AN ACTIVATED FORM OF TRANSFORMING GROWTH FACTOR-BETA IS PRODUCED BY COCULTURES OF ENDOTHELIAL-CELLS AND PERICYTES
AN ACTIVATED FORM OF TRANSFORMING GROWTH FACTOR-BETA IS PRODUCED BY COCULTURES OF ENDOTHELIAL-CELLS AND PERICYTES
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DOI：
10.1073/pnas.86.12.4544
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发表时间：
1989-06-01
影响因子：
11.1
通讯作者：
DAMORE, PA
中科院分区：
文献类型：
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作者：
ANTONELLIORLIDGE, A;SAUNDERS, KB;DAMORE, PA
使用体外共培养系统来模拟血管壁细胞之间的相互作用，我们以前已经表明，周细胞和平滑肌细胞（SMC）抑制毛细血管内皮细胞（EC）的生长。我们已经进行了研究，以确定这种抑制的机制。使用条件培养基和亲和纯化的转化生长因子β的抗体，（TGF-β），我们现在证明活化的TGF-β。在这些共培养物中产生介导EC生长抑制。没有检测到抑制活性时，由个别培养EC，SMC，或周细胞条件培养基进行了检查，其对EC生长的影响。与此相反，条件培养基的EC-SMC和EC-周细胞的共培养物抑制EC增殖到相同的程度作为共培养本身。共培养物衍生的条件培养基与TGF-β抗体的免疫吸附消除了抑制活性。单独培养的任何细胞条件的无血清培养基的酸活化产生抑制活性，而共培养条件培养基的活化不增加其抑制活性。添加抗TGF-β。共培养物的中和抗体阻断周细胞介导的EC生长抑制。这些结果表明，潜在的TGF-β是由这些细胞产生的，它是由一种需要两种细胞接触的机制激活的。
Using an in vitro coculture system to mimic the interactions between the cells of the vessel wall, we have previously shown that pericytes and smooth muscle cells (SMC) inhibit the growth of capillary endothelial cells (EC). We have undertaken studies to determine the mechanism of this inhibition. Using conditioned media and affinity-purified antibodies to transforming growth factor .beta. (TGF-.beta.), we now demonstrate that activated TGF-.beta. produced in these cocultures mediates EC growth inhibition. No inhibitory activity was detected when media conditioned by individual cultures of EC, SMC, or pericytes were examined for their effect on EC growth. In contrast, media conditioned by cocultures of EC-SMC and EC-pericytes inhibited EC proliferation to the same degree as the coculture itself. Immunoadsorption of coculture-derived conditioned media with antibodies to TGF-.beta. eliminated the inhibitory activity. Acid activation of serum-free media conditioned by any of the cells cultured alone yielded inhibitory activity, whereas activation of coculture conditioned media did not increase its inhibitory activity. Addition of anti-TGF-.beta. neutralizing antibodies to cocultures blocked the pericyte-mediated EC growth inhibition. These results indicate that latent TGF-.beta. is produced by these cells and it is activated by a mechanism that requires contact between the two cell types.