Giuseppe
Emeritus
[Source: Proceedings of the National Academy of Sciences of the United States of America, full page: (LINK). Abstract, edited.]
The adjuvant MF59 induces ATP release from muscle that potentiates response to vaccination
Maria Vono<SUP>a</SUP>,<SUP>b</SUP>, Marianna Taccone<SUP>b</SUP>, Paola Caccin<SUP>a</SUP>, Marilena Gallotta<SUP>b</SUP>, Giovanna Donvito<SUP>c</SUP>, Simonetta Falzoni<SUP>c</SUP>, Emiliano Palmieri<SUP>b</SUP>, Michele Pallaoro<SUP>b</SUP>, Rino Rappuoli<SUP>b</SUP>,<SUP>1</SUP>, Francesco Di Virgilio<SUP>c</SUP>, Ennio De Gregorio<SUP>b</SUP>,<SUP>1</SUP>, Cesare Montecucco<SUP>a</SUP>,<SUP>1</SUP>, and Anja Seubert<SUP>b</SUP>
<SUP></SUP>
Author Affiliations: <SUP>a</SUP>Dipartimento di Scienze Biomediche and Istituto di Neuroscienze del Consiglio Nazionale delle Ricerche, Universit? di Padova, 35131 Padova, Italy; <SUP>b</SUP>Research Center, Novartis Vaccines and Diagnostics, 53100 Siena, Italy; and <SUP>c</SUP>Dipartimento di Morfologia, Chirurgia e Medicina Sperimentale, Universit? di Ferrara, 44121 Ferrara, Italy
Contributed by Rino Rappuoli, November 4, 2013 (sent for review August 1, 2013)
Published online before print December 9, 2013, doi: 10.1073/pnas.1319784110 / <ABBR>PNAS</ABBR> December 9, 2013
Significance
Release of endogenous danger signals has been found to be a key mechanism of many conventional, broadly used adjuvants. Yet extracellular ATP has so far not been linked to vaccination success. Here we show that ATP release from muscle is crucial for the mechanism of action of the vaccine adjuvant MF59, leading to efficient CD4 T-cell priming and high antibody titers.
Abstract
Vaccines are the most effective agents to control infections. In addition to the pathogen antigens, vaccines contain adjuvants that are used to enhance protective immune responses. However, the molecular mechanism of action of most adjuvants is ill-known, and a better understanding of adjuvanticity is needed to develop improved adjuvants based on molecular targets that further enhance vaccine efficacy. This is particularly important for tuberculosis, malaria, AIDS, and other diseases for which protective vaccines do not exist. Release of endogenous danger signals has been linked to adjuvanticity; however, the role of extracellular ATP during vaccination has never been explored. Here, we tested whether ATP release is involved in the immune boosting effect of four common adjuvants: aluminum hydroxide, calcium phosphate, incomplete Freund?s adjuvant, and the oil-in-water emulsion MF59. We found that intramuscular injection is always associated with a weak transient release of ATP, which was greatly enhanced by the presence of MF59 but not by all other adjuvants tested. Local injection of apyrase, an ATP-hydrolyzing enzyme, inhibited cell recruitment in the muscle induced by MF59 but not by alum or incomplete Freund?s adjuvant. In addition, apyrase strongly inhibited influenza-specific T-cell responses and hemagglutination inhibition titers in response to an MF59-adjuvanted trivalent influenza vaccine. These data demonstrate that a transient ATP release is required for innate and adaptive immune responses induced by MF59 and link extracellular ATP with an enhanced response to vaccination.
vaccine adjuvants - danger associated molecular pattern ? DAMP ? inflammation
Footnotes
<SUP>1</SUP>To whom correspondence may be addressed. E-mail: rino.rappuoli@novartis.com, cesare.montecucco@gmail.com, or ennio.de_gregorio@novartis.com.
Author contributions: M.V., R.R., F.D.V., E.D.G., C.M., and A.S. designed research; M.V., M.T., P.C., M.G., G.D., S.F., and A.S. performed research and analyzed data; E.P. and M.P. contributed adjuvant formulations and analytic tools; and M.V., R.R., F.D.V., E.D.G., C.M., and A.S. wrote the paper.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1319784110/-/DCSupplemental.
Freely available online through the PNAS open access option.
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The adjuvant MF59 induces ATP release from muscle that potentiates response to vaccination
Maria Vono<SUP>a</SUP>,<SUP>b</SUP>, Marianna Taccone<SUP>b</SUP>, Paola Caccin<SUP>a</SUP>, Marilena Gallotta<SUP>b</SUP>, Giovanna Donvito<SUP>c</SUP>, Simonetta Falzoni<SUP>c</SUP>, Emiliano Palmieri<SUP>b</SUP>, Michele Pallaoro<SUP>b</SUP>, Rino Rappuoli<SUP>b</SUP>,<SUP>1</SUP>, Francesco Di Virgilio<SUP>c</SUP>, Ennio De Gregorio<SUP>b</SUP>,<SUP>1</SUP>, Cesare Montecucco<SUP>a</SUP>,<SUP>1</SUP>, and Anja Seubert<SUP>b</SUP>
<SUP></SUP>
Author Affiliations: <SUP>a</SUP>Dipartimento di Scienze Biomediche and Istituto di Neuroscienze del Consiglio Nazionale delle Ricerche, Universit? di Padova, 35131 Padova, Italy; <SUP>b</SUP>Research Center, Novartis Vaccines and Diagnostics, 53100 Siena, Italy; and <SUP>c</SUP>Dipartimento di Morfologia, Chirurgia e Medicina Sperimentale, Universit? di Ferrara, 44121 Ferrara, Italy
Contributed by Rino Rappuoli, November 4, 2013 (sent for review August 1, 2013)
Published online before print December 9, 2013, doi: 10.1073/pnas.1319784110 / <ABBR>PNAS</ABBR> December 9, 2013
Significance
Release of endogenous danger signals has been found to be a key mechanism of many conventional, broadly used adjuvants. Yet extracellular ATP has so far not been linked to vaccination success. Here we show that ATP release from muscle is crucial for the mechanism of action of the vaccine adjuvant MF59, leading to efficient CD4 T-cell priming and high antibody titers.
Abstract
Vaccines are the most effective agents to control infections. In addition to the pathogen antigens, vaccines contain adjuvants that are used to enhance protective immune responses. However, the molecular mechanism of action of most adjuvants is ill-known, and a better understanding of adjuvanticity is needed to develop improved adjuvants based on molecular targets that further enhance vaccine efficacy. This is particularly important for tuberculosis, malaria, AIDS, and other diseases for which protective vaccines do not exist. Release of endogenous danger signals has been linked to adjuvanticity; however, the role of extracellular ATP during vaccination has never been explored. Here, we tested whether ATP release is involved in the immune boosting effect of four common adjuvants: aluminum hydroxide, calcium phosphate, incomplete Freund?s adjuvant, and the oil-in-water emulsion MF59. We found that intramuscular injection is always associated with a weak transient release of ATP, which was greatly enhanced by the presence of MF59 but not by all other adjuvants tested. Local injection of apyrase, an ATP-hydrolyzing enzyme, inhibited cell recruitment in the muscle induced by MF59 but not by alum or incomplete Freund?s adjuvant. In addition, apyrase strongly inhibited influenza-specific T-cell responses and hemagglutination inhibition titers in response to an MF59-adjuvanted trivalent influenza vaccine. These data demonstrate that a transient ATP release is required for innate and adaptive immune responses induced by MF59 and link extracellular ATP with an enhanced response to vaccination.
vaccine adjuvants - danger associated molecular pattern ? DAMP ? inflammation
Footnotes
<SUP>1</SUP>To whom correspondence may be addressed. E-mail: rino.rappuoli@novartis.com, cesare.montecucco@gmail.com, or ennio.de_gregorio@novartis.com.
Author contributions: M.V., R.R., F.D.V., E.D.G., C.M., and A.S. designed research; M.V., M.T., P.C., M.G., G.D., S.F., and A.S. performed research and analyzed data; E.P. and M.P. contributed adjuvant formulations and analytic tools; and M.V., R.R., F.D.V., E.D.G., C.M., and A.S. wrote the paper.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1319784110/-/DCSupplemental.
Freely available online through the PNAS open access option.
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