title: Automated prognostic pattern detection shows favourable diffuse pattern of FOXP3(+) Tregs in follicular lymphoma.
abstract: BACKGROUND
  Histopathological prognostication relies on morphological pattern recognition, but as numbers of biomarkers increase, human prognostic pattern-recognition ability decreases. Follicular lymphoma (FL) has a variable outcome, partly determined by FOXP3 Tregs. We have developed an automated method, hypothesised interaction distribution (HID) analysis, to analyse spatial patterns of multiple biomarkers which we have applied to tumour-infiltrating lymphocytes in FL.
  
  
  METHODS
  A tissue microarray of 40 patient samples was used in triplex immunohistochemistry for FOXP3, CD3 and CD69, and multispectral imaging used to determine the numbers and locations of CD3(+), FOXP3/CD3(+) and CD69/CD3(+) T cells. HID analysis was used to identify associations between cellular pattern and outcome.
  
  
  RESULTS
  Higher numbers of CD3(+) (P=0.0001), FOXP3/CD3(+) (P=0.0031) and CD69/CD3(+) (P=0.0006) cells were favourable. Cross-validated HID analysis of cell pattern identified patient subgroups with statistically significantly different survival (35.5 vs 142 months, P=0.00255), a more diffuse pattern associated with favourable outcome and an aggregated pattern with unfavourable outcome.
  
  
  CONCLUSIONS
  A diffuse pattern of FOXP3 and CD69 positivity was favourable, demonstrating ability of HID analysis to automatically identify prognostic cellular patterns. It is applicable to large numbers of biomarkers, representing an unsupervised, automated method for identification of undiscovered prognostic cellular patterns in cancer tissue samples.
authors.affiliation: Medical School, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
authors.forename: Lilli S
authors.initials: LS
authors.lastname: Nelson
authors.affiliation: Perkin-Elmer, 68 Elm Street, Hopkinton, Massachusetts 01748, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Mansfield
authors.affiliation: Perkin-Elmer, 68 Elm Street, Hopkinton, Massachusetts 01748, USA.
authors.forename: Roslyn
authors.initials: R
authors.lastname: Lloyd
authors.affiliation: Institute of Cancer Sciences, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Oguejiofor
authors.affiliation: Department of Medical Oncology, The Christie Foundation NHS Trust, 550 Wilmslow Rd, Manchester M20 4BX, UK.
authors.forename: Zena
authors.initials: Z
authors.lastname: Salih
authors.affiliation: Department of Histopathology, The Christie Foundation NHS Trust, 550 Wilmslow Rd, Manchester M20 4BX, UK.
authors.forename: Lia P
authors.initials: LP
authors.lastname: Menasce
authors.affiliation: Institute of Cancer Sciences, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.|Department of Medical Oncology, The Christie Foundation NHS Trust, 550 Wilmslow Rd, Manchester M20 4BX, UK.
authors.forename: Kim M
authors.initials: KM
authors.lastname: Linton
authors.affiliation: Institute of Population Health, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.|Manchester Academic Health Science Centre, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Rose
authors.affiliation: Institute of Cancer Sciences, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.|Manchester Academic Health Science Centre, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.|Department of Histopathology, Manchester Royal Infirmary, Oxford Road, Manchester M13 9WL, UK.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Byers
mesh_terms: D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D005260:Female; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism; D016246:Lymphocytes, Tumor-Infiltrating / Q000378:metabolism / Q000473:pathology; D008224:Lymphoma, Follicular / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D014408:Biomarkers, Tumor; D017252:CD3 Complex; D051858:Forkhead Transcription Factors; D037181:Lectins, C-Type; D000099192:CD69 Antigens; C418974:FOXP3 protein, human