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BACKGROUND: CD8+ tumor-infiltrating lymphocytes (TILs) have emerged as a prognostic indicator in triple-negative breast cancer (TNBC). There is debate surrounding the prognostic value of hot spots for CD8+ TIL enumeration. METHODS: We compared hot spot versus whole-tumor CD8+ TIL enumeration in prognosticating TNBC using immunohistochemistry on whole tissue sections and quantification by digital image analysis (Halo imaging analysis software; Indica Labs, Corrales, NM). A wide range of clinically relevant hot spot sizes was evaluated. RESULTS: CD8+ TIL enumeration was independently statistically significant for all hot spot sizes and whole-tumor annotations for disease-free survival by multivariate analysis. A 10× objective (2.2 mm diameter) hot spot was found to correlate significantly with overall survival (P = .04), while the remaining hot spots and whole-tumor CD8+ TIL enumeration did not (P > .05). Statistical significance was not demonstrated when comparing between hot spots and whole-tumor annotations, as the groups had overlapping confidence intervals. CONCLUSION: CD8+ TIL hot spot enumeration is equivalent to whole-tumor enumeration for prognostication in TNBC and may serve as a good alternative methodology in future studies and clinical practice.

Original publication




Journal article


Clin Breast Cancer

Publication Date





451 - 458.e1


Carcinoma, TIL, Adult, Aged, Aged, 80 and over, Carcinoma, Ductal, Breast, Carcinoma, Lobular, Cohort Studies, Combined Modality Therapy, Female, Follow-Up Studies, Humans, Image Processing, Computer-Assisted, Lymphocytes, Tumor-Infiltrating, Middle Aged, Prognosis, Survival Rate, Triple Negative Breast Neoplasms