중증 혈소판감소증에서 망상혈소판 유세포분석을 통한 혈소판형성 평가
Estimate of Thrombopoiesis by Flow Cytometric Analysis of Reticulated Platelets in Patients with Severe Thrombocytopenia
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배경 : 망상혈소판(reticulated platelets, RPs) 분석은 혈소판감소증의 원인 감별과 혈소판형성을 추적하는 데에 유용하다. 이에 본 연구에서는 중증 혈소판감소증 환자군을 대상으로 하여 망상혈소판 비율(RPs%) 뿐만 아니라 유세포분석에서의 forwardscatter (FSC)와 thiazole orange (TO) 형광을 응용한 여러 지수들의 혈소판형성에 대한 감별능력을 평가해 보고자 하였다. 방법 : 혈소판형성 감소군 40명, 증가군 20명, 간경화군 20명을선정하였다. 혈소판 표지자인 CD41-PE 염색을 고려한 방법과 고려하지 않은 방법 두 가지 분석 방법에 의한 유세포 분석을 통해RPs%를 포함한 일차 지수들을 측정하였고 그로부터 응용 지수들을 산출하였다. 또 각 지수들의 진단 효율을 비교하였고 혈소판 광산란 gate내 혈소판 순도를 구하였다. 결과 : 혈소판 광산란 gate내 순도가 중증 혈소판감소증 환자군에서 정상인군에 비해 유의하게 감소되어 있어(P<10-6) 혈소판 gating시CD41-PE를 사용하여 RPs%의 진단 효율을 높일 수 있었다. 또한 일차 지수들보다는 RPs%와 FSC, TO 형광을 함께 고려한 응용 지수들이 감소군, 증가군 간 유의한 차이를 보였고 진단 효율도 더 높았다(RPs%: 55%, RPs%×sMFI: 80%). 결론 : 중증 혈소판감소증 환자에서 유세포분석을 통한 혈소판 형성 평가시 혈소판 표지자를 사용하고, RPs%와 더불어 TO 형광을 함께 고려하는 것이 혈소판형성 예측에 더 도움이 될 것으로 생각된다.
Abstract
Background : Analysis of reticulated platelets (RPs) is useful for discriminating the causes of thrombocytopenia and monitoring the thrombopoiesis. In the patients with severe thrombocytopenia, we evaluated the thrombopoiesis-discriminating ability of several indices applying forward scatter (FSC) and thiazole orange (TO) fluorescence in addition to the percentage of reticulated platelets (RPs%). Methods : Forty cases with decreased thrombopoiesis, twenty cases with increased thrombopoiesis and twenty cases with liver cirrhosis were selected. By flow cytometry with two analytic methods, dependent on or independent of the staining of CD41-PE as a platelet marker, the primary parameters including RPs% were measured and the applied parameters were calculated from them. And we compared the diagnostic efficiency of each parameter and analyzed the purity of platelet light scatter gate. Results : The purity of platelet light scatter gate was significantly lower in patients with severe thrombocytopenia than in healthy persons with normal platelet counts (P<10-6), so the use of CD41- PE for platelet gating improved the diagnostic efficiency of RPs%. Compared to the primary parameters, the applied parameters originated from RPs%, FSC and TO fluorescence improved diagnostic efficiency significantly (RPs%: 55%, RPs%×s MFI: 80%) between decreased and increased thrombopoiesis groups. Conclusions : In the patients with severe thrombocytopenia, the estimate of the thrombopoiesis by a flow cytometric analysis can be more predictable by using platelet markers and by considering the fluorescence intensity of TO together with the RPs%.
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