Toll-like receptors in tonsillar epithelial cells.
OBJECTIVE
The Waldeyer's ring, comprised of the nasopharyngeal tonsil, the paired tubal tonsils, the paired palatine tonsils, and the lingual tonsil, is arranged in a circular orientation around the wall of the throat. The location of the palatine tonsils, specifically, enables these structures to come in direct contact with potentially harmful inhaled and ingested material that exist in their native form since digestive enzymes are not present in the oral cavity. Thus, the tonsillar epithelium must not only serve a protective role but it must also function in an antigen-sampling role. Previous studies involving the tissues of the Waldeyer's ring have been focused on the adaptive immune system, with little consideration toward the innate immune system. Studies have demonstrated that the tonsils are capable of producing proinflammatory and antiviral cytokines and chemokines. In addition, other studies have highlighted the importance of epithelial cells in this response. Therefore, we postulate that toll-like receptors (TLRs), which recognize components of pathogenic organisms, may play a key role in the innate immune response in tonsillar epithelial cells. TLRs are innate pattern recognition receptors, which produce proinflammatory cytokines and chemokines upon ligation. In this study, we examine the expression and function of TLRs in the tonsillar epithelial cell lines, UT-SCC-60A and UT-SCC-60B. Additionally, we demonstrate successful isolation of primary tonsillar epithelial cells and examine TLR expression in these cells.


METHODS
We utilized endpoint RT-PCR, real time RT-PCR, and flow cytometric analysis to determine TLR expression. To assess TLR function, cells were stimulated with TLR ligands and supernatants were assayed for secretion of cytokines.


RESULTS
UT-SCC-60A and UTSCC-60B express TLR mRNA and TLR protein, and the observed responses to the TLR ligands, Pam3Cys and Poly I:C suggest that TLR2 and TLR3 are functional in these cells. Additionally, primary tonsillar epithelial cells express TLRs.


CONCLUSIONS
TLRs are expressed in human tonsillar epithelial cells and may play a vital role in the immunological outcomes in this tissue.
Lange
Margaret J
MJ
Department of Molecular Microbiology and Immunology, University of Missouri, School of Medicine, Columbia, MO 65212, USA.
Lasiter
John C
JC
Misfeldt
Michael L
ML
D000293:Adolescent; D002460:Cell Line; D018925:Chemokines / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D016207:Cytokines / Q000235:genetics / Q000276:immunology; D004586:Electrophoresis; D004847:Epithelial Cells / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology*; D007223:Infant; D018836:Inflammation Mediators / Q000032:analysis; D008297:Male; D014066:Palatine Tonsil / Q000276:immunology* / Q000378:metabolism; D011070:Poly I-C; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism*
D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators; D012333:RNA, Messenger; D051193:Toll-Like Receptors; D011070:Poly I-C