Reciprocal Interactions of Human Monocytes and Cancer Cells in Co-Cultures In Vitro.

The tumor microenvironment (TME) includes immune and stromal cells and noncellular extracellular matrix (ECM) components. Tumor-associated macrophages (TAMs) are the most important immune cells in TME and are crucial for carcinomas' progression. The purpose was to analyze direct and indirect interactions in co-culture of tumor cells with monocytes/macrophages and, additionally, to indicate which interactions are more important for cancer development. Cytokines, reactive oxygen species, nitric oxide level, tumor cell cycle and changes in tumor cell morphology after human tumor cells (Hep-2 and RK33 cell lines) with human monocyte/macrophage (THP-1 cell line) interactions were tested. Morphology and cytoskeleton organization of tumor cells did not change after co-culture with macrophages. In co-culture of tumor cells with human monocyte, changes in the percentage of tumor cells in cell cycle phases was observed. No significant changes in reactive oxygen species (ROS) were found in the co-culture as compared to the tumor cell mono-culture. Monocytes produced about three times higher ROS than tumor cells. In co-cultures, a lower nitric oxide (NOx) level was found as compared to the sum of the production by both mono-cultures. Co-culture conditions limited the production of cytokines (IL-4, IL-10 and IL-13) as compared to the sum of their level in mono-cultures. In conclusion, macrophages influence tumor cell growth and functions. Mutual (direct and paracrine) interactions between tumor cells and macrophages changed cytokine production and tumor cell cycle profile. The data obtained may allow us to initially indicate which kind of interactions may have a greater impact on cancer development processes.

ThymicPeptides Reverse Immune Exhaustion in Patients with Reactivated Human Alphaherpesvirus1 Infections.

Recurrent infection with human alphaherpesvirus 1 (HHV-1) may be associated with immune exhaustion that impairs virus elimination. Thymic peptides enhance immune function and thus could overcome immune exhaustion. In this study, we investigated whether reactivation of herpes infections was associated with immune exhaustion. Moreover, we examined the impact of treatment with thymostimulin on the expression of programmed cell death protein 1 (PD-1) and its ligand (PD-L1) on T and B lymphocytes in patients suffering from recurrent HHV-1 reactivation. We also assessed the effector function of peripheral blood mononuclear cells (PBMCs) after stimulation with thymic peptides. We enrolled 50 women with reactivated HHV-1 infections and healthy volunteers. We measured the expression of various activation and exhaustion markers on the surface of PBMCs using flow cytometry. In ex vivo experiments, we measured the secretion of inflammatory cytokines by PBMCs cultured with thymostimulin. Compared with controls, patients with reactivated HHV-1 infections had increased percentages of CD3+ co-expressing CD25, an activation marker (p < 0.001). Moreover, these patients had increased percentages of CD4+ and CD8+ cells co-expressing the inhibitory markers PD-1 and PD-L1. In cultures of PBMCs from the patients, thymostimulin increased the secretion of interferon gamma (p < 0.001) and interleukin (IL)-2 (p = 0.023), but not IL-4 or IL-10.Two-month thymostimulin therapy resulted in no reactivation of HHV-1 infection during this period and the reduction of PD-1 and PD-L1 expression on the surface of T and B lymphocytes (p < 0.001). In conclusion, reactivation of herpes infection is associated with immune exhaustion, which could be reversed by treatment with thymic peptides.

Imperatorin as a Promising Chemotherapeutic Agent Against Human Larynx Cancer and Rhabdomyosarcoma Cells.

Naturally occurring coumarins are bioactive compounds widely used in Asian traditional medicine. They have been shown to inhibit proliferation, induce apoptosis, and/or enhance the cytotoxicity of currently used drugs against a variety of cancer cell types. The aim of our study was to examine the antiproliferative activity of different linear furanocoumarins on human rhabdomyosarcoma, lung, and larynx cancer cell lines, and dissolve their cellular mechanism of action. The coumarins were isolated from fruits of Angelica archangelica L. or Pastinaca sativa L., and separated using high-performance counter-current chromatography (HPCCC). The identity and purity of isolated compounds were confirmed by HPLC-DAD and NMR analyses. Cell viability and toxicity assessments were performed by means of methylthiazolyldiphenyl-tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays, respectively. Induction of apoptosis and cell cycle progression were measured using flow cytometry analysis. qPCR method was applied to detect changes in gene expression. Linear furanocoumarins in a dose-dependent manner inhibited proliferation of cancer cells with diverse activity regarding compounds and cancer cell type specificity. Imperatorin (IMP) exhibited the most potent growth inhibitory effects against human rhabdomyosarcoma and larynx cancer cell lines owing to inhibition of the cell cycle progression connected with specific changes in gene expression, including CDKN1A. As there are no specific chemotherapy treatments dedicated to laryngeal squamous cell carcinoma and rhabdomyosarcoma, and IMP seems to be non-toxic for normal cells, our results could open a new direction in the search for effective anti-cancer agents.

Anterolateral thigh free flap in reconstruction of lateral skull base defects after oncological resection.

PURPOSE: Evaluation of the utility of the free anterolateral thigh flap reconstruction of the defects resulting from radical temporal bone resection in the management of lateral skull base malignancies in a single institution. METHODS: An analysis of 17 en bloc subtotal petrosectomies for removal of malignant tumours was performed. There were 12 squamous cell carcinomas, 4 basal cell carcinomas and 1 adenoid cystic carcinoma. The tumours were staged with the University of Pittsburgh TNM system. In all patients, the lateral temporal bone with the preservation of the petrous apex and carotid artery was performed. All patients had parotid gland resection. The post-resection defect was reconstructed with an ALT free flap. RESULTS: Tumour radical resection and defect reconstruction with an ALT flap was achieved in all patients without intraoperative complications. The transplants were harvested as fasciocutaneous flaps, 11 perfused by musculocutaneous and 6 by septocutaneous perforators. The ALT flaps had a mean pedicle length of 8 cm (6-12 cm), and the flap size ranged between 6 × 15 cm and 15 × 30 cm. The flaps were supplied by nine facial, five occipital and three maxillary arteries. Recipient-site veins included eight internal jugular, seven facial, one retromandibular and one external jugular vein. All arterial pedicles were anastomosed in an end-to-end manner. The veins were anastomosed with interrupted sutures and in 11 cases with Synovis-Coupler® devices. All the flap transfers were performed successfully. Three patients experienced postoperative complications. CONCLUSIONS: The ALT flap proved to be effective for covering large temporal skull base defects resulting from the radical removal of temporal bone malignancies. The functional and cosmetic results were acceptable with a low complication rate.

Toll-Like Receptors-2 and -4 in Graves' Disease-Key Players or Bystanders?

Graves' disease (GD) is an autoimmune disease that affects the thyroid. The development of autoimmunity is associated with innate immune responses where the prominent role plays Toll-like receptors (TLRs). The aim of our study was to assess the relationship between the expression levels of TLR-2 and TLR-4 on CD4+ and CD8+ T as well as CD19+ B lymphocytes in patients with GD and selected clinical parameters. The study group consisted of 32 women with GD, the control group consisted of 20 healthy women. Immunophenotyping was performed using the flow cytometry and cytokines concentrations were assessed using ELISA assay. The mean percentage of CD4+/TLR-2+ and CD8+/TLR-2+ T cells in patients with GD was higher than in the control group (p < 0.0001). After obtaining euthyroidism, the mean percentage of CD4+/TLR-2+ T cells in patients with GD decreased (p < 0.0001). The expression level of TLR-2 on CD4+ T lymphocytes correlated with serum FT3 concentration in patients with GD (r = 0.47, p = 0.007). The mean percentage of CD8+/TLR-2+ T cells in patients with GD before treatment compared to patients with GD after obtaining euthyroidism was higher (p = 0.0163). Similar findings were found for TLR-4. Thus the TLR-2 and TLR-4 can be a prognostic marker for Graves' disease.

Subpopulations of natural killer-T-like cells before and after surgical treatment of laryngeal cancer.

INTRODUCTION: Tumours connected with head and neck comprise about 5% of all tumours. The most frequent histological type of laryngeal carcinoma is squamous cell carcinoma. Different research projects suggest that the role of T lymphocytes might be significant in tumour development. iNKT cells are a new subpopulation of T cells and show cytotoxic activity against tumours. iNKT cells participate in modulating the function of other cells which have anti-tumour properties and secrete cytokines, which have pro-inflammatory and anti-inflammatory effects. In animal models the significance of iNKT cells in various diseases including cancer was shown. AIM OF THE STUDY: The aim of this study was to determine the percentages of iNKT cells, CD161+ cells, CD161- cells, iNKT CD4+ cells, and iNKT CD8+ cells, NK cells, NKT-like cells, and T cells subsets present in peripheral blood of patients with laryngeal cancer before and two months after the tumour resection, in comparison to healthy volunteers. MATERIALS AND METHODS: This study included material from laryngeal patients who were treated at the Department of Otolaryngology and Laryngological Oncology (Medical University of Lublin) between 2012 and 2013. A total of 50 patients (40 men and 10 women) aged between 45 and 77 years (median age: 60 years) were enrolled. Based on the TNM classification, the patients were classified as having stage I-IV laryngeal cancer. The control group was composed of 15 healthy volunteers (12 men and three women) aged between 43 and 82 years (median age: 61 years). The protocol of the study was approved by the Local Bioethical Committee at the Medical University of Lublin.Peripheral blood samples (15 ml) from the basilic vein were collected by venipuncture using sterile, sodium heparin-treated tubes (20 units per ml of blood) and used for cytometric analyses. RESULTS: iNKT cells were analysed among T CD3+ cells. The percentage of CD3+ and CD3+CD4+ T cells before tumour resection was higher than in the control group, but the increase of CD3+ T cells was not significant. The T CD3+CD4+ / T CD3+CD8+ cell ratio was significantly higher than in healthy donors. After tumour resection a decreased percentage of CD3+CD4+ T cells but an increased percentage of CD8+CD3+T cells was noted. The T CD3+CD4+ / T CD3+CD8+ cell ratio was significantly higher in patients before and after the surgery than in the control group. The amount of NKT-like cells increased after resection and was significantly higher than in the control group. CONCLUSIONS: Our study exhibited the change in percentage of iNKT, NK, NKT-like cells, and T lymphocytes after tumour resection in patients with laryngeal cancer. The research explains the contribution of those cells in immunological response against tumour.

Is immunohistochemical evaluation of p16 in oropharyngeal cancer enough to predict the HPV positivity?

AIM: Our goal was to determine the expression levels of p16 in the cohort of the OPSCC patients and evaluation of the pathological and clinical differences between these two groups including patients' survival. BACKGROUND: HPV infection is the main causative factor of oropharyngeal cancer (OPSCC). Identification of HPV status in OPSCC requires positive evaluation of viral DNA integration into host cell however, p16 accumulation in the proliferating cell layers has been accepted as an alternative marker for HPV infection. MATERIAL AND METHODS: The IHC staining for p16 has been performed in tumor tissue from 382 OPSCC patients. The sample was considered positive based on more than 70% of carcinoma tissue showing strong and diffused nuclear and cytoplasmic immunostaining. The clinicopathological characteristics of the patients including site, age, gender, tumor grade, tumor stage, the nodal status, smoking and survival have been analyzed when comparing p16 positive and p16 negative tumors. RESULTS: Out of our cohort in 38.2% cases positive staining for p16 has been recorded. Our analysis did not indicate significant differences in the distribution of the p16 positive patients and age of the patients, stage of the disease. Among the patients who have presented with the N+ neck, there were significantly more p16 positive tumors than in the group with N0 neck (p = 0.0062). There was highly significant correlation between the expression of p16 and smoking (p < 0.0001). The significant difference in survival (p < 0.0001) with more favorable prognosis in the p16 positive group has been observed. CONCLUSIONS: Overexpression of p16 is accepted as a surrogate diagnostic marker for detecting HPV infection in oropharyngeal cancer. However, one should remember about existence of the small subgroups of p16 positive but HPV negative tumors, with relatively worse prognosis. Immunostaining for p16, however useful on everyday basis, should be complemented with other techniques in terms of reliable identification of the HPV infection.

The percentage of iNKT cells among other immune cells at various clinical stages of laryngeal cancer.

INTRODUCTION: Invariant natural killer T (iNKT) cells constitute a small population of immune cells that share functional and phenotypic characteristics of T lymphocytes and NK cells. Due to their involvement in specific and non-specific immune responses, iNKT cells may represent an important component of antitumor and anti-infectious immunity. MATERIAL AND METHODS: Using flow cytometry, we analyzed the percentages of iNKT cells as well as T and B lymphocytes in peripheral blood of 50 laryngeal cancer patients at various clinical stages in comparison to healthy controls (n=15). Moreover, we determined the expression of CD25, CD69 and CD95 antigens on T lymphocytes. RESULTS: The percentage of CD4+/CD3+ T lymphocytes in the controls was higher than in laryngeal cancer patients, both with early and late stages of the disease. The percentage of CD8+/CD3+ T lymphocytes in healthy controls was lower than in patients with early and late clinical stages of laryngeal cancer. Patients with advanced laryngeal cancer showed a lower percentage of iNKT cells and higher frequencies of T regulatory cells (Tregs) than the controls. Advanced clinical stages of laryngeal cancer are associated with impaired activation of lymphocytes. CONCLUSIONS: Our study confirmed that laryngeal cancer cells exert a strong suppressor effect on the immune system of the host. This is reflected by a decrease in the percentage of iNKT cells that are capable of cancer cell elimination, and a concomitant increase in the percentage of Tregs. However, further studies are needed in order to explain the underlying mechanisms of immunosuppression and understand interactions between immune and cancer cells.

The Relationship between Cancer Stage, Selected Immunological Parameters, Epstein-Barr Virus Infection, and Total Serum Content of Iron, Zinc, and Copper in Patients with Laryngeal Cancer.

(1) Background: the purpose of the study was to assess the relationship between cancer stage, selected immunological parameters, Epstein-Barr virus (EBV) infection, and total serum content of iron, zinc, and copper in patients with laryngeal cancer (LC). (2) Methods: serum Fe, Zn, and Cu were measured in 40 LC patients and 20 controls. Immunophenotyping of peripheral blood lymphocytes was performed by flow cytometry using fluorescent antibodies against CD3, CD4, CD8, CD19, CD25, CD69, and PD-1. Tumor and lymph node lymphocytes were analyzed by flow cytometry. EBV DNA was quantified by real-time PCR, targeting the EBNA-1 gene. Associations between serum elements, immune markers, and cancer grade/stage were evaluated using ANOVA and appropriate nonparametric tests. (3) Results: levels of Fe, Cu, and Zn were lower, while Cu/Zn was statistically higher, in patients with LC than in the control group. Correlation analysis showed a statistically significant association between the levels of these elements and parameters of the TNM (Tumor, Node, Metastasis) staging system, immunophenotype, and the amount of EBV genetic material in patients with LC who survived for more than 5 years. (4) Conclusion: the results suggest that the total serum levels of the determined micronutrients may significantly affect the immunopathogenesis and progression of LC.

Expression of selected regulatory molecules on the CD83+ monocyte-derived dendritic cells generated from patients with laryngeal cancer and their clinical significance.

B7H1 and B7H4 overexpression is associated with inhibition of the immune system in many solid tumors, and altogether with CD200 molecule plays an important role in tumor invasion by promoting malignant transformation. However, there is no report about impact of these molecules on laryngeal squamous cell carcinoma. The objective of the present study was to assess by means of flow cytometry the expression of B7H1, B7H4, CD200, and CD200R on CD83+ monocyte-derived dendritic cells (Mo-DC), pulsed with autologous tumor cell lysates (aTCL) in patients who suffer from G1, G2, or G3 laryngeal carcinoma (LC, n = 60) in comparison to healthy donors (HD, n = 15). It has been demonstrated that median value of the percentages of CD83+ B7H1+, CD83+ B7H4+, and CD83+ CD200+ cells were higher in LC patients than HD (p = 0.041, p ≤ 0.0001, and p = 0.02, respectively). Mean fluorescence intensity (MFI) of CD200, CD200R, B7H1, and B7H4 on the Mo-DC pulsed with aTCL of the patients was also higher than on the Mo-DC of HD (p ≤ 0.0001, p ≤ 0.0001, p = 0.002, and p ≤ 0.0001, respectively). The highest MFI levels of all molecules were noted in grade 3 LC. The aforementioned results prove that there is a relation between the presence of laryngeal cancer and the expression of B7H1, B7H4, CD200, and CD200R regulatory molecules on the CD83+ Mo-DC pulsed with autologous cancer cell lysates. Strong association of LC grade and the tested antigens expression suggests a critical role for these proteins in LC biology.

Head and neck cancer: value of perfusion CT in depicting primary tumor spread.

BACKGROUND: The aim of this study was to assess head and neck squamous cell cancer and surrounding tissue in computed tomography contrast enhanced and perfusion studies, and to examine the role of perfusion imaging in depiction of tissue infiltration. MATERIAL/METHODS: We prospectively evaluated 43 primary malignant head and neck tumors, using standard CT followed by perfusion. Blood flow, blood volume, mean transit time, and permeability values were obtained using regions of interest (ROIs) over lesions and surrounding tissue. Results were compared with histological analysis of resected tissue. Sensitivity, specificity, accuracy, positive and negative predictive values were calculated for both methods. RESULTS: We found significant differences between infiltrated and non-infiltrated tissue, especially with regard to muscles. In case of bone and salivary gland infiltration, change in perfusion parameters did not allow proper diagnosis. CONCLUSIONS: CTP shows promise in depicting malignant infiltration. The combined use of CECT plus CTP results in correct staging of the majority of head and neck tumors.

[The evaluation of apoptosis in peripheral blood lymphocytes in laryngeal cancer patients]. / Ocena apoptozy limfocytów krwi obwodowej u chorych leczonych z powodu raka krtani.

UNLABELLED: The laryngeal cancer is the most often cancer among others in head and neck region. It occurs mostly among 55 and 69. Its development depends on immunological state of the body. Vitality of the immunological system cells was considered due to growth, treatment sensitivity and prognosis of some neoplasms. The aim of this work were estimation and comparison the phenomenon of lymphocytes T and B apoptosis in laryngeal cancer patients treated with surgery and radiotherapy. MATERIAL AND METHODS: The material were 30 patients hospitalized in The Department of Otolaryngology Medical University of Lublin. They all were treated with surgery or surgery and radiotherapy. Apoptosis was estimated on different stages of the treatment process. All samples were examined with the flow cytometry method. The control group were 21 patients hospitalized because of the suspicion of the apnea syndrome, which wasn't confirmed with polysomnographic examination. RESULTS: Results of this study show significantly increasing percentage of peripheral blood apoptotic B (CD19+) cells caused by surgical treatment. The results considering radiotherapy showed different influence on the phenomenon of immunological cells apoptosis, still those results weren't significant. CONCLUSIONS: The surgical treatment causes increased amount of apoptotic peripheral blood lymphocytes.

Determinants and disparities in access to paediatricians in Poland.

BACKGROUND: The purpose of this study was to identify the factors that determine the differences in the distribution and workload of paediatricians in Poland. This research, specific to conditions found within Poland, will help further advance knowledge in this area. Data were derived from the database of Statistics Poland. The level of convergence of the phenomenon studied was analysed. The paediatricians' accessibility index was ascertained and its spatial diversity examined. The level of correlation of patients treated per paediatrician was analysed in relation to indices of urbanisation, availability of paediatricians and disposable income. RESULTS: A moderate variation of patients treated per paediatrician was found and the conditional convergence of the investigated phenomenon observed. A close negative association between the number of patients treated and access to paediatricians (-0.686, p = 0.005) was revealed. CONCLUSIONS: The research suggests that socioeconomic factors may affect the uneven spatial distribution of the workload of paediatricians in Poland and cause differences between the provinces in the equal access to paediatricians. This research may thus provide implications for policy and practice as well as lead to a better understanding of the problem.

The Clinical, Pathological, and Prognostic Value of High PD-1 Expression and the Presence of Epstein-Barr Virus Reactivation in Patients with Laryngeal Cancer.

Due to the development of molecular diagnostic techniques, the latest research in the diagnosis of cancer diseases, including laryngeal cancer, has been focused on the occurrence of specific types of molecular patterns, including markers expressed on cells of the immune system (e.g., PD-1, PD-L1, and CTLA-4), which may be directly or indirectly involved in the development of neoplastic diseases. Laryngeal cancer is one of the diseases that is diagnosed more often in men than in women, and many factors are involved in its development, including environmental and lifestyle factors, viral infections (e.g., HPV, HHV-1, and EBV), and disorders of the immune system. In this study, we determined the level of PD-1 receptor expression on T and B lymphocytes and their relationships based on the classification of the grade and TNM scale, in turn based on blood, tumor, and lymph node samples from patients diagnosed with laryngeal cancer. In addition, we determined the presence of EBV genetic material in the tested biological materials as well as the degree of cancer advancement and its correlation with the level of PD-1 receptor expression. The results suggested that the level of PD-1 expression on T and B lymphocytes was significantly higher in the tumor samples as compared to the lymph node samples, and their comparison with the immunophenotype results from the blood samples provided statistically significant data on changes in the incidence of individual subpopulations of T and B lymphocytes and the level of PD-1 receptor expression. The analysis of the individual parameters of the TNM scale also showed significant changes between the PD-1 expression and the tested biological material in individual subgroups of the scale. We also found that the expression of PD-1 on the CD4+ T cells from the lymph node samples caused an almost 1.5-fold increase in the risk of death. In the analyses of the presence of EBV, the highest concentration was recorded in the tumor samples, then for the lymph node samples, and followed by the blood samples. Furthermore, we showed that the presence of EBV genetic material was positively correlated with the level of PD-1 expression in the tested biological materials.

Epistaxis in patients with hereditary hemorrhagic telangiectasia treated with selective arterial embolization.

BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) is a rare systemic fibrovascular dysplasia also known as Rendu-Osler-Weber disease. Epistaxis is often the first and foremost symptom of the disease and recurrent epistaxis is the main complaint. PURPOSE: Evaluation of effectiveness and safety of endovascular treatment of epistaxis in HHT patients. MATERIAL AND METHODS: Between June 2004 and January 2008, 14 patients with HHT underwent embolization of external carotid artery (ECA) branches due to severe epistaxis. There were eight men and six women aged 27-42 years. Patients were referred for endovascular treatment when primary management with electrocautery and nasal packing was ineffective. Arteries suspected of bleeding were catheterized super-selectively with micro-catheters to perform embolization. Polyvinyl alcohol particles (PVA) or embospheres in diameter of 500-700-µm were used as an embolic material. RESULTS: Embolization was technically successful in 11 (78.5%) patients. Immediate, complete control of bleeding was achieved in 12 (85%) patients. During long-term follow-up recurrent nasal bleeding occurred in six (43%) patients 6-24 months after treatment. There were no severe complications after embolization. Nine patients experienced mild facial pain which resolved within three days after the treatment. Three patients in whom arterial spasm was observed, suffered from facial paraesthesia lasting for 3-5 days after embolization. Out of 14 patients, 12 were available for 24-month follow-up evaluation. All except one claimed reduction in frequency and severity of epistaxis which had a positive impact on their quality of life. CONCLUSION: Endovascular embolization of epistaxis in patients with HHT is an effective and safe procedure although it cannot provide a long-term cure in most cases. Embolization reduces the frequency, severity and duration of nasal bleeding in patients with HHT, improving patients' quality of life.

[Evaluation of cytochrome concentration in peripheral blood lymphocytes of patients with laryngeal cancer]. / Ocena stezenia cytochromu c w limfocytach krwi obwodowej chorych na raka krtani.

The aim of the study was evaluation of cytochrome concentration in the peripheral blood lymphocytes of patients with laryngeal cancer. The study was conducted in a group of 62 patients presenting different clinical advancement of the disease. The study material consisted of the studied population's peripheral blood from which T lymphocytes were isolated and incubated with monoclonal antibodies. To evaluate antigens' expression, a FACS Galibur flow cytometre was used; the evaluated cells were labeled with fluorochrome-conjugated monoclonal antibodies. Next, mononuclear cells were rinsed with cold PBS and suspended in the lysis buffer. In the obtained cell lysate the c cytochrome concentration was determined with the use of an immunoenzymatic test, HumanCytochrome cELISA Kit (BenderMed Systems, Austria). Obtained results were compared with the measurements taken in 20 healthy individuals who constituted the control group. On the basis of conducted study it was found that the level of c cytochrome concentration was significantly increased in the T CD3+ lymphocytes in the peripheral blood of patients with stage IV of laryngeal cancer. A positive correlatio was also found between the cytochrome level in lymphocytes and the advancing stage of the disease. Our own observation give grounds to conjecture that together with the progress of laryngeal cancer the energetic potential of lymphocytes increases and so does the readiness of the lymphatic cells to undergo the redox processes, therefore the amount of c cytochrome in the cells increases.

T-Lymphocyte Activation Is Correlated With the Presence of Anti-EBV in Patients With Laryngeal Squamous Cell Carcinoma.

BACKGROUND/AIM: Chronic viral infection is an important risk factor in the development of cancer. Failure of immune response to clear the oncogenic infection can facilitate cancer progression. The aim of the present study was to analyze early and late activation of T-lymphocytes related to Epstein-Barr virus (EBV) infection by the expression of markers of activation (CD69, CD25) on the surface of T-lymphocytes (CD3+, CD4+, CD8+) in patients bearing laryngeal cancer according to absence/presence immunoglobulin G antibodies to EBV nuclear antigen (EBNA1). MATERIALS AND METHODS: Thirty-three patients with laryngeal squamous cell carcinoma (LC) and 20 volunteers without cancer (control group) were enrolled in the study. Peripheral blood samples were collected from every individual. The markers of activation of T-lymphocytes were determined by flow cytometry, whereas commercial immunoenzymatic assay kits were used for detection of anti-viral capsid antigen (VCA) IgM, anti-VCA IgG, and anti-EBNA1 IgG. RESULTS: Increased early activation of CD8+ and CD4+ T-lymphocytes was found in patients with LC. There was a significantly higher proportion of CD4+ and CD8+T-lymphocytes expressing CD69 antigen in patients with LC compared to the control group. The proportion of CD4+ CD25+ T-lymphocytes in patients with LC positive for anti-EBNA1 IgG and anti-VCA IgM was lower compared to patients without antibodies to VCA IgM. CONCLUSION: The dysfunction of immune response in larynx cancer patients could be associated with EBV infection.

Cyclooxygenase-2 Inhibition Enhances Proliferation of NKT Cells Derived from Patients with Laryngeal Cancer.

BACKGROUND/AIM: The aim of this study was to analyze whether inhibition of cyclooxygenase-2 by celecoxib and the subsequent enhancement in the proliferation of natural killer T (NKT) cells could play a role in dendritic cell (DC)-based laryngeal cancer (LC) immunotherapy. PATIENTS AND METHODS: Peripheral blood mononuclear cells were obtained from 48 male patients diagnosed with LC and 30 control patients without cancer disease. Neoplastic cell lysate preparations were made from cancer tissues obtained after surgery and used for in vitro DCs generation. NKT cells proliferation assay was performed based on 3H-thymidine incorporation assay. RESULTS: An increased proliferation of NKT cells was obtained from control patients compared to NKT cells obtained from LC patients regardless of the type of stimulation or treatment. In the patient group diagnosed with LC, COX-2 inhibition resulted in a significantly enhanced proliferation of NKT cells when stimulated with autologous DCs than NKT cells stimulated with DCs without COX-2 inhibition. These correlations were not present in the control group. Higher proliferation rate of NKT cells was also observed in non-metastatic and highly differentiated LC, which was independent of the type of stimulation or treatment. CONCLUSION: COX-2 inhibition could be regarded as immunotherapy-enhancing tool in patients with LC.

Multicenter experiences in temporal bone cancer surgery based on 89 cases.

OBJECTIVE: To present outcomes of extensive surgery: lateral, subtotal, total petrosectomies in patients with temporal bone invasion resulting from specific primary cancers. STUDY DESIGN: Retrospective case review. SETTING: Four tertiary referral centers. MATERIAL: 89 patients with cancer of the temporal bone treated between January 2006 and December 2010. INTERVENTION: Multidisciplinary team approach including surgical resection, reconstruction, and postoperative radiotherapy. MAIN OUTCOME MEASURE: Disease-specific survival, overall survival. RESULTS: In 27.0% of the patients, relapse was reported, with an average of 6.3 months after surgery; 31 patients (34.8%) died during the follow-up. The average mortality was 22.1 months. Fifty-four patients (58.7%) stayed alive during the time of observation. The average survival time was 42.0 months. The median time of survival with relapse was 12 months (range: 1-51 months). The three-year disease-free rate was 38.0% and the overall survival rate was 58.7%. CONCLUSIONS: Petrosectomy is an effective treatment for malignant temporal bone invasion. The probability of a good outcome was statistically decreased with a high T grade, positive margins, and salvage surgery. Younger age is connected with better prognosis. One of the major tasks remains to improve detection and to shorten the time to diagnosis, keeping in mind that symptoms are insidious and in younger people, the time before diagnosis was longer.