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Bacillus Calmette-Guérin vaccination at birth and in vitro cytokine responses to non-specific stimulation. A randomized clinical trial

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Abstract

Several studies have shown increased in vitro cytokine responses to non-related pathogens after Bacillus Calmette-Guérin (BCG) vaccination. A total of 158 infants (80 BCG administered within 7 days of birth; 78 controls) were bled 4 days post-randomization, and at age 3 and 13 months. Geometric mean concentrations of IL-1β, TNF-α, IL-6 (24 h stimulation) and IFN-γ, IL-10, IL-17, IL-22 (96 h stimulation) in response to in vitro stimulation with RPMI, LPS, PHA, Escherichia coli, Streptococcus pneumoniae, Candida albicans and BCG were compared among BCG vaccinated children and controls. BCG vaccination did not affect in vitro cytokine production, except IFN-γ and IL-22 response to BCG. Stratifying for ‘age at randomization’ we found a potentiating effect of BCG on cytokine production (TNF-α, IL-6, IL-10) in the 4 days post randomization stimulations, among children who were vaccinated at age 2–7 days versus age 0–1 days. BCG vaccination did not potentiate cytokine production to non-BCG antigens. At 4 days post randomization, BCG was associated with higher cytokine production in the later randomized children.

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Acknowledgements

Thanks to Monica Ladekarl for vaccinating most of the children; Linda Billetorp and Birgit Peitersen for examining and interviewing most of the participants; Vera Zingmark, Trine Mølbæk Jensen, Signe Kjeldgaard Jensen and Arild Ejsing for assisting the bleeding and lab work. We thank Heidi Lemmers and Helga Dijkstra for their help with the ELISAs. We thank the families and children for their participation, and The Danish Calmette Study staff for their commitment.

Funding

This work was supported by Copenhagen University Hospital, Hvidovre, and The Danish National Research Foundation (DNRF108). The funders did not have any role in the: study design, data collection, analysis and interpretation of data, writing the paper or in the decision to submit the article for publication.

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Authors and Affiliations

  1. Department of Pediatrics, 460, Copenhagen University Hospital, Kettegaard Allé 30, DK-2650, Hvidovre, Denmark

    T. N. Nissen, N. M. Birk, T. Hoffmann, D. L. Jeppesen & O. Pryds

  2. Division of Experimental Internal Medicine, Department of Internal Medicine, Radboud University Medical Center and Radboud Center for Infectious Diseases, Geert Grooteplein 10, 6525GA, Nijmegen, The Netherlands

    B. A. Blok, R. J. W. Arts & M. G. Netea

  3. Research Center for Vitamins and Vaccines (CVIVA), Bandim Health Project, Statens Serum Institut, Artillerivej 5, DK-2300, Copenhagen S, Denmark

    B. A. Blok, A. Andersen, P. Aaby & C. S. Benn

  4. Odense Patient data Explorative Network, Odense University Hospital/Institute of Clinical Research, University of Southern Denmark, Odense, Denmark

    B. A. Blok & C. S. Benn

  5. The Child and Adolescent Clinic 4072, Juliane Marie Centret, Rigshospitalet, Copenhagen University Hospital, Blegdamsvej 9, DK-2100, Copenhagen Ø, Denmark

    J. Kjærgaard & L. G. Stensballe

  6. Department of Pediatrics, Kolding Hospital, Skovvangen 2-8, DK-6000, Kolding, Denmark

    L. M. Thøstesen & P.-E. Kofoed

  7. Viro-immunology Research Unit, Department of Infectious Diseases, Rigshospitalet, Copenhagen University Hospital, Blegdamsvej 9, DK-2100, Copenhagen Ø, Denmark

    S. D. Nielsen

  8. Human Immunology, Infectious Disease Immunology, Division of Vaccine, Statens Serum Institut, DK-2300, Artillerivej 5, Denmark

    M. Ruhwald

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  1. T. N. Nissen
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Contributions

CB and MGN conceived the idea; TNN, MR, CB, OP and MGN designed the experiments. CB, PA, LGS, DLJ, PEK, TH, SN and OP supervised the collection of data in the overall Danish Calmette Study. JK and LMT participated in the overall data collection of The Danish Calmette Study. TNN and NMB collected the data for this sub-study. TNN and NBE initiated and managed the collection of biological samples of the cohort. BAB and RJWA participated and MGN supervised the ELISA analysis. TNN carried out data management, and analyzed the data with help from MGN, CB, MR, AA and OP. AA supervised the statistical analyses. TNN drafted the manuscript. All authors read and approved the final manuscript.

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Correspondence to T. N. Nissen.

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Conflict of interest

All authors have completed the Unified Competing Interest form at www.icmje.org/coi_disclosure.pdf (available on request from the corresponding author), and all authors declare to have no potential conflict of interest. MGN was supported by an ERC Consolidator Grant (#310372) and a Spinoza grant of the Netherlands Organization for Scientific Research. MR receives funding from European Commission H2020 program [grant number TBVAC2020 643,381] and Research Council Norway [GLOBVAC 248042/H10].

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Suppl. Figure 1

Dot plot of IFN-γ response to BCG stimulation in BCG vaccinated children and controls at 13 months. The dotted line displays the cut-off of 107 pg/ml and was defined as the value where 95% of the control children responded below. Of the BCG vaccinated children, 54% had IFN-γ response above the cut-off (GIF 4 kb)

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Nissen, T.N., Birk, N.M., Blok, B.A. et al. Bacillus Calmette-Guérin vaccination at birth and in vitro cytokine responses to non-specific stimulation. A randomized clinical trial. Eur J Clin Microbiol Infect Dis 37, 29–41 (2018). https://doi.org/10.1007/s10096-017-3097-2

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