Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) and Foxp3 (forkhead box P3) are receptors present on T cells which play a critical role in the down-regulation of antigen-activated immune responses. To evaluate the potential influences of CTLA-4 and Foxp3 on cancer invasiveness, a case-control study was conducted in 86 patients treated for squamous cell laryngeal carcinoma. The abundance of CTLA-4 and Foxp3 gene transcripts in the purified peripheral blood mononuclear cells (PBMCs) by quantitative real-time PCR (qRT-PCR) was determined. The analysis of proteins by Western blot was performed. The relationships between CTLA-4 and Foxp3 gene and protein expression as well as the aggressiveness of tumor determined on pT, type and depth of invasion were investigated. Our work revealed a significant dependence of mRNA CTLA-4 on tumor front grading (TFG) total score (p = 0.04) as well as CTLA-4 protein expression on pT (p = = 0.03) and type of invasion (p = 0.03). Advanced pT3-pT4 tumors with diffuse infiltration and > 14 TFG points were characterized by higher average values of CTLA-4 protein in PBMCs. Our data also demonstrated significant differences between Foxp3 protein levels in relation to pT (p = 0.04), depth of invasion (p = = 0.02) and type of invasion (p = 0.03). In tumors with the highest invasiveness identified by the pT3-pT4 status, deep invasion with involvement of cartilage and diffuse infiltration, the highest Foxp3 protein level was observed. In conclusion, these results suggest an impact of CTLA-4 and Foxp3 in determining proliferative and aggressive potential of laryngeal carcinoma, highlighting the significance of CTLA-4 and Foxp3 as potential predictive indicators.

Aim of the studyAim of the study was to evaluate the potential role of regulatory and proinflammatory cytokines IL-23 and IL-17 as Th17 lymphocyte activity markers in relation to invasiveness in laryngeal cancer.Material and methodsThe immunological analysis was conducted in 50 patients treated for squamous cell laryngeal carcinoma and 30 healthy volunteers as controls. The levels of IL-23 and IL-17 in supernatants of purified peripheral blood mononuclear cell cultures were determined by using the enzyme-linked immunosorbent assay (ELISA). The clinicomorphological criteria included pTNM, stage, G, and the total tumour front grading (TFG) score.ResultsOur data demonstrated higher concentrations of IL-23 in patients as compared to controls (p = 0.0001). No statistical difference for IL-17 in these groups was observed. Our study revealed significant dependences in IL-23 expression on pT (p = 0.04), histological differentiation (p = 0.04), and TFG total score (p = 0.02). Advanced tumours (pT3–pT4) with higher grade (G2–G3) and higher invasiveness (> 14 TFG points) were characterised by elevated IL-23 levels in PBMC supernatants. Our data did not indicate a relationship between cytokine levels and three- and five-year survival. However, a tendency towards lower content of IL-23 in PBMC cultures in patients who lived longer than five years after treatment was noted. The relationships between IL-17 level in PBMC cultures and clinicomorphological and prognostic parameters have not been disclosed.ConclusionsThe results of this study suggest the importance of regulatory cytokine IL-23 in determining the aggressive potential of laryngeal carcinomas.

Aim of the studyExpression of EP2 protein, the prostaglandin E2 (PGE2) receptor, produced by tumour microenvironment inflammatory cells as well as tumour cells, may promote cellular proliferation and growth in an autocrine and paracrine fashion. The phenomenon involving these proteins is regulated by interleukin 1β (IL-1β). Many researchers indicate a connection of EP2 and IL-1β in various types of neoplasms with higher tumour progression and poor prognosis. The aim of this study was to analyse the EP2 expression within laryngeal carcinoma tissue and IL-1β levels in peripheral blood mononuclear cell supernatants and to find relationships between clinicomorphological features.Material and methodsA group of 50 patients with verified squamous cell laryngeal carcinoma was analysed in this study. The pathological evaluation included pTNM depth of invasion according to tumour front grading criteria. Immunohistochemical analysis for membranous staining of EP2 in tumour tissues was used. The IL-1β expression was determined by enzyme-linked immunosorbent assay (ELISA).ResultsIncreased EP2 expression in carcinoma cells was confirmed for more advanced tumours (pT3-pT4 vs. pT1-pT2, p < 0.0001 and pN1-3 vs. pN0, p = 0.02). Tumours with the highest aggressiveness identified by deeper invasion of submucosa or cartilage were characterised by the highest expression of EP2 (p < 0.0001). In laryngeal carcinomas characterised by a lower differentiation the highest EP2 expression in tumour cells was noted (p = 0.009). A positive relationship between IL-1β expression and the presence of lymph node metastases was also confirmed (p = 0.04).ConclusionsThe study indicates the potential effect of EP2 receptor and IL-1β on tumour progression in laryngeal carcinoma.

Aim of the studyIntercellular adhesion molecules present in immunocompetent cells as well as endothelium and tumour cells can regulate cell migration, angiogenesis, apoptosis, proliferation, and metastases in solid tumours. The aim of this study was to analyse the sICAM-1 (soluble intercellular adhesion molecule 1) and sVCAM-1 (soluble vascular cell adhesion molecule 1) expression in peripheral blood mononuclear cell (PBMC) cultures, and to find their relationships with clinicomorphological characteristics in laryngeal cancer.Materials and methodsThe analysis included a group of 50 patients with verified squamous cell carcinoma of the larynx. The control group constituted 30 healthy volunteers. The pathological assessment included pTNM, stage, histological grade, and type of invasion according to the tumour front grading. The expression of adhesion molecules was assessed using the enzyme-linked immunosorbent assay (ELISA).ResultsIncreased expression of sICAM-1 and sVCAM-1 was an indicator of more aggressive laryngeal carcinomas. More advanced local changes evaluated on the pT feature were connected with a higher sVCAM-1 (p = 0.017), but not sICAM-1 level. The presence of lymph node metastases correlated with a higher expression of adhesion molecules (p = 0.012 and p = 0.003, for sICAM-1 and sVCAM-1, respectively). Tumours with more diffuse growth and infiltrating with small cell groups (< 15/hpf) was characterised by the highest level of adhesive proteins (p = 0.001 and p = 0.02 for sICAM and sVCAM, respectively). Moreover, lower levels of sICAM-1 and sVCAM-1 were observed more frequently in patients who lived longer than five years after treatment.ConclusionsThe study indicates the importance of the sICAM and sVCAM expression as indicators of advanced changes and prognosis in patients with laryngeal carcinoma.

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