Journal article
CD4+CD8β+ double-positive T cells in skin-draining lymph nodes respond to inflammatory signals from the skin
National Veterinary Institute, Technical University of Denmark1
Adaptive Immunology, Division of Immunology & Vaccinology, National Veterinary Institute, Technical University of Denmark2
T-cells & Cancer, Division of Immunology & Vaccinology, National Veterinary Institute, Technical University of Denmark3
University of Queensland4
University of Melbourne5
CD4+CD8+ double-positive (DP), mature, peripheral T cells are readily detectable in a variety of species and tissues. Despite a common association with autoimmune and malignant skin disorders, however, little is understood about their role or function. Herein, we show that DP T cells are readily detectable in the blood, spleen, and peripheral lymph nodes of naïve C57BL/6 mice.
DP T cells were also present in Jα-/- and CD1d-/- mice, indicating that these cells are not NK-T cells. After skin administration of CASAC adjuvant, but not Quil A adjuvant, both total DP T cells and skin-infiltrating DP T cells increased in number. We explored the possibility that DP T cells could represent aggregates between CD4+ and CD8+ single-positive T cells and found strong evidence that a large proportion of apparent DP T cells were indeed aggregates.
However, the existence of true CD4+CD8+ DP T cells was confirmed by Amnis Image-Stream (Millipore Sigma, Billerica, MA, USA) imaging. Multiple rounds of FACS sorting separated true DP cells from aggregates and indicated that conventional analyses may lead to ~10-fold overestimation of DP T cell numbers.
The high degree of aggregate contamination and overestimation of DP abundance using conventional analysis techniques may explain discrepancies reported in the literature for DP T cell origin, phenotype, and function.
Language: | English |
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Publisher: | Society for Leukocyte Biology |
Year: | 2017 |
Pages: | 837-844 |
ISSN: | 19383673 and 07415400 |
Types: | Journal article |
DOI: | 10.1189/jlb.1AB0217-065R |
ORCIDs: | Overgaard, Nana Haahr |