Safety and immunogenicity of boosting BCG vaccinated subjects with BCG: comparison with boosting with a new TB vaccine, MVA85A

Kathryn T Whelan, Ansar A Pathan, Clare R Sander, Helen A Fletcher, Ian Poulton, Nicola C Alder, Adrian V S Hill, Helen McShane, Kathryn T Whelan, Ansar A Pathan, Clare R Sander, Helen A Fletcher, Ian Poulton, Nicola C Alder, Adrian V S Hill, Helen McShane

Abstract

Objectives: To investigate the safety and immunogenicity of a booster BCG vaccination delivered intradermally in healthy, BCG vaccinated subjects and to compare with a previous clinical trial where BCG vaccinated subjects were boosted with a new TB vaccine, MVA85A.

Design: Phase I open label observational trial, in the UK. Healthy, HIV-negative, BCG vaccinated adults were recruited and vaccinated with BCG. The primary outcome was safety; the secondary outcome was cellular immune responses to antigen 85, overlapping peptides of antigen 85A and tuberculin purified protein derivative (PPD) detected by ex vivo interferon-gamma (IFN-gamma) ELISpot assay and flow cytometry.

Results and conclusions: BCG revaccination (BCG-BCG) was well tolerated, and boosting of pre-existing PPD-specific T cell responses was observed. However, when these results were compared with data from a previous clinical trial, where BCG was boosted with MVA85A (BCG-MVA85A), MVA85A induced significantly higher levels (>2-fold) of antigen 85-specific CD4+ T cells (both antigen and peptide pool responses) than boosting with BCG, up to 52 weeks post-vaccination (p = 0.009). To identify antigen 85A-specific CD8+ T cells that were not detectable by ex vivo ELISpot and flow cytometry, dendritic cells (DC) were used to amplify CD8+ T cells from PBMC samples. We observed low, but detectable levels of antigen 85A-specific CD8+ T cells producing IFNgamma (1.5% of total CD8 population) in the BCG primed subjects after BCG boosting in 1 (20%) of 5 subjects. In contrast, in BCG-MVA85A vaccinated subjects, high levels of antigen 85A-specific CD8+ T cells (up to 14% total CD8 population) were observed after boosting with MVA85A, in 4 (50%) of 8 subjects evaluated. In conclusion, revaccination with BCG resulted in modest boosting of pre-existing immune responses to PPD and antigen 85, but vaccination with BCG-MVA85A induced a significantly higher response to antigen 85 and generated a higher frequency of antigen 85A-specific CD8+ T cells.

Trial registration: ClinicalTrials.gov NCT00654316 NCT00427830.

Conflict of interest statement

Competing Interests: AAP, AVSH and HMcS are all co-inventors on a composition of matter patent for MVA85A and are shareholders in a Joint Venture established to develop this vaccine.

Figures

Figure 1. Consort Flowcharts for (a) BCG-BCG…
Figure 1. Consort Flowcharts for (a) BCG-BCG and (b) BCG-MVA85A Vaccination Regimens After 52 Weeks Post-Vaccination.
Figure 2. Median IFNγ ELISpot Responses to…
Figure 2. Median IFNγ ELISpot Responses to PPD, antigen 85A protein, and SPP, up to 52 Weeks Post-Vaccination.
Median IFNγ ELISpot responses were assessed for PBMC from subjects in the BCG-BCG and BCG-MVA85A vaccination regimens against: (A) Tuberculin PPD; (B) Purified antigen 85 protein; (c) summed pooled peptide (SPP) of antigen 85A peptides. (d) A dot plot of AUC0–52weeks values calculated for PPD, antigen 85A protein, and SPP, and statistical analyses performed using Mann-Whitney test to compare results from BCG-BCG and BCG-MVA85A subjects. The results demonstrate significantly higher responses to antigen 85A protein (p = 0.009) and SPP (p = 0.0001) in BCG-MVA subjects compared with BCG-BCG subjects.
Figure 3. Comparison of IFNγ ELISpot Responses…
Figure 3. Comparison of IFNγ ELISpot Responses in BCG-BCG and BCG-MVA85A Subjects.
IFNγ ELISpot responses for BCG-BCG and BCG-MVA85A vaccine regimens at (A) 1 week and (B) 52 weeks post-vaccination. Statistically significant differences between the two vaccination regimens were assessed using Mann-Whitney test. BCG-MVA85A subjects had significantly greater responses to antigen 85A protein and SPP compared with BCG-BCG subjects.
Figure 4. Dendritic Cell Amplification of TB-Specific,…
Figure 4. Dendritic Cell Amplification of TB-Specific, IFNγ Producing CD8 T Cells.
TB-specific CD8+ T cells producing IFNγ in response to an immunodominant peptide KLIANNTRV, were identified in HLA-A2 positive subjects in the BCG-BCG and BCG-MVA85A vaccine regimens at 2 and 4 weeks. TB-specific CD8+ T cells were not detectable above 0.5% TB-specific CD8+ T cells at the 0, 1, 8, and 24 week timepoints analysed (data not shown). Values are presented as mean (+SD) and represents triplicate samples.

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