title: Objective measurement and clinical significance of TILs in non-small cell lung cancer.
abstract: BACKGROUND
  Tumor-infiltrating lymphocytes (TILs) are usually measured using subjective methods. Studies suggest that TIL subtypes have independent roles in cancer and that they could support the use of novel immunostimulatory therapies. We simultaneously measured TIL subtypes in non-small cell lung cancer (NSCLC) samples using objective methods and determined their relationship with clinico-pathologic characteristics and survival.
  
  
  METHODS
  Using multiplexed quantitative fluorescence (QIF), we measured the levels of CD3, CD8, and CD20 in 552 NSCLC from two independent collections represented in tissue microarrays (YTMA79, n = 202 and YTMA140, n = 350). The level of TILs was obtained in different tumor compartments using cytokeratin stain to define tumor cells and 4',6-Diamidino-2-Phenylindole. Association of TILs with clinical parameters was determined using univariate and multivariable analyses. All statistical tests were two-sided.
  
  
  RESULTS
  In both NSCLC collections there was a low correlation between the three TIL markers (linear regression coefficients (R(2)) = 0.19-0.22, P < .001 for YTMA79 and R(2) = 0.23-0.32, P < .001 for YTMA140). No consistent association between the level of TIL subtypes and age, sex, smoking history, tumor size, stage, and histology type was found. In univariate analysis, an elevated CD3 or CD8 signal was statistically significantly associated with longer survival in both collections. However, only CD8 was independent from age, tumor size, histology, and stage in multivariable analysis. High CD20 was associated with longer survival in the YTMA79 cohort.
  
  
  CONCLUSIONS
  Increased levels of CD3 and CD8 + TILs are associated with better outcome in NSCLC, but only CD8 is independent from other prognostic variables. Objective measurement of TIL subpopulations could be useful to predict response or evaluate the local immune effect of anticancer immune checkpoint inhibitors.
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS). kurt.schalper@yale.edu.
authors: Kurt A
authors: KA
authors: Schalper
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Jason
authors: J
authors: Brown
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Daniel
authors: D
authors: Carvajal-Hausdorf
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Joseph
authors: J
authors: McLaughlin
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Vamsidhar
authors: V
authors: Velcheti
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Konstantinos N
authors: KN
authors: Syrigos
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: Roy S
authors: RS
authors: Herbst
authors: Department of Pathology (KAS, JB, DCH, DLR) and Medical Oncology (JM, RSH), Yale University School of Medicine, New Haven, CT; Solid Tumor Oncology, Cleveland Clinic, Cleveland, OH (VV); Oncology Unit, 3rd Department of Medicine, School of Medicine, University of Athens, Sotiria General Hospital, Athens, Greece (KNS).
authors: David L
authors: DL
authors: Rimm
mesh_terms: D018951:Antigens, CD20; D017252:CD3 Complex; D016827:CD8 Antigens; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology* / Q000401:mortality; D015986:Confounding Factors, Epidemiologic; D005456:Fluorescent Dyes; D006801:Humans; D007211:Indoles; D053208:Kaplan-Meier Estimate; D008175:Lung Neoplasms / Q000276:immunology* / Q000401:mortality; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008856:Microscopy, Fluorescence; D011237:Predictive Value of Tests; D012189:Retrospective Studies; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D018951:Antigens, CD20; D017252:CD3 Complex; D016827:CD8 Antigens; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI
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