Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Clin Oncol. Oct 10, 2016; 7(5): 387-394
Published online Oct 10, 2016. doi: 10.5306/wjco.v7.i5.387
Tumor infiltrating lymphocytes in triple negative breast cancer receiving neoadjuvant chemotherapy
Carlos A Castaneda, Elizabeth Mittendorf, Sandro Casavilca, Yun Wu, Miluska Castillo, Patricia Arboleda, Teresa Nunez, Henry Guerra, Carlos Barrionuevo, Ketty Dolores-Cerna, Carolina Belmar-Lopez, Julio Abugattas, Gabriela Calderon, Miguel De La Cruz, Manuel Cotrina, Jorge Dunstan, Henry L Gomez, Tatiana Vidaurre
Carlos A Castaneda, Miluska Castillo, Patricia Arboleda, Ketty Dolores-Cerna, Carolina Belmar-Lopez, Department of Research, Instituto Nacional de Enfermedades Neoplasicas, Lima 15038, Peru
Carlos A Castaneda, Henry L Gomez, Tatiana Vidaurre, Department of Medical Oncology, Instituto Nacional de Enfermedades Neoplasicas, Lima 15038, Peru
Elizabeth Mittendorf, Department of Surgical Oncology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, United States
Sandro Casavilca, Patricia Arboleda, Teresa Nunez, Henry Guerra, Carlos Barrionuevo, Department of Pathology, Instituto Nacional de Enfermedades Neoplasicas, Lima 15038, Peru
Yun Wu, Department of Pathology, Division of Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, United States
Julio Abugattas, Gabriela Calderon, Miguel De La Cruz, Manuel Cotrina, Jorge Dunstan, Department of Breast Cancer Surgery, Instituto Nacional de Enfermedades Neoplasicas, Lima 15038, Peru
Author contributions: Castaneda CA, Mittendorf E, Casavilca S and Wu Y contributed to the conception and design of the study and performed data analysis and interpretation; Castaneda CA, Casavilca S, Castillo M, Nunez T and Guerra H performed data acquisition, as well as providing administrative, technical, and material support; all authors drafted the article and made critical revisions related to the intellectual content of the manuscript, and approved the final version of the article to be published.
Institutional review board statement: This study was reviewed and approved by the Instituto Nacional de Enfermedades Neoplasicas Institutional Review Board.
Informed consent statement: Not needed due to the non-clinical study design.
Conflict-of-interest statement: To our knowledge, no conflict of interest exists.
Data sharing statement: No data were created so no data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Carlos A Castaneda, MD, MSc, Department of Medical Oncology, Instituto Nacional de Enfermedades Neoplasicas, Av. Angamos Este 2520 Surquillo, Lima 15038, Peru. carloscastanedaaltamirano@yahoo.com
Telephone: +51-1-2157220 Fax: +51-1-6204991
Received: April 18, 2016
Peer-review started: April 18, 2016
First decision: June 12, 2016
Revised: August 13, 2016
Accepted: September 21, 2016
Article in press: September 23, 2016
Published online: October 10, 2016
Processing time: 173 Days and 9.7 Hours
Abstract
AIM

To determine influence of neoadjuvant-chemotherapy (NAC) over tumor-infiltrating-lymphocytes (TIL) in triple-negative-breast-cancer (TNBC).

METHODS

TILs were evaluated in 98 TNBC cases who came to Instituto Nacional de Enfermedades Neoplasicas from 2005 to 2010. Immunohistochemistry staining for CD3, CD4, CD8 and FOXP3 was performed in tissue microarrays (TMA) sections. Evaluation of H/E in full-face and immunohistochemistry in TMA sections was performed in pre and post-NAC samples. STATA software was used and P value < 0.05 was considered statistically significant.

RESULTS

Higher TIL evaluated in full-face sections from pre-NAC tumors was associated to pathologic-complete-response (pCR) (P = 0.0251) and outcome (P = 0.0334). TIL evaluated in TMA sections showed low level of agreement with full-face sections (ICC = 0.017-0.20) and was not associated to pCR or outcome. TIL in post-NAC samples were not associated to response or outcome. Post-NAC lesions with pCR had similar TIL levels than those without pCR (P = 0.6331). NAC produced a TIL decrease in full-face sections (P < 0.0001). Percentage of TIL subpopulations was correlated with their absolute counts. Higher counts of CD3, CD4, CD8 and FOXP3 in pre-NAC samples had longer disease-free-survival (DFS). Higher counts of CD3 in pre-NAC samples had longer overall-survival. Higher ratio of CD8/CD4 counts in pre-NAC was associated with pCR. Higher ratio of CD4/FOXP3 counts in pre-NAC was associated with longer DFS. Higher counts of CD4 in post-NAC samples were associated with pCR.

CONCLUSION

TIL in pre-NAC full-face sections in TNBC are correlated to longer survival. TIL in full-face differ from TMA sections, absolute count and percentage analysis of TIL subpopulation closely related.

Keywords: Triple-negative breast cancer; Survival; Tumor-infiltrating lymphocytes; Neoadjuvant therapy

Core tip: We evaluated a series of 98 triple negative breast cancer cases who received neoadjuvant chemotherapy. Pre-neoadjuvant chemotherapy and post neoadjuvant chemotherapy samples were analyzed. We compared tumor-infiltrating lymphocytes evaluated in whole slides vs in tissue microarray slides. We also compared subsets of tumor-infiltrating-lymphocytes (TILs) evaluated through an absolute counting vs a percentage calculation. Our results confirm the predictive and prognostic role of tumor infiltrating lymphocytes, and evaluated the behavior of tumor infiltrating lymphocytes and lymphocyte subsets during chemotherapy in triple negative breast cancer. Our results increase the understanding of methodological issues for TIL evaluation as well as provide information about variation of the whole TIL population and TIL subpopulation during chemotherapy.