雏鸡免疫器官中chTLRLR4的表达与定位

Expression and Location of Chicken Toll-like Receptor 4 in Immune Organs of Chick

为研究鸡Toll样受体4（chTLR4）在雏鸡免疫器官（法氏囊、胸腺、脾脏）中的表达与定位，将50只健康海兰白蛋鸡随机分成10组，每组5只，分别于1、7、14、21、28、31、35、42、45、49日龄时随机扑杀一组，取法氏囊、胸腺及脾脏，应用实时荧光定量PCR、Westernblot及间接免疫荧光方法检测雏鸡免疫器官中chTLR4的表达情况。结果：①在各日龄雏鸡法氏囊、胸腺及脾脏内均检测到chTLR4转录及chTLR4蛋白的表达。②法氏囊淋巴滤泡的皮质和髓质内均可见免疫阳性细胞，尤其是在髓质部免疫阳性反应较强；胸腺小叶的皮质、髓质及脾脏的红髓和白髓细胞内均可观察到免疫阳性反应区。③法氏囊中chTLR4蛋白表达在1～4213龄均处于较高水平并存在小幅波动，42～49日龄表达量逐渐下降；各日龄雏鸡胸腺、脾脏中chTLR4蛋白表达趋势相似，1～14日龄表达量逐渐升高，14～35日龄呈波动变化，35～42日龄呈下降趋势。研究表明，在各日龄雏鸡免疫器官中均有chTLR4表达，且存在差异。研究结果为进一步探究chTLR4与雏鸡免疫功能相关性提供了依据。

In order to study the expression and location of chicken Toll-like receptor 4 （ehTLR4）in chick immune organs, five healthy chicks were euthanized for collection of bursa of Fabricius, thymus and spleen at 1,7,14,21, 28,3l, 35,42,45,49-day-age in fifty Hy-Line white hens. Quantitative real-time PCR, Western blot and IFA were used to detect the expression of chTLR4 in chick immune organs. The results showed that the chTLR4 transcription and chTLR4 protein expression were detected in the bursa of Fabricius, thymus and spleen at each age. ehTLR4~ cells were detected in the cortex and medulla of bursal lymphoid follicles, especially in medulla of bursal lymphoid follicles. The immune positive reaction zone could be detected in the cortex, medulla of the thymus, and spleen. The expression of chTLR4 protein in the bursa of Fabricius was at a high level with a slight fluctuation at 1 to 42 days of age,and gradually declined at 42 to 49 days of age. The expression pattern of chTLR4 protein in thymus and spleen was more consistent which showed an upregulation trend at 1 to 14 days of age and a fluctuation trend at 14 to 35days of age and went down at 55 to 42 days of age. The results indicated chTLR4 was expressed in the immune organs of chick with different ages, and there were differences in the expression. This study provided a scientific basis for the further research on the correlation between chTLR4 and the immune function of chick.