Mcp-1 Release Modulation Through Interaction of Pulmonary Endothelial Cells and Mesenchymal Stromal Cells

<p><strong>Background:</strong> Ischemic stroke is a leading cause of death and long-term disability</p> <p>around the world. Current treatment options are limited to the administration of</p> <p>tissue plasminogen activator (tPA) and/or endovascular therapy, administered within</p> <p>a limited time window. However, cell-based therapies such as mesenchymal stromal</p> <p>cells (MSCs) have increasingly shown great promise for ischemic stroke recovery</p> <p>with some therapies already in various stages of clinical trials. Intravenous (IV)</p> <p>administration of the MSCs leads to the entrapment of these MSCs in the lungs.</p> <p>These entrapped MSCs interact with the pulmonary endothelial cells (PECs) and</p> <p>could modulate the immune response through the release of cytokines and</p> <p>chemokines. Monocyte Chemoattractant Protein - 1 (MCP-1), is an important</p> <p>chemokine involved in the recruitment of monocytes and macrophages. In our study,</p> <p>we wanted to explore the interactions between MSCs with PECs and how this</p> <p>interaction changes the expression levels of MCP-1 and other cytokines after an</p> <p>inflammatory event such as stroke. We also wanted to see if MCP-1 released</p> <p>through the interaction between MSCs and PECs under inflammatory conditions,</p> <p>modulates the immune response through the modification of monocytes.</p> <p><strong>Methods:</strong> Cultured murine PECs were grown either alone or in a combination of</p> <p>murine MSCs, and were exposed to 1) a combination of IFN-γ and TNF-α</p> <p>inflammatory stimuli or 2) Anti MCP-1 antibody to neutralize any secreted MCP-1.</p> <p>The secretome release of IL-1β, IL-1ra, IL-6, MCP-1, and VEGF were analyzed</p> <p>using ELISA (BD Biosciences and R&D Systems). To further understand the</p> <p>immunomodulatory response, the collected media from the previous step was added</p> <p>to splenic immune cells (CD11b+) and splenic monocytes (CD115+). The secretome</p> <p>release was analyzed from these cells using ELISA.</p> <p><strong>Results:</strong> MCP-1 secretion levels were increased from PECs as well as co-cultures</p> <p>of PECs and MSCs when they were exposed to inflammatory stimuli. When</p> <p>cocultures of PECs and MSCs were exposed to recombinant MCP-1 or MCP-1</p> <p>neutralizing antibody, VEGF secretion levels decreased. In the presence of</p> <p>inflammatory stimuli, co-cultures of PECs and MSCs secreted elevated levels of</p> <p>VEGF. While under inflammation, we also observed that IL-6 levels were elevated</p> <p>and they remain elevated even when MCP-1 was neutralized. We did not observe</p> <p>any difference in secretome release from neither the splenic immune cells (CD11b+)</p> <p>nor the splenic monocytes (CD115+).</p> <p><strong>Conclusion:</strong> Our data show that MCP-1 release under stroke-like conditions is</p> <p>modulated through the interaction of PECs and MSCs. However, our study was</p> <p>unable to elucidate MCP-1’s role in the modification of monocytes.</p>

Name thesis:degree_name

Masters of Science (MS)

Level thesis:degree_level

Thesis (MS)

Year dc:date.available

2020

Authors dc:creator

• Giridhar, Kaavya
• <p>0000-0001-6912-1458</p>

Contributors dc:contributor

• Dr. Sean I. Savitz
• Dr. Jaroslaw Aronowski
• Dr. Matthew T. Harting

• Stroke
• Cell Therapy
• Mesenchymal Stromal Cells
• Pulmonary Endothelial Cells
• Immunomodulation
• Monocytes
• MCP-1
• IL-6
• VEGF
• Medicine and Health Sciences

Repository record dc:identifier

https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1000

OAI identifier oai:identifier

oai:digitalcommons.library.tmc.edu:utgsbs_dissertations-2050