Parallel assessment of Th17 cell frequencies by surface marker co-expression versus ex vivo IL-17 production in HIV-1 infection
| Autor: | Olaf Degen, Gabor Artur Dunay, Joachim Hauber, Eva Tolosa, Jan van Lunzen, Julian Schulze zur Wiesch, Ilona Tóth, Johanna M. Eberhard |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Histology medicine.diagnostic_test business.industry T cell hemic and immune systems Cell Biology CXCR3 Peripheral blood mononuclear cell Pathology and Forensic Medicine Flow cytometry 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure Immunology Medicine Interleukin 17 business Cytometry Viral load Ex vivo 030215 immunology |
| Zdroj: | Cytometry Part B: Clinical Cytometry. 90:486-492 |
| ISSN: | 1552-4949 |
| Popis: | INTRODUCTION Th17 cells can either be identified by co-staining of surface markers or by intracellular cytokine staining (ICS) for IL-17 production. Discrepancies regarding the published frequencies of Th17 cells in peripheral blood mononuclear cells (PBMC) of HIV patients may partly be due to the different methodologies used. METHODS Cryopreserved PBMC from healthy controls and HIV-infected subjects, including treated (cART) and viremic patients, were split and analyzed side-by-side by flow cytometry for expression of surface markers CCR6, CXCR3, CCR4, and CD161, or for intracellular expression of IL-17A and IFNγ after stimulation. RESULTS The characterization of Th17 cells as CXCR3 - CCR6 + CCR4 + CD161+ yielded considerably higher frequencies than the corresponding frequencies obtained by characterization via cytokines (IL-17 + IFNγ-), regardless of the HIV status. However, the overall frequencies delivered by the two methods significantly correlated. The relative frequency of Th17 cells within the CD4+ T cell compartment was preserved in HIV infection but there was a significant decrease in the absolute Th17 number, which was restored after initiation of cART, paralleling CD4+ T cell recovery. Absolute Th17 numbers inversely correlated with HIV viral load. CONCLUSION The definition of Th17 cells by surface markers might overestimate their frequency in comparison to functional assessment of IL-17 production by ICS, regardless of the HIV infection status. However, both methods yield proportionate results with reduced absolute numbers of Th17 cells in untreated HIV disease, reflecting the depletion of total CD4+ T cells in viremic HIV patients, and restoration with cART. © 2016 International Clinical Cytometry Society. |
| Databáze: | OpenAIRE |
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