Progresses in DNA-Based Heterologous Prime-Boost Immunization Strategies

Abstract

Although recombinant DNA and recombinant viral vectors expressing HIV antigens have yielded positive outcomes in animal models, these vaccines have not been effectively translated to humans. Despite this, there is still a high level of optimism that poxviral-based vaccine strategies could offer the best hope for developing an effective vaccine against not only HIV-1 but also other chronic diseases where good-quality T and B cell immunity is needed for protection. In this chapter we discuss step by step (1) how recombinant poxviral vectors co-expressing HIV antigens and promising mucosal/systemic adjuvants (e.g., IL-13Rα2) are constructed, (2) how these vectors can be used in alternative heterologous prime-boost immunization strategies, (3) how systemic and mucosal samples are prepared for analysis, followed by (4) two immunological assays: multicolor intracellular cytokine staining and tetramer/homing maker analysis that are used to evaluate effective systemic and mucosal T cell immunity.

Heterologous Prime-Boost Vaccination Enhances TsPmy’s Protective Immunity against Trichinella spiralis Infection in a Murine Model

“…In general, DNA vaccine and recombinant protein vaccine stimulates different antigen-specific immune response using different mechanisms, the former induce strong Th1 cellular immune responses and also prime antigen-specific memory B cells, and the latter induce predominant Th2 antibody responses (Jackson et al, 2014; Kardani et al, 2016). This different immunological mechanisms lead to the heterogeneous DNA-prime and protein-boost immunization regimen in order to enhance both cellular and humoral immune responses.…”

“…Heterologous prime-boost immunization regimen, primed by DNA vaccine and boosted by recombinant protein, has be proven to induce stronger cellular and humoral immunity (Jackson et al, 2014; Kardani et al, 2016) and the better protection against some infectious diseases such as Leishmania infantum (Shahbazi et al, 2015), Toxoplasma gondii (Han et al, 2017), Plasmodium yoelii (Cabrera-Mora et al, 2016), and Schistosoma japonicum (Dai et al, 2017). In present study we used Salmonella -delivered Ts Pmy DNA as prime immunization followed by two boost immunizations with expressed recombinant Ts Pmy protein (r Ts Pmy) in order to enhance Ts Pmy’s immunogenicity and protective efficacy against T. spiralis infection in a murine model.…”

Recombinant fowlpox virus vector-based vaccines: expression kinetics, dissemination and safety profile following intranasal delivery

“…Recombinant viruses were constructed by homologous recombination as described previously [ 50 ]. Parent rFPV091-HIVgagpol constructed previously [ 26 ] using the vector pKG10a [ 49 ] contains the insertion of the HIVgagpol (AE clade) genes into the intergenic region between ORFs FPV133 (VV J1R homologue) and FPV134 (J3R).…”

“…Fluorescent plaques were viewed using an IX71 inverted bright-field/fluorescence microscope (Olympus). Absence of parental virus contamination was confirmed by PCR, virus stocks were prepared and viral titre was determined [ 50 ]. Viral growth curve analysis also was performed on mCherry virus.…”

Mucosal and systemic SIV-specific cytotoxic CD4+ T cell hierarchy in protection following intranasal/intramuscular recombinant pox-viral vaccination of pigtail macaques

“…S8). FPV-SIV mac239 gag/pol/env parent virus, co-expressing the macaque IL-4C123 gene driven by a late promoter was constructed as described previously 47,76 . Similarly, MVA-SIV mac239 gag/pol parent vaccine and the MVA-SIV mac239 gag/pol co-expressing macaque IL-4C123 gene were constructed (see Supplementary Fig.…”