Nicholas D. Stafford

Serum IL10 and circulating CD4+CD25high regulatory T cell numbers as predictors of clinical outcome and survival in patients with head and neck squamous cell carcinoma

Patients with head and neck squamous cell carcinoma (HNSCC) commonly have an imbalance in T helper (Th)1/Th2‐type cytokines and elevated levels of CD4+CD25high regulatory T cells (Treg). Here, we investigated the association of circulating interleukin (IL)10, IL12, and Treg‐cells with clinical outcome in patients with HNSCC.

Unraveling the Chromosomal Aberrations of Head and Neck Squamous Cell Carcinoma: A Review

Information from the genetic analysis of head and neck cancer has grown enormously in the last 20 years. The advent of high-resolution genetic analysis techniques such as microarray technology will further expand this field in the future. Here we review the data on chromosomal aberrations of head and neck squamous cell carcinoma, focusing on the data generated by comparative genomic hybridization analysis, and suggest how such findings will be taken forward over the next decade. With the search engine PUBMED, the key words “comparative genomic hybridisation,” “head and neck,” “oral,” “hypopharyngeal,” “laryngeal,” and “squamous cell carcinoma” were used. Publications unavailable in English were excluded.

Increased frequency and suppressive activity of CD127low/− regulatory T cells in the peripheral circulation of patients with head and neck squamous cell carcinoma are associated with advanced stage and nodal involvement

The presence of regulatory T (Treg) cells is thought to be an important mechanism by which head and neck squamous cell carcinoma (HNSCC) successfully evades the immune system. Using multicolour flow cytometry, the frequency and functional capacity of two CD4+ CD127low/− Treg cell populations, separated on the basis of different levels of CD25 expression (CD25inter and CD25high), from the peripheral circulation of newly presenting HNSCC patients were assessed with regard to clinicopathological features and healthy controls. The frequency of circulating Treg cells was similar between HNSCC patients and healthy controls, and for patients with HNSCC developing from different subsites (laryngeal compared with oropharyngeal). However, patients with advanced stage tumours and those with nodal involvement had significantly elevated levels of CD4+ CD25high CD127low/− Treg cells compared with patients who had early stage tumours (P = 0·03) and those without nodal involvement (P = 0·03), respectively. CD4+ CD25high CD127low/− Treg cells from the entire HNSCC patient cohort and from patients whose tumours had metastasized to the lymph nodes were also shown to suppress the proliferation of effector T cells significantly more, compared with those from healthy controls (P = 0·04) or patients with no nodal involvement (P = 0·04). Additionally, CD4+ CD25inter CD127low/− Treg cells consistently induced greater suppressive activity than CD4+ CD25high CD127low/− Treg cells on the proliferation of the effector T‐cell populations (CD4+ CD25− CD127−/+ and CD4+ CD25+ CD127+). Peripheral Treg cells, identified by the CD127low/− phenotype, have been shown to be influenced by a patients tumour stage and/or nodal status in HNSCC; suggesting a role in tumour progression that could be manipulated by future immunotherapy.

Clinical relevance of immune parameters in the tumor microenvironment of head and neck cancers.

The tumour microenvironment is a highly complex region where multiple interactions occur between host and cancer cells. The host response is defined by the presence and function of different tumour infiltrating lymphocytes and cytokines. Head and neck squamous cell carcinomas comprise a subgroup of human cancers with significant morbidity and mortality.

Prognostic value of the neutrophil-to-lymphocyte ratio in patients with laryngeal squamous cell carcinoma.

The neutrophil/lymphocyte ratio (NLR) has been found to be predictive of survival outcome in a range of tumors. The purpose of this study was to investigate the prognostic value of pretreatment (NLR) in patients with laryngeal squamous cell carcinoma (SCC).

Analysis of Radiation-Induced Cell Death in Head and Neck Squamous Cell Carcinoma and Rat Liver Maintained in Microfluidic Devices

Objective The aim of this study was to investigate how head and neck squamous cell carcinoma (HNSCC) tissue biopsies maintained in a pseudo in vivo environment within a bespoke microfluidic device respond to radiation treatment. Study Design Feasibility study. Setting Tertiary referral center. Subjects and Methods Thirty-five patients with HNSCC were recruited, and liver tissue from 5 Wistar rats was obtained. A microfluidic device was used to maintain the tissue biopsy samples in a viable state. Rat liver was used to optimize the methodology. HNSCC was obtained from patients with T1-T3 laryngeal or oropharyngeal SCC; N1-N2 metastatic cervical lymph nodes were also obtained. Irradiation consisted of single doses of between 2 Gy and 40 Gy and a fractionated course of 5×2 Gy. Cell death was assessed in the tissue effluent using the soluble markers lactate dehydrogenase (LDH) and cytochrome c and in the tissue by immunohistochemical detection of cleaved cytokeratin18 (M30 antibody). Results A significant surge in LDH release was demonstrated in the rat liver after a single dose of 20 Gy; in HNSCC, it was seen after 40 Gy compared with the control. There was no significant difference in cytochrome c release after 5 Gy or 10 Gy. M30 demonstrated a dose-dependent increase in apoptotic index for a given increase in single-dose radiotherapy. There was a significant increase in apoptotic index between 1×2 Gy and 5×2 Gy. Conclusion M30 is a superior method compared with soluble markers in detecting low-dose radiation-induced cell death. This microfluidic technique can be used to assess radiation-induced cell death in HNSCC and therefore has the potential to be used to predict radiation response.

Assessment of dendritic cell number and radiosensitivity in laryngeal tumours

Objectives:u2002 Radiotherapy is one of the principal treatment modalities for many types of head and neck tumour; what effects the dendritic cell (DC) population may have on treatment outcome have not been critically evaluated in laryngeal cancer.

Measuring the response of human head and neck squamous cell carcinoma to irradiation in a microfluidic model allowing customized therapy

Radiotherapy is the standard treatment for head and neck squamous cell carcinoma (HNSCC), however, radioresistance remains a major clinical problem despite significant improvements in treatment protocols. Therapeutic outcome could potentially be improved if a patients tumour response to irradiation could be predicted ex vivo before clinical application. The present study employed a bespoke microfluidic device to maintain HNSCC tissue whilst subjecting it to external beam irradiation and measured the responses using a panel of cell death and proliferation markers. HNSCC biopsies from five newly-presenting patients [2xa0lymph nodexa0(LN); 3xa0primary tumour (PT)] were divided into parallel microfluidic devices and replicates of each tumour were subjected to single-dose irradiation (0, 5, 10, 15 and 20xa0Gy). Lactate dehydrogenasexa0(LDH) release was measured and tissue sections were stained for cytokeratin (CK), cleaved-CK18 (cCK18), phosphorylated-H2AX (γH2AX) and Ki‑67 by immunohistochemistry. In addition, fragmented DNA was detected using terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). Compared with non‑irradiated controls, higher irradiation doses resulted in elevated CK18-labelling index in two lymph nodes [15xa0Gy; 34.8% on LN1 and 31.7% on LN2 (p=0.006)] and a single laryngeal primary tumour (20xa0Gy; 31.5%; p=0.014). Significantly higher levels of DNA fragmentation were also detected in both lymph node samples and one primary tumour but at varying doses of irradiation, i.e.,xa0LN1 (20xa0Gy; 27.6%; p=0.047), LN2 (15xa0Gy; 15.3%; p=0.038) and PT3 (10xa0Gy; 35.2%; p=0.01). The γH2AX expression was raised but not significantly in the majority of samples. The percentage of Ki‑67 positive nuclei reduced dose-dependently following irradiation. In contrast no significant difference in LDH release was observed between irradiated groups and controls. There is clear inter- and intra-patient variability in response to irradiation when measuring a variety of parameters, which offers the potential for the approach to provide clinically valuable information.