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Abstract
DNA vaccine approaches have been applied to generate the protective immunity against various pathogens. However, the strength of immune responses induced by DNA vaccine is weak compared with conventional vaccines. The prime-boost vaccination using DNA vaccine and other viral vector has been suggested as one way to circumvent this limitation. In the present study, we used in vivo CTL activity assay to determine CD8+ T cell-mediated immunity induced by prime-boost vaccination with a DNA vaccine (gB498–505 DNA) and recombinant vaccinia virus (VVgB498–505) expressing gB498–505 epitope peptide (SSIEFARL) of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB). The most potent in vivo CTL activity was induced in mice received VVgB498–505 when both gB498–505 and VVgB498–505 were used at priming step and boosted with the alternative vaccine vector expressing whole antigen protein (gBw). Priming with vaccine vector expressing gBw followed by the use of VVgB498–505 at boosting step also induced strong in vivo CTL activity. We also examined in vivo CTL activity after immunization of mice with epitope-expressing vaccine vector at both priming and boosting step. Curiously, in vivo CTL activity mediated by CD8+ T cells was strongly elicited at memory stage when animals were primed with VVgB498–505 and subsequently boosted with gB498–505 DNA. Because the use of VVgB498–505 at priming followed by boosting with gB498–505 DNA induced most optimal immunity, these results suggest that the order of vaccine type should be carefully considered when used vaccine type expressing only epitope for prime-boost vaccination.
References
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Figure 1.
(A) Diagram for prime-boost immunization. C57BL/6 (n=6∼7) were immunized i.m. with VVgB498–505, gB498–505 DNA, VVgBw, or gBw DNA, and boosted via the same route with the alternative vaccine type. Two (acute phase) and five (memory phase) weeks later, in vivo CTL killing activity was determined. (B) In vivo CTL killing assay for determining immunity mediated by epitope-specific CD8+ T cells. Splenocytes from naïve C57BL/6 mice (n=6∼7) were evenly split into two populations. One was plused with gB498–505 peptide (1 μg/ml) and then labeled with a high concentration (5.0 μM) of CFSE (CFSEhigh/gB498–505). The other was incubated without epitope peptide and labeled with with a low concentration (0.5 μM) of CFSE (CFSElow/No peptide). An equal number of cells from each population were mixed together and adoptively transferred into mice immunized with prime-boost protocols. Both CFSEhigh/gB498–505 and CFSElow/No peptide cells were analyzed with the flow cytometry 5 h after adoptive transfer. The histogram shows the reduced peak of CFSEhigh/gB498–505 due to cytolysis by epitope-specific CD8+ T cells.
Figure 2.
In vivo CTL killing activity of C57BL/6 mice immunized with prime-boost protocols using recombinant vaccinia virus (VVgB498–505) and DNA vaccine (gB498–505) expressing gB498–505 epitope peptide at priming. C57BL/6 (n=6∼7) were immunized i.m. with VVgB498–505, gB498–505 DNA, VVgBw, or gBw DNA and boosted via the same route with either gBw DNA or VVgBw. Two (A; acute phase) and five (B; memory phase) weeks later, in vivo CTL killing activity was determined. The graphs show the average and standard deviation (SD) of three to four mice per experiment.
Figure 3.
In vivo CTL killing activity of C57BL/6 mice immunized with prime-boost protocols using recombinant vaccinia virus (VVgB498–505) and DNA vaccine (gB498–505) expressing gB498–505 epitope peptide at boosting. C57BL/6 (n=6∼7) were immunized i.m. with either VVgBw or gBw DNA, and boosted via the same route with VVgB498–505, gB498–505 DNA, VVgBw, or gBw DNA. Two (A; acute phase) and five (B; memory phase) weeks later, in vivo CTL killing activity was determined. The graphs show the average and standard deviation (SD) of three to four mice per experiment.
Figure 4.
In vivo CTL killing activity of C57BL/6 mice immunized with prime-boost protocols using recombinant vaccinia virus (VVgB498–505) and DNA vaccine (gB498–505) expressing gB498–505 epitope peptide at priming and boosting. C57BL/6 (n=6∼7) were immunized i.m. with VVgB498–505, gB498–505 DNA, VVgBw, or gBw DNA, and boosted via the same route with the alternative vaccine type. Two (A; acute phase) and five (B; memory phase) weeks later, in vivo CTL killing activity was determined. The graphs show the average and standard deviation (SD) of three to four mice per experiment.
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