Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining

Joshua Saylor, Zhaoxuan Ma, Helen S. Goodridge, Fangjin Huang, Anne E Cress, Stephen J. Pandol, Stephen L. Shiao, Adriana C. Vidal, Lily Wu, Nicholas G. Nickols, Arkadiusz Gertych, Beatrice S. Knudsen

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

An array of phenotypically diverse myeloid cells and macrophages (MC&M) resides in the tumor microenvironment, requiring multiplexed detection systems for visualization. Here we report an automated, multiplexed staining approach, named PLEXODY, that consists of five MC&M-related fluorescently-tagged antibodies (anti - CD68, - CD163, - CD206, - CD11b, and - CD11c), and three chromogenic antibodies, reactive with high- and low-molecular weight cytokeratins and CD3, highlighting tumor regions, benign glands and T cells. The staining prototype and image analysis methods which include a pixel/area-based quantification were developed using tissues from inflamed colon and tonsil and revealed a unique tissue-specific composition of 14 MC&M-associated pixel classes. As a proof-of-principle, PLEXODY was applied to three cases of pancreatic, prostate and renal cancers. Across digital images from these cancer types we observed 10 MC&M-associated pixel classes at frequencies greater than 3%. Cases revealed higher frequencies of single positive compared to multi-color pixels and a high abundance of CD68+/CD163+ and CD68+/CD163+/CD206+ pixels. Significantly more CD68+ and CD163+ vs. CD11b+ and CD11c+ pixels were in direct contact with tumor cells and T cells. While the greatest percentage (~70%) of CD68+ and CD163+ pixels was 0-20 microns away from tumor and T cell borders, CD11b+ and CD11c+ pixels were detected up to 240 microns away from tumor/T cell masks. Together, these data demonstrate significant differences in densities and spatial organization of MC&M-associated pixel classes, but surprising similarities between the three cancer types.

Original languageEnglish (US)
Number of pages1
JournalFrontiers in Immunology
Volume9
DOIs
StatePublished - Jan 1 2018

Fingerprint

Myeloid Cells
Macrophages
Staining and Labeling
T-Lymphocytes
Neoplasms
Tumor Microenvironment
Kidney Neoplasms
Palatine Tonsil
Masks
Keratins
Pancreatic Neoplasms
Anti-Idiotypic Antibodies
Prostatic Neoplasms
Colon
Color
Molecular Weight
Antibodies

Keywords

  • FFPE
  • immunofluorescence
  • immunohistochemistry
  • macrophage
  • multiplex
  • myeloid
  • spatial profiling

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining. / Saylor, Joshua; Ma, Zhaoxuan; Goodridge, Helen S.; Huang, Fangjin; Cress, Anne E; Pandol, Stephen J.; Shiao, Stephen L.; Vidal, Adriana C.; Wu, Lily; Nickols, Nicholas G.; Gertych, Arkadiusz; Knudsen, Beatrice S.

In: Frontiers in Immunology, Vol. 9, 01.01.2018.

Research output: Contribution to journalArticle

Saylor, J, Ma, Z, Goodridge, HS, Huang, F, Cress, AE, Pandol, SJ, Shiao, SL, Vidal, AC, Wu, L, Nickols, NG, Gertych, A & Knudsen, BS 2018, 'Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining', Frontiers in Immunology, vol. 9. https://doi.org/10.3389/fimmu.2018.02925
Saylor, Joshua ; Ma, Zhaoxuan ; Goodridge, Helen S. ; Huang, Fangjin ; Cress, Anne E ; Pandol, Stephen J. ; Shiao, Stephen L. ; Vidal, Adriana C. ; Wu, Lily ; Nickols, Nicholas G. ; Gertych, Arkadiusz ; Knudsen, Beatrice S. / Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining. In: Frontiers in Immunology. 2018 ; Vol. 9.
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