Y consists of V5+ T cells, when the Junctional Adhesion Molecule A (JAM-A) Proteins custom synthesis dermal compartment comprises high frequencies of V4+ and V6+ T cells (Fig. 107). It follows that an more counterstaining of 17D1+ skin T cells with a distinct anti-V5 mAb clone 536, see Table 21, would further aid to discriminate in between dermal and and TCRhigh epidermal T cells (Fig. 107B and not shown). In contrast, peripheral lymph nodes lack V5+ T cells. Even though V6+ T cells only represent a tiny population in peripheral lymph nodes, a big proportion of T cells are V4+ T cells and V6-V4- T cells (primarily V1+ T cells).Author Manuscript 1.1.eight.Murine NKT cellsOverview Murine organic killer T (NKT) cells have been initially defined by their co-expression of surface markers characteristic for T cells (i.e., the TCR) and NK cells (e.g., NK1.1 in C57BL/6 mice) [815, 816]. This chapter focuses on the phenotypic characterization of so-called murine invariant iNKT cells, which express an invariant V14J18 TCR chain plus a restricted set of TCR chains using a preference for V8, V7, and V2 [817, 818]. iNKT cells recognize lipids, which include -galactosyl ceramide (GalCer), within the context from the nonclassical MHC molecule CD1d [819]. As a consequence, iNKT cells is often unambiguously identified by surface staining applying CD1d tetramers loaded with GalCer or its derivatives, including PBS-57 [820, 821] (Fig. 108). Subphenotyping of developmental stages within the thymus and effector subsets determined by surrogate surface markers and key transcription things is described.Author Manuscript Author Manuscript Author Manuscript1.eight.Introduction Development of iNKT cells diverges at the CD4+CD8+ double-positive stage of T-cell development. Choice of iNK T cells is mediated by cortical thymocytes instead of epithelial cells. Comparable to other unconventional T cells, iNKT cells are selected by sturdy TCR signals in a procedure known as agonist selection [822]. iNKT cells, using the notable exception of some tissue-resident subsets, express and are dependent on the prototypical transcription issue for innate-like T cells, PLZF (encoded by Zbtb16) [823, 824]. Intrathymic development of iNKT cells has initially been described to progress by way of 4 phenotypically distinct stages (stage 0), characterized by differential expression in the surface markers CD24, CD44, and NK1.1 (in C57BL/6 mice) too as cell size [825827] (Fig. 109A). Far more recent research showed that stage three iNKT cells represent long-term resident cells within the thymus [828, 829]. The thymus of young adult C57BL/6 mice consists of around 3 105 iNKT cells, corresponding to an all round frequency of 0.three.5 of all thymocytes. Additional recently, iNKT cells have already been IL-12 alpha Proteins Source categorized into functional subsets based on expression of sort 1, two, or 17 cytokines [830] (Fig. 109B). Like their standard T-cell counterparts,Eur J Immunol. Author manuscript; obtainable in PMC 2020 July 10.Cossarizza et al.PageNKT1 cells are characterized by expression with the transcription element T-bet, NKT17 cells express RORt, whereas NKT2 cells are most frequently characterized by absence of expression of each transcription things while simultaneously expressing very high levels of PLZF (See Chapter VI Section 1.1 Murine T cells). The prototypic sort two transcription factor GATA-3 is variably expressed in all iNKT cells and cannot be employed for discrimination of NKT2 cells. As a consequence, in the thymus PLZFhi NKT cells include both, precursors (NKTp) and NKT2 cells. These cells is usually further distinguis.