Abstract P5-01-08: Changes induced by neoadjuvant chemotherapy (NCT) in breast cancer tumor infiltrating lymphocytes (TIL) subpopulations are associated with chemo-sensitivity and prognosis
Authors: García-Martínez E , Luengo-Gil G , Chaves A , Gonzalez-Billalabeitia E , García García T , Vicente Conesa MA , García Garre E , de la Morena P , Vicente V , Ayala de la Peña F 
Published: August 2014

Abstract
Background: Recent data support the modulation of immune response in breast cancer microenvironment by neoadjuvant chemotherapy. This immune modulation might favor treatment response and modify post-treatment tumor immunosurveillance. The association of TIL with outcome after breast cancer (BC) NCT has also been suggested by several studies. Thereby, the differences in NCT-induced immune changes could reflect different profiles of immune modulation and translate into diverse prognosis. The objective of our study was to evaluate the predictive and prognostic value of NCT-induced TIL changes in patients with BC.
Methods: We analyzed a series of consecutive breast cancer patients treated with sequential anthracyclines and taxanes NCT (80.4% sequential AC-docetaxel). Pre- and post-chemotherapy biopsies were included in a tissue microarray and immunohistochemical (IHC) staining was performed for CD3, CD4, CD8, CD20, FOXP3 and CD68. After slide scanning and acquisition of digital images, computerized morphometric analysis (TIL count/mm2) was performed with ImageJ (NIH) software. Variations on pre- and post-NCT TIL infiltration were quantified and expressed as the absolute difference in number of cells/mm2 (postNCT TIL – preNCT TIL); median values were used as cut-point. Association of TIL changes with pCR was evaluated by logistic regression models. Kaplan-Meier analysis and Cox proportional hazard models were used for survival
Results: 121 patients with a diagnosis of invasive BC were included. Stages: IIB (28%), IIIA-C (56.4%). Phenotype was determined by immunohistochemistry: 50.4% Her2- hormone-sensitive (HS), 13.2% Her2+ HS, 10.7% Her2+ non-HS, and 21.5% triple negative. Pathologic complete response (pCR) rate was 17.4%. NCT produced a decrease in tumor infiltration by CD4 (p = 0.01), CD20 (p = 0.04) and CD68 (p = 0.03) and an increase in CD8 infiltrating lymphocytes (p = 0.0001). NCT-induced decreases of TIL infiltration above the median level of change were associated with pCR: CD3 (p = 0.01; odds ratio [OR]: 0.12, 95% confidence interval [95%CI] 0.02-0.63), CD4 (p = 0.02; OR 0.2; 95%CI 0.05-0.85) and CD20 (p = 0.01; OR 0.15; 95%CI 0.03-0.64). With a median follow-up of 60 months, median overall survival (OS) and disease free survival (DFS) has not been reached. A NCT-induced CD3 decrease higher than the median cut-off independently predicted higher DFS (hazard ratio [HR]:6.56; 95%CI 1.3-33.2; p = 0,02) and OS (HR:9.85; 95%CI 1.03-93.7, p = 0.04).
Conclusions: TIL variations induced by chemotherapy are related to pRC and prognosis in patients with invasive BC receiving anthracyclines and taxanes. These results suggest that the degree of immune modulation by NCT has an impact on survival and might be useful for prognostic classification in the setting of BC treated with NCT [Supported by GEICAM-Beca Ana Balil].
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-08.