Dozmorov I M, Lutsenko G V, Sidorov L A, Miller R A
Department of Immunology, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
J Immunol Methods. 1996 Feb 5;189(2):183-96. doi: 10.1016/0022-1759(95)00241-3.
Limiting dilution (LD) cultures are often used to study cellular heterogeneity in responses of murine splenocytes to specific or polyclonal activation. LD titration curves often reveal a nonlinear dependence of response on input cell dose. Although 'zigzag' shaped curves of this kind are often interpreted and analyzed as resulting from interactions among three distinct cell types, we observe that a more parsimonious two cell model, including a cell type that can generate both positive and negative effects, provides better fit to a wide range of experimental data. We have developed mathematical models for the accurate estimation of the frequencies of both interacting cell types and of the parameters for their multi-hit interaction. We show examples of LD cultures in which specific experimental manipulations alter the frequency of only one of the two cell types, or alter the interaction parameters without a change in responder frequency. We also provide a simplified method for approximation of the model parameters using graphical approaches and simple algebra. Lastly, we present an improved method for calculation of the effect generated per responder cell in microclonal cultures.
有限稀释（LD）培养常用于研究小鼠脾细胞对特异性或多克隆激活反应中的细胞异质性。LD滴定曲线通常显示反应对输入细胞剂量的非线性依赖性。尽管这种“锯齿形”曲线常被解释和分析为由三种不同细胞类型之间的相互作用导致，但我们观察到一个更简约的双细胞模型，包括一种能产生正负两种效应的细胞类型，能更好地拟合广泛的实验数据。我们已经开发出数学模型，用于准确估计两种相互作用细胞类型的频率及其多击相互作用的参数。我们展示了LD培养的例子，其中特定的实验操作仅改变两种细胞类型之一的频率，或改变相互作用参数而不改变反应细胞频率。我们还提供了一种使用图形方法和简单代数近似模型参数的简化方法。最后，我们提出了一种改进的方法，用于计算微克隆培养中每个反应细胞产生的效应。
