Multiplexed analysis of serum cytokines as biomarkers in squamous cell carcinoma of the head and neck patients

Objective: Our objective was to characterize cytokine profiles in the systemic circulation of patients with active squamous cell carcinoma of the head and neck (SCCHN) compared with long-term tobacco smokers as controls. Furthermore, we hypothesized that persistent immune dysregulation in patients cured of their disease may be reflected in altered cytokine profiles in the systemic circulation. Study Design: Retrospective, case-control study. Methods: Using three well-defined clinical cohorts of SCCHN patients and smoker controls, we analyzed 100 μL of serum for a panel of 10 cytokines to determine whether serum cytokine profiles could distinguish clinically defined groups of SCCHN patients. Statistical analysis of multiplexed cytokine profiles was applied to classify three clinically defined groups: active SCCHN patients, treated SCCHN patients with no evidence of disease for over 3 years, and matched disease-free controls. Discrimination of outcome status was accomplished using classification trees, and 10-fold cross-validation was implemented to assess classification accuracy using independent data. Results: We show that multiplexed cytokine and chemokine profiling may be performed to reflect the immune status of SCCHN patients. Selected cytokine profiles indicate that immunologic responses to carcinogenesis may not normalize even in the absence of tumor for over 3 years. Conclusion: Multiplexed serum cytokine profiles may be applicable to early detection, for screening those at high risk for SCCHN, and as clinically predictive biomarkers of disease status in successfully treated patients.