Kidney Week

Abstract: SA-PO358

A Cross-Talk Between HIV-Injured Podocytes and Parietal Epithelial Cells (PECs) Results in Proliferation of PECs

Session Information

• Glomerular Diseases: Immunology and Inflammation - III
  October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

• 1202 Glomerular Diseases: Immunology and Inflammation

Authors

• Kumar, Vinod, Fienstine Institute for Medical Research, NEW YORK, New York, United States

Vinod Kumar,

• Lan, Xiqian, Feinstein Institute for Medical Research, Manhasset, New York, United States

Xiqian Lan,

• Ayasolla, Kamesh R., Feinstein Institute for Medical Research, Manhasset, New York, United States

Kamesh R. Ayasolla,

• Chowdhary, Sheetal, Feinstein Institute of Medical Research, New Hyde Park, New York, United States

Sheetal Chowdhary,

• Marashi Shoshtari, Seyedeh Shadafarin, The Feinstein Institute for Medical Research, Manhasset, New York, United States

Seyedeh Shadafarin Marashi Shoshtari,

• Malhotra, Ashwani, Feinstein Institute Medical Research and NSLIJ, MANHASSET, New York, United States

Ashwani Malhotra,

• Meyer-Schwesinger, Catherine, University of Hamburg, Hamburg, Germany

Catherine Meyer-Schwesinger,

• Skorecki, Karl, Rambam Health Care Campus, Haifa, Israel

Karl Skorecki,

• Singhal, Pravin C., North Shore LIJ Health System, Great Neck, New York, United States

Pravin C. Singhal,

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. Interleukin (IL)-1 β has been reported to stimulate PECs proliferation. We have recently reported that HIV infection stimulates generation of IL-1β by PDs. We now hypothesize that a cross-talk between HIV-infected PDs and PECs would promote PECs proliferation.

Methods

Immortalized differentiated human PDs (DPDs) were transduced with either vector (DPDV) or HIV (NL4-3, DPDHIV) and assayed for pyroptosis (morphologic assay). Control DPDs and DPDHIV 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 (DPDV) and experimental (DPDHIV) 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 growth assay. To confirm the role of cross-talk, PECs were grown in outer wells and DPDV/DPDHIVs were seeded into inner wells (Trans-well plates). After 48 hours, cells in outer wells were assayed for proliferation. Cellular lysates/incubation media of DPDVs and DPDHIVs 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

DPDHIV displayed a higher percentage of cells with pyroptosis (P<0.01 vs respective controls). Cellular lysates and incubation media of DPDHIV showed increased (P<0.05 vs. DPDV) content of IL-1β. Conditioned media of DPDHIV promoted PECs proliferation; however, anti-IL-1β antibody partially inhibited DPDHIV-conditioned media-mediated proliferation. PECs growing in outer wells of trans-well plates containing DPDHIV also displayed enhanced proliferation. PECs treated with DPDHIV conditioned media exhibited a higher percentage (P<0.01 vs. DPDV) of PCNA/Ki67 +ve cells.

Conclusion

A cross-talk between PDs to PECs promotes PECs proliferation in HIV milieu.

Funding

• NIDDK Support