Madsen M, Johnsen H E, Hansen P W, Christiansen S E
J Immunol Methods. 1980;33(4):323-36. doi: 10.1016/0022-1759(80)90003-4.
Some methodological aspects of E-rosette gradient centrifugation for separation of human B and T lymphocytes are described. Treatment of sheep red blood cells (SRBC) with 2-aminoethylisothiouronium bromide (AET) accelerates and enhances E-rosette formation providing an effective and time saving method which takes less than 2 h to perform. Depletion of E-rosette forming cells (E-RFC) is almost complete, leaving less than 1% of the initial E-RFC at the interphase layer. By use of the lymphocyte donor's autologous serum containing naturally occurring anti-SRBC antibody and complement the SRBC in the rosetted T lymphocyte fraction are easily and completely lysed without damage to the lymphocytes. This procedure is shown to have little effect on the properties of the isolated T cells. The isolated lymphocyte subpopulations have been characterized by tests for lymphocyte and monocyte markers.
本文描述了用于分离人B淋巴细胞和T淋巴细胞的E花环梯度离心法的一些方法学方面。用2-氨基乙基异硫脲溴化物（AET）处理绵羊红细胞（SRBC）可加速并增强E花环的形成，提供了一种有效且省时的方法，该方法操作时间不到2小时。E花环形成细胞（E-RFC）的去除几乎是完全的，在中间层仅留下不到初始E-RFC的1%。通过使用含有天然抗SRBC抗体和补体的淋巴细胞供体自体血清，可轻松且完全地裂解玫瑰花结T淋巴细胞部分中的SRBC，而不会损伤淋巴细胞。结果表明该方法对分离的T细胞特性影响很小。已通过淋巴细胞和单核细胞标志物检测对分离的淋巴细胞亚群进行了表征。
