School of Medicine and Health Sciences Poster Presentations

Title

The Role of SUMOylation in human Th1 and Th2 responses

Poster Number

282

Document Type

Poster

Status

Medical Student

Abstract Category

Immunology/Infectious Diseases

Keywords

SUMOylation, Th1, Th2, Immunology, HIV

Publication Date

Spring 2018

Abstract

In order to study this phenomenon, naïve human CD4 T cells will be isolated from PBMCs of health donors and activated with IL-12 or IL-4 in order to induce a TH1 or a TH2 response respectively (objective #1). This activation will be done in the presence of 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one (HODHBt), a benzotriazole which was previously been characterized by the Bosque lab to have the highest activity blocking the SUMOylation of STAT5. Levels of phosphorylation of STAT4 and STAT6 will be measured by flow cytometry and western blot using phospohspecific antibodies (objective #2). TH1 and TH2 responses will be measured by intracellular cytokine staining (TH1: staining for IFN-gamma, TH2: staining for IL-4) after stimulation with PMA and Ionomycin (objective #3). Further characterization of the cytokine expression profiles of these cells will be done by a TH Cytokine Panel from Biolegend® and subsequent flow cytometry (Objective #4). The panel which includes IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-21, IL-22, IFN-γ and TNF-α, will enable us to distinguish between the production of a TH1, TH2, TH9, TH17, TH22 and TFH response. A detailed analysis of the data collected from this study will contribute to a greater understanding of the role of SUMOylation in the control TH1 and TH2 responses with respect to STAT4 and STAT6 activation (Objective #5)

The results of my research have yet to be fully elucidated, however, preliminary analysis seems to be inconclusive.

This study may require further work in order to understand the role of HODHBT in STAT4 and STAT6 Sumoylation, however, as previously stated, the results of my research have yet to be fully elucidated.

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The Role of SUMOylation in human Th1 and Th2 responses

In order to study this phenomenon, naïve human CD4 T cells will be isolated from PBMCs of health donors and activated with IL-12 or IL-4 in order to induce a TH1 or a TH2 response respectively (objective #1). This activation will be done in the presence of 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one (HODHBt), a benzotriazole which was previously been characterized by the Bosque lab to have the highest activity blocking the SUMOylation of STAT5. Levels of phosphorylation of STAT4 and STAT6 will be measured by flow cytometry and western blot using phospohspecific antibodies (objective #2). TH1 and TH2 responses will be measured by intracellular cytokine staining (TH1: staining for IFN-gamma, TH2: staining for IL-4) after stimulation with PMA and Ionomycin (objective #3). Further characterization of the cytokine expression profiles of these cells will be done by a TH Cytokine Panel from Biolegend® and subsequent flow cytometry (Objective #4). The panel which includes IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-21, IL-22, IFN-γ and TNF-α, will enable us to distinguish between the production of a TH1, TH2, TH9, TH17, TH22 and TFH response. A detailed analysis of the data collected from this study will contribute to a greater understanding of the role of SUMOylation in the control TH1 and TH2 responses with respect to STAT4 and STAT6 activation (Objective #5)

The results of my research have yet to be fully elucidated, however, preliminary analysis seems to be inconclusive.

This study may require further work in order to understand the role of HODHBT in STAT4 and STAT6 Sumoylation, however, as previously stated, the results of my research have yet to be fully elucidated.