Abstract A120: The infiltration of Th17 cells in tumor microenvironment negatively correlate with the prognosis of the patients with head and neck squamous cell carcinoma

Abstract Introduction: Head and Neck squamous cell carcinoma (HNSCC) remains to the malignancies with the unsure prognosis. Regarding the etiology, we are facing different clinical manifestation, behavior and overall survival. This fact can be explained by the differences in the immune response of the patients. Understanding of the basic principles of the immune system of HNSCC patients would help us to better combine the therapeutical strategies with the respect to the immune cell subpopulations and their regulation. One of the most important cells are Th17 lymphocytes, which are increased in the peripheral blood and the microenvironment of many types of cancers. Methods: 178 patients with primary HNSCC were included to the study. Samples of the peripheral blood and from the primary tumors, metastatic lymph nodes and non-affected lymph nodes were collected at the moment of surgical therapy and measured by flow cytometry. The data were analyzed by the Cox regression and compared with the group of healthy blood donors. Sixty six tumors out of 178 primary tumors were localized in the tonsillar region, 50 in the root of the tongue, 33 in the larynx and 8 in hypopharynx. The average follow-up was 40 months. Results: For the prognosis of the patients the most important oncological parameters were T and N stadium of the disease (p = 0.02, resp. p = 0.03), radicality of the surgery (p = 0.006), presence of the adverse prognostic risk factors (p = 0.01). From the analyzed parameters of the immune system we found negative prognostic correlation of the level of B lymphocytes (CD19+; p = 0.03) and the positive prognostic correlation of the level of naïve CD4+ T cells (CD4+CD45RA+, p = 0.02) in the peripheral blood of the patients. We found strong negative prognostic correlation of the Th17 cells (CD4+CD161+) in the peripheral blood (p = 0.04) and also in the tumor microenvironment (p = 0.005). Conclusion: We can conclude that the examination of parameters of immune system could be helpful for better understanding of biological behavior and interaction of the HNSCC. The level of Th17 cells in both periphery and in the tumor microenvironment correlate with the worst survival of the patients with the HNSCC. The analyses of the Th17 level in the peripheral blood before the therapy could help us with the prediction of the cause of the disease. The future studies should be focused on the changes in immune cells infiltration in response to the different therapeutical strategies in HNSCC oncology. Acknowledgments: The research was supported by AZV MZ CR (grant No. NV16-28594A and NV16-28600A). Note: This abstract was not presented at the conference. Citation Format: Jan Boucek, Michal Zabrodsky. The infiltration of Th17 cells in tumor microenvironment negatively correlate with the prognosis of the patients with head and neck squamous cell carcinoma [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr A120.</jats:p

Abstract B85: The relation between Treg and cancer stem cells in microenvironment of head and neck squamous cell carcinoma.

Abstract Head and neck squamous cell carcinoma (HNSCC) remains one of the most difficult problems in the current oncology. Overall survival of the patients have not improved in more than 30 years and have not reached approximately 50% at 5 years for all stages. The combination of surgery with radiation or chemotherapy resulted in limited improvement in survival rates, although there was shown the efficacy of concomitant chemo-radiation in organ preservation. On the other hand, identification of some biological or immunological markers (e.g. HPV, Treg) allowed us to better understand the behavior of the disease. Cancer stem-cell (CSC) theory of tumorigenesis has been established during the last decade. HNSCC is histologically heterogeneous and in tumor microenvironment plays important role also diverses cancer cells, stromal cells and infiltrating inflammatory cells. But only a small subpopulation of CSCs has been identified to be responsible to development and the growth of solid tumors. Several surface epitopes have been published as distinguishing markers for CSCs identification. The most often mentioned are CD44+, CD29+, CD24+ and CD133+ markers. Regulatory T cells (Treg, CD4+CD25+Foxp3+) were described as one of the critical factors for regulation or inhibition of effective anti-cancer immune response and, consequently, the prognosis and survival for different types of tumors, including HNSCC. There were 119 patients with HNSCC included in the study from September 2010. Out of 119 primary tumors 50 were localized in the tonsillar region, 42 in the root of the tongue, 20 in the larynx and 7 in hypopharynx. All patients underwent primary surgical therapy, in dependence on the final pathological staging 15 underwent adjuvant chemo-radiation, 73 adjuvant radiation and 31 had no adjuvant therapy. Follow-up of all of them has been provided on author´s department. The presence of the immunocompetent cells in the tumor microenvironment of specimens from primary tumors, from metastases to the neck lymph nodes and from control lymph nodes was correlated with the numbers of HNSCC cancer stem cells (CD44+, CD29+, CD133+) in a subgroup of patients (68), where samples were taken during the surgery. The number of Treg in specimens from primary tumors was significantly higher than from metastatic neck lymph nodes or in neck lymph nodes without metastatic extension (5.53%, SD 4.12 vs. 3.96%, SD 2.64 vs. 4.9%, SD 3.31, p=0.01). CD44+ or CD29+ or CD133+ cancer stem cells were detected in the tumor microenvironment and the numbers of all of these cells were dramatically higher in samples from primary tumors than from metastases or from control lymph nodes (CD44+: 47.1%, SD 26.4 vs. 28.3%, SD 30.2 vs. 6.5%, SD 16.4, p=0.0002; CD29+: 18.3%, SD 21.7 vs. 12.0%, SD 14.1 vs. 4.4%, SD 12.1, p=0.0028; CD133+: 6.1%, SD 8.7 vs. 3.6%, SD 4.8 vs. 0.6%, SD 0.8, p=0.0002). There was a significant correlation of Treg infiltration with percentage of CD44+ cancer cells (Treg high vs. Treg low group, CD44+ 54.4%, SD 31.3 vs. 40.4%, SD 20.4, p= 0.04) in the samples of primary tumors. We can conclude that examination of these parameters could be helpful for better understanding of biological behavior of the HNSCC and could help us with prognostication and choice of treatment strategy in HNSCC patients. Acknowledgments: The research was supported by IGA MZ CR (grant No. 11542). Citation Format: Jan Boucek, Michal Zabrodsky, Jan Betka, Tomas Eckschlager, Blanka Rihova. The relation between Treg and cancer stem cells in microenvironment of head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B85.</jats:p

The Role of Narrow Band Imaging in the Detection of Recurrent Laryngeal and Hypopharyngeal Cancer after Curative Radiotherapy

Narrow band imaging is considered a significant improvement in the possibility of detecting early mucosal lesion of the upper aerodigestive tract. Early detection of mucosal neoplastic lesions is of utmost importance for patients survival. There is evidence that, especially in patients previously treated by means of curative radiotherapy or chemoradiotherapy, the early detection rate of recurrent disease is quite low. The aim of this study was to prove whether the videoendoscopy coupled with NBI might help detect recurrent or secondary tumors of the upper aerodigestive tract. 66 patients previously treated by means of RT or CRT with curative intent were enrolled in the study. All patients underwent transnasal flexible videoendoscopy with NBI mode under local anesthesia. When a suspicious lesion was identified in an ambulatory setting, its nature was proved histologically. Many of these changes were not identifiable by means of conventional white light (WL) endoscopy. The accuracy, sensitivity, specificity, and positive and negative predictive value of the method are very high (88%, 92%, 76%, 96%, and 91%, resp.). Results demonstrate that outpatient transnasal endoscopy with NBI is an excellent method for the follow-up of patients with carcinomas of the larynx and the hypopharynx primarily treated with radiotherapy

Abstract B94: The role of Th17 cells in the head and neck squamous cell carcinoma evolution.

Abstract Head and neck squamous cell carcinoma (HNSCC) is the most frequent histotype of the mucosal malignant tumor in this region. Many new diagnostic and therapeutic modalities have been implemented in the diagnostic and treatment protocols however all of them failed to prove a significantly improved overall survival of patient. Surgery combined with radiotherapy and chemotherapy remains a mainstay of the current treatment approaches. A better understanding of the tumor behavior and its immunological profile and microenvironment (e.g. HPV, Treg) allowed us to modify the treatment strategy. Regulatory T cells (Treg, CD4+CD25+Foxp3+) were described as one of the critical factors for regulation or inhibition of effective anti-cancer immune response. Increased Treg frequency in the peripheral circulation was proven to correlate with the progression HNSCC. In many cancers a synergy of Treg and Th17 (CD4+161+) has been shown. So far, little is known about the importance of Tc17 (CD8+161+) in HNSCC. Particularly, the relationship between Tc17 and tonsillar cancers could be very interesting, because the CCL20 receptor (ligand for CCR6 overexpressed on Tc17) was also frequently found in tonsillar epithelia. There were 119 patients with HNSCC included in the study from September 2010. Out of 119 primary tumors 50 were localized in the tonsillar region, 42 in the root of the tongue, 20 in the larynx and 7 in hypopharynx. All patients underwent primary surgical therapy, in dependence on the final pathological staging 15 underwent adjuvant chemo-radiation, 73 adjuvant radiation and 31 had no adjuvant therapy. We examined periphery blood from all patients with the focus on lymphocyte subpopulation (CD3+, CD4+CD25+, CD4+/CD8+, CD19+, CD4+CD45RA+, CD8+CD28-, CD3-CD16+CD56+, CD4+CD25+Foxp3+, CD4+161+, CD8+161+) before the start of anti-tumor therapy. The infiltration of immunocompetent cells in the tumor microenvironment of specimens from primary tumors, from metastases to the neck lymph nodes and from control lymph nodes was measured in a subgroup of patients (68), where samples were taken during the surgery. The correlation of Treg and Th17 resp. Tc17, was analyzed with a special interest. The percentage of Th17 cells (CD4+CD161+) in peripheral blood does not differ in patients with HNSCC (5.8%, SD 2.5) and in healthy controls (5.6%, SD 3.5, p = 0.46) and also the presence of Tc17 (CD8+CD161+) in peripheral blood shows the same pattern in both groups (7.0%, SD 5.5 vs. 8.0%, SD 4.3, p = 0.31). The percentage of Th17 cells detected in primary tumors was significantly much higher than in metastatic neck lymph nodes or in neck lymph nodes without metastatic extension (7.1%, SD 7.2 vs. 4.2%, SD 3.0 vs. 4.8%, SD 2.0, p=0.01 resp. p=0.03). There was no difference in the infiltration of Tc17 cells, nor was also observed increased infiltration by Tc17 tonsillar carcinomas compared with other localizations of HNSCC. Very interesting and highly significant was the correlation of Treg infiltration with percentage of Th17 (Treg high vs. Treg low group, CD4+161+: 5.14%, SD 4.63 vs. 8.99%, SD 8.6, p= 0.03) in the samples of primary tumors. Our results show that Th17 are increasingly detected in the primary HNSCC. Their importance for the prognosis of patients could be analyzed over a longer period of time. Acknowledgments: The research was supported by IGA MZ CR (Grant No. 11544 and 11542). Citation Format: Michal Zabrodsky, Jan Betka, Tomas Eckschlager, Blanka Rihova, Jan Boucek. The role of Th17 cells in the head and neck squamous cell carcinoma evolution. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B94.</jats:p

Group-level Regional Cerebral Uptake Quantification in Micro PET-CT

Abstract The lack of soft tissue CT contrast is an impediment to accurate quantification of regional cerebral uptake on micro PET-CT. Co-registration to an atlas can aid with quantitative analysis, particularly when MRI is not available. Methods: A subject CT is cropped to the skull, the brain void is then filled to create a “skull cavity mask”. An MRI mouse brain volumetric template is then co-registered to the skull cavity mask via affine transformation. Each subsequent subject CT is then skull cropped and co-registered to the first, these transformations are then applied to each corresponding PET for group level co-registration. Results: The method was tested on eight mice with each possible registration permutation; resulting in small variations in regional cerebral PET counts. Conclusion: We present a semi-automated method that allows for quantification of regional cerebral PET uptake from micro PET-CT images without subject MRI data.</jats:p