THE PLANT HORMONE ZEATIN RIBOSIDE INHIBITS T LYMPHOCYTE ACTIVITY Rebecca R. Wise, Cody J. Lloyd and Courtney M. Lappas Lebanon Valley College, Annville, PA Results Background Cytokinins are a class of phytohormones that are involved in multiple aspects of plant growth and development. Naturally occurring cytokinins are predominantly adenine derivatives, and almost all cytokinins exist in plants as both a free base and corresponding nucleosides and nucleotides. Although cytokinin-binding proteins have been identified in mammalian sera, the biological functions of cytokinins and underlying mechanisms of action in mammalian systems remain largely uncharacterized. Interestingly, the cytokinin zeatin riboside has been shown to activate adenosine A 2A receptor (A 2A R) signaling in a mammalian neuronal cell line, eliciting subsequent PKA-dependent neuroprotective effects. Given the adenosine-based Objectives In this study we sought to determine if zeatin riboside effects the pro- inflammatory activities and/or viability of murine T lymphocytes Conclusions We show that concentrations of Zeatin Riboside under 1000 µM are not lethal to T lymphocytes. Overall, the production of cytokines TNF-α, IFN-γ, and IL-2 is Methods T lymphocytes were isolated from C57BL/6 mice and used to look at cell viability and cytokine production in presence of Zeatin Riboside. To find percent viability, Trypan Blue was added after a 24 hour incubation of T lymphocytes along with various concentrations of Zeatin Riboside. Detecting the production of cytokines TNF-α, IFN-γ, and IL-2 in T lymphocytes was accomplished through first activating the cells via Anti-CD3. An ELISA assay was then used to monitor the amount produced when concentration of Zeatin Riboside is varied. Percent Viability of Murine T Lymphocytes in Presence of Zeatin Riboside: Various concentrations of Zeatin Riboside cultured with T lymphocytes aided in finding a nonlethal concentration of Zeatin Riboside. After 24 hours, Trypan Blue was added to determine percentage of cells still alive. Concentration of TNF-α Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti- CD3. An ELISA assay was utilized to detect the amount of TNF-α produced. Concentration of IFN-γ Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti- CD3. An ELISA assay was utilized to detect the amount of IFN-γ produced. Concentration of IL-2 Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti-CD3. An ELISA assay was utilized to detect the amount of IL-2 produced.

Zeatin Riboside- inquiry 2014

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THE PLANT HORMONE ZEATIN RIBOSIDE INHIBITS T LYMPHOCYTE ACTIVITY

Rebecca R. Wise, Cody J. Lloyd and Courtney M. LappasLebanon Valley College, Annville, PA

ResultsBackground

Cytokinins are a class of phytohormones that are involved in multiple aspects of plant growth and development. Naturally occurring cytokinins are predominantly adenine derivatives, and almost all cytokinins exist in plants as both a free base and corresponding nucleosides and nucleotides. Although cytokinin-binding proteins have been identified in mammalian sera, the biological functions of cytokinins and underlying mechanisms of action in mammalian systems remain largely uncharacterized. Interestingly, the cytokinin zeatin riboside has been shown to activate adenosine A2A receptor (A2AR) signaling in a mammalian neuronal cell line, eliciting subsequent PKA-dependent neuroprotective effects. Given the adenosine-based structure of zeatin riboside, its activity as an A2AR agonist is not entirely surprising; however, it does endow the compound with significant therapeutic potential because the activity of virtually all inflammatory cells is modulated by the selective activation of the Gs-coupled A2AR, with multiple manifestations of inflammatory cell activation inhibited by A2AR agonist treatment.

Objectives In this study we sought to determine if zeatin riboside effects the pro-inflammatory activities and/or viability of murine T lymphocytes

ConclusionsWe show that concentrations of Zeatin Riboside under 1000 µM are not lethal to T lymphocytes. Overall, the production of cytokines TNF-α, IFN-γ, and IL-2 is inhibited as concentration of Zeatin Riboside is increased.

MethodsT lymphocytes were isolated from C57BL/6 mice and used to look at cell viability and cytokine production in presence of Zeatin Riboside. To find percent viability, Trypan Blue was added after a 24 hour incubation of T lymphocytes along with various concentrations of Zeatin Riboside. Detecting the production of cytokines TNF-α, IFN-γ, and IL-2 in T lymphocytes was accomplished through first activating the cells via Anti-CD3. An ELISA assay was then used to monitor the amount produced when concentration of Zeatin Riboside is varied.

Percent Viability of Murine T Lymphocytes in Presence of Zeatin Riboside: Various concentrations of Zeatin Riboside cultured with T lymphocytes aided in finding a nonlethal concentration of Zeatin Riboside. After 24 hours, Trypan Blue was added to determine percentage of cells still alive.

Concentration of TNF-α Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti-CD3. An ELISA assay was utilized to detect the amount of TNF-α produced.

Concentration of IFN-γ Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti-CD3. An ELISA assay was utilized to detect the amount of IFN-γ produced.

Concentration of IL-2 Produced by Murine T Lymphocytes after Activation and Addition of Zeatin Riboside: T Lymphocytes were activated through the use of Anti-CD3. An ELISA assay was utilized to detect the amount of IL-2 produced.