Gene expression, immunolocalization, and secretion of human defensin-5 in human female reproductive tract.
Gene expression, immunolocalization, and secretion of human defensin-5 in human female reproductive tract.
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发表时间：
1998-05
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通讯作者：
A. Quayle;E. Porter;Aimee A. Nussbaum;Yun-Mei Wang;C. Brabec;K. Yip;S. Mok
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作者：
A. Quayle;E. Porter;Aimee A. Nussbaum;Yun-Mei Wang;C. Brabec;K. Yip;S. Mok
本研究描述了抗菌肽人肠防御素-5（HD-5）在女性生殖道上皮中的新定位。使用cDNA末端的3'快速扩增（RACE）方案，从阴道上皮细胞RNA制备物中克隆HD-5，并通过测序确认其身份。随后通过逆转录聚合酶链反应筛选来自多个供体的组织样品的HD-5表达。HD-5在正常阴道、子宫颈外端和炎症输卵管中表达不变，但在正常子宫颈内端、子宫内膜和输卵管中表达不稳定（分别为60%、64%和29%的标本）。子宫内膜中的表达在月经周期的早期分泌期最高。使用免疫组化和共聚焦显微镜，HD-5肽定位于阴道和宫颈的复层鳞状上皮的上半部分，细胞染色的强度向管腔增加。在阳性的宫颈管、子宫内膜和输卵管标本中，HD-5位于顶端定向的颗粒中和部分柱状上皮细胞的顶端表面上。使用蛋白质印迹分析，分泌的HD-5中检测到宫颈阴道灌洗液，在月经周期的分泌期发现的最高浓度。我们假设HD-5是女性泌尿生殖道先天免疫防御系统的内在组成部分，其表达可能受激素和促炎因子的调节。
This study describes the novel localization of the antimicrobial peptide human intestinal defensin-5 (HD-5) in female genital tract epithelia. Using a 3' rapid amplification of cDNA ends (RACE) protocol, HD-5 was cloned from a vaginal epithelial cell RNA preparation, and its identity was confirmed by sequencing. Tissue samples from multiple donors were subsequently screened for HD-5 expression by reverse transcription polymerase chain reaction. HD-5 message was invariantly expressed by normal vagina and ectocervix and inflamed fallopian tube, but variably expressed by normal endocervix, endometrium, and fallopian tube (60, 64, and 29% of specimens, respectively). Expression in endometrium was the highest during the early secretory phase of the menstrual cycle. Using immunohistochemistry and confocal microscopy, HD-5 peptide was localized in the upper half of the stratified squamous epithelium of the vagina and ectocervix, with the intensity of cellular staining increasing toward the lumen. In positive endocervix, endometrium, and fallopian tube specimens, HD-5 was located in apically oriented granules and on the apical surface of a proportion of columnar epithelial cells. Using Western blot analysis, secreted HD-5 was detected in cervicovaginal lavages, with the highest concentrations found during the secretory phase of the menstrual cycle. We hypothesize that HD-5 is an intrinsic component of the female urogenital innate immune defense system and that its expression may be modulated by hormonal and proinflammatory factors.