A novel method for isolation of human lung T cells from lung resection tissue reveals increased expression of GAPDH and CXCR6

C.E. Day, Shu-Dong Zhang, J. Riley, T. Gant, A.J. Wardlaw, C. Guillen

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Lung T lymphocytes are important in pulmonary immunity and inflammation. it has been difficult to study these cells due to contamination with other cell types, mainly alveolar macrophages. We have developed a novel method for isolating lung T cells from lung resection tissue, using a combination of approaches. Firstly the lung tissue was finely chopped and filtered through a nylon mesh. Lymphocytic cells were enriched by Percoll density centrifugation and the T cells purified using human CD3 microbeads, resulting in 90.5% +/- 1.9% (n = 11) pure lymphocytes. The T cell yield from the crude cell preparation was 10.8 +/- 2.1% and viability, calculated using propidium iodide (PI) staining and trypan blue, was typically over 95%. The purification process did not affect expression of CD69 or CD103, nor was there a difference in the proportion of CD4 and CD8 cells between the starting population and the purified cells. Microarray analysis and real time RT-PCR revealed upregulation of GAPDH and CXCR6 of the lung T cells as compared to blood-derived T cells. This technique highly enriches lung T cells to allow detailed investigation of the biology of these cells. (C) 2008 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)91-97
Number of pages7
JournalJournal of Immunological Methods
Volume342
Issue number1-2
DOIs
Publication statusPublished - 15 Mar 2009

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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