Production of inflammatory mediators and cytokine responsiveness of an SV40-transformed human proximal tubular epithelial cell line.
Proximal tubular epithelial cells (PTEC) play a central role in the physiology of the renal tubulointerstitium. To be able to study the relationship between tubular cells and inflammatory renal diseases the availability of cultured cells is of importance. This study describes an immortalized proximal tubular epithelial cell line which was generated using SV40 DNA. To determine whether the transformation altered the cell line, the transformed cell line was characterized phenotypically using different monoclonal antibodies directed against peptidases, which are characteristic of PTEC, such as adenosine deaminase binding protein (CD26), leucine amino peptidase and carboxy peptidase M by immunofluorescent staining and FACS analysis. All peptidases were clearly present on the parental cell line and the transformed cell line. However, the level of expression of the peptidases was lower on the transformed cell line as compared to the parental nontransfected cells. The morphology of the transformed cell line, determined using a transwell culture system and electron microscopy, showed a polarized morphology of the tubular cells, tight junctions and microvilli. The transformed cell line was compared with the parental proximal tubular epithelial cells in its ability to respond to inflammatory cytokines such as IL- 1alpha TNF-alpha, IFN-gamma. Stimulation with these cytokines resulted in enhanced production of complement components C2, C3, C4 and factor H, IL-6 and the chemokines IL-8 and MCP-1. The transformed cell line responded in a similar fashion as the parental cell line, although the amount of the different proteins produced was significantly higher in the transformed cell line. Overall, the transformed tubular cell line seems to be a suitable model to study different effects on tubular cells in relation to inflammatory kidney diseases.
Gerritsma
J S
JS
Department of Nephrology, Leiden University Hospital, The Netherlands.
van Kooten
C
C
Gerritsen
A F
AF
Mommaas
A M
AM
van Es
L A
LA
Daha
M R
MR
D015139:Blotting, Southern; D002461:Cell Line, Transformed / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D018932:Chemokine CCL2 / Q000378:metabolism; D003175:Complement C2 / Q000378:metabolism; D003176:Complement C3 / Q000378:metabolism; D003181:Complement C4 / Q000378:metabolism; D017242:Complement Factor H / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology*; D004279:DNA, Viral; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D008854:Microscopy, Electron; D015183:Restriction Mapping; D013539:Simian virus 40; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C493495:CFH protein, human; D018932:Chemokine CCL2; D003175:Complement C2; D003176:Complement C3; D003181:Complement C4; D016207:Cytokines; D004279:DNA, Viral; D018836:Inflammation Mediators; D007375:Interleukin-1; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D017242:Complement Factor H; D007371:Interferon-gamma