彩色毛线虫排泄/分泌产物对HT29-D4细胞生长的刺激。

Epithelial cell biology Pub Date : 1995-01-01
H Hoste, J L Nano, S Mallet, F Huby, S Fournel, P Rampal
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引用次数: 0

摘要

小肠中存在的毛状毛线虫寄生虫与上皮更新的增加有关。为了评估该蠕虫的排泄/分泌产物对细胞增殖的可能作用,我们在Dulbecco's Modified Eagle's培养基中体外培养成虫色毛线虫24 h,并将条件培养基添加到转化的上皮细胞系HT29-D4的培养基中。通过细胞计数、MTT法和氚化胸腺嘧啶的掺入,确定0.25-1.0微克蛋白/ml浓度对HT29-D4细胞生长的刺激作用。结果表明,黄颡鱼第四期幼虫的排泄/分泌产物中氚化胸腺嘧啶的掺入量在1.0微克/毫升时也有所增加。经蠕虫调节的培养基透析表明,该因子的分子量大于8000道尔顿。条件培养基的热处理、酸水解和三氯乙酸沉淀导致增殖作用消失,而胰蛋白酶处理部分减少了刺激活性。上述结果提示,绿梭菌可产生促进宿主小肠上皮再生的蛋白因子。
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Stimulation of HT29-D4 cell growth by excretory/secretory products of the parasite nematode Trichostrongylus colubriformis.

The presence of the nematode parasite Trichostrongylus colubriformis in the small intestine is associated with an increase in epithelial renewal. To assess the possible role of excretory/secretory products from the worm on cell proliferation, adult Trichostrongylus colubriformis were incubated in vitro in Dulbecco's Modified Eagle's Medium for 24 h and the conditioned medium was added to the culture medium of the transformed epithelial cell line HT29-D4. A stimulation of the HT29-D4 cell growth was ascertained at concentrations of 0.25-1.0 micrograms protein/ml using counts of cell numbers, the MTT method and incorporation of tritiated thymidine. An increased incorporation of tritiated thymidine was also observed with the excretory/secretory products from T. colubriformis fourth stage larvae at 1.0 microgram/ml. Dialysis of the medium conditioned by the worms indicated that the molecular weight of the factor is greater than 8000 Daltons in size. Heat treatment, acid hydrolysis and precipitation by trichloracetic acid of the conditioned medium resulted in the disappearance of the proliferative effect while treatment with trypsin partially depleted the stimulative activity. These results suggest that T. colubriformis produce some protein factor which could increase the epithelial regeneration in the host small intestine.

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