Ittenson A, Mansfeld H W, Ansorge S
Allerg Immunol (Leipz). 1987;33(1):53-62.
Several methods for enumeration of Fc receptor bearing T lymphocytes and mononuclear cells from human peripheral blood were compared. The detection of Fc receptors is based on the formation of EA rosettes by using bovine erythrocytes and purified rabbit IgG or IgM antibodies. As alternative method the mixed rosette assay (EA rosettes plus sheep erythrocyte rosettes) (3) was applied for determining TG lymphocytes without the need of T cell separation. Independent of the method used for T cell separation (preparative rosetting with sheep erythrocytes stabilized by AET or HSA) the number of TG and TM lymphocytes was found to be identical. TG values obtained by use of the mixed rosette assay were significant lower (10 +/- 2%) than those obtained with the classical test (18 +/- 5%) (EA rosettes after T cell separation). Obviously this difference is due to a contamination of T lymphocyte preparations by non-T cells. On freshly isolated T lymphocytes without overnight culture we obtained 29% and 35%, respectively TM lymphocytes after separation of T cells using sheep erythrocyte rosettes stabilized with AET or HSA. The expression of FcIgM receptors was found to be strongly dependent on the composition and pH value of the culture medium. In the presence of human AB serum the maximum of FcIgM receptor expression on isolated T cells was obtained at pH 8.5. Under optimum conditions we found 63% and 66% respectively TM lymphocytes after T cell separation using AET or HSA stabilized sheep erythrocytes.
比较了几种从人外周血中计数携带Fc受体的T淋巴细胞和单核细胞的方法。Fc受体的检测基于使用牛红细胞和纯化的兔IgG或IgM抗体形成EA花环。作为替代方法，混合花环试验（EA花环加绵羊红细胞花环）（3）被用于测定TG淋巴细胞，而无需分离T细胞。无论使用何种方法分离T细胞（用AET或HSA稳定的绵羊红细胞进行制备性花环形成），TG和TM淋巴细胞的数量都相同。使用混合花环试验获得的TG值（10±2%）明显低于经典试验（18±5%）（T细胞分离后的EA花环）获得的值。显然，这种差异是由于T淋巴细胞制剂被非T细胞污染所致。在未经过夜培养的新鲜分离的T淋巴细胞上，使用用AET或HSA稳定的绵羊红细胞分离T细胞后，我们分别获得了29%和35%的TM淋巴细胞。发现FcIgM受体的表达强烈依赖于培养基的组成和pH值。在人AB血清存在的情况下，分离的T细胞上FcIgM受体表达的最大值在pH 8.5时获得。在最佳条件下，使用AET或HSA稳定的绵羊红细胞分离T细胞后，我们分别发现63%和66%的TM淋巴细胞。
