INNATE INTRAEPITHELIAL DEFENSE AGAINST ORAL INFECTION

针对口腔感染的先天上皮内防御

• 批准号: 2634143
• 负责人: MARK C HERZBERG
• 金额: $ 18.58万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-06 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2634143
• 关键词: Actinobacillus actinomycetemcomitans; Bacteroides gingivalis; Escherichia coli; HeLa cells; antibacterial agents; bacterial cytopathogenic effect; clinical research; complementary DNA; disease /disorder etiology; epithelium; gene expression; gingiva; human subject; hybridomas; keratinocyte; laboratory mouse; monoclonal antibody; oligonucleotides; periodontium disorder; protein purification; transfection

DESCRIPTION (Adapted from investigator's Abstract): In the pathogenesis of periodontal disease, invasion of the gingival epithelium by putative pathogens appears to be critical. While many mechanisms exist to protect the mucosal surface against colonization, it is unclear if gingival epithelial cells can defend against invading bacteria. Gingival keratinocytes do express calprotectin, a cytoplasmic antibacterial protein complex. the Principal Investigator hypothesizes that calprotectin serves as an innate intraepithelial antimicrobial mechanism. To test this novel hypothesis, calprotectin will be expressed and purified and complexes will be used to determine the subunit requirements for antimicrobial activity. Normal human gingival keratinocytes will be screened for expression of calprotectin. Calprotectin-negative and calprotectin-positive normal gingival keratinocytes will be compared for resistance to colonization and invasion by putative periodontal pathogens, including Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. To show that the antimicrobial effect is restricted to calprotectin, HeLa cells will be stably transfected with calprotectin-specific cDNA and compared to control negative transfections for resistance to colonization and invasion. To demonstrate the change in innate antimicrobial activity, immortalized keratinocyte cell lines will be screened for expression of calprotectin. Specific expression of calprotectin in selected immortalized lines and normal gingival keratinocytes will be abrogated with antisense DNA and cells will then be tested for altered antimicrobial properties. These studies will define if calprotectin serves an innate defense against bacterial invasion in vitro and provide strong evidence for a fundamental role in resistance to infections.

描述（根据研究者的摘要改编）： 牙周疾病，推定牙龈上皮的入侵 病原体似乎很关键。 虽然存在许多保护 粘膜表面反对定殖，目前尚不清楚牙龈是否存在 上皮细胞可以防御入侵细菌。 牙龈 角质形成细胞确实表达钙染色素，一种细胞质抗菌蛋白 复杂的。 首席研究者假设钙骨蛋白酶服务 作为先天的上皮内抗菌机制。 测试这本小说 假设，钙染色素将被表达和纯化，复合物将 用于确定抗菌活性的亚基要求。 正常的人类牙龈角质形成细胞将被筛选以表达 钙骨蛋白。 钙染色蛋白阴性和钙染色蛋白阳性正常 将比较牙龈角质形成细胞的抗性和定殖和 推定的牙周病原体的侵袭，包括卟啉念珠菌 牙龈和放线杆菌放线菌。 表明 抗菌作用仅限于钙骨蛋白，HeLa细胞将是 稳定地用钙粘蛋白特异性cDNA转染并与对照 抗性和侵袭的负转染。 到 证明先天抗菌活性的变化，永生 角质形成细胞系将被筛选以表达钙染色蛋白。 钙特染色素在选定的永生线中的特异性表达和 正常的牙龈角质形成细胞将用反义DNA和细胞消除 然后将测试改变的抗菌特性。 这些研究 将定义钙染色蛋白是否为抗细菌提供先天的防御 体外入侵，并为在 对感染的抵抗力。