Characterisation of lymphocyte subpopulations in infantile haemangioma

J Clin Pathol. 2015 Oct;68(10):812-8. doi: 10.1136/jclinpath-2015-203073. Epub 2015 Jun 11.

Abstract

Aims: Interstitial CD45+ cells and T lymphocytes have previously been demonstrated within infantile haemangioma (IH). This study investigated the expression of B and T lymphocyte markers by the CD45+ population, and the expression of Thy-1, a marker of thymocyte progenitors, which have the ability to give rise to both B and T cells.

Methods: Immunohistochemical (IHC) staining was performed on proliferating and involuted IHs for the expression of CD45, CD3, CD20, CD79a, Thy-1 and CD34. The presence of mRNA corresponding to CD45, CD3G, CD20 and Thy-1 was confirmed by reverse transcriptase-polymerase chain reaction in snap-frozen IH tissues. Cell counting of 3,3-diaminobenzidine IHC-stained slides was performed on CD45+ only cells and dually stained CD45+/CD3+ cells or CD45+/CD20+ cells and analysed statistically. In situ hybridisation and mass spectrometry were also performed to confirm the presence and abundance of Thy-1, respectively.

Results: IHC staining showed a subpopulation of CD45+ interstitial cells that expressed the T lymphocyte marker, CD3, and another subpopulation that expressed the B lymphocyte marker, CD20, in proliferating and diminished in involuted IHs. The abundant expression of Thy-1 on the endothelium of proliferating, but not involuted IH, was demonstrated by IHC staining and confirmed by in situ hybridisation and mass spectrometry.

Conclusions: Both B and T lymphocytes are present within the interstitium of proliferating and involuted IH. The expression of Thy-1 by the endothelium suggests that B and T cells in IH may have originated from within the lesion, rather than migrating from the peripheral circulation.

Keywords: HAEMATOLOGY; IMMUNOHISTOCHEMISTRY; LYMPHOCYTES; PAEDIATRIC PATHOLOGY; VASCULAR TUMOURS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD20 / analysis
  • B-Lymphocyte Subsets / immunology*
  • B-Lymphocyte Subsets / pathology
  • Biomarkers, Tumor / analysis*
  • Biomarkers, Tumor / genetics
  • CD3 Complex / analysis
  • Cell Lineage
  • Cell Proliferation
  • Endothelial Cells / immunology
  • Endothelial Cells / pathology
  • Hemangioma / genetics
  • Hemangioma / immunology*
  • Hemangioma / pathology
  • Humans
  • Immunohistochemistry
  • Immunophenotyping
  • In Situ Hybridization
  • Infant
  • Leukocyte Common Antigens / analysis
  • Leukocyte Common Antigens / genetics
  • Phenotype
  • RNA, Messenger / analysis
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / pathology
  • Tandem Mass Spectrometry
  • Thy-1 Antigens / analysis
  • Thy-1 Antigens / genetics

Substances

  • Antigens, CD20
  • Biomarkers, Tumor
  • CD3 Complex
  • RNA, Messenger
  • Thy-1 Antigens
  • Leukocyte Common Antigens
  • PTPRC protein, human