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Issue 7, 2015
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An infrared spectral signature of human lymphocyte subpopulations from peripheral blood

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Abstract

Metastatic melanomas are frequently refractory to most adjuvant therapies such as chemotherapies and radiotherapies. Recently, immunotherapies have shown good results in the treatment of some metastatic melanomas. Immune cell infiltration in the tumor has been associated with successful immunotherapy. More generally, tumor infiltrating lymphocytes (TILs) in the primary tumor and in metastases of melanoma patients have been demonstrated to correlate positively with favorable clinical outcomes. Altogether, these findings suggest the importance of being able to identify, quantify and characterize immune infiltration at the tumor site for a better diagnostic and treatment choice. In this paper, we used Fourier Transform Infrared (FTIR) imaging to identify and quantify different subpopulations of T cells: the cytotoxic T cells (CD8+), the helper T cells (CD4+) and the regulatory T cells (T reg). As a proof of concept, we investigated pure populations isolated from human peripheral blood from 6 healthy donors. These subpopulations were isolated from blood samples by magnetic labeling and purities were assessed by Fluorescence Activated Cell Sorting (FACS). The results presented here show that Fourier Transform Infrared (FTIR) imaging followed by supervised Partial Least Square Discriminant Analysis (PLS-DA) allows an accurate identification of CD4+ T cells and CD8+ T cells (>86%). We then developed a PLS regression allowing the quantification of T reg in a different mix of immune cells (e.g. Peripheral Blood Mononuclear Cells (PBMCs)). Altogether, these results demonstrate the sensitivity of infrared imaging to detect the low biological variability observed in T cell subpopulations.

Graphical abstract: An infrared spectral signature of human lymphocyte subpopulations from peripheral blood

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Supplementary files

Article information


Submitted
08 Dec 2014
Accepted
15 Dec 2014
First published
15 Dec 2014

Analyst, 2015,140, 2257-2265
Article type
Paper

An infrared spectral signature of human lymphocyte subpopulations from peripheral blood

N. Wald, A. Legat, C. Meyer, D. E. Speiser and E. Goormaghtigh, Analyst, 2015, 140, 2257
DOI: 10.1039/C4AN02247E

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