Pre-B cell generation potentiated by soluble factors from a bone marrow stromal cell line.
Pre-B cell generation potentiated by soluble factors from a bone marrow stromal cell line.
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DOI：
10.4049/jimmunol.140.3.845
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发表时间：
1988-02
影响因子：
4.4
通讯作者：
K. Landreth;K. Dorshkind
中科院分区：
文献类型：
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作者：
K. Landreth;K. Dorshkind
从Dexter型长期骨髓培养物的粘附层分离的两种骨髓基质细胞系在其造血支持能力上明显不同。S17支持骨髓生成和早期B细胞前体分化为B淋巴细胞，而S10支持骨髓细胞分化而不是B淋巴细胞生成。鉴定具有B细胞支持能力的基质细胞系促使研究S17的作用是否通过可溶性因子介导。本文提供的结果表明，由S17而不是S10条件化的培养基含有可以诱导220，000m.w. 14.8抗原和IG的细胞质μ H链，所述细胞质μ H链在尚未表达这些标志物的B淋巴细胞祖细胞中。在抗体包被的皮氏培养皿上，骨髓细胞耗尽14.8+，细胞质mu+前B细胞。在24小时液体培养后，通过免疫荧光将新产生的前B细胞计数为表达IG的细胞质μ H链但不表达IG L链的细胞。通过14.8抗体结合监测Ly 5（220）的表达。这种前B细胞分化活性通过用链霉蛋白酶、氨肽酶或羧肽酶消化而被废除。等电聚焦数据显示活性具有5.9至6.2的等电点。使用HPLC分级分离S17条件培养基，并测试每个级分的前B细胞生成活性。发现从Superose 12凝胶过滤柱收集的级分具有与表观m.w.大约6万到1万人。当将异质基质细胞培养物调节的培养基分级分离时，观察到几乎相同的活性峰。S10条件培养基的分离显示无隐蔽活性。通过阴离子交换色谱进一步表征S17条件培养基，并且显示大部分前B细胞生成活性与从MonoQ柱洗脱的空隙体积相关。将这些级分在Superose上再次色谱分离，再次发现活性与对应于表观m.w. 60,000和10000。S17前B细胞分化活性似乎是由于一种新分子的存在，因为其他充分表征的介质在这种短期液体培养系统中没有活性。当向培养物中加入IL-1或含有IL-2、IL-3或IL-4（B细胞刺激因子1）的条件培养基时，未观察到前B细胞生成活性。（400字处截断摘要）
Two bone marrow stromal cell lines isolated from the adherent layer of a Dexter-type long term bone marrow culture differ markedly in their hemopoietic support capacity. S17 supports myelopoiesis and the differentiation of early B cell precursors into B lymphocytes while S10 supports myeloid cell differentiation and not B lymphopoiesis. The identification of a stromal cell line with B cell support capacity prompted an investigation of whether the effects of S17 were mediated via soluble factors. Results presented herein indicate that medium conditioned by S17 but not S10 contains an activity that can induce the expression of the 220,000 m.w. 14.8 antigen and cytoplasmic mu H chain of Ig in B lymphocyte progenitors that have not yet expressed these markers. Bone marrow cells were depleted of 14.8+, cytoplasmic mu+ pre-B cells on antibody-coated petri dishes. After 24-h liquid culture newly generated pre-B cells were enumerated as cells that expressed cytoplasmic mu H chain of Ig but not Ig L chains by immunofluorescence. Expression of Ly5(220) was monitored by 14.8 antibody binding. This pre-B cell differentiation activity was abrogated by digestion with pronase, aminopeptidase, or carboxypeptidase. Isoelectric focusing data revealed the activity to have isoelectric point of 5.9 to 6.2. S17-conditioned medium was fractionated using HPLC and each fraction tested for pre-B cell-generating activity. Fractions collected from a Superose 12 gel filtration column were found to have two peaks of activity associated with molecules of apparent m.w. of approximately 60,000 and 10,000. Virtually identical peaks of activity were observed when medium conditioned by heterogeneous stromal cell cultures was fractionated. Separation of S10-conditioned medium revealed no cryptic activity. S17-conditioned medium was further characterized by anion exchange chromatography and the majority of the pre-B cell generating activity shown to be associated with the void volume that eluted from a MonoQ column. These fractions were rechromatographed on Superose and the activity again found to be associated with two fractions corresponding to apparent m.w. of 60,000 and 10,000. The S17 pre-B cell differentiation activity appears to result from the presence of a novel molecule because other well characterized mediators had no activity in this short-term liquid culture system. No pre-B cell-generating activity was observed when IL-1 or conditioned medium containing IL-2, IL-3, or IL-4 (B cell stimulatory factor 1) were added to cultures.(ABSTRACT TRUNCATED AT 400 WORDS)