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Comparative Study
. 2011 Jul 26;29(33):5399-406.
doi: 10.1016/j.vaccine.2011.05.083. Epub 2011 Jun 7.

Different patterns of expansion, contraction and memory differentiation of HIV-1 Gag-specific CD8 T cells elicited by adenovirus type 5 and modified vaccinia Ankara vaccines

Vinod Kumar Bhaskara Pillai  1 , Sunil Kannanganat, Pablo Penaloza-Macmaster, Lakshmi Chennareddi, Harriet L Robinson, Jerry Blackwell, Rama Rao Amara
Affiliations
Comparative Study

Different patterns of expansion, contraction and memory differentiation of HIV-1 Gag-specific CD8 T cells elicited by adenovirus type 5 and modified vaccinia Ankara vaccines

Vinod Kumar Bhaskara Pillai et al. Vaccine. .

Abstract

The magnitude and functional quality of antiviral CD8 T cell responses are critical for the efficacy of T cell based vaccines. Here, we investigate the influence of two popular viral vectors, adenovirus type 5 (Ad5) and modified vaccinia Ankara (MVA), on expansion, contraction and memory differentiation of HIV-1 Gag insert-specific CD8 T cell responses following immunization and show different patterns for the two recombinant viral vectors. The Ad5 vector primed 6-fold higher levels of insert-specific CD8 effector T cells than the MVA vector. The Ad5-primed effector cells also underwent less contraction (<2-fold) than the MVA-primed cells (>5-fold). The Ad5-primed memory cells were predominantly CD62L negative (effector memory) whereas the MVA-primed memory cells were predominantly CD62L positive (central memory). Consistent with their memory phenotype, MVA-primed CD8 T cells underwent higher fold expansion than Ad5-primed CD8 T cells following a homologous or heterologous boost. Impressively, the Ad5 boost changed the quality of MVA-primed memory response such that they undergo less contraction with effector memory phenotype. However, the MVA boost did not influence the contraction and memory phenotype of Ad5-primed response. In conclusion, our results demonstrate that vaccine vector strongly influences the expansion, contraction and the functional quality of insert-specific CD8 T cell responses and have implications for vaccine development against infectious diseases.

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Figures

Figure 1
Figure 1
Magnitude of Gag tetramer-specific CD8 T cells in blood and spleen following priming with recombinant Ad5 and MVA vaccines. (A) Representative FACS plots demonstrating the frequency of Gag-tetramer-specific CD8 T cells raised by MVA/HIV (106 pfu, i.m.) or Ad5/HIV (106 pfu, i.m.) vaccines at 2 weeks following the prime in blood. PBMC were stained for CD8, CD4, CD20, CD11a and tetramer. CD8 T cells (CD4−, CD20−, CD8+) that were positive for tetramer and CD11ahi were scored as tetramer positive cells. The numbers on the FACS plots represent the frequency of tetramer positive cells as a percent of total CD8 T cells. (B) Temporal frequency of tetramer positive cells following the prime with 106 pfu of either vector in blood. Data represents mean values with standard error of mean (SEM) for a group of 5-10 mice. (C) Frequency of tetramer positive cells following prime with 106 pfu of either vector in spleen at the indicated time points. Data represent mean values with SEM for a group of 3-5 mice. (D) Frequency of tetramer positive cells following the prime with 107 pfu of either vector in blood. Data represents mean values with SEM for a group of 5 mice.
Figure 2
Figure 2
Phenotype and magnitude of the Gag tetramer-specific memory cells following priming with recombinant Ad5 and MVA vectors. (A) Representative FACS plots showing the frequency of Gag tetramer-specific CD8 T cells expressing either CD127 or CD62L raised by MVA/HIV (106 pfu, i.m.) or Ad5/HIV (106 pfu, i.m.) vaccines at 2 and 8 weeks following prime. Splenocytes were stained for CD8, CD3, tetramer and CD62L or CD127. CD8 T cells (CD3+, CD8+) were analyzed for expression of CD62L or CD127 and tetramer binding. Open black histograms represent tetramer positive cells and closed gray histograms represent total CD8 T cells. The numbers on the FACS plots represent the frequency of tetramer-positive cells that express CD62L or CD127 expressed as a percent of total tetramer-positive cells. (B) Temporal frequency of tetramer positive cells expressing either CD127 or CD62L following prime in spleen. The frequency of tetramer positive cells that express CD62L or CD127 expressed as a percent of total tetramer positive cells is shown. Data represent mean values for a group of 3-4 mice. (C) Magnitude of CD62L positive and negative cells in spleen at 8 weeks following prime. Frequency of tetramer+ CD62L+ or tetramer+ CD62L− cells were expressed as a percent of total CD8 T cells is shown. Data represents mean values with SEM for a group of 3-4 mice.
Figure 3
Figure 3
Cytokine production patterns of Gag-specific CD8 T cells following prime with Ad5 and MVA. (A) Representative FACS plots demonstrating the co-expression of IFN-γ and IL-2 by CD8 T cells raised by MVA and Ad5 vaccines at 12 weeks following prime. Splenocytes were stimulated with media alone or Gag immunodominant peptide and stained for CD8, CD3, IFN-γ and IL-2. CD8 T cells (CD3+, CD8+) were analyzed for expression of IFN-γ and IL-2. The numbers on the FACS plots represent the frequency of IL-2 positive cells that co-express IFN-γ, expressed as a percent of total IFN-γ positive cells. (B) Proportion of tetramer positive cells expressing either IFN-γ or IL-2 at 12 weeks following prime in spleen. The frequency of IFN-γ or IL-2 positive cells expressed as a percent of tetramer positive cells is shown. (C) Summary of the frequency of IL-2 positive cells that co-express IFN-γ expressed as a percent of total IFN-γ positive cells. (D) Magnitude of IFN-γ and IFN-γ plus IL-2 positive cells in spleen. Frequency of IFN-γ or IFN-γ plus IL-2 positive cells expressed as a percent of total CD8 T cells is shown. Data represents mean values with SEM for a group of 4 mice. * - Significantly different, p<0.05.
Figure 4
Figure 4
Expansion of Gag-specific CD8 T cells following boost in blood. MVA or Ad5 primed mice were boosted with either MVA or Ad5 at 8-10 weeks after the prime. (A) Expansion of MVA-primed Gag tetramer-specific CD8 T cells following MVA or Ad5 boost. (B) Expansion of Ad5-primed Gag tetramer-specific CD8 T cells following MVA or Ad5 boost. (C) Fold increase in the frequency of Gag tetramer-specific CD8 T cells following boost. Fold increase was calculated as a ratio of tetramer positive cells at week 1 post boost (post) and pre boost (pre). (D) Fold contraction of the frequency of Gag tetramer-specific CD8 T cells following boost. Fold contraction was calculated as a ratio of tetramer positive cells at week 1 post boost and week 10 post boost. (E) CD62L expression by Gag tetramer positive cells at 10 weeks post boost. ***, p<0.001; **, p<0.01; *, p<0.05; p values indicate significantly lower or higher responses compared to MVA/MVA group.
Figure 5
Figure 5
Persistence of (A) Ad5 viral vector but not (B) MVA viral vector in spleen at 12 weeks following prime. DNA was isolated from splenocytes and analyzed for viral genomes using semi-quantitative PCR as described in methods. Analyses for Ad5 genome were performed in duplicates. MVA standard was generated as described in materials and methods.
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