Miyamoto N, Bakaletz L O
Department of Otolaryngology, College of Medicine, Ohio State University, Columbus 43210, USA.
Microb Pathog. 1996 Nov;21(5):343-56. doi: 10.1006/mpat.1996.0067.
Frozen sections of chinchilla Eustachian tube (ET) and middle ear mucosa were incubated with either FITC-labeled non-typeable Haemophilus influenzae (NTHi) or Bordetella pertussis. The number of bacteria adherent to "roof" vs "floor" regions was compared for each of three anatomic portions of the ET and for middle ear epithelium noting whether bacteria adhered to mucus or to epithelial cells. NTHi strains adhered significantly greater to mucus in the ET lumen whereas B. pertussis preferentially adhered to epithelial cells lining the ET (P < or = 0.05). A non-fimbriated isogenic mutant of NTHi adhered significantly less to mucus than the parental isolate at all sites of the ET floor (P < or = 0.05). Isolated fimbrin protein adhered to ET mucus and blocked adherence of whole organisms. Treatment with the mucolytic agent N-acetyl-L-cysteine resulted in significantly reduced adherence of NTHi to mucus (P < or = 0.001) and eliminated the ability to detect binding of isolated fimbrin protein. N-acetyl-L-cysteine treatment did not affect adherence of either B. pertussis or NTHi to epithelial cells. These data indicated that NTHi may mediate ascension of the ET from the nasopharynx primarily via adherence to and growth in mucus overlying the floor region of the tubal lumen. The OMP P5-homologous fimbriae were shown to contribute to this binding.
将豚鼠咽鼓管（ET）和中耳黏膜的冰冻切片与异硫氰酸荧光素标记的不可分型流感嗜血杆菌（NTHi）或百日咳博德特氏菌一起孵育。比较了ET三个解剖部位以及中耳上皮的“顶部”与“底部”区域附着的细菌数量，记录细菌是附着于黏液还是上皮细胞。NTHi菌株在ET管腔内对黏液的附着明显更多，而百日咳博德特氏菌则优先附着于ET内衬的上皮细胞（P≤0.05）。NTHi的非菌毛同基因突变体在ET底部的所有部位对黏液的附着均明显少于亲本菌株（P≤0.05）。分离出的菌毛蛋白附着于ET黏液并阻断了完整生物体的附着。用黏液溶解剂N-乙酰-L-半胱氨酸处理导致NTHi对黏液的附着显著减少（P≤0.001），并消除了检测分离出的菌毛蛋白结合的能力。N-乙酰-L-半胱氨酸处理不影响百日咳博德特氏菌或NTHi对上皮细胞的附着。这些数据表明，NTHi可能主要通过附着于管腔底部区域上方的黏液并在其中生长来介导ET从鼻咽部向上蔓延。已证明OMP P5同源菌毛有助于这种结合。
