Angarano G, Carlone A, Ieva R, Luchena N, Pastore G
Boll Soc Ital Biol Sper. 1984 May 30;60(5):949-53.
To evaluate the behaviour of the Theophylline-sensitive T lymphocytes subpopulation some modifications of the standard procedure are proposed. Lymphoprep purified lymphocytes were counted in a Neubauer hemocytometer after Acridine Orange stain, viability was evaluated by Ethidium Bromide counterstain and monocytes contamination was evaluated by the peroxidase stain. Sheep red blood cells were treated with AET, Theophylline was used at 3 mM (final concentration) and the results compared with untreated lymphocytes; the enumeration of the rosetting lymphocytes was facilitated by adding Acridine Orange prior to the resuspension. The modifications described were able to increase the % of rosetting T lymphocytes, to eliminate differences depending by different lots of sheep red blood cells and to decrease differences depending by subjective evaluation of the rosetting T lymphocytes.
为评估对茶碱敏感的T淋巴细胞亚群的行为，本文提出了一些对标准程序的修改。吖啶橙染色后，在血细胞计数板中对淋巴细胞分离液纯化的淋巴细胞进行计数，用溴化乙锭复染评估细胞活力，用过氧化物酶染色评估单核细胞污染情况。绵羊红细胞用AET处理，茶碱使用浓度为3 mM（终浓度），并将结果与未处理的淋巴细胞进行比较；在重悬前加入吖啶橙有助于对形成花环的淋巴细胞进行计数。所描述的修改能够提高形成花环的T淋巴细胞百分比，消除因不同批次绵羊红细胞导致的差异，并减少因对形成花环的T淋巴细胞主观评估导致的差异。
