High-sensitivity immunofluorescence for detection of the pro- and anti-inflammatory cytokines gamma interferon and interleukin-10 on the surface of cytokine-secreting cells

Expression of the integrin αEβ7identifies unique subsets of CD25+as well as CD25−regulatory T cells

“…However, we cannot exclude that surfacebound TGF␤ might be resistant to antibody blockade, as has been described recently (27). This might also apply for the blockade of IL-10, because this cytokine can be detected on the surface of CD4 ϩ T cells as well (56). Indeed, suppressive effects of ␣ E ϩ T cells were found to be cell contact-dependent, as has been described for other regulatory populations (13).…”

How αβ T cells deal with induced TCRα ablation

“…However, the TCR␣-deficient cells did not contain detectable mRNA for the pT␣ chain as assessed by reverse transcription-PCR and are thus unlikely to express pre-TCR (data not shown; see also Materials and Methods). At low levels of antigen expression (in the range of 100 molecules per cell), liposome-based staining is no longer quantitative in that only a fraction of the cells may be hit by a liposomeconjugated antibody (15). The staining data in Fig.…”

“…Spleen or lymph node cells were stained in PBS containing 2% BSA and 0.2% of NaN 3 and analyzed on a FACS Calibur by using CELLQUEST software (Becton Dickinson). To increase sensitivity of TCR␤ staining, we used magnetofluorescent liposomes, as described previously (14)(15)(16). Briefly, cells were incubated with FITC-conjugated anti-TCR␤ mAbs (PharMingen) followed by digoxigenin (DIG)-conjugated anti-FITC mAbs and, finally, anti-DIG mAbs coupled with magnetofluorescent liposomes.…”

“…Although the staining of these cells was 2.5-3 orders of magnitude below that of wild-type cells, indicating loss of the vast majority of TCR complexes from the cell surface, conventional staining methods do not permit the detection of a small number of such complexes that might persist on the cell surface for extended periods of time. We therefore used a more sensitive staining method that makes use of antibodies conjugated to magnetofluorescent liposomes, increases sensitivity by 1-2 orders of magnitude, and has previously allowed the detection of membrane-bound proteins, which are below, or just at, the limit of detection by standard methods (15,16).…”

Ci8 short, a novel LPS-induced peptide from the ascidian Ciona intestinalis, modulates responses of the human immune system

“…The TCR-driven pathways that lead to proliferation appear separable to the secretion of cytokines, another event that by immune cells defines a significant part of their functional activity (Guy et al, 2013). Our hypothesis is that Ci8 short cytokines production is mediated by Effector Memory T cells (T EM ) (Farber, 2014) that can produce these immediately after antigen challenge with a rapid recall mechanism (Scheffold et al, 2000;Han et al, 2012;Campbell et al, 2010). These events are also mediated by TCR engagement and high ZAP-70 tyrosine kinase expression levels.…”