0.01) in contrast with their control,and the percentage of cells in the region live cells(R1) was 88.1%(P>0.05) and 7.8%(P<0.01) of the ones of their control;whereas the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(500 μmol/L) increased by 71.9% and 73.8%(P<0.01) in contrast with their control,and the percentage of cells in R1 was 36.5% and 2.7%(P<0.01) of the ones of their control.Conclusions: The apoptosis of γδT cells and αβT cells can be induced by ceramide;it needs higher concentration of C2 Cer to induce the apoptosis of γδT cells than to induce the apoptosis of αβT cells.
- apoptosis /
- ceramide /
- γδT cells /
- αβT cells /
- Mycobacterium tuberculosis antigen

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神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用

通讯作者: 李柏青, 教授(bb_bqli@yahoo.com)

作者简介: 梅传忠(1972-),男,安徽宿州人,硕士研究生,讲师,研究方向:细胞免疫与细胞凋亡。

1. 安徽医科大学病原生物学教研室, 安徽合肥 230032;
2. 蚌埠医学院生化与分子生物学教研室, 安徽蚌埠 233003;
3. 蚌埠医学院免疫学教研室, 安徽蚌埠 233003

收稿日期: 2003-11-25

基金项目: 国家自然科学基金(NO30070721)资助项目

关键词:

摘要: 目的: 观察神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用。方法: 用结核杆菌低分子多肽抗原(Mtb-Ag)和CD3单抗(CD3mAb)分别刺激正常人外周血单个核细胞,获得Mtb-Ag激活的T细胞(γδT细胞为主)及CD3mAb激活的T细胞(αβT细胞为主);用不同浓度的神经酰胺(C2-Cer)分别作用于γδT细胞和αβT细胞,培养3 h后收获细胞,应用Annexin-V-FITC/PI染色,流式细胞术检测不同亚群T细胞的凋亡,应用四甲基噻唑氮蓝(MTT)比色法检测T细胞的活性。结果: Mtb-Ag激活的T细胞中γδT细胞比例可达70%~90%,CD3mAb激活的T细胞中αβT细胞比例>90%;与对照组比较,低浓度的C2-Cer (≤ 10 μmol/L)对人γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用均不明显(P>0。05);而高浓度的C2-Cer对γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用存在显著差异:100 μmol/L的C2-Cer诱导γδT细胞、αβT细胞凋亡比例分别增加10。4%(P>0。05)及73。5%(P<0。01),活细胞区(R1区)的细胞比例分别为对照组的88。1%(P>0。05)和7。8%(P<0。01);而500 μmol/L时诱导γδT细胞和αβT细胞凋亡比例分别增加71。9%和73。8%(P<0。01),R1区的细胞比例分别为对照组的36。5%和2。7%(P<0。01)。结论: 神经酰胺能诱导活化的人γδT细胞和αβT细胞发生凋亡,诱导γδT细胞发生凋亡所需的C2-Cer浓度明显高于诱导αβT细胞所需浓度。

English Abstract

Apoptosis of γδT cells and αβT cells induced by ceramide

Corresponding author: LI Bai-qing, 教授(bb_bqli@yahoo.com)

Department of Microbiology and Parasitology, Anhui Medical University, Hefei, Anhui 230032, China

Received Date: 2003-11-25

Abstract: Objective: To investigate the role of apoptosis of γδT cells and αβT cells induced by ceramide.Methods: The peripheral blood mononuclear lymphocytes of healthy donors were stimulated respectively with the low molecular peptide antigen derived from Mycobacterium tuberculosis antigen(Mtb Ag) and CD3mAb to generate Mtb Ag activated T cells(γδT cells were predominent) and CD3mAb activated T cells(αβT cells were predominent).These two T cell subsets were treated with Ceramide(C2 Cer) at different concentration;after 3hours of cultivation,the apoptosis of different subsets of T cells was measured by flowcytometry(FCM),using Annexin V FITC/PI staining;and the availability of T cells was detected by methl thiazolyl tetrazolium(MTT) colorimetric assay.Results: The percentage of γδT cells in Mtb Ag activated T cells and αβT cells in CD3mAb activated T cells were 70% to 90% and >90%,respectively.The apoptosis and the inhibition of the availability of γδT cells and αβT cells induced by lower concentration of C2 Cer(≤ 10 μmol/L)appeared weak in contrast to their control.There were significant differences in the role of apoptosis of γδT cells and αβT cells induced by higher concentration of C2 Cer;the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(100 μmol/L) increased by 10.4%(P>0.05) and 73.5%(P<0.01) in contrast with their control,and the percentage of cells in the region live cells(R1) was 88.1%(P>0.05) and 7.8%(P<0.01) of the ones of their control;whereas the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(500 μmol/L) increased by 71.9% and 73.8%(P<0.01) in contrast with their control,and the percentage of cells in R1 was 36.5% and 2.7%(P<0.01) of the ones of their control.Conclusions: The apoptosis of γδT cells and αβT cells can be induced by ceramide;it needs higher concentration of C2 Cer to induce the apoptosis of γδT cells than to induce the apoptosis of αβT cells.