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A Single Dose Respiratory Recombinant Adenovirus-Based Vaccine Provides Long-Term Protection for Non-Human Primates from Lethal Ebola Infection.

Choi JH, Jonsson-Schmunk K, Qiu X, Shedlock DJ, Strong J, Xu JX, Michie KL, Audet J, Fernando L, Myers MJ, Weiner D, Bajrovic I, Tran LQ, Wong G, Bello A, Kobinger GP, Schafer SC, Croyle MA - Mol. Pharm. (2014)

Bottom Line: Three primates were then given the vaccine in a formulation that improved the immune response to Ebola in rodents.Three primates were immunized with 2.0×10(10) ivp/kg of vaccine by the SL route.To our knowledge, this is the first time that durable protection from a single dose respiratory adenovirus-based Ebola vaccine has been demonstrated in primates.

View Article: PubMed Central - PubMed

Affiliation: †Division of Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, Texas 78712, United States.

ABSTRACT
As the Ebola outbreak in West Africa continues and cases appear in the United States and other countries, the need for long-lasting vaccines to preserve global health is imminent. Here, we evaluate the long-term efficacy of a respiratory and sublingual (SL) adenovirus-based vaccine in non-human primates in two phases. In the first, a single respiratory dose of 1.4×10(9) infectious virus particles (ivp)/kg of Ad-CAGoptZGP induced strong Ebola glycoprotein (GP) specific CD8+ and CD4+ T cell responses and Ebola GP-specific antibodies in systemic and mucosal compartments and was partially (67%) protective from challenge 62 days after immunization. The same dose given by the SL route induced Ebola GP-specific CD8+ T cell responses similar to that of intramuscular (IM) injection, however, the Ebola GP-specific antibody response was low. All primates succumbed to infection. Three primates were then given the vaccine in a formulation that improved the immune response to Ebola in rodents. Three primates were immunized with 2.0×10(10) ivp/kg of vaccine by the SL route. Diverse populations of polyfunctional Ebola GP-specific CD4+ and CD8+ T cells and significant anti-Ebola GP antibodies were present in samples collected 150 days after respiratory immunization. The formulated vaccine was fully protective against challenge 21 weeks after immunization. While diverse populations of Ebola GP-specific CD4+ T cells were produced after SL immunization, antibodies were not neutralizing and the vaccine was unprotective. To our knowledge, this is the first time that durable protection from a single dose respiratory adenovirus-based Ebola vaccine has been demonstrated in primates.

No MeSH data available.


Related in: MedlinePlus

Study 1: Respiratory immunization induces strong antigen-specificT cell responses after administration of a single dose of a formulatedadenovirus-based Ebola vaccine. (A) Quantitative analysis of Ebolaglycoprotein-specific CD4+ T cells in BALfluid. Cells were isolated from whole blood 20 days after immunizationand stimulated with a peptide library for Ebola glycoprotein or peptidesspecific for the MHC class II associated invariant chain peptide thatbinds the MHC class II groove of cells (h-Clip, negative control).Positive control cells were stimulated with PMA and ionomycin. Eachcell population was stimulated for 5 h, stained for phenotypic markers,and analyzed by flow cytometry. (B) Quantitative analysis of Ebolaglycoprotein-specific CD8+ T cells in BALfluid. Cells were treated as described for Panel A. (C) Magnitudeof the antigen-specific response of mononuclear cells isolated fromwhole blood of macaques. PBMCs were isolated 20 days after immunizationfrom whole blood and evaluated for IFN-γ secretion after stimulationwith an Ebola GP-specific peptide library by ELISpot. (D) Magnitudeof the antigen-specific response in mononuclear cells isolated fromiliac lymph nodes (ILNs) of primates. MNCs were isolated 20 days afterimmunization from ILNs and evaluated for IFN-γ secretion afterstimulation with an Ebola GP-specific peptide library by ELISpot.(E) Proliferative capacity of Ebola GP-specific T cells collected38 days after immunization of naive primates by various routes. Theproliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells isolated from whole blood was evaluatedfor each animal by stimulation for 5 days with an Ebola GP-specificpeptide library and subsequent staining for Ki-67, an intracellularmarker for proliferation.59 (F) Proliferativecapacity of adenovirus serotype 5-specific T cells after immunizationby various routes. Cells were isolated from whole blood 38 days afterimmunization and stimulated for 5 days with a first generation adenovirusthat does not contain a transgene cassette (AdNull, MOI 1:1,000).The proliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells was determined by intracellular stainingfor Ki-67. Animal numbers displayed in each panel and their correspondingtreatments are summarized in Table 1.
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fig4: Study 1: Respiratory immunization induces strong antigen-specificT cell responses after administration of a single dose of a formulatedadenovirus-based Ebola vaccine. (A) Quantitative analysis of Ebolaglycoprotein-specific CD4+ T cells in BALfluid. Cells were isolated from whole blood 20 days after immunizationand stimulated with a peptide library for Ebola glycoprotein or peptidesspecific for the MHC class II associated invariant chain peptide thatbinds the MHC class II groove of cells (h-Clip, negative control).Positive control cells were stimulated with PMA and ionomycin. Eachcell population was stimulated for 5 h, stained for phenotypic markers,and analyzed by flow cytometry. (B) Quantitative analysis of Ebolaglycoprotein-specific CD8+ T cells in BALfluid. Cells were treated as described for Panel A. (C) Magnitudeof the antigen-specific response of mononuclear cells isolated fromwhole blood of macaques. PBMCs were isolated 20 days after immunizationfrom whole blood and evaluated for IFN-γ secretion after stimulationwith an Ebola GP-specific peptide library by ELISpot. (D) Magnitudeof the antigen-specific response in mononuclear cells isolated fromiliac lymph nodes (ILNs) of primates. MNCs were isolated 20 days afterimmunization from ILNs and evaluated for IFN-γ secretion afterstimulation with an Ebola GP-specific peptide library by ELISpot.(E) Proliferative capacity of Ebola GP-specific T cells collected38 days after immunization of naive primates by various routes. Theproliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells isolated from whole blood was evaluatedfor each animal by stimulation for 5 days with an Ebola GP-specificpeptide library and subsequent staining for Ki-67, an intracellularmarker for proliferation.59 (F) Proliferativecapacity of adenovirus serotype 5-specific T cells after immunizationby various routes. Cells were isolated from whole blood 38 days afterimmunization and stimulated for 5 days with a first generation adenovirusthat does not contain a transgene cassette (AdNull, MOI 1:1,000).The proliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells was determined by intracellular stainingfor Ki-67. Animal numbers displayed in each panel and their correspondingtreatments are summarized in Table 1.

Mentions: Twenty days after immunization,PBMCs were isolated from whole blood and incubated with peptides specificfor Ebola glycoprotein (GP). Cells were then subjected to intracellularcytokine staining for CD8+ and CD4+ surfaceantigens and IFN-γ and sorted by flow cytometry. At this timepoint, few cells responsive to Ebola glycoprotein could be detectedin PBMCs obtained from any of the animals (data not shown). A similartrend was observed in samples taken from iliac lymph nodes (ILNs)of animals. Profound responses were seen in samples obtained fromthe BAL fluid of animals given the vaccine by the IN/IT route. Thestrongest response was seen in CD4+ cells with 12.5% ofthe population obtained from primate 52945 and 3.03% of the populationfrom primate 50459 responding (Figure 4A).Although the response from the third primate in this treatment group(52483) was small in comparison (0.71%), it was significantly higherthan that observed in animals given the vaccine by IM injection. TheCD8+ T cell response followed a similar trend (Figure 4B).


A Single Dose Respiratory Recombinant Adenovirus-Based Vaccine Provides Long-Term Protection for Non-Human Primates from Lethal Ebola Infection.

Choi JH, Jonsson-Schmunk K, Qiu X, Shedlock DJ, Strong J, Xu JX, Michie KL, Audet J, Fernando L, Myers MJ, Weiner D, Bajrovic I, Tran LQ, Wong G, Bello A, Kobinger GP, Schafer SC, Croyle MA - Mol. Pharm. (2014)

Study 1: Respiratory immunization induces strong antigen-specificT cell responses after administration of a single dose of a formulatedadenovirus-based Ebola vaccine. (A) Quantitative analysis of Ebolaglycoprotein-specific CD4+ T cells in BALfluid. Cells were isolated from whole blood 20 days after immunizationand stimulated with a peptide library for Ebola glycoprotein or peptidesspecific for the MHC class II associated invariant chain peptide thatbinds the MHC class II groove of cells (h-Clip, negative control).Positive control cells were stimulated with PMA and ionomycin. Eachcell population was stimulated for 5 h, stained for phenotypic markers,and analyzed by flow cytometry. (B) Quantitative analysis of Ebolaglycoprotein-specific CD8+ T cells in BALfluid. Cells were treated as described for Panel A. (C) Magnitudeof the antigen-specific response of mononuclear cells isolated fromwhole blood of macaques. PBMCs were isolated 20 days after immunizationfrom whole blood and evaluated for IFN-γ secretion after stimulationwith an Ebola GP-specific peptide library by ELISpot. (D) Magnitudeof the antigen-specific response in mononuclear cells isolated fromiliac lymph nodes (ILNs) of primates. MNCs were isolated 20 days afterimmunization from ILNs and evaluated for IFN-γ secretion afterstimulation with an Ebola GP-specific peptide library by ELISpot.(E) Proliferative capacity of Ebola GP-specific T cells collected38 days after immunization of naive primates by various routes. Theproliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells isolated from whole blood was evaluatedfor each animal by stimulation for 5 days with an Ebola GP-specificpeptide library and subsequent staining for Ki-67, an intracellularmarker for proliferation.59 (F) Proliferativecapacity of adenovirus serotype 5-specific T cells after immunizationby various routes. Cells were isolated from whole blood 38 days afterimmunization and stimulated for 5 days with a first generation adenovirusthat does not contain a transgene cassette (AdNull, MOI 1:1,000).The proliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells was determined by intracellular stainingfor Ki-67. Animal numbers displayed in each panel and their correspondingtreatments are summarized in Table 1.
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Related In: Results  -  Collection

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fig4: Study 1: Respiratory immunization induces strong antigen-specificT cell responses after administration of a single dose of a formulatedadenovirus-based Ebola vaccine. (A) Quantitative analysis of Ebolaglycoprotein-specific CD4+ T cells in BALfluid. Cells were isolated from whole blood 20 days after immunizationand stimulated with a peptide library for Ebola glycoprotein or peptidesspecific for the MHC class II associated invariant chain peptide thatbinds the MHC class II groove of cells (h-Clip, negative control).Positive control cells were stimulated with PMA and ionomycin. Eachcell population was stimulated for 5 h, stained for phenotypic markers,and analyzed by flow cytometry. (B) Quantitative analysis of Ebolaglycoprotein-specific CD8+ T cells in BALfluid. Cells were treated as described for Panel A. (C) Magnitudeof the antigen-specific response of mononuclear cells isolated fromwhole blood of macaques. PBMCs were isolated 20 days after immunizationfrom whole blood and evaluated for IFN-γ secretion after stimulationwith an Ebola GP-specific peptide library by ELISpot. (D) Magnitudeof the antigen-specific response in mononuclear cells isolated fromiliac lymph nodes (ILNs) of primates. MNCs were isolated 20 days afterimmunization from ILNs and evaluated for IFN-γ secretion afterstimulation with an Ebola GP-specific peptide library by ELISpot.(E) Proliferative capacity of Ebola GP-specific T cells collected38 days after immunization of naive primates by various routes. Theproliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells isolated from whole blood was evaluatedfor each animal by stimulation for 5 days with an Ebola GP-specificpeptide library and subsequent staining for Ki-67, an intracellularmarker for proliferation.59 (F) Proliferativecapacity of adenovirus serotype 5-specific T cells after immunizationby various routes. Cells were isolated from whole blood 38 days afterimmunization and stimulated for 5 days with a first generation adenovirusthat does not contain a transgene cassette (AdNull, MOI 1:1,000).The proliferative capacity of CD4+ (white bars) and CD8+ (black bars) T cells was determined by intracellular stainingfor Ki-67. Animal numbers displayed in each panel and their correspondingtreatments are summarized in Table 1.
Mentions: Twenty days after immunization,PBMCs were isolated from whole blood and incubated with peptides specificfor Ebola glycoprotein (GP). Cells were then subjected to intracellularcytokine staining for CD8+ and CD4+ surfaceantigens and IFN-γ and sorted by flow cytometry. At this timepoint, few cells responsive to Ebola glycoprotein could be detectedin PBMCs obtained from any of the animals (data not shown). A similartrend was observed in samples taken from iliac lymph nodes (ILNs)of animals. Profound responses were seen in samples obtained fromthe BAL fluid of animals given the vaccine by the IN/IT route. Thestrongest response was seen in CD4+ cells with 12.5% ofthe population obtained from primate 52945 and 3.03% of the populationfrom primate 50459 responding (Figure 4A).Although the response from the third primate in this treatment group(52483) was small in comparison (0.71%), it was significantly higherthan that observed in animals given the vaccine by IM injection. TheCD8+ T cell response followed a similar trend (Figure 4B).

Bottom Line: Three primates were then given the vaccine in a formulation that improved the immune response to Ebola in rodents.Three primates were immunized with 2.0×10(10) ivp/kg of vaccine by the SL route.To our knowledge, this is the first time that durable protection from a single dose respiratory adenovirus-based Ebola vaccine has been demonstrated in primates.

View Article: PubMed Central - PubMed

Affiliation: †Division of Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, Texas 78712, United States.

ABSTRACT
As the Ebola outbreak in West Africa continues and cases appear in the United States and other countries, the need for long-lasting vaccines to preserve global health is imminent. Here, we evaluate the long-term efficacy of a respiratory and sublingual (SL) adenovirus-based vaccine in non-human primates in two phases. In the first, a single respiratory dose of 1.4×10(9) infectious virus particles (ivp)/kg of Ad-CAGoptZGP induced strong Ebola glycoprotein (GP) specific CD8+ and CD4+ T cell responses and Ebola GP-specific antibodies in systemic and mucosal compartments and was partially (67%) protective from challenge 62 days after immunization. The same dose given by the SL route induced Ebola GP-specific CD8+ T cell responses similar to that of intramuscular (IM) injection, however, the Ebola GP-specific antibody response was low. All primates succumbed to infection. Three primates were then given the vaccine in a formulation that improved the immune response to Ebola in rodents. Three primates were immunized with 2.0×10(10) ivp/kg of vaccine by the SL route. Diverse populations of polyfunctional Ebola GP-specific CD4+ and CD8+ T cells and significant anti-Ebola GP antibodies were present in samples collected 150 days after respiratory immunization. The formulated vaccine was fully protective against challenge 21 weeks after immunization. While diverse populations of Ebola GP-specific CD4+ T cells were produced after SL immunization, antibodies were not neutralizing and the vaccine was unprotective. To our knowledge, this is the first time that durable protection from a single dose respiratory adenovirus-based Ebola vaccine has been demonstrated in primates.

No MeSH data available.


Related in: MedlinePlus