Dengue 2 PreM-E/LAMP chimera targeted to the MHC class II compartment elicits long-lasting neutralizing antibodies

Vaccine. 2003 May 16;21(17-18):2178-89. doi: 10.1016/s0264-410x(03)00009-4.


A dengue 2 plasmid DNA vaccine (pD2) expressing the pre-membrane and envelope proteins (preM-E) was modified by replacing the dengue transmembrane and cytoplasmic sequences with those of the mouse lysosome-associated membrane protein (pD2/LAMP). Immunofluorescence and confocal microscopy of human 293, NIH 3T3, and macrophage IC21 cell lines transfected with pD2/LAMP showed that the preM-E/LAMP protein chimera was present in vesicles containing endogenous LAMP and major histocompatability complex class II (MHC II), in contrast to the non-vesicular localization of native preM-E protein lacking the LAMP targeting sequence. Mice immunized with pD2 showed an antigen-specific immunoglobulin response but the neutralizing antibodies titers (plaque reduction neutralization test, PRNT(50)) elicited by the native protein were minimal. In contrast, vaccination with pD2/LAMP resulted in PRNT(50) of 270, 320 and 160 at approximately 1, 3 and 8 months after two immunizations with 50 microg DNA, and approached 100% neutralization at 1:20 dilution. Additional immunization with pD2/LAMP, after 8 months, increased the neutralizing antibody titers to >640. Comparable neutralizing antibody responses were induced by two vector backbones, pVR1012 and pVax-1, at 5 and 50 microg of DNA. The neutralizing responses to the pD2/LAMP chimera were greatly superior to those elicited by pD2 in all conditions. These results underscore the importance of MHC class II presentation of DNA-encoded dengue-virus envelope protein for production of neutralizing antibodies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Viral / immunology
  • COS Cells
  • Cell Adhesion Molecules, Neuronal / immunology
  • Cell Line
  • Chlorocebus aethiops
  • Dengue Virus / immunology*
  • GPI-Linked Proteins
  • Histocompatibility Antigens Class II / immunology*
  • Immunoglobulin G / immunology
  • Injections, Subcutaneous
  • Mice
  • Mice, Inbred BALB C
  • Vaccines, DNA / administration & dosage
  • Vaccines, DNA / immunology*
  • Vero Cells
  • Viral Vaccines / administration & dosage
  • Viral Vaccines / immunology*


  • Antibodies, Viral
  • Cell Adhesion Molecules, Neuronal
  • GPI-Linked Proteins
  • Histocompatibility Antigens Class II
  • Immunoglobulin G
  • Vaccines, DNA
  • Viral Vaccines
  • limbic system-associated membrane protein