Protective immunity to DENV2 after immunization with a recombinant NS1 protein using a genetically detoxified heat-labile toxin as an adjuvant

Vaccine. 2012 Jan 20;30(5):837-45. doi: 10.1016/j.vaccine.2011.12.034. Epub 2011 Dec 15.

Abstract

The dengue virus non-structural 1 (NS1) protein contributes to evasion of host immune defenses and represents a target for immune responses. Evidences generated in experimental models, as well as the immune responses elicited by infected individuals, showed that induction of anti-NS1 immunity correlates with protective immunity but may also result in the generation of cross-reactive antibodies that recognize platelets and proteins involved in the coagulation cascade. In the present work, we evaluated the immune responses, protection to type 2 dengue virus (DENV2) challenges and safety parameters in BALB/c mice vaccinated with a recombinant NS1 protein in combination with three different adjuvants: aluminum hydroxide (alum), Freund's adjuvant (FA) or a genetically detoxified derivative of the heat-labile toxin (LT(G33D)), originally produced by some enterotoxigenic Escherichia coli (ETEC) strains. Mice were subcutaneously (s.c.) immunized with different vaccine formulations and the induced NS1-specific responses, including serum antibodies and T cell responses, were measured. Mice were also subjected to lethal challenges with the DENV2 NGC strain. The results showed that maximal protective immunity (50%) was achieved in mice vaccinated with NS1 in combination with LT(G33D). Analyses of the NS1-specific immune responses showed that the anti-virus protection correlated mainly with the serum anti-NS1 antibody responses including higher avidity to the target antigen. Mice immunized with LT(G33D) elicited a prevailing IgG2a subclass response and generated antibodies with stronger affinity to the antigen than those generated in mice immunized with the other vaccine formulations. The vaccine formulations were also evaluated regarding induction of deleterious side effects and, in contrast to mice immunized with the FA-adjuvanted vaccine, no significant hepatic damage or enhanced C-reactive protein levels were detected in mice immunized with NS1 and LT(G33D.) Similarly, no detectable alterations in bleeding time and hematological parameters were detected in mice vaccinated with NS1 and LT(G33D). Altogether, these results indicate that the combination of a purified recombinant NS1 and a nontoxic LT derivative is a promising alternative for the generation of safe and effective protein-based anti-dengue vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / administration & dosage*
  • Adjuvants, Immunologic / adverse effects
  • Adjuvants, Immunologic / genetics
  • Aluminum Hydroxide / administration & dosage
  • Animals
  • Antibodies, Viral / blood
  • Bacterial Toxins / administration & dosage*
  • Bacterial Toxins / adverse effects
  • Bacterial Toxins / genetics
  • Dengue / mortality
  • Dengue / pathology
  • Dengue Vaccines / administration & dosage
  • Dengue Vaccines / adverse effects
  • Dengue Vaccines / immunology*
  • Dengue Virus / genetics
  • Dengue Virus / immunology*
  • Enterotoxins / administration & dosage*
  • Enterotoxins / adverse effects
  • Enterotoxins / genetics
  • Escherichia coli Proteins / administration & dosage*
  • Escherichia coli Proteins / adverse effects
  • Escherichia coli Proteins / genetics
  • Freund's Adjuvant / administration & dosage
  • Humans
  • Injections, Subcutaneous
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Survival Analysis
  • T-Lymphocytes / immunology
  • Toxoids / administration & dosage*
  • Toxoids / adverse effects
  • Toxoids / genetics
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / adverse effects
  • Vaccines, Synthetic / immunology
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / immunology*

Substances

  • Adjuvants, Immunologic
  • Antibodies, Viral
  • Bacterial Toxins
  • Dengue Vaccines
  • Enterotoxins
  • Escherichia coli Proteins
  • NS1 protein, Dengue virus type 2
  • Recombinant Proteins
  • Toxoids
  • Vaccines, Synthetic
  • Viral Nonstructural Proteins
  • heat-labile enterotoxin, E coli
  • Aluminum Hydroxide
  • Freund's Adjuvant