Woodward T A, McNiece I K, Witte P L, Bender P, Crittenden R, Temeles D S, Robinson B E, Baber G B, Deacon D H, Isakson P C
Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville 22908.
Blood. 1990 Jun 1;75(11):2130-6.
Adherent murine stromal cells support long-term in vitro lymphopoiesis or myelopoiesis dependent on the culture conditions used. A cell line, TC-1, isolated from long-term liquid murine marrow cultures under conditions approaching those permissive for lymphoid growth, has been found to produce an activity that acts synergistically with interleukin-3 (IL-3) or colony-stimulating factor-1 (CSF-1) to stimulate in vitro myeloid colonies, but which has no intrinsic colony-stimulating activity. We report here the presence of multiple growth factors in conditioned medium (CM) from the TC-1 line, including granulocyte-macrophage colony-stimulating factor (GM-CSF) (bioassay with antibody blocking and messenger RNA [mRNA] analysis), granulocyte CSF (G-CSF) and IL-4 (factor-dependent cell line bioassay), and CSF-1 (radioimmunoassay, mRNA) along with a pre-B cell inducing activity, which appears separate from these CSFs and segregates with the myeloid synergizing activity through anion exchange, sizing, and Conconavalin A chromatography. Because these activities are not yet purified to homogeneity, their identity or lack of identity remains an open question. Assays of TC-1 CM or cellular mRNA analysis have given negative results for IL-1, IL-2, IL-3, IL-6, and IL-7, and IL-6 does not stimulate pre-B cells in this assay. However, IL-4 and G-CSF do stimulate in vitro induction of pre-B cells from pre-B and B-cell-depleted Balb/C marrow and are present in CM by selective cell line assay. A monoclonal antibody to IL-4 that inhibited its pre-B inducing activity did not inhibit pre-B inducing activity of TC-1 CM. These data suggest the existence of a unique synergizing and pre-B inducing factor(s) in TC-1 CM. Given the known capacity of subliminal levels of growth factors to act synergistically, an alternate possibility is that these biologic phenomena represent the actions of low concentrations of growth factors acting synergistically and possibly associated with some core protein.
贴壁的小鼠基质细胞能够根据所使用的培养条件，在体外支持长期淋巴细胞生成或髓细胞生成。从接近允许淋巴细胞生长条件的长期液体小鼠骨髓培养物中分离出的细胞系TC-1，已发现其能产生一种活性物质，该物质与白细胞介素-3（IL-3）或集落刺激因子-1（CSF-1）协同作用，刺激体外髓系集落形成，但本身没有集落刺激活性。我们在此报告，来自TC-1细胞系的条件培养基（CM）中存在多种生长因子，包括粒细胞-巨噬细胞集落刺激因子（GM-CSF）（通过抗体阻断生物测定和信使核糖核酸 [mRNA] 分析）、粒细胞集落刺激因子（G-CSF）和IL-4（因子依赖细胞系生物测定），以及CSF-1（放射免疫测定、mRNA），同时还有一种前B细胞诱导活性，它似乎与这些集落刺激因子不同，并通过阴离子交换、尺寸排阻和伴刀豆球蛋白A层析与髓系协同活性分离。由于这些活性尚未纯化至同质，它们的同一性或非同一性仍然是一个悬而未决的问题。对TC-1 CM的检测或细胞mRNA分析对IL-1、IL-2、IL-3、IL-6和IL-7均给出阴性结果，并且在该检测中IL-6不刺激前B细胞。然而，IL-4和G-CSF确实能刺激体外从前B细胞和B细胞缺失的Balb/C骨髓中诱导前B细胞，并且通过选择性细胞系检测存在于CM中。一种抑制IL-4前B诱导活性的单克隆抗体并未抑制TC-1 CM的前B诱导活性。这些数据表明TC-1 CM中存在一种独特的协同和前B诱导因子。鉴于已知生长因子的阈下水平具有协同作用的能力，另一种可能性是这些生物学现象代表低浓度生长因子协同作用的结果，并且可能与某种核心蛋白相关。
