GENE EXPRESSION PROFILE OF BKV INFECTED WI-38CELLS.

 

Parmjeet S Randhawa, Jian-Hua Luo, Debbie Zygmunt, Uma Chandran, Songhui Li, John Gilbertson

Divisions of Molecular-Cellular Pathology,Transplantation Pathology & Informatics, Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA.

 

 

BACKGROUND: BK virus nephropathy (BKVN) is a complication of excessive immunosuppression that is increasingly recognized after kidney transplantation. We have analyzed virus induced changes in host gene expression with a view to enhance our understanding of the pathogenesis of BKVN.

 

DESIGN: WI-38 cells were grown in DMEM medium supplemented by 10% fetal bovine serum. Each 25 square cm flask containing 300,000 cells was infected with 3x 107 BKV particles, & incubated at 37 degrees C, till marked viral cytopathic effect was evident. At this stage, cells were harvested for total RNA extraction, cDNA synthesis, & production of biotinylated cRNA, followed by hybridization to Affymetrix HG-U133 DNA microarray chips. Changes in gene expression seen consistently in 2 different experiments were reported using Affymetrix MAS 5.0 software

 

RESULTS: Statistically significant up-regulation of mRNA’s was seen in 705 genes including the following groups: (a) interleukins (IL- 1, 6, 8, 11), (b) interleukin receptors (IL- 4R, IL- 13R alpha 2), (c) Cell adhesion molecules (ICAM-1), (d) growth factors (VEGF), (e) chemokines (MCP-1, RANTES) (f) oncogenes (R-binding protein-8), (g) MHC class I molecules (HLA –B, C, F), and (h) cell cycle proteins (PCNA, Ki67, cyclin H). Genes marginally down-regulated by BKV infection belonged to the following categories: (a) cell adhesion molecules (VCAM-1), (b) growth factors (FGF-5), (c) oncogenes of the RAS family (RAB-2,4), (d) extracellular matrix proteins (collagen type 1 alpha 2, TGF- beta receptor II, fibronectin, laminin, thrombospondin), and (e) cell cycling proteins (cyclin G1).

 

CONCLUSIONS: (a) The upregulation of cell cycle proteins illustrates the dependence of BKV on host cellular factors that may not be present in quiescent cells. (b) Influx of inflammatory cells in BKVN is likely mediated by cytokines IL-1, IL-6, and chemokines IL- 8, MCP-1, and RANTES. (c) The upregulation of MHC class I proteins, provides a potential explanation for changes mimicking acute rejection that are frequently biopsies with BKVN.

 

 

 

 

GENE EXPRESSION PROFILE OF BKV INFECTED WI-38CELLS.

 

Parmjeet S Randhawa, Jian-Hua Luo, Debbie Zygmunt, Uma Chandran, Songhui Li, and John Gilbertson

 

Divisions of Molecular-Cellular Pathology,Transplantation Pathology & Informatics, Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA.

 

 

Address for correspondence:

Parmjeet Randhawa, M.D.,

Associate Professor, Division of Transplantation Pathology,

Department of Pathology, University of Pittsburgh,

Room C903.1 UPMC-Presbyterian Hospital,

200 Lothrop Street, Pittsburgh, PA 15213.

Phone: 412 647 7646

Fax:     412 647 5237