布病在我区是最重要的人蓄共患传染病，发病率逐年升高，危害极大。关于布病的研究也成为热点，但布鲁菌所引起人体免疫功能损害的研究仍有很多不足，本研究以确诊为急、慢性期布病患者为研究对象，通过对外周血NK细胞、CD8+、CD4+、γδT细胞及相关细胞因子的测定评价布病患者机体细胞免疫功能状态。从而试图揭示人外周血NK细胞、T淋巴细胞对布鲁菌抗感染免疫机制中的作用。具体方法包括根据伦理学要求与受试者充分的沟通和签署知情同意后抽血，分离提纯NK细胞并培养后流式细胞仪检测NK细胞的亚群及表型；定量RT-PCR方法检测NK细胞穿孔素蛋白、颗粒酶B基因表达量；通过竞争性LDH释放检测NK细胞的杀伤活性；

Brucellosis in our region is the most important person in storage and zoonotic diseases, the incidence rate increases year by year with Great harm. Research on brucellosis has become a hot spot, but there are still some shortcomings in the study of Brucella caused immune function damage. This study confirmed the diagnosis of acute, and saw chronic phase of brucellosis patients as the research object. To determine and evaluate organism cellular immunity function in patients with Brucella state through NK cells, CD8+, CD4+, γδT and related cytokines. It attempts to reveal the human peripheral blood NK cells, T lymphocyte immunity to infection of Brucella mechanism. Specific methods including in accordance with the requirements of ethics and the subjects fully communication and blood after signing of the informed consent, separation, purification of NK cells and cells after training the detecting subgroup and NK cell phenotype; Quantitative rt-pcr method for detecting NK cell perforin protein, grain enzyme B gene expression; Through a competitive LDH release testing the killing activity of NK cells;

背景：布鲁菌病是内蒙古自治区重要的人蓄共患传染病，发病率逐年升高，危害极大。目的：探究布鲁菌病患者外周血NK细胞免疫功能状态。方法：本研究以布鲁菌病患者为研究对象，根据SAT结果分为SAT阴性组，SAT1:50+组，SAT1:100+组，利用流式细胞术测定外周血NK细胞表型、NK细胞穿孔素蛋白及颗粒酶B表达量；分离纯化NK细胞通过LDH实验测定NK细胞杀伤活性。结果：布鲁菌病患者平均年龄48.84岁，职业以农民为主，主要与羊接触。布病患者（SAT1：100+组）外周血NK细胞比例与SAT阴性组比较无统计学差异（p>0.05）；布病患者（SAT1：100组）外周血CD56dimCD16+NK细胞和CD56brightCD16+/-NK比例与SAT阴性组比较无统计学差异（p>0.05）；颗粒酶B在布病患者（SAT1：100+组）外周血NK细胞的表达率与SAT阴性组比较无统计学差异（p>0.05），穿孔素蛋白在布病患者（SAT1：100+组）外周血NK细胞的表达率与SAT阴性组比较差异具有统计学差异（p<0.01）。结论：布鲁菌病患者（SAT1：100+~++++）外周血NK细胞表型及颗粒酶素B表达无明显变化，穿孔素蛋白表达下降。