We stained a large panel of human PBMCs with various markers, including CD3, CD4, CD8, CD25, CD38, CD45RA, CD127, CD161, as well as with several chemokine receptors CD183 (CXCR3), CD185 (CXCR5), CD194 (CCR4), CD196 (CCR6), CD197 (CCR7) and CCR10. Our original aim was to analyze the CD4+ T helper subsets, for which the expression of these markers are well defined and the gating tree is clear. Now, we would like to analyze also the CD8+ T cell subsets. However, we have a very hard time finding solid information about the expression of above markers on various Tc-subsets as most publications apparently focus on cytokine expression. All I got so far is Tc1 (CD183+ CD195+), Tc9 (CD196+), Tc17 (CD161+, CD195+ CD196+ and CD194-neg), nothing for Tc2.
Any suggestions on the gating strategy and any related references would be greatly appreciated!
The subsets found among CD8+ T cells do not compare directly to those found in CD4+ T cells. Tc1, of which I presume you mean type 1 cells (granzyme B, T-bet, IFNy etc.) would be the major line of differentiation, Tc2 (IL-4 production, GATA3 etc.) are found among CD8+ T cells but in low frequencies, which is also the case for type 17 CD8+ T cells. The subset division of CD8+ T cells is based on different definitions. Perhaps our review 'Blood and Beyond' could be of help. Good luck!
See below
BLOOD, 1 JULY 2002 VOLUME 100, NUMBER 1
Trafficking machinery of NKT cells: shared and differential chemokine receptor expression among Vα24+Vβ11+ NKT cell subsets with distinct cytokine-producing capacity
Chang H. Kim, Brent Johnston, and Eugene C. Butcher
Abstract
Natural killer T (NKT) cells are important regulators of the immune system, but their trafficking machinery, including expression of chemokine receptors, has been poorly defined. Unlike other conventional T-cell populations, we show that most NKT cells express receptors for extralymphoid tissue or inflammation-related chemokines (CCR2, CCR5, and CXCR3), while few NKT cells express lymphoid tissue–homing chemokine receptors (CCR7 and CXCR5). A population with homing potential for lymph nodes (L selectin+ CCR7+) exists only within a small subset of CD4 NKT cells. We show differential expression of chemokine receptors among NKT cell subsets: CCR4 is mainly expressed by a high cytokine (interleukin-4/interleukin-2)–producing (CD4) NKT subset, while CCR1, CCR6, and CXCR6 are preferentially expressed by the low cytokine-producing CD8 and CD4−CD8− subsets. In line with this, TARC/CCL17 (a CCR4 ligand) induces preferential chemotaxis of the CD4 NKT subset, while chemotactic activities of LARC/CCL20 (a CCR6 ligand) and MIP-1α/CCL3 (a CCR1 ligand) are focused on the CD8 and CD4−CD8− NKT cells. We conclude that, unlike conventional naive, memory, or effector T cells, the entire NKT cell population expresses nonlymphoid tissue homing chemokine receptors, yet NKT cell subsets differ considerably from each other by displaying distinct and reciprocal expression patterns of some chemokine receptors. Our results identify chemokine receptors that are potentially important for trafficking of human blood NKT cell subsets and reveal their function (cytokine production capacity)–dependent differential trafficking potentials.
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