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Immune response at birth, long-term immune memory and 2 years follow-up after in-utero anti-HBV DNA immunization

Gene Therapy volume 11, pages 544–551 (2004)Cite this article

Abstract

Infections occurring at the end of pregnancy, during birth or by breastfeeding are responsible for the high toll of death among first-week infants. In-utero DNA immunization has demonstrated the effectiveness in inducing specific immunity in newborns. A major contribution to infant immunization would be achieved if a vaccine proved able to be protective as early as at the birth, preventing the typical ‘first-week infections’. To establish its potential for use in humans, in-utero DNA vaccination efficiency has to be evaluated for short- and long-term safety, protection at delivery, efficacy of boosts in adults and effective window/s for modulation of immune response during pregnancy, in an animal model suitable with human development. Here we show that a single intramuscular in-utero anti-HBV DNA immunization at two-thirds of pig gestation produces, at birth, antibody titers considered protective in humans. The boost of antibody titers in every animal following recall at 4 and 10 months demonstrates the establishment of immune memory. The safety of in-utero fetus manipulation is guaranteed by short-term (no fetus loss, lack of local alterations, at-term spontaneous delivery, breastfeeding) and long-term (2 years) monitoring. Treatment of fetuses closer to delivery results in immune ignorance without induction of tolerance. This result highlights the repercussion of selecting the appropriate time point when this approach is used to deliver therapeutic genes. All these findings illustrate the relevance of naked DNA-based vaccination technology in therapeutic efforts aimed to prevent the high toll of death among first-week infants.

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References

  1. Zimmerman RK, Ruben FL, Ahwesh ER . Hepatitis B virus infection, hepatitis B vaccine, and hepatitis B immune globulin. J Fam Pract 1997; 45: 295–315.

    CAS  PubMed  Google Scholar 

  2. Baker DA . Issues and management of herpes in pregnancy. Int J Fertil Womens Med 2002; 47: 129–135.

    PubMed  Google Scholar 

  3. Ahdieh L . Pregnancy and infection with human immunodeficiency virus. Clin Obstet Gynecol 2001; 44: 154–166.

    Article  CAS  Google Scholar 

  4. Berner R . Group B streptococci during pregnancy and infancy. Curr Opin Infect Dis 2002; 15: 307–313.

    Article  Google Scholar 

  5. Revello MG, Gerna G . Diagnosis and management of human cytomegalovirus infection in the mother, fetus, and newborn infant. Clin Microbiol Rev 2002; 15: 680–715.

    Article  Google Scholar 

  6. Aujard Y, Fauroux B . Risk factors for severe respiratory syncytial virus infection in infants. Respir Med 2002; 96 (Suppl B): S9–S14.

    Article  Google Scholar 

  7. Syrjanen S, Puranen M . Human papillomavirus infections in children: the potential role of maternal transmission. Crit Rev Oral Biol Med 2000; 11: 259–274.

    Article  CAS  Google Scholar 

  8. Wright PF, Wright PF . Infectious diseases in early life in industrialized countries. Vaccine 1998; 16: 1355–1359.

    Article  CAS  Google Scholar 

  9. Towers CV, Asrat T, Rumney P . The presence of hepatitis B surface antigen and deoxyribonucleic acid in amniotic fluid and cord blood. Am J Obstet Gynecol 2001; 184: 1514–1518.

    Article  CAS  Google Scholar 

  10. Komatsu H et al. Sequence analysis of hepatitis B virus genomes from an infant with acute severe hepatitis and a hepatitis B e antigen-positive carrier mother. J Med Virol 2001; 65: 457–462.

    Article  CAS  Google Scholar 

  11. Sterneck M et al. Neonatal fulminant hepatitis B: structural and functional analysis of complete hepatitis B virus genomes from mother and infant. J Infect Dis 1998; 177: 1378–1381.

    Article  CAS  Google Scholar 

  12. Prevention of perinatal hepatitis B through enhanced case management – Connecticut, 1994–95, and the United States, 1994. MMWR Morb Mortal Wkly Rep 1996; 45: 584–587.

  13. Ranger-Rogez S, Alain S, Denis F . Hepatitis viruses: mother-to-child transmission. Pathol Biol (Paris) 2002; 50: 568–575.

    Article  CAS  Google Scholar 

  14. Vranckx R, Alisjahbana A, Meheus A . Hepatitis B virus vaccination and antenatal transmission of HBV markers to neonates. J Viral Hepat 1999; 6: 135–139.

    Article  CAS  Google Scholar 

  15. Siegrist CA . Neonatal and early life vaccinology. Vaccine 2001; 19: 3331–3346.

    Article  CAS  Google Scholar 

  16. Gerdts V et al. Fetal immunization by a DNA vaccine delivered into the oral cavity. Nat Med 2000; 6: 929–932.

    Article  CAS  Google Scholar 

  17. Prud'homme GJ et al. Immunotherapeutic gene transfer into muscle. Trends Immunol 2001; 22: 149–155.

    Article  CAS  Google Scholar 

  18. Spiegelberg HL, Raz E . DNA-based approaches to the treatment of allergies. Curr Opin Mol Ther 2002; 4: 64–71.

    CAS  PubMed  Google Scholar 

  19. Ashmore CR, Addis PB, Doerr L . Development of muscle fibers in the fetal pig. J Anim Sci 1973; 36: 1088–1093.

    Article  CAS  Google Scholar 

  20. Swatland HJ, Cassens TG . Prenatal development, histochemistry and innervation of porcine muscle. J Anim Sci 1973; 36: 343–354.

    Article  CAS  Google Scholar 

  21. Tlaskalova-Hogenova L et al. Development of immune responses in early pig ontogeny. Vet Immunol Immunopathol 1994; 43: 135–142.

    Article  CAS  Google Scholar 

  22. West DJ, Calandra GB . Vaccine induced immunologic memory for hepatitis B surface antigen: implications for policy on booster vaccination. Vaccine 1996; 14: 1019–1027.

    Article  CAS  Google Scholar 

  23. del Canho R et al. Ten-year neonatal hepatitis B vaccination program, The Netherlands, 1982-1992: protective efficacy and long-term immunogenicity. Vaccine 1997; 15: 1624–1630.

    Article  CAS  Google Scholar 

  24. Binns RM . Bone marrow and lymphoid cell injection of the pig foetus resulting in transplantation tolerance or immunity, and immunoglobulin production. Nature 1967; 214: 179–181.

    Article  CAS  Google Scholar 

  25. Swain WE et al. Tolerability and immune responses in humans to a PowderJect DNA vaccine for hepatitis B. Dev Biol (Basel) 2000; 104: 115–119.

    CAS  Google Scholar 

  26. Davis HL, Michel ML, Whalen RG . DNA-based immunization for hepatitis B induces continuous secretion of antigen and high levels of circulating antibody. Hum Mol Genet 1993; 2: 1847–1851.

    Article  CAS  Google Scholar 

  27. Gerdts V et al. Oral DNA vaccination in-utero induces mucosal immunity and immune memory in the neonate. J Immunol 2002; 168: 1877–1885.

    Article  CAS  Google Scholar 

  28. Ichino M et al. Factors associated with the development of neonatal tolerance after the administration of a plasmid DNA vaccine. J Immunol 1999; 162: 3814–3818.

    CAS  PubMed  Google Scholar 

  29. Randall GC . Changes in the concentrations of corticosteroids in the blood of fetal pigs and their dams during late gestation and labor. Biol Reprod 1983; 29: 1077–1084.

    Article  CAS  Google Scholar 

  30. Fevre J . Maternal and fetal corticosteroids in the sow at the end of gestation. C R Acad Sci Hebd Seances Acad Sci D 1975; 281: 2009–2012.

    CAS  PubMed  Google Scholar 

  31. Murphy BE . Human fetal serum cortisol levels related to gestational age: evidence of a midgestational fall and a steep late gestational rise, independent of sex or mode of delivery. Am J Obstet Gynecol 1982; 144: 276–282.

    Article  CAS  Google Scholar 

  32. Ishii KJ, Veiss WR, Klinman DM . Prevention of neonatal tolerance by a plasmid encoding granulocyte-macrophage colony stimulating factor. Vaccine 1999; 18: 703–710.

    Article  CAS  Google Scholar 

  33. Rinaldi M et al. Safe fetal gene transfer by transuterine injection of naked DNA into pig muscle and localized perinatal expression of different transgenes. (submitted for publication).

  34. Everett RSR, Gerrard DE, Grant AL . Factors affecting production of luciferase and epitope-tagged IGF-I in porcine muscle after DNA injection. J Endocrinol 2000; 166: 255–263.

    Article  CAS  Google Scholar 

  35. Parker SE et al. Plasmid DNA malaria vaccine: tissue distribution and safety studies in mice and rabbits. Hum Gene Ther 1999; 10: 741–758.

    Article  CAS  Google Scholar 

  36. Le TP et al. Safety, tolerability and humoral immune responses after intramuscular administration of a malaria DNA vaccine to healthy adult volunteers. Vaccine 2000; 18: 1893–1901.

    Article  CAS  Google Scholar 

  37. Martin T et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Hum Gene Ther 1999; 10: 759–768.

    Article  CAS  Google Scholar 

  38. Davis HL, Millan CL, Watkins SC . Immune-mediated destruction of transected muscles fibers direct gene transfer with antigen-expressing plasmid DNA. Gene Therapy 1997; 4: 181–188.

    Article  CAS  Google Scholar 

  39. Mor G et al. Do DNA vaccines induce autoimmune disease? Hum Gene Ther 1997; 8: 293–300.

    Article  CAS  Google Scholar 

  40. Wang G et al. Ultrasound-guided gene transfer to hepatocytes in-utero. Fetal Diagn Ther 1998; 13: 197–205.

    Article  Google Scholar 

  41. Resti M et al. Ten-year follow-up study of neonatal hepatitis B immunization: are booster injections indicated? Vaccine 1997; 15: 1338–1340.

    Article  CAS  Google Scholar 

  42. Watts AM et al. Fetal immunization of baboons induces a fetal-specific antibody response. Nat Med 1999; 5: 427–430.

    Article  CAS  Google Scholar 

  43. Ciafrè SA et al. A plasmid family containing two different expression cassettes suitable for immunomodulation and genetic immunization. Plasmid 1998; 40: 84–89.

    Article  Google Scholar 

  44. Rinaldi M et al. Treatment of severe hypercholesterolemia in apolipoprotein E-deficient mice by intramuscular injection of plasmid DNA. Gene Therapy 2000; 7: 1795–1801.

    Article  CAS  Google Scholar 

  45. Alexander JM et al. Risk of hepatitis B transmission after amniocentesis in chronic hepatitis B carriers. Infect Dis Obstet Gynecol 1999; 7: 283–286.

    Article  CAS  Google Scholar 

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Acknowledgements

This study was supported in part by grants from the Italian Ministry of Health (IRCCS-CSS, RC99). We are particularly grateful to all the technical staff of the ‘Stabilimento di Allevamento e di Utilizzazione Animali’ of the Catholic University of Rome for their skilful help and advise in every procedure with animals, their warm attention to the animal care and their tireless assistance. We thank Dr Francesca Berlinguer for English mothertongue proof-reading and Miss Simona Virga for secretarial assistance and artwork. This work is dedicated to Professors Domenico Fazio and Franca Mastroviti-Fazio for their lifelong and loving contribution to Culture and Education, and to Professor ‘Nino’ Gaetano Salvatore for his high and timeless contribution to the scientific knowledge and University formation.

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Authors and Affiliations

  1. Interdisciplinary Center for Biomedical Research, Laboratory for Molecular Medicine and BioTechnology, Università ‘Campus Bio-Medico’, Rome and Laboratory for Gene Therapy and Oncology, IRCCS H ‘Casa Sollievo della Sofferenza’, San Giovanni Rotondo (FG), Italy

    VM Fazio & E Signori

  2. Institutes of Patologia Generale, Università Cattolica del Sacro Cuore (UCSC), Rome, Italy

    F Ria

  3. Department of Sanità Pubblica e Biologia Cellulare, Università ‘Tor Vergata’, Rome, Italy

    E Franco & L Zaratti

  4. Ostetricia e Ginecologia, Università Cattolica del Sacro Cuore (UCSC), Rome, Italy

    P Rosati & G Monego

  5. Anestesiologia e Rianimazione, Università Cattolica del Sacro Cuore (UCSC), Rome, Italy

    G Cannelli & E Iannace

  6. Divisione Gene-Medicine, Istituto Neurobiologia e Medicina Molecolare, Sezione Medicina Molecolare, CNR, Rome, Italy

    E Signori, D Fioretti, S Iurescia & M Rinaldi

  7. Laboratory for Gene Therapy and Oncology, IRCCS H. ‘Casa Sollievo della Sofferenza’, San Giovanni Rotondo (FG), Italy

    P Parrella, D Fioretti & S Iurescia

  8. Servizio Stabulario, Università Cattolica del Sacro Cuore (UCSC), Rome, Italy

    S Blogna & R Filippetti

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  1. VM Fazio
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  2. F Ria
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  3. E Franco
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  5. G Cannelli
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  6. E Signori
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  8. L Zaratti
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  9. E Iannace
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  10. G Monego
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  11. S Blogna
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  12. D Fioretti
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  13. S Iurescia
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  14. R Filippetti
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  15. M Rinaldi
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Fazio, V., Ria, F., Franco, E. et al. Immune response at birth, long-term immune memory and 2 years follow-up after in-utero anti-HBV DNA immunization. Gene Ther 11, 544–551 (2004). https://doi.org/10.1038/sj.gt.3302179

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  • DOI: https://doi.org/10.1038/sj.gt.3302179

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