Fig. 1

Long-term clinical and translational studies from CAVE-Lung trial patients. A. Among the patients enrolled in CAVE-Lung trial, 3 patients had received a previous line of therapy with single agent anti-PD-1 mAb: patient #2, pembrolizumab; patients #9 and #11, nivolumab. Left panel. Response to previous anti-PD-1 therapy: patient #2 obtained partial response (PR) as best radiological response with PFS of 6 months; patients #9 and #11 had stable disease (SD) with PFS of 7 and 7 months, respectively. Right panel. Patients were enrolled in CAVE-Lung trial and received cetuximab plus avelumab treatment, all obtaining SD as best radiologic response with PFS of 15, 19, and 34+ months, respectively. For these 3 patients, collection of PMBC during treatment for analysis of NK cell activation was done at 10, 12, and 15 months. B. LDH release assay in vitro was performed to assess NK cell activation from patient-derived PBMC samples. For the patient with the longest response (34 months, ongoing), LDH levels were constantly higher as compared to baseline, while in the other two patients, LDH levels were high and started to decline at the time (15 months) of progression of disease (PD) or close to PD. C, D. FACS analysis was performed to further evaluate the effects on immune cell populations, that could be induced by cetuximab plus avelumab treatment. At the time of clinical response, CD107A+ cells increased, suggesting NK cell degranulation (C); while TIM3+ and PD-L1+ immune-suppressive cells decreased, suggesting T cell activation (D). Results were inverted at the time of PD. P-values were calculated by ANOVA test. **p < 0.01, *p < 0.05