硫代黄素荧光染色法显示周围血液中的淋巴细胞异质性

THE HETEROGENEITY OF LYMPHOCYTES DEMONSTRATED BY THIOFLAVINE FLUORESCENT STAINING IN HUMAN PERIPHERAL BLOOD

本文对人周围血液内的淋巴细胞进行了E-玫瑰花环和EAC玫瑰花环试验及硫代黄素荧光染色。实验证明人周围血液中的大部分淋巴细胞是蓝色核蓝色荧光胞质的小淋巴细胞和蓝色荧光核胞质无荧光而含有蓝白色块状荧光的淋巴细胞。其次是深蓝色核鲜蓝色荧光胞质的淋巴细胞和桔黄色核或桔红色核、黄色荧光胞质的淋巴细胞。此外,有时还可以看到灰蓝色——灰黄色核淡蓝绿色荧光胞质和淡黄色荧光的淋巴细胞。偶而可见桔红色核几乎看不到胞质的淋巴细胞。蓝色较小的细胞核境界不清楚的淋巴细胞和含有蓝白色块状荧光的淋巴细胞及桔黄色核黄色荧光胞质的淋巴细胞与羊红细胞能形成E-玫瑰花环,是T细胞。另一种较小的蓝色荧光的淋巴细胞和深蓝色核、鲜蓝色荧光胞质的淋巴细胞及桔红色核、黄色荧光胞质的淋巴细胞与特异性抗体和补体致敏的羊红细胞能形成EAC玫瑰花环,是B细胞。

In previous studies,we demonstrated 10 types of lymphocytes in lymph nodes,each exhibited a different fluorescent color by our thioflavine staining method.Among them,4 types were able to differentiate into plasma cells of the same fluorescent colors.In the present study,different types of lymphocytes were demonstrated in human peripheral blood by their different fluorescent colors after thioflavine staining.The lymphocytes from the venous blood of 50 healthy persons were isolated with Ficoll-Conray solution and E-rosette and EAC-rosette tests and fluorescent staining with thioflavine were performed.Most of the lymphocytes in peripheral blood are small ones with nuclei and cytoplasm showing blue fluorescence and the blue fluorescence of the cytoplasm is paler than that of the nuclei.The nuclei in a part of these lymphocytes have distinct boundaries.The nuclei in another part of these lymphocytes are smaller and with indistinct boundaries and indentation on one side and show dim fluorescence.Other lymphocytes show different fluorescence.Some show blue round nuclei with distinct nuclear membrane,and no color of fluorescence in cytoplasm,but with blue white patches on one side of the nuclei.Some show dark blue nuclei and bright blue cytoplasm and others show orange yellow or orange red nuclei and yellow cytoplasm.In addition,lymphocytes of grayish blue or grayish yellow nuclei and bluish green cytoplasm or lymphocytes of yellowish fluorescence may be seen at times.Very few lymphoeytes of orange red nuclei with nearly no cytoplasm may be seen occasionally.The lymphocytes with blue fluorescence and indentation on one side of nucleus,those with blue nuclei and blue white patches in the cytoplasm as well as those with orange yellow nuclei and yellow cytoplasm can form E-rosettes with sheep erythrocytes.They are T cells.The lymphocytes with distinct boundaries of nucleus,small size and blue fluorescence those with dark blue nuclei and bright blue cytoplasm as well as those with orange red nuclei and yellow cytoplasm can form EAC-rosettes with sheep erythrocytes sensitized by specific antibody and complement.They are B cells.The lymphocytes with blue nuclei and blue white patches may transform into lymphocytes with orange yellow nuclei and yellow cytoplasm under ultra-violet light irradiation.The latter are few in number in the blood but may be progressively increased in number on prolonged observation.They belong to GroupⅢof lymphocytes and are mainly located in the paracortical thymus-dependent zone of lymph nodes.The sma ller lymphocytes with blue fluorescence and distinct nuclear boundary may transform into lymphocytes with orange red nuclei and yellow cytoplasm,which are also very few in number in the blood and are also progressively increased in number on prolonged observation.They belong to GroupⅡof lymphocytes and constitute the main component of lymph nodules in lymph nodes.