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Merck
  • Modulation of hepatitis C virus core DNA vaccine immune responses by co-immunization with CC-chemokine ligand 20 (CCL20) gene as immunoadjuvant.

Modulation of hepatitis C virus core DNA vaccine immune responses by co-immunization with CC-chemokine ligand 20 (CCL20) gene as immunoadjuvant.

Molecular biology reports (2014-06-29)
Christine Hartoonian, Zargham Sepehrizadeh, Mehdi Mahdavi, Arash Arashkia, Yon Suk Jang, Maasoumeh Ebtekar, Mojtaba Tabatabai Yazdi, Babak Negahdari, Azita Nikoo, Kayhan Azadmanesh
ABSTRACT

Plasmid DNA vaccination is a promising vaccine platform for prevention and treatment of infectious disease. Enhancement of the DNA vaccine potency by co-inoculation of immunoadjuvant has been shown to be an effective strategy. Modulation of dendritic cells and T-cells locomotion and trafficking to prime an immune response is mediated by distinct chemokines. The recent study was designed to elucidate the adjuvant activity of plasmid expressing CC-chemokine ligand 20 (pCCL20) in co-inoculation with hepatitis C virus (HCV) core DNA vaccine immunization. pCCL20 was constructed and evaluated for its functional expression. Sub-cutaneous inoculation of pCCL20 with HCV core DNA vaccine was performed via electroporation in BALB/c mice on day 0 and 14 and a HCV core protein booster was applied on day 28. On week after final immunization, both humoral and cell-mediated immune responses were assessed by indirect ELISA for core specific antibodies, lymphocyte proliferation, cytokine ELISA/ELISpot and cytotoxic Grenzyme B (GrzB) release assays. Mice were co-immunized with pCCL20 developed higher levels of core specific IFN-γ/IL-4 ratio and IL-2 release, IFN-γ producing cells, lymphocyte proliferation and cytotoxic Grenzyme B release in both draining lymph nodes and spleen cells of immunized mice. The core-specific serum total IgG and IgG2a/IgG1 ratio were significantly higher when the pCCL20 was co-inoculated. These results suggest the potential of CCL20 chemokine as vaccine adjuvant to enhance Th1 mediated cellular and humoral immune responses in HCV core DNA immunization.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Rat IL-4 ELISA Kit, for serum, plasma and cell culture supernatant
Sigma-Aldrich
Rat IL-4 ELISA Kit, for cell and tissue lysates
Sigma-Aldrich
Rat Ccl20 / C-C Motif Chemokine 20 ELISA Kit
Sigma-Aldrich
Cotton Rat IL4 / Interleukin 4 ELISA Kit
Sigma-Aldrich
Mouse IL-4 ELISA Kit, for cell and tissue lysates
Sigma-Aldrich
Mouse IL-4 ELISA Kit, for serum, plasma and cell culture supernatant
Sigma-Aldrich
Human IL-4 ELISA Kit, for serum, plasma, cell culture supernatant and urine