title: Immunogenicity and Protective Efficacy of Brugia malayi Heavy Chain Myosin as Homologous DNA, Protein and Heterologous DNA/Protein Prime Boost Vaccine in Rodent Model.
abstract: We earlier demonstrated the immunoprophylactic efficacy of recombinant heavy chain myosin (Bm-Myo) of Brugia malayi (B. malayi) in rodent models. In the current study, further attempts have been made to improve this efficacy by employing alternate approaches such as homologous DNA (pcD-Myo) and heterologous DNA/protein prime boost (pcD-Myo+Bm-Myo) in BALB/c mouse model. The gene bm-myo was cloned in a mammalian expression vector pcDNA 3.1(+) and protein expression was confirmed in mammalian Vero cell line. A significant degree of protection (79.2%±2.32) against L3 challenge in pcD-Myo+Bm-Myo immunized group was observed which was much higher than that exerted by Bm-Myo (66.6%±2.23) and pcD-Myo (41.6%±2.45). In the heterologous immunized group, the percentage of peritoneal leukocytes such as macrophages, neutrophils, B cells and T cells marginally increased and their population augmented further significantly following L3 challenge. pcD-Myo+Bm-Myo immunization elicited robust cellular and humoral immune responses as compared to pcD-Myo and Bm-Myo groups as evidenced by an increased accumulation of CD4+, CD8+ T cells and CD19+ B cells in the mouse spleen and activation of peritoneal macrophages. Though immunized animals produced antigen-specific IgG antibodies and isotypes, sera of mice receiving pcD-Myo+Bm-Myo or Bm-Myo developed much higher antibody levels than other groups and there was profound antibody-dependent cellular adhesion and cytotoxicity (ADCC) to B. malayi infective larvae (L3). pcD-Myo+Bm-Myo as well as Bm-Myo mice generated a mixed T helper cell phenotype as evidenced by the production of both pro-inflammatory (IL-2, IFN-γ) and anti-inflammatory (IL-4, IL-10) cytokines. Mice receiving pcD-Myo on contrary displayed a polarized pro-inflammatory immune response. The findings suggest that the priming of animals with DNA followed by protein booster generates heightened and mixed pro- and anti-inflammatory immune responses that are capable of providing high degree of protection against filarial larval invasion.
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, BS 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, India.|Academy of scientific and Innovative Research, New Delhi, India.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Gupta
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, BS 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, India.
authors.forename: Manisha
authors.initials: M
authors.lastname: Pathak
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, BS 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, India.|Academy of scientific and Innovative Research, New Delhi, India.
authors.forename: Sweta
authors.initials: S
authors.lastname: Misra
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, BS 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, India.|Academy of scientific and Innovative Research, New Delhi, India.
authors.forename: Shailja
authors.initials: S
authors.lastname: Misra-Bhattacharya
mesh_terms: D000818:Animals; D017178:Brugia malayi / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology; D002448:Cell Adhesion; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D016207:Cytokines / Q000378:metabolism; D005368:Filariasis / Q000517:prevention & control*; D005786:Gene Expression Regulation; D016753:Interleukin-10 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology; D017737:Macrophages, Peritoneal / Q000166:cytology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018995:Myosin Heavy Chains / Q000276:immunology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013154:Spleen / Q000166:cytology; D018418:Th2 Cells / Q000166:cytology; D019444:Vaccines, DNA / Q000276:immunology*; D014709:Vero Cells
chemical_list: D016207:Cytokines; D017382:Reactive Oxygen Species; D019444:Vaccines, DNA; D016753:Interleukin-10; D015847:Interleukin-4; D018995:Myosin Heavy Chains