Glomerular T cell V beta repertoire in experimental autoimmune glomerulonephritis

Giles Walters¹ Anne-Marie Habib² John Reynolds² Huiling Wu¹, John Knight¹, Charles Pusey².

1. Centre for Kidney Research, Children's Hospital at Westmead, Sydney, NSW.
2. Renal Section, Division of Medicine, Imperial College School of Medicine, London UK.

Experimental Autoimmune Glomerulonephritis (EAG) is a model of human Goodpasture's disease induced in susceptible strains of rats by a single intramuscular injection of collagenase-solubilised rat glomerular basement membrane in adjuvant. This induces severe proteinuria and crescentic nephritis at 4 weeks. Previous work has shown this to be a T cell dependent disease (1). The aim of this work was to investigate the T cells infiltrating the glomeruli in this disease model using reverse transcription PCR, Complementarity Determining Region 3 (CDR3) spectratyping and sequencing. EAG was induced in Wistar Kyoto rats by standard methods. The kidneys were perfused with saline and the glomeruli separated by a sieving method. RNA was extracted and reverse transcribed to cDNA. Polymerase chain reaction using primers for 20 T cell receptor V beta genes was used to further examine the cDNA. This showed the overexpression of an average of two V beta families in each kidney analysed. Overexpressed families varied from animal to animal. CDR3 spectratyping of Fam-labelled PCR product showed widespread restriction of multiple V beta families compared to matching splenic samples. Two V beta families (BV16 and BV10) showed a common spectratype band across six of seven animals. Three families showed a further common band across five of seven animals. Sequencing is being performed to further investigate these spectratypes. Restricted spectratypes shared between animals suggest that oligoclonal T cells are infiltrating the glomeruli and that similar clones may be present in different animals. These results suggest that T cells recognising specific antigens infiltrate the glomerulus in the course of EAG.

1. Reynolds J, Tam FWK, Chandraker A, Smith J, Karkar AM, Cross J, Peach R, Sayegh MH, Pusey CD. CD28-B7 blockade prevents the development of experimental autoimmune glomerulonephritis. J. Clin. Invest. 105:643-651,2000.

Presented at the Annual Scientific Meeting of the Australian and New Zealand Society of Nephrology, Darwin, September 2001
Presented at the ASN/ISN World Congress of Nephrology, San Francisco, October 2001

Correspondence:
Dr Giles Walters
GilesW@chw.edu.au