The DNA co-vaccination using Sm antigen and IL-10 as prophylactic experimental therapy ameliorates nephritis in a model of lupus induced by pristane

by Beatriz Teresita Martín-Márquez, Minoru Satoh, Rogelio Hernández-Pando, Erika Aurora Martínez-García, Marcelo Heron Petri, Flavio Sandoval-García, Oscar Pizano-Martinez, Trinidad García-Iglesias, Fernanda Isadora Corona-Meraz, Monica Vázquez-Del Mercado

Introduction

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies such as anti-Sm. Studies in patients with SLE and murine models of lupus reveal that the most critical anti-Sm autoantibodies are predominantly direct against D1_(83–119), D2, and B´/B epitopes.

Objectives

The present study aimed to analyze the induction of antigen-specific tolerance after prophylactic immunization with a DNA vaccine encoding the epitopes: D1_83-119, D2, B´/B, and B´/B_COOH in co-vaccination with IFN-γ or IL-10 in a murine model of lupus induced by pristane.

Material and methods

To obtain endotoxin-free DNA vaccines, direct cloning techniques using pcDNA were performed: D1_83-119, D2, B´/B, B´/B_COOH, IFN-γ, or IL-10. Lupus was induced by 0.5 mL of pristane via intraperitoneal in BALB/c female mice. Immunoprecipitation with K562 cells was metabolically labeled with ³⁵S and ELISA to detect serum antibodies or mice IgG1, IgG2a isotypes. ELISA determined IL-10 and IFN-γ from splenocytes supernatants. Proteinuria was assessed monthly, and lupus nephritis was evaluated by immunofluorescence, and electron microscopy.

Results

The prophylactic co-vaccination with D2/IL-10 reduced the expression of kidney damage observed by electron microscopy, direct immunofluorescence, and H & E, along with reduced level of anti-nRNP/Sm antibodies (P = 0.048).

Conclusion

The prophylactic co-vaccination of IL-10 with D2 in pristane-induced lupus ameliorates the renal damage maybe by acting as prophylactic DNA tolerizing therapy.