Wattrang Eva, Dalgaard Tina S, Norup Liselotte R, Kjærup Rikke B, Lundén Anna, Juul-Madsen Helle R
National Veterinary Institute, Uppsala SE-75189, Sweden.
Department of Animal Science, Aarhus University, Blichers Allé 20, P.O. box 50, DK-8830 Tjele, Denmark.
J Immunol Methods. 2015 Apr;419:35-47. doi: 10.1016/j.jim.2015.02.011. Epub 2015 Mar 3.
The study aimed to evaluate cell surface mobilisation of CD107a as a general activation marker on chicken cytotoxic T cells (CTL). Experiments comprised establishment of an in vitro model for activation-induced CD107a mobilisation and design of a marker panel for the detection of CD107a mobilisation on chicken CTL isolated from different tissues. Moreover, CD107a mobilisation was analysed on CTL isolated from airways of infectious bronchitis virus (IBV)-infected birds direct ex vivo and upon in vitro stimulation. Results showed that phorbol 12-myristate 13-acetate (PMA) in combination with ionomycin was a consistent inducer of CD107a cell surface mobilisation on chicken CTL in a 4h cell culture model. In chickens experimentally infected with IBV, higher frequencies of CTL isolated from respiratory tissues were positive for CD107a on the cell surface compared to those from uninfected control chickens indicating in vivo activation. Moreover, upon in vitro PMA+ ionomycin stimulation, higher proportions of CTL isolated from the airways of IBV-infected chickens showed CD107a mobilisation compared to those from uninfected control chickens. Monitoring of CD107a cell surface mobilisation may thus be a useful tool for studies of chicken CTL cytolytic potential both in vivo and in vitro.
该研究旨在评估CD107a在鸡细胞毒性T细胞(CTL)上作为一般激活标志物的细胞表面动员情况。实验包括建立激活诱导的CD107a动员的体外模型,以及设计用于检测从不同组织分离的鸡CTL上CD107a动员的标志物组合。此外,对从感染传染性支气管炎病毒(IBV)的鸡的气道直接离体分离的CTL以及体外刺激后的CTL进行了CD107a动员分析。结果表明,在4小时细胞培养模型中,佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)与离子霉素联合使用是鸡CTL上CD107a细胞表面动员的一致诱导剂。在实验感染IBV的鸡中,与未感染对照鸡相比,从呼吸组织分离的CTL细胞表面CD107a阳性频率更高,表明体内激活。此外,在体外PMA +离子霉素刺激后,与未感染对照鸡相比,从IBV感染鸡的气道分离的CTL中显示CD107a动员的比例更高。因此,监测CD107a细胞表面动员可能是研究鸡CTL体内和体外溶细胞潜力的有用工具。