Iuchi Hiroyuki, Ohori Junichiro, Kyutoku Takayuki, Ito Kotoko, Kurono Yuichi
Department of Otolaryngology, Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Department of Otolaryngology, Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Auris Nasus Larynx. 2019 Aug;46(4):513-519. doi: 10.1016/j.anl.2018.11.003. Epub 2018 Nov 29.
Phosphorylcholine (PC) is a structural component of Streptococcus pneumoniae (Spn) and nontypeable Haemophilus influenzae (NTHi), and is known to be associated with adherence through the platelet activating factor receptor (PAF-R). Furthermore, high PC expression is considered to be involved in Spn and NTHi virulence. In this study, we examined the influence of PC expression on the adherence of Spn and NTHi to epithelial cells in order to clarify the potential effectiveness of a vaccine targeting PC.
Twenty-seven strains of Spn and twenty-two strains of NTHi were used, cultured overnight, and PC expression was evaluated by fluorescence activated cell sorting; the strains were divided into two groups: PC low expression (PC-low) and PC high expression (PC-high) groups. Bacterial adherence was then examined using Detroit 562 cells and BALB/c mice. Bacterial invasion was then examined in Detroit 562 cells.
The adherence of Spn and NTHi and invasion of NTHi in the PC-high group was significantly reduced by pretreatment with a monoclonal anti-PC antibody (TEPC-15), PAF-R antagonist (ABT-491), and PC-keyhole limpet hemocyanin (PC-KLH). However, such findings were not observed in the PC-low group.
The present study suggests that PC is involved in the mucosal adhesion of Spn and NTHi, and the mucosal invasion of NTHi with PC-high strains, but not PC-low strains. These results suggest that a PC-targeting mucosal vaccine only affects PC-high Spn and NTHi strains and does not disturb commensal bacterial flora in the upper respiratory tract, which comprises nonpathogenic PC-low bacteria.
磷酰胆碱(PC)是肺炎链球菌(Spn)和不可分型流感嗜血杆菌(NTHi)的结构成分,已知其通过血小板活化因子受体(PAF-R)参与黏附过程。此外,高PC表达被认为与Spn和NTHi的毒力有关。在本研究中,我们检测了PC表达对Spn和NTHi黏附上皮细胞的影响,以阐明靶向PC的疫苗的潜在有效性。
使用27株Spn和22株NTHi,过夜培养,通过荧光激活细胞分选评估PC表达;将菌株分为两组:PC低表达(PC-low)组和PC高表达(PC-high)组。然后使用底特律562细胞和BALB/c小鼠检测细菌黏附情况。接着在底特律562细胞中检测细菌侵袭情况。
用单克隆抗PC抗体(TEPC-15)、PAF-R拮抗剂(ABT-491)和PC-钥孔血蓝蛋白(PC-KLH)预处理后,PC-high组中Spn和NTHi的黏附以及NTHi的侵袭显著降低。然而,在PC-low组中未观察到此类结果。
本研究表明,PC参与Spn和NTHi的黏膜黏附以及PC-high菌株(而非PC-low菌株)的NTHi黏膜侵袭。这些结果表明,靶向PC的黏膜疫苗仅影响PC-high Spn和NTHi菌株,不会干扰上呼吸道中由非致病性PC-low细菌组成的共生菌群。