Abstract: SA-PO621
HIV-Induced Podocyte Pyroptosis Contributes to Proliferation of Parietal Epithelial Cells
Session Information
- Glomerular Diseases: Immunology, Inflammation - II
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1202 Glomerular Diseases: Immunology and Inflammation
Authors
- Adnani, Harsha, Feinstein Institute of Research, Manhasset, New York, United States
- Kumar, Vinod, Feinstein Institute of Research, Manhasset, New York, United States
- Vashistha, Himanshu, Ochsner Health System, New Orleans, Louisiana, United States
- Ayasolla, Kamesh R., Feinstein Institute for Medical Research, Manhasset, New York, United States
- Lan, Xiqian, Feinstein Institute for Medical Research, Manhasset, New York, United States
- Qayyum, Maleeha, Northwell health , Hicksville, New York, United States
- Chinnapaka, Sushma, Northwell health , Hicksville, New York, United States
- Malhotra, Ashwani, Feinstein Institute Medical Research and NSLIJ, Manhasset, New York, United States
- Skorecki, Karl, Rambam Health Care Campus, Haifa, Israel
- Singhal, Pravin C., North Shore LIJ Health System, Great Neck, New York, United States
Background
The lack of or abundance of proliferating parietal epithelial cells (PECs) in Bowman's space determines the glomerular phenotype in focal segmental glomerulosclerosis. Proliferating PECs in Bowman's space characterize the HIV-associated nephropathy. The involved mechanism of PECs proliferation in HIV milieu is not understood. Interleukin (IL)-1 β has been reported to stimulate PECs proliferation. We have recently demonstrated that HIV infection stimulates the generation of IL-1β by podocytes. We hypothesize that massive injury of HIV-infected podocytes would stimulate PECs proliferation.
Methods
Immortalized human podocytes were differentiated and transduced with either vector (V-PDs) or HIV (NL4-3, HIV-PDs) and evaluated for pyroptosis (morphologic assay). V-PDs and HIV-PDs were incubated in serum-free media for 24 hours. Incubation (conditioned, C) media was collected and stored at -80°C. PECs were incubated in serum-free media containing 10% of control (V-PDs), and experimental (HIV-PDs) conditioned media for 48 hours. In another set of experiments, PECs were incubated in serum-free media containing 10% control and experimental media with or without IL-1β (neutralizing) antibodies for 48 hours. Cells were evaluated for proliferation by MTT cell viability assay. To establish an interaction, PECs were grown in outer wells, and V-PDs/HIV-PDs were seeded into inner wells (Trans-well plates). After 48 hours,-PDs were assayed for IL-1β by ELISA. Additionally, PECs grown on coverslips were treated with 10% control and experimental media for 48 hours, followed by immunolabeling for either PCNA or Ki67.
Results
HIV-PDs showed a higher percentage of pyroptosed cells (P<0.01 vs. V-PDs). Cellular lysates and incubation media of HIV-PDs showed increased (P<0.05 vs.V-PDs) generation of IL-1β. Conditioned media of HIV-PDs stimulated PECs proliferation; however, anti-IL-1β antibody partially inhibited HIV-PDs conditioned media-mediated proliferation. PECs growing in outer wells of trans-well plates containing HIV-PDs showed increased proliferation. PECs treated with HIV-PDs conditioned media showed a higher percentage (P<0.01 vs. V-PDs) of PCNA/Ki67 +ve cells.
Conclusion
HIV-induced podocyte pyroptosis contributes to PECs proliferation.
Funding
- NIDDK Support