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Entrapment of H1N1 Influenza Virus Derived Conserved Peptides in PLGA Nanoparticles Enhances T Cell Response and Vaccine Efficacy in Pigs.

Hiremath J, Kang KI, Xia M, Elaish M, Binjawadagi B, Ouyang K, Dhakal S, Arcos J, Torrelles JB, Jiang X, Lee CW, Renukaradhya GJ - PLoS ONE (2016)

Bottom Line: Biodegradable polylactic-co-glycolic acid (PLGA) nanoparticle (PLGA-NP) based vaccine delivery system enhances cross-presentation of antigens by the professional antigen presenting cells.Immunologically, PLGA-NP peptides vaccination (without adjuvant) significantly increased the frequency of antigen-specific IFNγ secreting CD4 and CD8 T cells response in the lung lymphocytes, despite not boosting the antibody response both at pre- and post-challenge.In summary, our data indicated that nanoparticle-mediated delivery of conserved H1N1 influenza peptides induced the virus specific T cell response in the lungs and reduced the challenged heterologous virus load in the airways of pigs.

View Article: PubMed Central - PubMed

Affiliation: Food Animal Health Research Program, Ohio Agricultural Research and Development Center, 1680 Madison Avenue, Wooster, Ohio, 44691, United States of America, and Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, 43210, United States of America.

ABSTRACT
Pigs are believed to be one of the important sources of emerging human and swine influenza viruses (SwIV). Influenza virus conserved peptides have the potential to elicit cross-protective immune response, but without the help of potent adjuvant and delivery system they are poorly immunogenic. Biodegradable polylactic-co-glycolic acid (PLGA) nanoparticle (PLGA-NP) based vaccine delivery system enhances cross-presentation of antigens by the professional antigen presenting cells. In this study, Norovirus P particle containing SwIV M2e (extracellular domain of the matrix protein 2) chimera and highly conserved two each of H1N1 peptides of pandemic 2009 and classical human influenza viruses were entrapped in PLGA-NPs. Influenza antibody-free pigs were vaccinated with PLGA-NPs peptides cocktail vaccine twice with or without an adjuvant, Mycobacterium vaccae whole cell lysate, intranasally as mist. Vaccinated pigs were challenged with a virulent heterologous zoonotic SwIV H1N1, and one week later euthanized and the lung samples were analyzed for the specific immune response and viral load. Clinically, pigs vaccinated with PLGA-NP peptides vaccine had no fever and flu symptoms, and the replicating challenged SwIV was undetectable in the bronchoalveolar lavage fluid. Immunologically, PLGA-NP peptides vaccination (without adjuvant) significantly increased the frequency of antigen-specific IFNγ secreting CD4 and CD8 T cells response in the lung lymphocytes, despite not boosting the antibody response both at pre- and post-challenge. In summary, our data indicated that nanoparticle-mediated delivery of conserved H1N1 influenza peptides induced the virus specific T cell response in the lungs and reduced the challenged heterologous virus load in the airways of pigs.

No MeSH data available.


Related in: MedlinePlus

Clinical outcome and virus load in the lungs of vaccinated and SwIV challenged pigs.(A) Rectal temperature was recorded everyday post-challenge (day -1 to 7). SwIV titer in BAL fluid at DPC 7 was detected by (B) viral RNA load by qRT-PCR, and (C) replicating infectious virus load using MDCK cells by immunofluorescence assay. The ratio indicated above each bar represents the number of pigs positive for virus and contributed to the data out of 6 or 7 animals. The viral titer is expressed in tissue culture infective dose 50 (TCID50) in each ml of the BAL fluid. Each data point or bar represents the average value of 6 to 7 pigs ± SEM. Asterisk denotes statistically significant difference at P<0.05 (*) between MKC and NPP or NPA pig groups determined by One way ANOVA followed by Tukey Post-hoc test.
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pone.0151922.g004: Clinical outcome and virus load in the lungs of vaccinated and SwIV challenged pigs.(A) Rectal temperature was recorded everyday post-challenge (day -1 to 7). SwIV titer in BAL fluid at DPC 7 was detected by (B) viral RNA load by qRT-PCR, and (C) replicating infectious virus load using MDCK cells by immunofluorescence assay. The ratio indicated above each bar represents the number of pigs positive for virus and contributed to the data out of 6 or 7 animals. The viral titer is expressed in tissue culture infective dose 50 (TCID50) in each ml of the BAL fluid. Each data point or bar represents the average value of 6 to 7 pigs ± SEM. Asterisk denotes statistically significant difference at P<0.05 (*) between MKC and NPP or NPA pig groups determined by One way ANOVA followed by Tukey Post-hoc test.

Mentions: In the mock and unentrapped peptides vaccinated and virulent SwIV H1N1 challenged pigs, the rectal temperature was 1.5 to 2 °F higher at DPC 1 to 3 compared to mock animals (Fig 4A). The pigs which had fever were anorexic and lethargic during those initial days of viral challenge. Similarly challenged, but NPP and NPA vaccinated pig groups did not suffer from hyperthermia (Fig 4A). To associate the body temperature to viral load, we measured the viral RNA copies in BAL fluid and observed comparable RNA load among all the virus-challenged pig groups (Fig 4B). But the replicating SwIV was detected in ≥ 50% of MKC and PEP pig groups, and it was undetectable in the NPP and NPA vaccinated animals (Fig 4C). The assay was repeated three times to confirm the live SwIV titer in the BAL fluid samples. Nasal swabs collected at DPC 4 and 7 did not show any detectable replicating virus in any of the virus challenged pig groups (data not shown). In summary, clinical signs of flu and replicating detectable lung viral load were detected only in SwIV challenged control (MCK and PEP), but not in PLGA-NP peptides (NPP and NPA) vaccinated pig groups.


Entrapment of H1N1 Influenza Virus Derived Conserved Peptides in PLGA Nanoparticles Enhances T Cell Response and Vaccine Efficacy in Pigs.

Hiremath J, Kang KI, Xia M, Elaish M, Binjawadagi B, Ouyang K, Dhakal S, Arcos J, Torrelles JB, Jiang X, Lee CW, Renukaradhya GJ - PLoS ONE (2016)

Clinical outcome and virus load in the lungs of vaccinated and SwIV challenged pigs.(A) Rectal temperature was recorded everyday post-challenge (day -1 to 7). SwIV titer in BAL fluid at DPC 7 was detected by (B) viral RNA load by qRT-PCR, and (C) replicating infectious virus load using MDCK cells by immunofluorescence assay. The ratio indicated above each bar represents the number of pigs positive for virus and contributed to the data out of 6 or 7 animals. The viral titer is expressed in tissue culture infective dose 50 (TCID50) in each ml of the BAL fluid. Each data point or bar represents the average value of 6 to 7 pigs ± SEM. Asterisk denotes statistically significant difference at P<0.05 (*) between MKC and NPP or NPA pig groups determined by One way ANOVA followed by Tukey Post-hoc test.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4836704&req=5

pone.0151922.g004: Clinical outcome and virus load in the lungs of vaccinated and SwIV challenged pigs.(A) Rectal temperature was recorded everyday post-challenge (day -1 to 7). SwIV titer in BAL fluid at DPC 7 was detected by (B) viral RNA load by qRT-PCR, and (C) replicating infectious virus load using MDCK cells by immunofluorescence assay. The ratio indicated above each bar represents the number of pigs positive for virus and contributed to the data out of 6 or 7 animals. The viral titer is expressed in tissue culture infective dose 50 (TCID50) in each ml of the BAL fluid. Each data point or bar represents the average value of 6 to 7 pigs ± SEM. Asterisk denotes statistically significant difference at P<0.05 (*) between MKC and NPP or NPA pig groups determined by One way ANOVA followed by Tukey Post-hoc test.
Mentions: In the mock and unentrapped peptides vaccinated and virulent SwIV H1N1 challenged pigs, the rectal temperature was 1.5 to 2 °F higher at DPC 1 to 3 compared to mock animals (Fig 4A). The pigs which had fever were anorexic and lethargic during those initial days of viral challenge. Similarly challenged, but NPP and NPA vaccinated pig groups did not suffer from hyperthermia (Fig 4A). To associate the body temperature to viral load, we measured the viral RNA copies in BAL fluid and observed comparable RNA load among all the virus-challenged pig groups (Fig 4B). But the replicating SwIV was detected in ≥ 50% of MKC and PEP pig groups, and it was undetectable in the NPP and NPA vaccinated animals (Fig 4C). The assay was repeated three times to confirm the live SwIV titer in the BAL fluid samples. Nasal swabs collected at DPC 4 and 7 did not show any detectable replicating virus in any of the virus challenged pig groups (data not shown). In summary, clinical signs of flu and replicating detectable lung viral load were detected only in SwIV challenged control (MCK and PEP), but not in PLGA-NP peptides (NPP and NPA) vaccinated pig groups.

Bottom Line: Biodegradable polylactic-co-glycolic acid (PLGA) nanoparticle (PLGA-NP) based vaccine delivery system enhances cross-presentation of antigens by the professional antigen presenting cells.Immunologically, PLGA-NP peptides vaccination (without adjuvant) significantly increased the frequency of antigen-specific IFNγ secreting CD4 and CD8 T cells response in the lung lymphocytes, despite not boosting the antibody response both at pre- and post-challenge.In summary, our data indicated that nanoparticle-mediated delivery of conserved H1N1 influenza peptides induced the virus specific T cell response in the lungs and reduced the challenged heterologous virus load in the airways of pigs.

View Article: PubMed Central - PubMed

Affiliation: Food Animal Health Research Program, Ohio Agricultural Research and Development Center, 1680 Madison Avenue, Wooster, Ohio, 44691, United States of America, and Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, 43210, United States of America.

ABSTRACT
Pigs are believed to be one of the important sources of emerging human and swine influenza viruses (SwIV). Influenza virus conserved peptides have the potential to elicit cross-protective immune response, but without the help of potent adjuvant and delivery system they are poorly immunogenic. Biodegradable polylactic-co-glycolic acid (PLGA) nanoparticle (PLGA-NP) based vaccine delivery system enhances cross-presentation of antigens by the professional antigen presenting cells. In this study, Norovirus P particle containing SwIV M2e (extracellular domain of the matrix protein 2) chimera and highly conserved two each of H1N1 peptides of pandemic 2009 and classical human influenza viruses were entrapped in PLGA-NPs. Influenza antibody-free pigs were vaccinated with PLGA-NPs peptides cocktail vaccine twice with or without an adjuvant, Mycobacterium vaccae whole cell lysate, intranasally as mist. Vaccinated pigs were challenged with a virulent heterologous zoonotic SwIV H1N1, and one week later euthanized and the lung samples were analyzed for the specific immune response and viral load. Clinically, pigs vaccinated with PLGA-NP peptides vaccine had no fever and flu symptoms, and the replicating challenged SwIV was undetectable in the bronchoalveolar lavage fluid. Immunologically, PLGA-NP peptides vaccination (without adjuvant) significantly increased the frequency of antigen-specific IFNγ secreting CD4 and CD8 T cells response in the lung lymphocytes, despite not boosting the antibody response both at pre- and post-challenge. In summary, our data indicated that nanoparticle-mediated delivery of conserved H1N1 influenza peptides induced the virus specific T cell response in the lungs and reduced the challenged heterologous virus load in the airways of pigs.

No MeSH data available.


Related in: MedlinePlus