Human peripheral blood mononuclear cell culture for flow cytometric analysis of phosphorylated mitogen-activated protein kinases
Lymphocyte activation and fine tuning of downstream signaling circuits for the regulation of cytokine expression are critical for a successful immune response. Hence, technical protocols permitting simultaneous testing of these attributes in peripheral blood lymphocytes are of paramount importance. Phospho-specific flow cytometry is a novel methodology that detects phosphorylation of signaling effectors in multiple, rare cellular populations within peripheral blood. In addition, it allows the quantification of phosphorylation levels for signaling proteins within each single cell, and therefore is superior compared to traditional biochemical approaches, such as Western blotting. One such important signaling pathway within immune cells is the p38 MAPK pathway involved in the regulation of cytokine expression, cell proliferation and apoptosis. In this chapter, we provide technical instructions for culturing human peripheral blood lymphocytes for simultaneous monitoring of p38 MAPK phosphorylation and associated cytokine expression, especially in rare cell populations, such as NK and NKT. © 2012 Springer Science+Business Media, LLC.
Showing items related by title, author, creator and subject.
The creative potential of medium-sized and small Greek cities: Reflections on contemporary cultural strategies Konsola, D.; Karachalis, N. (2010)As the creative city discourse has come to the centre of cultural policy debates, its rhetoric often neglects smaller cities and emphasises the cases of large cities that employ culture as a driver for economic growth. ...
Hepper, E. G.; Wildschut, T.; Sedikides, C.; Ritchie, T. D.; Yung, Y. F.; Hansen, N.; Abakoumkin, G.; Arikan, G.; Cisek, S. Z.; Demassosso, D. B.; Gebauer, J. E.; Gerber, J. P.; González, R.; Kusumi, T.; Misra, G.; Rusu, M.; Ryan, O.; Stephan, E.; J. Vingerhoets, A. J.; Zhou, X. (2014)Nostalgia is a frequently experienced complex emotion, understood by laypersons in the United Kingdom and United States of America to (a) refer prototypically to fond, self-relevant, social memories and (b) be more pleasant ...
Efforts to create an artificial testis: Culture systems of male germ cells under biochemical conditions resembling the seminiferous tubular biochemical environment Soffikitis, N.; Pappas, E.; Kawatani, A.; Baltogiannis, D.; Loutradis, D.; Kanakas, N.; Giannakis, D.; Dimitriadis, F.; Tsoukanelis, K.; Georgiou, I.; Makrydimas, G.; Mio, Y.; Tarlatzis, V.; Melekos, M.; Miyagawa, I. (2005)Induction of meiotic and post-meiotic alterations of male germ cells in vitro has been the target of several research efforts since 1960. However, to date, the establishment of an ideal culture system in which spermatogonial ...