Background: HPV-related HNSCC has been associated with an improved prognosis in patients treated with RT +/- CT; however, no information is available regarding patients treated with RT + EGFR inhib. Our purpose was to detect differences in outcome in patients treated with RT+EGFR inhib according to p16 tumor status. Methods: We retrospectively determined DNA of HPV 16 and immunohistochemical expression of p16 from formalin-fixed, paraffin-embedded (FFPE) tissue tumors from 78 patients with newly diagnosed locally advanced (LA)-HNSCC treated with RT + CT (37) and RT + EGFR inhib (41), entered in clinical trials. Tumors were classified dichotomously as either p16-positive (strong, diffuse staining) or negative, and DNA of HPV 16 was determined by multiplex polymerase chain reaction (PCR) methods. Disease free survival (DFS) and overall survival (OS) were analysed by Kaplan-Meyer method. Results: DNA of HPV 16 was found in 12 of 78 tumors (15%) and p16 positivity in 11 tumors (14%) with similar rates in both arms. p16 positive patients were less likely to be smokers than p16 negative patients (p < 0.01). After a median follow-up of 22 months (2-108), p16 positive patients showed a trend for better OS compared with p16-negative (median OS 58 vs 39 months; p = 0.4; HR 0.7; 95% CI 0.3 to 1.7). p16 positive patients treated with RT + EGFR inhib showed an improved survival compared with those treated with CT + RT (2-year OS 80% vs 60%; p = 0.03; HR 0.15; 95% CI 0.02 to 0.77 and 2–year DFS 80% vs 20%; p = 0.006; HR 0.11; 95%CI 0.02 to 0.61). However these differences were not observed in p16 negative patients (2-year OS 61% vs 57%; p = 0.9; HR 0.99; 95% CI 0.5 to 1.9 and 2-year DFS 41% vs 44%; p = 0.7; HR 1.07; 95% CI 0.6 to 2.0). Conclusions: This is the first study to report that p16 positive patients may benefit most from RT + EGFR inhibitors than conventional CT + RT. These results are hypothesis-generating and should be confirmed in prospective trials.