Background: Ischemic stroke is a leading cause of death and long-term disability
around the world. Current treatment options are limited to the administration of
tissue plasminogen activator (tPA) and/or endovascular therapy, administered within
a limited time window. However, cell-based therapies such as mesenchymal stromal
cells (MSCs) have increasingly shown great promise for ischemic stroke recovery
with some therapies already in various stages of clinical trials. Intravenous (IV)
administration of the MSCs leads to the entrapment of these MSCs in the lungs.
These entrapped MSCs interact with the pulmonary endothelial cells (PECs) and
could modulate the immune response through the release of cytokines and
chemokines. Monocyte Chemoattractant Protein - 1 (MCP-1), is an important
chemokine involved in the recruitment of monocytes and macrophages. In our study,
we wanted to explore the interactions between MSCs with PECs and how this
interaction changes the expression levels of MCP-1 and other cytokines after an
inflammatory event such as stroke. We also wanted to see if MCP-1 released
through the interaction between MSCs and PECs under inflammatory conditions,
modulates the immune response through the modification of monocytes.
Methods: Cultured murine PECs were grown either alone or in a combination of
murine MSCs, and were exposed to 1) a combination of IFN-γ and TNF-α
inflammatory stimuli or 2) Anti MCP-1 antibody to neutralize any secreted MCP-1.
The secretome release of IL-1β, IL-1ra, IL-6, MCP-1, and VEGF were analyzed
using ELISA (BD Biosciences and R&D Systems). To further understand the
immunomodulatory response, the collected media from the previous step was added
to splenic immune cells (CD11b+) and splenic monocytes (CD115+). The secretome
release was analyzed from these cells using ELISA.
Results: MCP-1 secretion levels were increased from PECs as well as co-cultures
of PECs and MSCs when they were exposed to inflammatory stimuli. When
cocultures of PECs and MSCs were exposed to recombinant MCP-1 or MCP-1
neutralizing antibody, VEGF secretion levels decreased. In the presence of
inflammatory stimuli, co-cultures of PECs and MSCs secreted elevated levels of
VEGF. While under inflammation, we also observed that IL-6 levels were elevated
and they remain elevated even when MCP-1 was neutralized. We did not observe
any difference in secretome release from neither the splenic immune cells (CD11b+)
nor the splenic monocytes (CD115+).
Conclusion: Our data show that MCP-1 release under stroke-like conditions is
modulated through the interaction of PECs and MSCs. However, our study was
unable to elucidate MCP-1’s role in the modification of monocytes.
","abstract_html":"<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>","abstract_has_math":false,"creators":["Giridhar, Kaavya","0000-0001-6912-1458
"],"institution":null,"degree_name":"Masters of Science (MS)","degree_level":"Thesis (MS)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Dr. Sean I. Savitz","Dr. Jaroslaw Aronowski","Dr. Matthew T. Harting"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-05-01T07:00:00Z","date_published":"2020-05-01T07:00:00Z","updated_at":"2026-07-24T05:50:02Z","subjects":["Stroke","Cell Therapy","Mesenchymal Stromal Cells","Pulmonary Endothelial Cells","Immunomodulation","Monocytes","MCP-1","IL-6","VEGF","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1000","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Dr. Sean I. Savitz","Dr. Jaroslaw Aronowski","Dr. Matthew T. Harting"]},{"key":"dc:creator","label":"Author","values":["Giridhar, Kaavya","0000-0001-6912-1458
"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2021-04-27T07:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis (MS)"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Masters of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Stroke","Cell Therapy","Mesenchymal Stromal Cells","Pulmonary Endothelial Cells","Immunomodulation","Monocytes","MCP-1","IL-6","VEGF","Medicine and Health Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1000"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Background: Ischemic stroke is a leading cause of death and long-term disability
around the world. Current treatment options are limited to the administration of
tissue plasminogen activator (tPA) and/or endovascular therapy, administered within
a limited time window. However, cell-based therapies such as mesenchymal stromal
cells (MSCs) have increasingly shown great promise for ischemic stroke recovery
with some therapies already in various stages of clinical trials. Intravenous (IV)
administration of the MSCs leads to the entrapment of these MSCs in the lungs.
These entrapped MSCs interact with the pulmonary endothelial cells (PECs) and
could modulate the immune response through the release of cytokines and
chemokines. Monocyte Chemoattractant Protein - 1 (MCP-1), is an important
chemokine involved in the recruitment of monocytes and macrophages. In our study,
we wanted to explore the interactions between MSCs with PECs and how this
interaction changes the expression levels of MCP-1 and other cytokines after an
inflammatory event such as stroke. We also wanted to see if MCP-1 released
through the interaction between MSCs and PECs under inflammatory conditions,
modulates the immune response through the modification of monocytes.
Methods: Cultured murine PECs were grown either alone or in a combination of
murine MSCs, and were exposed to 1) a combination of IFN-γ and TNF-α
inflammatory stimuli or 2) Anti MCP-1 antibody to neutralize any secreted MCP-1.
The secretome release of IL-1β, IL-1ra, IL-6, MCP-1, and VEGF were analyzed
using ELISA (BD Biosciences and R&D Systems). To further understand the
immunomodulatory response, the collected media from the previous step was added
to splenic immune cells (CD11b+) and splenic monocytes (CD115+). The secretome
release was analyzed from these cells using ELISA.
Results: MCP-1 secretion levels were increased from PECs as well as co-cultures
of PECs and MSCs when they were exposed to inflammatory stimuli. When
cocultures of PECs and MSCs were exposed to recombinant MCP-1 or MCP-1
neutralizing antibody, VEGF secretion levels decreased. In the presence of
inflammatory stimuli, co-cultures of PECs and MSCs secreted elevated levels of
VEGF. While under inflammation, we also observed that IL-6 levels were elevated
and they remain elevated even when MCP-1 was neutralized. We did not observe
any difference in secretome release from neither the splenic immune cells (CD11b+)
nor the splenic monocytes (CD115+).
Conclusion: Our data show that MCP-1 release under stroke-like conditions is
modulated through the interaction of PECs and MSCs. However, our study was
unable to elucidate MCP-1’s role in the modification of monocytes.
"]},{"key":"dc:title","label":"Title","values":["Mcp-1 Release Modulation Through Interaction of Pulmonary Endothelial Cells and Mesenchymal Stromal Cells"]}]}],"canonical_facts":{"dc:contributor":["Dr. Sean I. Savitz","Dr. Jaroslaw Aronowski","Dr. Matthew T. Harting"],"dc:creator":["Giridhar, Kaavya","0000-0001-6912-1458
"],"dc:date.available":["2021-04-27T07:00:00Z"],"dc:description.abstract":["Background: Ischemic stroke is a leading cause of death and long-term disability
around the world. Current treatment options are limited to the administration of
tissue plasminogen activator (tPA) and/or endovascular therapy, administered within
a limited time window. However, cell-based therapies such as mesenchymal stromal
cells (MSCs) have increasingly shown great promise for ischemic stroke recovery
with some therapies already in various stages of clinical trials. Intravenous (IV)
administration of the MSCs leads to the entrapment of these MSCs in the lungs.
These entrapped MSCs interact with the pulmonary endothelial cells (PECs) and
could modulate the immune response through the release of cytokines and
chemokines. Monocyte Chemoattractant Protein - 1 (MCP-1), is an important
chemokine involved in the recruitment of monocytes and macrophages. In our study,
we wanted to explore the interactions between MSCs with PECs and how this
interaction changes the expression levels of MCP-1 and other cytokines after an
inflammatory event such as stroke. We also wanted to see if MCP-1 released
through the interaction between MSCs and PECs under inflammatory conditions,
modulates the immune response through the modification of monocytes.
Methods: Cultured murine PECs were grown either alone or in a combination of
murine MSCs, and were exposed to 1) a combination of IFN-γ and TNF-α
inflammatory stimuli or 2) Anti MCP-1 antibody to neutralize any secreted MCP-1.
The secretome release of IL-1β, IL-1ra, IL-6, MCP-1, and VEGF were analyzed
using ELISA (BD Biosciences and R&D Systems). To further understand the
immunomodulatory response, the collected media from the previous step was added
to splenic immune cells (CD11b+) and splenic monocytes (CD115+). The secretome
release was analyzed from these cells using ELISA.
Results: MCP-1 secretion levels were increased from PECs as well as co-cultures
of PECs and MSCs when they were exposed to inflammatory stimuli. When
cocultures of PECs and MSCs were exposed to recombinant MCP-1 or MCP-1
neutralizing antibody, VEGF secretion levels decreased. In the presence of
inflammatory stimuli, co-cultures of PECs and MSCs secreted elevated levels of
VEGF. While under inflammation, we also observed that IL-6 levels were elevated
and they remain elevated even when MCP-1 was neutralized. We did not observe
any difference in secretome release from neither the splenic immune cells (CD11b+)
nor the splenic monocytes (CD115+).
Conclusion: Our data show that MCP-1 release under stroke-like conditions is
modulated through the interaction of PECs and MSCs. However, our study was
unable to elucidate MCP-1’s role in the modification of monocytes.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1000"],"dc:subject":["Stroke","Cell Therapy","Mesenchymal Stromal Cells","Pulmonary Endothelial Cells","Immunomodulation","Monocytes","MCP-1","IL-6","VEGF","Medicine and Health Sciences"],"dc:title":["Mcp-1 Release Modulation Through Interaction of Pulmonary Endothelial Cells and Mesenchymal Stromal Cells"],"thesis:degree_level":["Thesis (MS)"],"thesis:degree_name":["Masters of Science (MS)"]},"updated_at":"2026-07-24T05:50:02Z"}