γ干扰素和肿瘤坏死因子-α对少突胶质前体细胞存活和分化的影响。

Effects of interferon-gamma and tumor necrosis factor-alpha on survival and differentiation of oligodendrocyte progenitors.

作者信息

Feldhaus Beatrix, Dietzel Irmgard D, Heumann Rolf, Berger Richard

机构信息

Departments ofDepartment of Obstetrics and Gynecology, University of Bochum, Bochum, Germany.

出版信息

J Soc Gynecol Investig. 2004 Feb;11(2):89-96. doi: 10.1016/j.jsgi.2003.08.004.

DOI:10.1016/j.jsgi.2003.08.004

PMID:14980310

Abstract

OBJECTIVE

There is strong evidence from recent clinical studies that ascending intrauterine infection is associated with an increased incidence of periventricular leukomalacia in very premature fetuses. Periventricular leukomalacia is characterized by disrupted myelination from a loss of oligodendrocyte progenitors. We investigated the effects of proinflammatory cytokines on the survival and differentiation of this cell type.

METHODS

Cultures of more than 90% A2B5-positive progenitors were prepared from neonatal rats and kept for 3 days in medium supplemented with factors that stimulate cell proliferation. After 1 day in proliferation medium, cells were treated with interferon-gamma (100 U/mL) and tumor necrosis factor-alpha (100 ng/mL) for 48 hours triggering an increase in apoptotic A2B5 progenitor cells from 3.2 +/- 2.3% to 11.0 +/- 2.6%. After cytokine treatment cultures were transferred to medium containing factors to promote differentiation of progenitors into the myelinating phenotype.

RESULTS

In cytokine pretreated cultures, only 2.6 +/- 1.1% of total cells survived after a total of 9 days in vitro, whereas in untreated cultures most cells differentiated as shown by expression of myelin basic protein, myelin-associated glycoprotein, 2,3-cyclic nucleotide 3-phosphodiesterase, and myelin oligodendrocyte-specific protein. Using ten-fold reduced concentrations of combined interferon-gamma (10 U/mL) and tumor necrosis factor-alpha (10 ng/mL) pretreatment resulted in a survival to 11.2 +/- 4.9% of total cells with 36.3 +/- 11.6% A2B5-positive cells at day 9. This indicates a major enrichment of undifferentiated cells compared with untreated controls which harbored only 1.0 +/- 0.3% A2B5-positive cells.

CONCLUSION

Inflammatory cytokines not only induced apoptotic cell death but also prevented the differentiation of immature A2B5 oligodendrocyte progenitors into the myelinating phenotype.

摘要

目的

近期临床研究有强有力的证据表明，上行性宫内感染与极早产胎儿脑室周围白质软化症发病率增加有关。脑室周围白质软化症的特征是少突胶质前体细胞丢失导致髓鞘形成中断。我们研究了促炎细胞因子对这种细胞类型存活和分化的影响。

方法

从新生大鼠制备超过90% A2B5阳性前体细胞的培养物，并在添加刺激细胞增殖因子的培养基中培养3天。在增殖培养基中培养1天后，用γ干扰素（100 U/mL）和肿瘤坏死因子-α（100 ng/mL）处理细胞48小时，使凋亡的A2B5前体细胞从3.2±2.3%增加到11.0±2.6%。细胞因子处理后，将培养物转移到含有促进前体细胞分化为髓鞘形成表型因子的培养基中。

结果

在细胞因子预处理的培养物中，体外培养9天后，仅2.6±1.1%的细胞存活，而在未处理的培养物中，大多数细胞分化，表现为髓鞘碱性蛋白、髓鞘相关糖蛋白、2,3-环核苷酸3-磷酸二酯酶和髓鞘少突胶质细胞特异性蛋白的表达。使用浓度降低10倍的联合γ干扰素（10 U/mL）和肿瘤坏死因子-α（10 ng/mL）预处理，在第9天总细胞存活率为11.2±4.9%，A2B5阳性细胞为36.3±11.6%。这表明与未处理的对照相比，未分化细胞显著富集，未处理对照仅含有1.0±0.3%的A2B5阳性细胞。