Construction of, and T-helper (Th)1/Th2 immune responses to, a herpes simplex virus type 2 glycoprotein D-cytotoxic T-lymphocyte epitope DNA vaccine

Clin Exp Dermatol. 2010 Jul;35(5):537-42. doi: 10.1111/j.1365-2230.2009.03673.x. Epub 2010 May 20.

Abstract

Background: Recombinant cytotoxic T lymphocyte (CTL) epitope DNA vaccines offer an attractive approach for the induction of robust cellular and humoral immune responses directed against human pathogen target antigens.

Aim: To construct a pVAX-gD-CTL vector expressing the glycoprotein (g)D and gB CTL epitopes from herpes simplex virus type 2 (HSV2) and evaluate it in mice for immunogenicity and protective efficacy against intraperitoneal challenge with the HSV2 strain Sav.

Methods: The gD gene transcript and gB CTL epitope were inserted into the pVAX vector to obtain the recombinant plasmid pVAX-gD-CTL. An in vitro study was then conducted to detect the expression of gD by immunocytochemistry and western blotting. BALB/c mice were immunized with this DNA vaccine, then the CTL activity and expression of anti-HSV2 gD IgG, interferon-gamma and interleukin-4 by lactate dehydrogenase release assay and ELISA, respectively. The protection given to the mice was assayed by a fatal dose of virus.

Results: The pVAX-gD-CTL vector was successfully constructed and could express gD in COS-7 cells. Immunogenicity of gD, anti-HSV2 gD neutralizing serum IgG antibody and robust Th1-polarized immune responses were found. Furthermore, mice were prophylactically protected from challenge with a high dose of HSV2.

Conclusions: In summary, pVAX-gD-CTL vector was successfully used to elicit potent Th1-like cellular and humoral immune responses that were protective against HSV2 disease in mice.

MeSH terms

  • Animals
  • Antibody Formation / genetics
  • Antibody Formation / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology*
  • Herpes Simplex Virus Vaccines / genetics
  • Herpes Simplex Virus Vaccines / immunology*
  • Humans
  • Interferon-gamma / genetics
  • Interferon-gamma / immunology
  • Interleukin-4 / genetics
  • Interleukin-4 / immunology
  • Mice
  • Mice, Inbred BALB C
  • Models, Animal
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology*

Substances

  • Epitopes, T-Lymphocyte
  • Herpes Simplex Virus Vaccines
  • Vaccines, DNA
  • Viral Envelope Proteins
  • glycoprotein D-herpes simplex virus type 2
  • Interleukin-4
  • Interferon-gamma