Integrin-mediated interactions between primary/T-sv40 immortalized human glomerular epithelial cells and type IV collagen

Uma Krishnamurti, Yong Chen, Alfred Michael, Youngki Kim, Wei Wei Fan, Jörgen Wieslander, Charlotte Brunmark, Eric Rondeau, Jean Daniel Sraer, Francoise Delarue, Effie C. Tsilibary

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

The use of human glomerular epithelial cells (HGEC) in research has been severely restricted by several obstacles, which have been circumvented by the generation of T-SV40 immortalized human visceral glomerular epithelial cells (Delarue et al, 1991). In this work, we compared the primary and immortalized HGEC for expression of integrin and some nonintegrin surface receptors. We also studied the adhesion of both types of HGEC to glomerular basement membrane (GBM), type IV collagen (tIV), and its major noncollagenous NC1 domain. The integrins mediating adhesion of HGEC to tIV were also examined. Expression of integrin and some nonintegrin cell surface receptors was analyzed by flow cytometry. Adhesion to GBM, tIV, and its major noncollagenous NC1 domain was studied by direct solid phase cell adhesion assays. Identification of integrins mediating adhesion of HGEC to tIV was achieved by inhibition of cell adhesion using monoclonal antibodies to integrin subunits. The primary and immortalized HGEC share phenotypic characteristics, and α3β1 appeared to be the major integrin present on both HGEC types. The kinetics of binding to GBM, tIV, and its noncollagenous NC1 domain were similar in both the primary and immortalized HGEC, although the latter displayed a somewhat weaker binding. Both the primary and immortalized HGEC displayed significantly better adhesion to NC1-α3 compared with NC1-α1, α3β1 appears to be the major integrin mediating the adhesion of HGEC to tIV. Our studies suggest that α3β1 is the major integrin present on HGEC. This has been confirmed by flow cytometric analysis. In addition, we demonstrated a functional role for this integrin in mediating attachment of HGEC to tIV. Our data also demonstrate a preference in binding of HGEC to α3 chains of NC1 compared with α1 chains of NC1. These findings were seen in both the primary and immortalized HGEC. The T- SV40 immortalized HGEC can therefore serve as a very useful tool to study glomerular visceral cell biology.

Original languageEnglish (US)
Pages (from-to)650-657
Number of pages8
JournalLaboratory Investigation
Volume74
Issue number3
StatePublished - Mar 1996

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