International Centre for Diarrheal Disease Research, Bangladesh, Dhaka, Bangladesh.
Microbes Infect. 2011 Nov;13(12-13):1111-20. doi: 10.1016/j.micinf.2011.06.014. Epub 2011 Jul 18.
Patients with acute watery diarrhea caused by Vibrio cholerae O1 or enterotoxigenic Escherichia coli (ETEC) were analyzed for innate immune factors produced by the epithelium during the disease process. Duodenal biopsies were obtained from study participants at the acute (day 2) and convalescent (day 21) stages of disease. Levels of α-defensin (HD-5 and -6), β-defensin (hBD-1-4), and cathelicidin (LL-37) mRNAs were determined by real-time qRT-PCR. hBD-2, HD-5, LL-37 peptides were analyzed in duodenal epithelium by immunomorphometry. Concentration of hBD-2 in stool was determined by ELISA. Specimens from healthy controls were also analyzed. hBD-2 mRNA levels were significantly increased at acute stage of diarrhea; hBD-2 peptide was detected in fecal specimens but barely in duodenal epithelium at acute stage. Immunomorphometry analysis showed that Paneth cells contain significantly higher amounts of HD-5 pre/propeptide at convalescence (P<0.01) and in healthy controls (P<0.001) compared to acute stage, LL-37 peptide levels also decreased at acute stage while mRNA levels remained unchanged. mRNA expression levels of the other antimicrobial peptides remained unchanged with higher levels of α-defensins than β-defensins. V. cholerae induced an innate immune response at the acute stage of disease characterized by increased expression of hBD-2, and continued expression of hBD-1, HD-5-6, and LL-37.
对由霍乱弧菌 O1 或肠产毒性大肠杆菌 (ETEC) 引起的急性水样腹泻患者,分析了疾病过程中上皮细胞产生的固有免疫因子。在疾病的急性期(第 2 天)和恢复期(第 21 天),从研究参与者的十二指肠活检中获得。通过实时 qRT-PCR 测定α防御素(HD-5 和 -6)、β防御素(hBD-1-4)和抗菌肽(LL-37)mRNA 的水平。通过免疫形态计量法分析十二指肠上皮细胞中的 hBD-2、HD-5、LL-37 肽。通过 ELISA 测定粪便中 hBD-2 的浓度。还分析了健康对照者的标本。腹泻急性期 hBD-2 mRNA 水平显著升高;hBD-2 肽在粪便标本中检测到,但在急性期十二指肠上皮中几乎检测不到。免疫形态计量学分析表明,与急性期相比,恢复期(P<0.01)和健康对照组(P<0.001)中潘氏细胞中 HD-5 前/原肽的含量明显更高,LL-37 肽水平在急性期下降,而 mRNA 水平保持不变。其他抗菌肽的 mRNA 表达水平不变,α防御素的水平高于β防御素。霍乱弧菌在疾病的急性期诱导固有免疫反应,其特征是 hBD-2 表达增加,hBD-1、HD-5-6 和 LL-37 持续表达。