Background: Pre-operative neutrophil-to-lymphocyte ratio (NLR) has been found to be associated with adverse pathological results and poor long-term outcomes in patients treated with radical cystectomy (RC) for urothelial carcinoma (UC). We aimed to evaluate the predictive utility of NLR in patients treated with neoadjuvant chemotherapy (NAC) and RC for high-risk UC. Methods: We reviewed the records of 585 patients treated with NAC and RC at our institution from 2000-2013. We calculated NLR before initiation of NAC (pre-chemo NLR) and during the recovery window between NAC and RC (post-chemo NLR).. We excluded patients with concomitant infection, blood disorder, or second malignancy. We used univariate and multivariate CART models to determine the optimal NLR cut-off for survival outcomes. We estimated disease-specific (DSS) and overall survival (OS) using the Kaplan-Meier method. We used Cox proportional hazards regression to explore the association of NLR with DSS and OS. Results: 584 patients had NLR information in our cohort. The median follow-up among survivors was 4.9 years (IQR 2.4 – 8.8 years). We identified optimal NLR cut-points of 7.1 for pre-chemo, 4.9 for post-chemo, and 1.9 for change in NLR [(post-chemo) – (pre-chemo)]. Post-chemo NLR showed the strongest association with OS and DSS. Patients with a post-chemo NLR ≥ 4.9 (n = 103) had a 5-yr DSS and OS of 42% and 33% respectively, compared to 69% and 58% for patients with an NLR < 4.9 (n = 481). In the multivariable analysis, post-chemo NLR ≥ 4.9 was an independent predictor of DSS (HR = 2.5 [95% CI:1.8, 3.6] p < 0.001 ), and OS (HR = 2.1 [95% CI:1.6, 2.8] p < 0.001). Conclusions: A post-chemo NLR ≥ 4.9 is associated with poor DSS and OS in patients treated with NAC and RC. These findings may help guide treatment planning for adjuvant therapy following RC in patients with high-risk clinically localized bladder cancer.