再生障碍性贫血患者穿孔素和颗粒酶B表达及意义

Expressions and significances of perforin and granzyme B in patients with aplastic anemia

目的探讨再生障碍性贫血(aplastic anemia, AA)患者外周血穿孔素及颗粒酶B的表达及其临床意义。方法 AA患者60例,其中重型AA(severe AA, SAA)患者22例为SAA组,非SAA患者38例为非SAA组,选取60例体检健康者为正常对照组;采用流式细胞仪检测3组外周血T淋巴细胞亚群及穿孔素和颗粒酶B表达情况,采用ELISA法检测3组血清穿孔素和颗粒酶B水平,并进行比较。结果 SAA组及非SAA组外周血CD4^+细胞比率、CD4^+/CD8^+和自然杀伤(natural killing, NK)细胞比率均低于正常对照组(P<0.05),CD8^+细胞比率高于正常对照组(P<0.05)。SAA组和非SAA组CD8^+T淋巴细胞中穿孔素[(17.72±5.53)%、(15.95±6.11)%]和颗粒酶B[(76.91±17.44)%、(69.89±16.69)%]阳性表达率均高于正常对照组[(8.83±4.76)%、(32.57±15.22)%](P<0.05),SAA组NK细胞穿孔素[(78.82±9.41)%]和颗粒酶B[(87.66±10.53)%]阳性表达率高于正常对照组[(62.33±5.76)%、(73.15±8.94)%](P<0.05);SAA组血清穿孔素[(6 195.26±1 168.35)ng/L]和颗粒酶B[(7.726±2.611)ng/L]水平高于非SAA组[(3 875.77±1 094.68)ng/L、(3.152±1.787)ng/L]和正常对照组[(3 324.92±1 283.51)ng/L、(2.878±0.997)ng/L](P<0.05),非SAA组与正常对照组比较差异无统计学意义(P>0.05)。结论 CD8^+T细胞参与了AA的免疫应答,CD8^+T淋巴细胞、NK细胞中穿孔素和颗粒酶B在AA发病中可能起一定作用。

Objective To investigate the expressions and clinical significances of perforin and granzyme B in the peripheral blood of patients with aplastic anemia(AA). Methods In 60 AA patients, there were 22 patients with severe AA(SAA group) and 38 patients without SAA(non-SAA group). Another 60 healthy volunteers were as controls(control group). The expressions of T lymphocyte subsets, perforin and granzyme B were detected by flow cytometry, and the levels of perforin and granzyme B were detected by ELISA. Results The percentage of CD4^+, CD4^+/CD8^+ ratio and percentage of natural killing(NK) cells were lower in SAA group and non-SAA group than those in control group(P<0.05), and the percentage of CD8^+ was larger in SAA group and non-SAA group than that in control group(P<0.05). The positive rates of perforin and granzyme B in CD8^+T lymphocytes were higher in SAA group((17.72±5.53)%,(76.91±17.44)%) and non-SAA group((15.95±6.11)%,(69.89±16.69)%) than those in control group((8.83±4.76)%,(32.57±15.22)%)(P<0.05). The positive rates of perforin and granzyme B in NK cells were higher in SAA group((78.82±9.41)%,(87.66±10.53)%)than those in control group((62.33±5.76)%,(73.15±8.94)%)(P<0.05).The serum levels of perforin and granzyme B were higher in SAA group((6 195.26±1 168.35),(7.726±2.611)ng/L)than those in non-SAA group((3 875.77±1 094.68),(3.152±1.787)ng/L)and control group((3 324.92±1 283.51),(2.878±0.997)ng/L)(P<0.05),and showed no significant differences between non-SAA group and control group(P>0.05).Conclusion CD8+T lymphocytes participates in the immune response of AA.Both perforin and granzyme B in CD8+T lymphocytes and NK cells are involved in the pathogenesis of AA.