University of Louisville
Variable TLR Expression in Unstimulated Gingival Fibroblasts and Epithelial Cells
Institution
University of Louisville
Faculty Advisor/ Mentor
Denis Kinane
Abstract
Objectives: Toll-like receptors (TLRs) play a critical role in the detection of microbial insult. Inter-subject susceptibility of periodontal disease varies. This variation may result from different profiles of TLRs expression between subjects. To address how expression patterns of TLRs differ among subjects, the present study examined the mRNA expression of TLR-1 to 10 among cultured human gingival epithelial cells and fibroblasts derived from different subjects. Methods: Human gingival epithelial cells (HGEC-1, -2 and -3) were prepared from three healthy gingival tissues obtained from three subjects in a protocol approved by the IRB at the University of Louisville and maintained separately. Human gingival fibroblasts (HGF-1 and -2) prepared separately from the same gingival tissues and immortalized human gingival epithelial cell line (OBA-9), kindly provided by Dr. Shinya Murakami (Osaka University, Japan) were used for comparison with HGECs. The mRNA levels of TLR-1 to TLR-10 were examined by Real-time PCR. Results: Nine human TLRs mRNAs with the exception of TLR-8 mRNA are expressed in HGEC-1, -2, and -3 and HGF-1 and -2, albeit that some are at low copy numbers. On the other hand, all ten TLRs were detected in OBA-9. HGECs and OBA-9 showed less mRNA expression of TLR-1 than HGFs. HGECs showed much less mRNA expressions of TLR-4 than HGFs and OBA-9. Furthermore, each cell had their own individual expression pattern of TLRs. Conclusion: TLR-1 and -4 mRNA expression in HGECs are markedly different from those in HGFs and we consistently found that all unstimulated cells had unique TLR expression patterns. This variation in TLR expression may be a crucial element of inter-subject variation in periodontal disease susceptibility.
Variable TLR Expression in Unstimulated Gingival Fibroblasts and Epithelial Cells
Objectives: Toll-like receptors (TLRs) play a critical role in the detection of microbial insult. Inter-subject susceptibility of periodontal disease varies. This variation may result from different profiles of TLRs expression between subjects. To address how expression patterns of TLRs differ among subjects, the present study examined the mRNA expression of TLR-1 to 10 among cultured human gingival epithelial cells and fibroblasts derived from different subjects. Methods: Human gingival epithelial cells (HGEC-1, -2 and -3) were prepared from three healthy gingival tissues obtained from three subjects in a protocol approved by the IRB at the University of Louisville and maintained separately. Human gingival fibroblasts (HGF-1 and -2) prepared separately from the same gingival tissues and immortalized human gingival epithelial cell line (OBA-9), kindly provided by Dr. Shinya Murakami (Osaka University, Japan) were used for comparison with HGECs. The mRNA levels of TLR-1 to TLR-10 were examined by Real-time PCR. Results: Nine human TLRs mRNAs with the exception of TLR-8 mRNA are expressed in HGEC-1, -2, and -3 and HGF-1 and -2, albeit that some are at low copy numbers. On the other hand, all ten TLRs were detected in OBA-9. HGECs and OBA-9 showed less mRNA expression of TLR-1 than HGFs. HGECs showed much less mRNA expressions of TLR-4 than HGFs and OBA-9. Furthermore, each cell had their own individual expression pattern of TLRs. Conclusion: TLR-1 and -4 mRNA expression in HGECs are markedly different from those in HGFs and we consistently found that all unstimulated cells had unique TLR expression patterns. This variation in TLR expression may be a crucial element of inter-subject variation in periodontal disease susceptibility.