Rhodamine 123: a useful probe for monitoring T cell activation

Cytometry
C FerliniA Fattorossi

Abstract

The T cell activation pathway involves an increase in mitochondrial activity. This can be evaluated in individual cells using the fluorescent probe rhodamine 123 (Rh123) and flow cytometry. Peripheral blood mononuclear cells (PBMC) were stimulated with optimal concentrations of phytohaemagglutinin (PHA), superantigens (Sag) SEA and SEC2, and allogeneic cells. Activation kinetics were followed at days 1, 2, 4 and 7. In all activation conditions, Rh123 uptake was augmented with the CD25 expression, cell size, and DNA synthesis. Rh123 uptake reflected an increase in mitochondrial activity and mass, as assessed by experiments in which Rh123 was substituted for by the 10-nonyl acridine orange, which stains mitochondria in an energy-independent manner. The spectral characteristics of Rh123 allowed us to double stain cells with Rh123 and phycoerythrin-conjugated monoclonal antibodies. In PHA-activated cultures, CD4+ and CD8+ cells incorporated essentially the same amount of Rh123 at all time points, suggesting that the two subsets did not differ in their activation kinetics. Accordingly, after 1 week of culture, no significant modification in the CD4/CD8 ratio was observed. Sag-activated CD4+ cells incorporated a higher amount of Rh12...Continue Reading

References

Jan 1, 1992·Cytometry·Z DarzynkiewiczF Traganos
Aug 1, 1992·European Journal of Immunology·R NisiniR D'Amelio
Feb 1, 1989·Immunological Reviews·C A JanewayS Buxser
Nov 1, 1980·Cytometry·J T ThornthwaiteW J Temple
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·Z DarzynkiewiczM R Melamed
Jan 1, 1982·Cold Spring Harbor Symposia on Quantitative Biology·L B ChenT J Lampidis
Feb 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·L V JohnsonL B Chen
May 1, 1981·The Journal of Cell Biology·T W James, R Bohman
Jun 3, 1994·Journal of Immunological Methods·W G TelfordP J Fraker
Feb 1, 1993·Seminars in Immunology·N R Gascoigne

❮ Previous
Next ❯

Citations

Jun 1, 1997·International Journal of Immunopharmacology·M VioraA Fattorossi
Dec 15, 2007·Nature Protocols·Cristiano Ferlini, Giovanni Scambia
Oct 29, 2000·European Journal of Biochemistry·B D FollstadG Stephanopoulos
Sep 6, 2000·Proceedings of the National Academy of Sciences of the United States of America·M HothR S Lewis
May 1, 2004·Environmental Toxicology and Pharmacology·Francesco TomeiEnrico Tomao
Feb 27, 1999·Experimental Cell Research·C FerliniA Fattorossi
Oct 25, 2017·Parasite Immunology·K Donskow-ŁysoniewskaM Doligalska
Sep 1, 2006·Current Protocols in Toxicology·Stacey E AndersonBarbara J Meade
May 4, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·A PazirandehS Okret
Jan 9, 2021·Journal of Cellular Biochemistry·Wojciech TrybusGrzegorz Król
Mar 7, 2021·Plants·Taras PasternakSerhii Kondratenko

❮ Previous
Next ❯

Related Concepts

Related Feeds

Allogenic & Autologous Therapies

Allogenic therapies are generated in large batches from unrelated donor tissues such as bone marrow. In contrast, autologous therapies are manufactures as a single lot from the patient being treated. Here is the latest research on allogenic and autologous therapies.