Date of Award

2008

Degree Type

Thesis

Degree Name

Master of Science

Program

Microbiology and Immunology

Supervisor

Dr. Anthony Jevnikar

Second Advisor

Dr. Bhagirath Singh

Third Advisor

Dr. Ewa Cairns

Abstract

Human serpin protease inhibitor 9 (PI-9) inhibits Granzyme B (GrB) function. However, its role in protection from renal injury is unknown. Tubular epithelial cells (TEC) express serine protease inhibitor 6 (SPI-6), the murine homolog of human PI-9. SPI-6 protein expression increased in response to IFNγ or LPS and detachment stress. This occurred without increased mRNA expression suggesting SPI-6 is post transcriptionally regulated. Cloned TEC were resistant to GrB killing. However in SPI-6 gene silenced TEC, death significantly increased from baseline levels (10.7% + 2.4 to 21.22% + 1, p= 0.0002) when assessed by Annexin V∕7AAD. Kidney SPI-6 protein expression by immuno-histochemistry and western blotting, increased in TEC in vivo following ischemia-reperfusion injury (IRI). Expression of SPI-6 by TEC may provide protection from GrB producing cytotoxic CD8+T and NK cells, which are involved in IRI and rejection. Therefore, augmenting or maintaining SPI-6∕PI-9 expression by TEC may promote long term allograft survival.

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