TLR receptors in laryngeal carcinoma - immunophenotypic, molecular and functional studies.

Abstract

Toll-like receptors (TLRs) have been shown to play crucial role in the recognition of unicellular pathogens. We have shown the expression of three TLRs on tumor cells of human laryngeal carcinoma by means of immunohistochemistry. In the current study we searched presence of TLR1-10 on protein and molecular level in larynx carcinoma cell lines and the impact of respective TLR ligands on TLR expression. Larynx carcinoma cell lines have been used. Cell were subjected to immunocytochemistry. RNA isolated from the cells was tested by RT-PCR. Cells were cultured in the presence of respective TLR ligands. Cells than were harvested and subjected to flow cytometry, using anti TLR1-10 Moabs. The cells were evaluated of membrane and cytoplasmic cell staining. TLR reactivity varied in individual cell lines. RT-PCR allowed to show mRNA for all TLRs tested. After short-term cell culture each cell line exhibited distinct pattern of expression of TLRs following interaction with respective ligand. Cytoplasmic TLR staining had usually higher MFI value than membrane one, but after culture with ligand it became reversed. TLRs 7 and 9 showed highest expression in the majority of tumor cells tested. In conclusion, larynx carcinoma cell lines exhibit rather universal expression of TLRs, both on protein and molecular level. Culture of TLR expressing tumor cells with ligands points out for potential reactivity of tumor cells with TLR agonists, what may have therapeutic implications.

Hepatic stellate cells (HSC) cell line (LX-2) and Toll-like receptors – phenotypic/mRNA expression and in vitro functional studies

“…It suggests, that FBS cell stimulation enhances TLR expression and that the cells may absorb TLR molecules to cell interior. Such phenomenon was observed by us previously, in the case laryngeal carcinoma cell lines [10].…”

“…RNA isolation and RT-PCR reaction was carried out similarly to that described elsewhere [10]. GAPDH as a housekeeping gene was used.…”

Experimental immunology Determination of mRNA three Toll-like receptors in human carcinoma cell lines

“…We were able to demonstrate mRNA in head and neck cancer cell lines for all TLRs studied (TLR-1 to TLR-10) by rt PCR qualitative assay. We noticed, however, differences in the intensity of individual mRNA lines [8]. Such differences may have significant impact on cell-cell interactions in the attempts of immunotherapy involving TLRs.…”

“…The same head and neck carcinoma cell lines as studied previously [8] have been used. They originated from Pittsburgh Cancer Institute (USA) generously provided by professor Theresa Whiteside (PCI).…”

Expression of pattern recognition receptors in liver biopsy specimens of children chronically infected with HBV and HCV

“…Primary goat anti-PRRs antibodies used were all from Santa Cruz Biotechnology, CA, USA, and were the same as used previously [12]. The next day, sections were washed thrice with cold PBS and thereafter subjected to biotinylated donkey anti-goat IgG for 30 min.…”