Vaccination of chickens with recombinant Salmonella expressing M2e and CD154 epitopes increases protection and decreases viral shedding after low pathogenic avian influenza challenge

Avian influenza (AI) is a significant public health concern and serious economic threat to the commercial poultry industry worldwide. Previous research demonstrates that antibodies against M2e confer protection against influenza challenge. Using the Red recombinase system in combination with overlap...

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Veröffentlicht in:Poultry science 2009-11, Vol.88 (11), p.2244-2252
Hauptverfasser: Layton, S.L, Kapczynski, D.R, Higgins, S, Higgins, J, Wolfenden, A.D, Liljebjelke, K.A, Bottje, W.G, Swayne, D, Berghman, L.R, Kwon, Y.M, Hargis, B.M, Cole, K
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container_end_page 2252
container_issue 11
container_start_page 2244
container_title Poultry science
container_volume 88
creator Layton, S.L
Kapczynski, D.R
Higgins, S
Higgins, J
Wolfenden, A.D
Liljebjelke, K.A
Bottje, W.G
Swayne, D
Berghman, L.R
Kwon, Y.M
Hargis, B.M
Cole, K
description Avian influenza (AI) is a significant public health concern and serious economic threat to the commercial poultry industry worldwide. Previous research demonstrates that antibodies against M2e confer protection against influenza challenge. Using the Red recombinase system in combination with overlapping extension PCR, we recently developed several novel attenuated Salmonella Enteritidis strains that express a protective M2e epitope in combination with a potential immune-enhancing CD154 peptide sequence on the Salmonella outer membrane protein lamB. Commercial Leghorn chicks were orally immunized (immunization dose: 10⁶ to 10⁸ cfu/chick) with saline (negative control) or one of the recombinant Salmonella strains [ΔaroA M2e-CD154, ΔhtrA M2e-CD154, ΔaroA-ΔhtrA M2e(4)-CD154] on day of hatch and 21 d posthatch. These candidate vaccine strains were evaluated for their ability to invade, colonize, and persist in tissues and elicit an M2e-specific antibody response. The vaccine candidate strain ΔaroA M2e-CD154 exhibited significantly greater organ invasion in the liver and spleen at d 7 (P > 0.05); however, no marked differences in colonization of the cecal tonsils were observed. Vaccinated chickens exhibited significantly increased M2e-specific IgG responses, which were further enhanced by simultaneous expression of CD154 (P < 0.05). Virus neutralization assays gave neutralizing indices of 6.6, 6.3, and 6.3 for ΔaroA M2e-CD154, ΔhtrA M2e-CD154, and ΔaroA-ΔhtrA M2e(4)-CD154 seven days post booster immunization, respectively, indicating effective neutralization of AI by serum IgG of vaccinated chickens. In a subsequent direct challenge study, specific-pathogen-free Leghorn chicks immunized with ΔaroA-ΔhtrA M2e(4)-CD154 offered significant protection against direct challenge with low pathogenic AI H7N2, but not highly pathogenic H5N1 AI. Taken together, these data suggest that these Salmonella-vectored vaccines expressing M2e in association with CD154 are effective at protecting chickens against low pathogenic AI.
doi_str_mv 10.3382/ps.2009-00251
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Previous research demonstrates that antibodies against M2e confer protection against influenza challenge. Using the Red recombinase system in combination with overlapping extension PCR, we recently developed several novel attenuated Salmonella Enteritidis strains that express a protective M2e epitope in combination with a potential immune-enhancing CD154 peptide sequence on the Salmonella outer membrane protein lamB. Commercial Leghorn chicks were orally immunized (immunization dose: 10⁶ to 10⁸ cfu/chick) with saline (negative control) or one of the recombinant Salmonella strains [ΔaroA M2e-CD154, ΔhtrA M2e-CD154, ΔaroA-ΔhtrA M2e(4)-CD154] on day of hatch and 21 d posthatch. These candidate vaccine strains were evaluated for their ability to invade, colonize, and persist in tissues and elicit an M2e-specific antibody response. 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Previous research demonstrates that antibodies against M2e confer protection against influenza challenge. Using the Red recombinase system in combination with overlapping extension PCR, we recently developed several novel attenuated Salmonella Enteritidis strains that express a protective M2e epitope in combination with a potential immune-enhancing CD154 peptide sequence on the Salmonella outer membrane protein lamB. Commercial Leghorn chicks were orally immunized (immunization dose: 10⁶ to 10⁸ cfu/chick) with saline (negative control) or one of the recombinant Salmonella strains [ΔaroA M2e-CD154, ΔhtrA M2e-CD154, ΔaroA-ΔhtrA M2e(4)-CD154] on day of hatch and 21 d posthatch. These candidate vaccine strains were evaluated for their ability to invade, colonize, and persist in tissues and elicit an M2e-specific antibody response. 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Previous research demonstrates that antibodies against M2e confer protection against influenza challenge. Using the Red recombinase system in combination with overlapping extension PCR, we recently developed several novel attenuated Salmonella Enteritidis strains that express a protective M2e epitope in combination with a potential immune-enhancing CD154 peptide sequence on the Salmonella outer membrane protein lamB. Commercial Leghorn chicks were orally immunized (immunization dose: 10⁶ to 10⁸ cfu/chick) with saline (negative control) or one of the recombinant Salmonella strains [ΔaroA M2e-CD154, ΔhtrA M2e-CD154, ΔaroA-ΔhtrA M2e(4)-CD154] on day of hatch and 21 d posthatch. These candidate vaccine strains were evaluated for their ability to invade, colonize, and persist in tissues and elicit an M2e-specific antibody response. The vaccine candidate strain ΔaroA M2e-CD154 exhibited significantly greater organ invasion in the liver and spleen at d 7 (P &gt; 0.05); however, no marked differences in colonization of the cecal tonsils were observed. Vaccinated chickens exhibited significantly increased M2e-specific IgG responses, which were further enhanced by simultaneous expression of CD154 (P &lt; 0.05). Virus neutralization assays gave neutralizing indices of 6.6, 6.3, and 6.3 for ΔaroA M2e-CD154, ΔhtrA M2e-CD154, and ΔaroA-ΔhtrA M2e(4)-CD154 seven days post booster immunization, respectively, indicating effective neutralization of AI by serum IgG of vaccinated chickens. In a subsequent direct challenge study, specific-pathogen-free Leghorn chicks immunized with ΔaroA-ΔhtrA M2e(4)-CD154 offered significant protection against direct challenge with low pathogenic AI H7N2, but not highly pathogenic H5N1 AI. Taken together, these data suggest that these Salmonella-vectored vaccines expressing M2e in association with CD154 are effective at protecting chickens against low pathogenic AI.</abstract><cop>England</cop><pub>Poultry Science Association</pub><pmid>19834072</pmid><doi>10.3382/ps.2009-00251</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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subjects Animals
Antigens, Viral - genetics
Antigens, Viral - immunology
Antigens, Viral - metabolism
avian influenza
Chickens
Epitopes - genetics
Epitopes - metabolism
immune response
immunoglobulin G
Influenza A virus
Influenza in Birds - prevention & control
Influenza Vaccines - immunology
liver
membrane proteins
microbial colonization
molecular sequence data
neutralization tests
nucleotide sequences
oral administration
pathogenicity
polymerase chain reaction
recombinant antigens
recombinant vaccines
Salmonella - genetics
Salmonella - metabolism
Salmonella enteritidis
spleen
strains
tonsils
vaccination
viral antigens
Virus Shedding
title Vaccination of chickens with recombinant Salmonella expressing M2e and CD154 epitopes increases protection and decreases viral shedding after low pathogenic avian influenza challenge
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