DNA vaccination with naked DNA encoding MCP-1 and RANTES protects against renal injury in adriamycin nephropathy.
BACKGROUND
We have previously shown that monocyte chemoattractant protein-1 (MCP-1) and regulated upon activation, normal T-cell expressed and secreted (RANTES) are significantly increased in renal cortex in adriamycin nephropathy. In this study, we tested the effect of DNA vaccination encoding the C-C chemokines MCP-1 and RANTES in a rat model of adriamycin nephropathy.


METHODS
Both reverse transcription-polymerase chain reaction (RT-PCR) products of MCP-1 and RANTES used as constructs were cloned into a pTarget vector for naked DNA vaccination. Two hundred micrograms of DNA was injected into the tibialis anterior muscle four times at weekly intervals. One week after the last DNA vaccination, rats received adriamycin. All animals were sacrificed 4 weeks after adriamycin administration. Changes in renal function and histologic features were assessed. Enzyme-linked immunosorbent assay (ELISA) and Western blot were used for autoantibody determination. Antibody specificity was assessed in in vitro transmigration assays.


RESULTS
Chemokine DNA vaccination significantly reduced proteinuria (P < 0.05) and ameliorated creatinine clearance (P < 0.05) at 2, 3, and 4 weeks after adriamycin administration. Morphometric analysis showed less glomerular sclerosis (P < 0.001) and interstitial infiltrates (P < 0.005) in chemokine DNA vaccination group compared with control groups. Anti-MCP-1 and RANTES autoantibodies were detected in higher concentrations in chemokine DNA vaccinated rats than in control rats (P < 0.001) and serum from vaccinated rats blocked T-cell transmigration to MCP-1 and RANTES.


CONCLUSIONS
In this study, we have shown that naked DNA vaccination against MCP-1 and RANTES ameliorates the progression of renal disease in the rat adriamycin nephropathy model of chronic proteinuric renal disease. The protective mechanism may involve the production of autoantibodies against MCP-1 and RANTES.
Wu
Huiling
H
Centre for Kidney Research, The Children's Hospital at Westmead, Westmead, New South Wales, Australia. huilingw@med.usyd.edu.au
Wang
Yiping
Y
Tay
Yuet-Ching
YC
Zheng
Guoping
G
Zhang
Chun
C
Alexander
Stephen I
SI
Harris
David C H
DC
D000818:Animals; D000906:Antibodies / Q000097:blood; D018932:Chemokine CCL2 / Q000235:genetics / Q000276:immunology*; D018946:Chemokine CCL5 / Q000235:genetics / Q000276:immunology*; D002633:Chemotaxis; D004317:Doxorubicin / Q000633:toxicity*; D007668:Kidney / Q000187:drug effects*; D008297:Male; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D014611:Vaccination; D019444:Vaccines, DNA / Q000276:immunology*
D000906:Antibodies; C491766:Ccl2 protein, rat; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D012333:RNA, Messenger; D019444:Vaccines, DNA; D004317:Doxorubicin