Kees U, Kynast G, Weber E, Krammer P H
J Immunol Methods. 1984 Apr 27;69(2):215-27. doi: 10.1016/0022-1759(84)90320-x.
This paper describes a system for determining the frequency and fine specificity of influenza A virus-immune memory cytotoxic T cell (CTL) clones from limiting dilution (LD) microcultures. We found that such experiments can only be performed (1) by analyzing the clonal response of CTL from wells of replica plates containing fractions of one original plate, (2) when it has been ascertained that splitting is possible at the clonal level, and that each fraction of a microculture well gives an identical response on target cells infected with the stimulating virus. With these requirements fulfilled we found that short term CTL clones from LD microcultures from influenza A virus (A/X-31)-immune mice (C57BL/6) occur at a frequency of f = 1:546 to f = 1:6303. The effector cells carry the Lyt-2.2 marker and are specific for target cells infected with the immunizing virus (influenza virus A/X-31). They do not lyse NDV (Newcastle disease virus) or HSV (herpes simplex virus)-infected or NK (YAC) target cells.
本文描述了一种用于从有限稀释（LD）微培养物中确定甲型流感病毒免疫记忆细胞毒性T细胞（CTL）克隆的频率和精细特异性的系统。我们发现，此类实验只能在以下条件下进行：（1）通过分析来自包含一个原始平板各部分的复制平板孔中CTL的克隆反应；（2）当已确定在克隆水平上可以进行分割，且微培养孔的每个部分对感染刺激病毒的靶细胞产生相同反应时。满足这些要求后，我们发现来自甲型流感病毒（A/X-31）免疫小鼠（C57BL/6）的LD微培养物中的短期CTL克隆出现频率为f = 1:546至f = 1:6303。效应细胞携带Lyt-2.2标记，并且对感染免疫病毒（甲型流感病毒A/X-31）的靶细胞具有特异性。它们不裂解新城疫病毒（NDV）或单纯疱疹病毒（HSV）感染的靶细胞或NK（YAC）靶细胞。
