Abstract: FR-PO684

Podocyte Cross Talk with Parietal Epithelial Cells (PECs) Stimulates PECs Proliferation in HIV Milieu

Session Information

Category: Glomerular

  • 1001 Glomerular: Basic/Experimental Immunology and Inflammation

Authors

  • Kumar, Vinod, Fienstine Institute for Medical Research, New York, New York, United States
  • Lan, Xiqian, Feinstein Institute for Medical Research, Great Neck, New York, United States
  • Aslam, Rukhsana, Feinstein Institute for medical research, Glenoaks, New York, United States
  • Hussain, Ali, Feinstein Institute of Medical Research, New York, New York, United States
  • Marashi Shoshtari, Seyedeh Shadafarin, The Feinstein Institute for Medical Research, Manhasset, New York, United States
  • Meyer-Schwesinger, Catherine, University of Hamburg, Hamburg, Germany
  • Malhotra, Ashwani, Feinstein Inst.Med research and NSLIJ, Manhasset, New York, United States
  • Singhal, Pravin C., North Shore LIJ Health System, Great Neck, New York, United States
Background

HIV-associated nephropathy is characterized by an abundance of proliferating PECs in Bowman’s space. The involved mechanism of PECs proliferation in HIV milieu is not clear. Recently, we demonstrated that HIV stimulates IL-1β generation by PDs. We now hypothesize that cross talk between PDs to PECs and PECs to PECs promotes PECs proliferation.

Methods

Immortalized PECs and differentiated PDs were transduced with either vector (PECV/PDV) or HIV (PECHIV/PDHIV, NL4-3) and assayed for pyroptosis (morphologic assay). Control PECs/PDs, PECV/PDV, and PECHIV/PDHIV 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 (PECV/PDV) and experimental (PECHIV/PDHIV) 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 cellular viability assay. To confirm the role of cross talk, PECs were grown in outer wells and PECHIVs/PDHIVs were seeded into inner wells (Trans-well plates). After 48 hours, cells in outer wells were assayed for proliferation. Cellular lysates/incubation media of PECs/PDs and PECHIV/PDHIV 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

Both PDHIV and PECHIV displayed a higher percentage of pyroptosed cells (P<0.01 vs respective controls). Cellular lysates and incubation media of PDHIV and PECHIV displayed enhanced (P<0.05 vs. PDHIV and PECHIV) generation of IL-1β. Conditioned media of PDHIV and PECHIV promoted PECs proliferation; however, anti-IL-1β antibody partially inhibited PDHIV/PECHIV-conditioned media-mediated proliferation. PECs growing in outer wells of trans-well plates containing PDHIV/PECHIV also displayed enhanced proliferation. PECs treated with PECHIV/PDHIV conditioned media higher percentage (P<0.01 vs. PECV/PDV) of PCNA/Ki67 +ve cells.

Conclusion

Cross talks between PDs to PECs and PECs to PECs promote PECs proliferation in HIV milieu.

Funding

  • NIDDK Support