Baldwin C L, Sathiyaseelan T, Naiman B, White A M, Brown R, Blumerman S, Rogers A, Black S J
Paige Laboratory, Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA.
Vet Immunol Immunopathol. 2002 Sep 10;87(3-4):251-9. doi: 10.1016/s0165-2427(02)00091-0.
Bovine peripheral blood gammadelta T cells have been evaluated for effector function (IFN-gamma production) and clonal expansion in a variety of systems including following activation by mitogens, IL-12, and stimulation, through the T cell receptor (TCR) with anti-CD3 monoclonal antibody (mAb), a cell-bound molecule and a soluble antigenic extract. To evaluate cell division, carboxyfluorescein succinimidyl ester (CFSE) loading of cells and flow cytometric analysis were used, while IFN-gamma production was evaluated by intracytoplasmic staining. It was found that bovine gammadelta T cells produced IFN-gamma and clonally expanded when stimulated through the TCR/CD3 complex by a cell-associated autologous molecule on monocyte, by bacterial components following in vivo sensitization of gammadelta T cells with a leptospira vaccine or by anti-CD3 mAb. In addition, gammadelta T cells were activated efficiently for effector function but not clonal expansion by culturing with IL-12. In contrast, stimulation by Con A or PMA/ionomycin induced efficient replication but only low level IFN-gamma production which was not enhanced by the presence of IL-12. In several systems the amount of IFN-gamma produced per cell by gammadelta T cells was less than that produced by CD4 T cells in the same cultures.
牛外周血γδ T细胞已在多种系统中评估了效应功能(产生γ干扰素)和克隆扩增情况,这些系统包括经丝裂原、白细胞介素-12激活后,以及通过T细胞受体(TCR)用抗CD3单克隆抗体(mAb)、一种细胞结合分子和一种可溶性抗原提取物刺激后。为评估细胞分裂,使用了羧基荧光素琥珀酰亚胺酯(CFSE)标记细胞并进行流式细胞术分析,而γ干扰素的产生则通过胞内染色进行评估。结果发现,当通过单核细胞上的细胞相关自体分子、经钩端螺旋体疫苗体内致敏γδ T细胞后的细菌成分或抗CD3 mAb经TCR/CD3复合物刺激时,牛γδ T细胞会产生γ干扰素并发生克隆扩增。此外,通过与白细胞介素-12培养,γδ T细胞可有效激活效应功能,但不会发生克隆扩增。相比之下,刀豆蛋白A或佛波酯/离子霉素刺激可诱导高效复制,但仅产生低水平的γ干扰素,且白细胞介素-12的存在不会增强其产生。在几个系统中,γδ T细胞每细胞产生的γ干扰素量少于同一培养物中CD4 T细胞产生的量。