Quantifying soluble inflammatory factors in common marmoset (Callithrix jacchus) plasma
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Biology
Other
Biology
Advisor Information
Biology, Assistant Professor
Location
MBSC Ballroom Poster # 605 - U
Presentation Type
Poster
Start Date
24-3-2023 10:30 AM
End Date
24-3-2023 11:45 AM
Abstract
Quantifying soluble inflammatory factors in common marmoset (Callithrix jacchus) plasma
Donald DJ Rogers1, Aliyah Jabenis1, Mackenzie Conrin1,2, Shivdeep S. Hayer1,2, Haley R. Hassenstab2, Noel D. Johnson3,
Jonathan B. Clayton1,2,4, and Paul W. Denton1
1Department of Biology, University of Nebraska at Omaha
2Callitrichid Research Center, University of Nebraska at Omaha
3Department of Comparative Medicine, University of Nebraska Medical Center
4Nebraska Food for Health Center, University of Nebraska-Lincoln
Background
The complex interaction between the gut, the brain, and the immune system is believed to influence depression. Thus, this gut/brain/immune axis is important and should be carefully examined, particularly in the context of clinical interventions that can alter one or more components of this axis. Specifically, antibiotics can change the gut microbiome which can, in turn, change the brain’s chemical state and, potentially, lead to onset of depression. To study this in humans is more difficult than in animal models. For this reason, we are working with the UNO marmoset colony to decipher impacts of antibiotics on emotional states. Because immune communication is a bedrock of the gut/brain/immune axis and many immune communication molecules are inflammatory signals, we needed to develop an approach that can show the range and intensity of expression. This project focuses on developing a reliable approach for measuring soluble marmoset inflammatory molecules (e.g., cytokines, chemokines).
Methods
To detect and quantify immune analytes present in marmoset blood samples (UNO Callitrichid Research Center), we are utilizing electrochemiluminescent multiplex analysis according to the manufacturer’s instruction (Mesoscale Diagnostics). To obtain sample for analyses, we first isolate peripheral blood mononuclear cells (PBMCs) from whole blood. Next, we stimulate the PBMCs to produce inflammatory analytes. For this, we employed multiple stimulation strategies: cell activation cocktail (CAC), mixed lymphocyte reaction (MLR), or lipopolysaccharide stimulation (LPS). Culture supernatants were evaluated in the multiplex assay. The data was then analyzed through Microsoft Excel using techniques provided by Mesoscale Diagnostics.
Results
Our preliminary data showed that the assay we used, which was designed for human analyte detection, was cross-reactive for 5 out of 10 marmoset inflammatory analytes analyzed. The marmoset analytes that can be detected are IFN-g, IL-13, IL-1B, IL-4, and IL-8. These are our focus going forward.
Discussion
Our main goal in this project is to be able to link behavioral observations to inflammatory analyte levels. To help achieve this goal, our next steps are to validate that marmoset inflammatory analyte detection remains consistent when the cytokines are present in marmoset plasma as the biological matrix. When that is complete, we will have an experimental framework that will help us achieve our project goal.
Scheduling
10:45 a.m.-Noon, 1-2:15 p.m., 2:30 -3:45 p.m.
Quantifying soluble inflammatory factors in common marmoset (Callithrix jacchus) plasma
MBSC Ballroom Poster # 605 - U
Quantifying soluble inflammatory factors in common marmoset (Callithrix jacchus) plasma
Donald DJ Rogers1, Aliyah Jabenis1, Mackenzie Conrin1,2, Shivdeep S. Hayer1,2, Haley R. Hassenstab2, Noel D. Johnson3,
Jonathan B. Clayton1,2,4, and Paul W. Denton1
1Department of Biology, University of Nebraska at Omaha
2Callitrichid Research Center, University of Nebraska at Omaha
3Department of Comparative Medicine, University of Nebraska Medical Center
4Nebraska Food for Health Center, University of Nebraska-Lincoln
Background
The complex interaction between the gut, the brain, and the immune system is believed to influence depression. Thus, this gut/brain/immune axis is important and should be carefully examined, particularly in the context of clinical interventions that can alter one or more components of this axis. Specifically, antibiotics can change the gut microbiome which can, in turn, change the brain’s chemical state and, potentially, lead to onset of depression. To study this in humans is more difficult than in animal models. For this reason, we are working with the UNO marmoset colony to decipher impacts of antibiotics on emotional states. Because immune communication is a bedrock of the gut/brain/immune axis and many immune communication molecules are inflammatory signals, we needed to develop an approach that can show the range and intensity of expression. This project focuses on developing a reliable approach for measuring soluble marmoset inflammatory molecules (e.g., cytokines, chemokines).
Methods
To detect and quantify immune analytes present in marmoset blood samples (UNO Callitrichid Research Center), we are utilizing electrochemiluminescent multiplex analysis according to the manufacturer’s instruction (Mesoscale Diagnostics). To obtain sample for analyses, we first isolate peripheral blood mononuclear cells (PBMCs) from whole blood. Next, we stimulate the PBMCs to produce inflammatory analytes. For this, we employed multiple stimulation strategies: cell activation cocktail (CAC), mixed lymphocyte reaction (MLR), or lipopolysaccharide stimulation (LPS). Culture supernatants were evaluated in the multiplex assay. The data was then analyzed through Microsoft Excel using techniques provided by Mesoscale Diagnostics.
Results
Our preliminary data showed that the assay we used, which was designed for human analyte detection, was cross-reactive for 5 out of 10 marmoset inflammatory analytes analyzed. The marmoset analytes that can be detected are IFN-g, IL-13, IL-1B, IL-4, and IL-8. These are our focus going forward.
Discussion
Our main goal in this project is to be able to link behavioral observations to inflammatory analyte levels. To help achieve this goal, our next steps are to validate that marmoset inflammatory analyte detection remains consistent when the cytokines are present in marmoset plasma as the biological matrix. When that is complete, we will have an experimental framework that will help us achieve our project goal.