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Dual neonate vaccine platform against HIV-1 and M. tuberculosis.

Hopkins R, Bridgeman A, Joseph J, Gilbert SC, McShane H, Hanke T - PLoS ONE (2011)

Bottom Line: The first vaccine the majority of infants born in Africa receive is Mycobacterium bovis bacillus Calmette-Guérin (BCG) as a prevention against TB.While sMVA.HIVA.85A expressed higher levels of the immunogens, it was less immunogenic than mMVA.HIVA.85A in BALB/c mice.Induction of immune responses against these pathogens soon after birth is highly desirable and may provide a basis for lifetime protection maintained by boosts later in life.

View Article: PubMed Central - PubMed

Affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.

ABSTRACT
Acquired immunodeficiency syndrome and tuberculosis (TB) are two of the world's most devastating diseases. The first vaccine the majority of infants born in Africa receive is Mycobacterium bovis bacillus Calmette-Guérin (BCG) as a prevention against TB. BCG protects against disseminated disease in the first 10 years of life, but provides a variable protection against pulmonary TB and enhancing boost delivered by recombinant modified vaccinia virus Ankara (rMVA) expressing antigen 85A (Ag85A) of M. tuberculosis is currently in phase IIb evaluation in African neonates. If the newborn's mother is positive for human immunodeficiency virus type 1 (HIV-1), the baby is at high risk of acquiring HIV-1 through breastfeeding. We suggested that a vaccination consisting of recombinant BCG expressing HIV-1 immunogen administered at birth followed by a boost with rMVA sharing the same immunogen could serve as a strategy for prevention of mother-to-child transmission of HIV-1 and rMVA expressing an African HIV-1-derived immunogen HIVA is currently in phase I trials in African neonates. Here, we aim to develop a dual neonate vaccine platform against HIV-1 and TB consisting of BCG.HIVA administered at birth followed by a boost with MVA.HIVA.85A. Thus, mMVA.HIVA.85A and sMVA.HIVA.85A vaccines were constructed, in which the transgene transcription is driven by either modified H5 or short synthetic promoters, respectively, and tested for immunogenicity alone and in combination with BCG.HIVA(222). mMVA.HIVA.85A was produced markerless and thus suitable for clinical manufacture. While sMVA.HIVA.85A expressed higher levels of the immunogens, it was less immunogenic than mMVA.HIVA.85A in BALB/c mice. A BCG.HIVA(222)-mMVA.HIVA.85A prime-boost regimen induced robust T cell responses to both HIV-1 and M. tuberculosis. Therefore, proof-of-principle for a dual anti-HIV-1/M. tuberculosis infant vaccine platform is established. Induction of immune responses against these pathogens soon after birth is highly desirable and may provide a basis for lifetime protection maintained by boosts later in life.

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Protein expression from recombinant MVAs in CEF cells.CEF cells were infected with indicated viruses at MOI 1 for 16 h.                                (A) Cell were stained either for Gag p24 or Pk tag (on                            HIVA and Ag85A) expression as shown above and analyzed under fluorescent                            microscope. Also, the relative levels of protein expression were                            assessed using flow cytometry and shown either as a (B)                            histogram for parental MVA (black), 7.5MVA.HIVA (red), mMVA.HIVA.85A                            (blue) or sMVA.HIVA.85A (green) viruses or (C) expressed as                            mean fluorescent intensity (MFI) ± SD (C). The                            figure shows representative data of three independent experiments.
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pone-0020067-g002: Protein expression from recombinant MVAs in CEF cells.CEF cells were infected with indicated viruses at MOI 1 for 16 h. (A) Cell were stained either for Gag p24 or Pk tag (on HIVA and Ag85A) expression as shown above and analyzed under fluorescent microscope. Also, the relative levels of protein expression were assessed using flow cytometry and shown either as a (B) histogram for parental MVA (black), 7.5MVA.HIVA (red), mMVA.HIVA.85A (blue) or sMVA.HIVA.85A (green) viruses or (C) expressed as mean fluorescent intensity (MFI) ± SD (C). The figure shows representative data of three independent experiments.

Mentions: The mMVA.HIVA.85A and sMVA.HIVA.85A vaccines were first characterized as for the levels of the two transgene product expression. Both HIVA and Ag85A proteins contain a C-terminal epitope Pk recognized by monoclonal antibody SV5-P-k [23], which was attached to facilitate the immunogen detection. In addition, a mAb against p24 was employed to detect specifically the HIVA protein. Thus, monolayers of CEF cells were infected at MOI 1 with either the 7.5MVA.HIVA, mMVA.HIVA.85A or sMVA.HIVA.85A vaccines and the expression of the transgene products using the anti-Pk and p24 mAbs were readily detectable for the ssp and mH5 promoters, while for the P7.5 promoter, the immunofluorescence signal was fainter for anti-p24 mAb and almost undetectable for the Pk tag (Fig. 2A); this is similar to our previous experience [24]. To obtain more quantitative expression data, CEF cells were infected with empty parental or rMVAs and subjected to analysis by flow cytometry. The number and median fluorescent intensity (MFI) of cells expressing the Pk-epitope confirmed superior transgene expression from the sMVA.HIVA.85A ssp promoter (Fig. 2B and C).


Dual neonate vaccine platform against HIV-1 and M. tuberculosis.

Hopkins R, Bridgeman A, Joseph J, Gilbert SC, McShane H, Hanke T - PLoS ONE (2011)

Protein expression from recombinant MVAs in CEF cells.CEF cells were infected with indicated viruses at MOI 1 for 16 h.                                (A) Cell were stained either for Gag p24 or Pk tag (on                            HIVA and Ag85A) expression as shown above and analyzed under fluorescent                            microscope. Also, the relative levels of protein expression were                            assessed using flow cytometry and shown either as a (B)                            histogram for parental MVA (black), 7.5MVA.HIVA (red), mMVA.HIVA.85A                            (blue) or sMVA.HIVA.85A (green) viruses or (C) expressed as                            mean fluorescent intensity (MFI) ± SD (C). The                            figure shows representative data of three independent experiments.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3094449&req=5

pone-0020067-g002: Protein expression from recombinant MVAs in CEF cells.CEF cells were infected with indicated viruses at MOI 1 for 16 h. (A) Cell were stained either for Gag p24 or Pk tag (on HIVA and Ag85A) expression as shown above and analyzed under fluorescent microscope. Also, the relative levels of protein expression were assessed using flow cytometry and shown either as a (B) histogram for parental MVA (black), 7.5MVA.HIVA (red), mMVA.HIVA.85A (blue) or sMVA.HIVA.85A (green) viruses or (C) expressed as mean fluorescent intensity (MFI) ± SD (C). The figure shows representative data of three independent experiments.
Mentions: The mMVA.HIVA.85A and sMVA.HIVA.85A vaccines were first characterized as for the levels of the two transgene product expression. Both HIVA and Ag85A proteins contain a C-terminal epitope Pk recognized by monoclonal antibody SV5-P-k [23], which was attached to facilitate the immunogen detection. In addition, a mAb against p24 was employed to detect specifically the HIVA protein. Thus, monolayers of CEF cells were infected at MOI 1 with either the 7.5MVA.HIVA, mMVA.HIVA.85A or sMVA.HIVA.85A vaccines and the expression of the transgene products using the anti-Pk and p24 mAbs were readily detectable for the ssp and mH5 promoters, while for the P7.5 promoter, the immunofluorescence signal was fainter for anti-p24 mAb and almost undetectable for the Pk tag (Fig. 2A); this is similar to our previous experience [24]. To obtain more quantitative expression data, CEF cells were infected with empty parental or rMVAs and subjected to analysis by flow cytometry. The number and median fluorescent intensity (MFI) of cells expressing the Pk-epitope confirmed superior transgene expression from the sMVA.HIVA.85A ssp promoter (Fig. 2B and C).

Bottom Line: The first vaccine the majority of infants born in Africa receive is Mycobacterium bovis bacillus Calmette-Guérin (BCG) as a prevention against TB.While sMVA.HIVA.85A expressed higher levels of the immunogens, it was less immunogenic than mMVA.HIVA.85A in BALB/c mice.Induction of immune responses against these pathogens soon after birth is highly desirable and may provide a basis for lifetime protection maintained by boosts later in life.

View Article: PubMed Central - PubMed

Affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.

ABSTRACT
Acquired immunodeficiency syndrome and tuberculosis (TB) are two of the world's most devastating diseases. The first vaccine the majority of infants born in Africa receive is Mycobacterium bovis bacillus Calmette-Guérin (BCG) as a prevention against TB. BCG protects against disseminated disease in the first 10 years of life, but provides a variable protection against pulmonary TB and enhancing boost delivered by recombinant modified vaccinia virus Ankara (rMVA) expressing antigen 85A (Ag85A) of M. tuberculosis is currently in phase IIb evaluation in African neonates. If the newborn's mother is positive for human immunodeficiency virus type 1 (HIV-1), the baby is at high risk of acquiring HIV-1 through breastfeeding. We suggested that a vaccination consisting of recombinant BCG expressing HIV-1 immunogen administered at birth followed by a boost with rMVA sharing the same immunogen could serve as a strategy for prevention of mother-to-child transmission of HIV-1 and rMVA expressing an African HIV-1-derived immunogen HIVA is currently in phase I trials in African neonates. Here, we aim to develop a dual neonate vaccine platform against HIV-1 and TB consisting of BCG.HIVA administered at birth followed by a boost with MVA.HIVA.85A. Thus, mMVA.HIVA.85A and sMVA.HIVA.85A vaccines were constructed, in which the transgene transcription is driven by either modified H5 or short synthetic promoters, respectively, and tested for immunogenicity alone and in combination with BCG.HIVA(222). mMVA.HIVA.85A was produced markerless and thus suitable for clinical manufacture. While sMVA.HIVA.85A expressed higher levels of the immunogens, it was less immunogenic than mMVA.HIVA.85A in BALB/c mice. A BCG.HIVA(222)-mMVA.HIVA.85A prime-boost regimen induced robust T cell responses to both HIV-1 and M. tuberculosis. Therefore, proof-of-principle for a dual anti-HIV-1/M. tuberculosis infant vaccine platform is established. Induction of immune responses against these pathogens soon after birth is highly desirable and may provide a basis for lifetime protection maintained by boosts later in life.

Show MeSH
Related in: MedlinePlus