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Publication : CD4<sup>+</sup>CD8β<sup>+</sup> double-positive T cells in skin-draining lymph nodes respond to inflammatory signals from the skin.

First Author  Overgaard NH Year  2017
Journal  J Leukoc Biol Volume  102
Issue  3 Pages  837-844
PubMed ID  28637895 Mgi Jnum  J:254712
Mgi Id  MGI:5925745 Doi  10.1189/jlb.1AB0217-065R
Citation  Overgaard NH, et al. (2017) CD4+CD8beta+ double-positive T cells in skin-draining lymph nodes respond to inflammatory signals from the skin. J Leukoc Biol 102(3):837-844
abstractText  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 naive C57BL/6 mice. DP T cells were also present in Jalpha18-/- 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 ImageStream (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 approximately 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.
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