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

By C.E. Day, S.D. Zhang, J. Riley, T. Gant, A.J. Wardlaw and C. Guillen

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

Topics: Research Paper
Publisher: Elsevier
OAI identifier: oai:pubmedcentral.nih.gov:2666793
Provided by: PubMed Central
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