title: Quantitative, architectural analysis of immune cell subsets in tumor-draining lymph nodes from breast cancer patients and healthy lymph nodes.
abstract: BACKGROUND
  To date, pathological examination of specimens remains largely qualitative. Quantitative measures of tissue spatial features are generally not captured. To gain additional mechanistic and prognostic insights, a need for quantitative architectural analysis arises in studying immune cell-cancer interactions within the tumor microenvironment and tumor-draining lymph nodes (TDLNs).
  
  
  RESULTS
  We present a novel, quantitative image analysis approach incorporating 1) multi-color tissue staining, 2) high-resolution, automated whole-section imaging, 3) custom image analysis software that identifies cell types and locations, and 4) spatial statistical analysis. As a proof of concept, we applied this approach to study the architectural patterns of T and B cells within tumor-draining lymph nodes from breast cancer patients versus healthy lymph nodes. We found that the spatial grouping patterns of T and B cells differed between healthy and breast cancer lymph nodes, and this could be attributed to the lack of B cell localization in the extrafollicular region of the TDLNs.
  
  
  CONCLUSIONS
  Our integrative approach has made quantitative analysis of complex visual data possible. Our results highlight spatial alterations of immune cells within lymph nodes from breast cancer patients as an independent variable from numerical changes. This opens up new areas of investigations in research and medicine. Future application of this approach will lead to a better understanding of immune changes in the tumor microenvironment and TDLNs, and how they affect clinical outcomes.
authors.affiliation: Department of Medicine, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: A Francesca
authors.initials: AF
authors.lastname: Setiadi
authors.forename: Nelson C
authors.initials: NC
authors.lastname: Ray
authors.forename: Holbrook E
authors.initials: HE
authors.lastname: Kohrt
authors.forename: Adam
authors.initials: A
authors.lastname: Kapelner
authors.forename: Valeria
authors.initials: V
authors.lastname: Carcamo-Cavazos
authors.forename: Edina B
authors.initials: EB
authors.lastname: Levic
authors.forename: Sina
authors.initials: S
authors.lastname: Yadegarynia
authors.forename: Chris M
authors.initials: CM
authors.lastname: van der Loos
authors.forename: Erich J
authors.initials: EJ
authors.lastname: Schwartz
authors.forename: Susan
authors.initials: S
authors.lastname: Holmes
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lee
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016175:B-Lymphocyte Subsets / Q000378:metabolism*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000276:immunology* / Q000473:pathology*; D016022:Case-Control Studies; D002452:Cell Count; D006801:Humans; D008198:Lymph Nodes / Q000276:immunology*; D008207:Lymphatic Metastasis; D008875:Middle Aged; D057054:Molecular Imaging / Q000379:methods*; D011379:Prognosis; D016176:T-Lymphocyte Subsets / Q000378:metabolism*
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