Rajam Gowrisankar, Jackson Delois, Pilishvili Tamara, Whitney Cynthia G, Facklam Richard R, Carlone George M, Romero-Steiner Sandra
Division of Bacterial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Microbiol Methods. 2007 Aug;70(2):219-26. doi: 10.1016/j.mimet.2007.04.008. Epub 2007 Apr 24.
Pneumococcal conjugate vaccine (PCV7) reduces invasive disease and carriage caused by vaccine serotypes (VS). An increase in carriage and disease with non-vaccine serotypes (NVS) has been observed. We have developed an in vitro model with human nasopharyngeal (NP) epithelial cells (Detroit 562) to assess the adherence capacity of Streptococcus pneumoniae to NP cells in the presence or absence of a competing Pnc strain. Two hundred and fifty pneumococcal (Pnc) strains (10 strains per serotype for 7 VS and 18 NVS) were tested for their opacity phenotype. Strains exhibiting (> or =50%) the transparent phenotype (n=72) were evaluated for their adherence capacity to Detroit 562 cells. Mean adherence capacity (> or =129 CFU/well) to NP cells was high for VS 18C, 4, and 9V and for NVS 16F, 10A, and 6A. In the in vitro competition experiments, VS strains out-competed (42/108) or co-existed (43/108) with NVS strains for adherence to NP cells in most co-inoculations. By contrast, NVS (15C, 16F, 31, and 35B) out-competed with VS in only 9 of 108 co-inoculations. Serotype 16F out-competed or co-existed with some VS and NVS strains. This model may be used to identify Pnc strains of a given serotype with competitive potentials for replacement of VS in the nasopharynx and to screen Pnc strains for animal colonization models.
肺炎球菌结合疫苗(PCV7)可减少由疫苗血清型(VS)引起的侵袭性疾病和携带。已观察到非疫苗血清型(NVS)的携带和疾病有所增加。我们开发了一种用人鼻咽(NP)上皮细胞(底特律562)的体外模型,以评估肺炎链球菌在存在或不存在竞争性肺炎球菌菌株的情况下对NP细胞的黏附能力。对250株肺炎球菌(Pnc)菌株(7种VS和18种NVS各血清型10株)进行了其不透明表型检测。对表现出(≥50%)透明表型的菌株(n = 72)进行了对底特律562细胞的黏附能力评估。VS 18C、4和9V以及NVS 16F、10A和6A对NP细胞的平均黏附能力(≥129 CFU/孔)较高。在体外竞争实验中,在大多数共接种中,VS菌株在与NP细胞的黏附方面胜过(42/108)或与NVS菌株共存(43/108)。相比之下,NVS(15C、16F、31和35B)在108次共接种中仅9次胜过VS。血清型16F在与一些VS和NVS菌株的竞争中胜过或共存。该模型可用于识别给定血清型具有在鼻咽部替代VS竞争潜力的肺炎球菌菌株,并用于筛选用于动物定植模型的肺炎球菌菌株。