恒河猴胃肠道相关淋巴组织中CCL25、CCR9、CCL20及CCR6转录水平的定量分析

Quantitative analysis of CCL25,CCR9,CCL20 and CCR6 transcription levels in gut-associated lymphoid tissues of rhesus macaque

目的:阐明恒河猴胃肠道相关淋巴组织中CCL25、CCR9、CCL20和CCR6转录产物的分布和丰度。方法:建立CCL25、CCR9、CCL20和CCR6转录水平的Taqman探针实时定量PCR检测方法。提取胃肠道相关淋巴组织和非胃肠道相关淋巴组织的总细胞RNA,检测CCL25、CCR9、CCL20和CCR6的mRNA的存在和表达水平。结果:(1)在恒河猴的所有检测组织中CCL25、CCR9、CCL20和CCR6的mRNA均有表达。在胃肠道相关淋巴组织中,CCL25和CCR9的mRNA表达主要在小肠而不是大肠,而CCL20和CCR6的mRNA则表达在胃肠道所有节段中。在非胃肠道相关淋巴组织中,CCL25和CCR9的mRNA在胸腺中的表达水平最高;而CCR6 mRNA在脾脏和腹股沟淋巴结中含量最丰富。(2)相关分析结果显示CCL25与CCR9mRNA的表达具有显著的相关性(P=0.002 0,r2=0.480 2),CCR9与CCR6 mRNA的表达也具有相关性(P=0.003 9,r2=0.436 4)。结论:与人相似,CCL25、CCR9、CCL20和CCR6 mRNA在恒河猴的胃肠道相关淋巴组织中高度且有区别地表达,表明恒河猴适宜用于与这些分子相关的黏膜免疫及人类疾病研究。

Objective：To find out the distribution and abundance of CCL25, CCR9, CCL20 and CCR6 transcripts in gut-asso- ciated lymphoid tissues （GALT） of rhesus macaques. Methods： TaqMan probe real-time RT-PCR methods for CCL25, CCR9, CCL20 and CCR6 mRNA quantification were established. Total tissue RNA was extracted from GALT and some other non-GALT tissues and the presence and levels of CCL25, CCR9, CCL20 and CCR6 mRNA were examined. Results： （ 1 ） CCL25, CCR9, CCL20 and CCR6 mRNA were detected in all the tissues examined including both GALT and non-GALT tissues. Within GALT, CCL25 and CCR9 mRNA levels were much higher in the small intestine than in the large intestine, while CCL20 and CCR6 mRNA levels were high at all seg- ments of the gastrointestinal tract. For non-GALT tissues, the highest levels of CCL25 and CCR9 mRNA were in the thymus ; while CCR6 mRNA was most abundant in spleen and inguinal lymph nodes. （2） The correlation analysis showed that the expression of CCL25 and CCR9 mRNA significantly correlated （P = 0. 002 O, r2 = 0. 480 2 ）, while the expression of CCR9 and CCR6 mRNA was also correlated （P =0. 003 9, r2 =0. 436 4）. Conclusion： Similar to that of humans, CCL25, CCR9 and CCL20, CCR6 mRNA are highly and differentially expressed in the GALT of rhesus macaques, which indicates that rhesus macaques are suitable for studies on human mucosal immunology and related diseases.