Chronic tonsillar hypertrophy (CTH) is an idiopathic, obstructive lymphoproliferative disease, probably due to continuous antigenic stimulation of an unknown source. The hypothesis to be tested is that CTH is caused by bacterial antigens from metacolonies. This hypothesis is based on our preliminary data: i. Review of limited samples of archival materials found 3 types of bacterial metacolonies, in total, in 80% of CTH cases but only in 7% of non-inflammatory tonsillar biopsies; ii. Metacolonies (n=3) are highly organized polybacterial communities consisting of ~100 species. Members of three genera are possible metacolonycommon species (MC-species); iii. CTH tonsils contain high levels of antibodies to the putative MC-species. We will test the hypothesis by the following 3 Specific Aims.
Specific Aim 1 is designed to determine whether there is a significant correlation between the presence of metacolonies and CTH. We will determine the true occurrence rate of each type of metacolony and inflammatory reaction to them, in adequately sampled CTH tonsils and controls.
Specific Aim 2 is designed to examine whether there are MC-species among metacolonies. Besides cultivation, metacolonies with unique banding patterns on Denaturing Gradient Gel Electrophoresis will be selected for analysis by 16S PCR-based sequencing and in situ examinations to identify putative MC-species. Their conservation in the remaining metacolonies will be validated by species-specific, quantitative real time PCR.
Specific Aim 3 is designed to determine whether B cell proliferation in CTH is a response to antigens from MC-species. Initially, CD19+ B cells will be pulled out by labelling with antigens of the MC-species and single cell sorting, and tested for their ability to produce antibodies to the antigens. Later, we will develop a simple ELISA capable of screening a large number of patients for tonsillar antibodies to the MC-species. Addressing the three aims will provide evidence to examine the hypothesis of a bacterial etiology of CTH. ? ?
|Pei, Anna; Li, Hongru; Oberdorf, William E et al. (2012) Diversity of 5S rRNA genes within individual prokaryotic genomes. FEMS Microbiol Lett 335:11-8|
|Yang, Liying; Francois, Fritz; Pei, Zhiheng (2012) Molecular pathways: pathogenesis and clinical implications of microbiome alteration in esophagitis and Barrett esophagus. Clin Cancer Res 18:2138-44|
|Ahn, Jiyoung; Yang, Liying; Paster, Bruce J et al. (2011) Oral microbiome profiles: 16S rRNA pyrosequencing and microarray assay comparison. PLoS One 6:e22788|
|Pei, Anna Y; Oberdorf, William E; Nossa, Carlos W et al. (2010) Diversity of 16S rRNA genes within individual prokaryotic genomes. Appl Environ Microbiol 76:3886-97|
|Sturt, Amy S; Yang, Liying; Sandhu, Kuldip et al. (2010) Streptococcus gallolyticus subspecies pasteurianus (biotype II/2), a newly reported cause of adult meningitis. J Clin Microbiol 48:2247-9|
|Nossa, Carlos W; Oberdorf, William E; Yang, Liying et al. (2010) Design of 16S rRNA gene primers for 454 pyrosequencing of the human foregut microbiome. World J Gastroenterol 16:4135-44|
|Yang, Liying; Lu, Xiaohua; Nossa, Carlos W et al. (2009) Inflammation and intestinal metaplasia of the distal esophagus are associated with alterations in the microbiome. Gastroenterology 137:588-97|
|Pei, Anna; Nossa, Carlos W; Chokshi, Pooja et al. (2009) Diversity of 23S rRNA genes within individual prokaryotic genomes. PLoS One 4:e5437|
|Gao, Zhan; Tseng, Chi-hong; Strober, Bruce E et al. (2008) Substantial alterations of the cutaneous bacterial biota in psoriatic lesions. PLoS One 3:e2719|
|Ouyang, Jie; Pei, Zhiheng; Lutwick, Larry et al. (2008) Case report: Paenibacillus thiaminolyticus: a new cause of human infection, inducing bacteremia in a patient on hemodialysis. Ann Clin Lab Sci 38:393-400|
Showing the most recent 10 out of 12 publications