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A Recombinant Raccoon Poxvirus Vaccine Expressing both Yersinia pestis F1 and Truncated V Antigens Protects Animals against Lethal Plague.

Rocke TE, Kingstad-Bakke B, Berlier W, Osorio JE - Vaccines (Basel) (2014)

Bottom Line: Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens.This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis.The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

View Article: PubMed Central - PubMed

Affiliation: National Wildlife Health Center, U.S. Geological Survey, 6006 Schroeder Rd., Madison, WI 53711, USA. trocke@usgs.gov.

ABSTRACT
In previous studies, we demonstrated in mice and prairie dogs that simultaneous administration of two recombinant raccoon poxviruses (rRCN) expressing Yersinia pestis antigens (F1 and V307-a truncated version of the V protein) provided superior protection against plague challenge compared to individual single antigen constructs. To reduce costs of vaccine production and facilitate implementation of a sylvatic plague vaccine (SPV) control program for prairie dogs, a dual antigen construct is more desirable. Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens. This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis. The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of dual antigen plague vaccine construct, RCN-F1/V307, versus individual constructs, RCN-F1 and RCN-V307. Vero cell monolayers were infected with RCN constructs at an MOI of 1 and pellet (pt) and supernatant (sup) fractions were harvested 24 h post infection. Blot A was probed with anti-V antibody, and blot B was probed with anti-F1 antibody. Arrows indicate Vt and F1 proteins. Lanes for blot A are: (1) ladder, (2) RCN-V307 pt, (3) RCN-V307 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-F1 pt, (7) RCN-F1 sup. Lanes for Blot B are: (1) ladder, (2) RCN-F1 pt, (3) RCN-F1 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-V307 pt, (7) RCN-V307 sup.
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vaccines-02-00772-f002: Western blot analysis of dual antigen plague vaccine construct, RCN-F1/V307, versus individual constructs, RCN-F1 and RCN-V307. Vero cell monolayers were infected with RCN constructs at an MOI of 1 and pellet (pt) and supernatant (sup) fractions were harvested 24 h post infection. Blot A was probed with anti-V antibody, and blot B was probed with anti-F1 antibody. Arrows indicate Vt and F1 proteins. Lanes for blot A are: (1) ladder, (2) RCN-V307 pt, (3) RCN-V307 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-F1 pt, (7) RCN-F1 sup. Lanes for Blot B are: (1) ladder, (2) RCN-F1 pt, (3) RCN-F1 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-V307 pt, (7) RCN-V307 sup.

Mentions: A dual antigen RCN-vectored vaccine was successfully constructed containing both F1 and a truncated form of the lcrV gene (V307). The genetic stability of the RCN-F1/V307 construct was assessed following ten blind passages in Vero cells. No changes in PCR fragment size and sequence were observed for the F1 and V307 inserts [12]. The in vitro expression of RCN-F1/V307 was examined by western blot analyses at 24 h PI in comparison to RCN-F1 and RCN-V307 (Figure 2). Samples were treated identically and to ensure equal loading, the same samples were used for both blots A and B. No differences were detected in the migration between proteins expressed from the RCN-F1/V307 construct and the individual RCN-F1 and RCN-V307 constructs. For the RCN-F1/V307 construct, Y. pestis antigens were detected in the cell pellet and supernatant of Vero-infected cells. Similar levels of V307 expression were observed in the RCN-V307 and RCN-F1/V307 constructs. In contrast, F1 expression was significantly lower in the RCN-F1/V307 than the RCN-F1 construct. Multiple-sized fragments are due to both glycosylated and non-glycosylated forms of the protein being expressed [8].


A Recombinant Raccoon Poxvirus Vaccine Expressing both Yersinia pestis F1 and Truncated V Antigens Protects Animals against Lethal Plague.

Rocke TE, Kingstad-Bakke B, Berlier W, Osorio JE - Vaccines (Basel) (2014)

Western blot analysis of dual antigen plague vaccine construct, RCN-F1/V307, versus individual constructs, RCN-F1 and RCN-V307. Vero cell monolayers were infected with RCN constructs at an MOI of 1 and pellet (pt) and supernatant (sup) fractions were harvested 24 h post infection. Blot A was probed with anti-V antibody, and blot B was probed with anti-F1 antibody. Arrows indicate Vt and F1 proteins. Lanes for blot A are: (1) ladder, (2) RCN-V307 pt, (3) RCN-V307 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-F1 pt, (7) RCN-F1 sup. Lanes for Blot B are: (1) ladder, (2) RCN-F1 pt, (3) RCN-F1 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-V307 pt, (7) RCN-V307 sup.
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Related In: Results  -  Collection

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vaccines-02-00772-f002: Western blot analysis of dual antigen plague vaccine construct, RCN-F1/V307, versus individual constructs, RCN-F1 and RCN-V307. Vero cell monolayers were infected with RCN constructs at an MOI of 1 and pellet (pt) and supernatant (sup) fractions were harvested 24 h post infection. Blot A was probed with anti-V antibody, and blot B was probed with anti-F1 antibody. Arrows indicate Vt and F1 proteins. Lanes for blot A are: (1) ladder, (2) RCN-V307 pt, (3) RCN-V307 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-F1 pt, (7) RCN-F1 sup. Lanes for Blot B are: (1) ladder, (2) RCN-F1 pt, (3) RCN-F1 sup, (4) RCN-F1/V307 pt, (5) RCN-F1/V307 sup, (6) RCN-V307 pt, (7) RCN-V307 sup.
Mentions: A dual antigen RCN-vectored vaccine was successfully constructed containing both F1 and a truncated form of the lcrV gene (V307). The genetic stability of the RCN-F1/V307 construct was assessed following ten blind passages in Vero cells. No changes in PCR fragment size and sequence were observed for the F1 and V307 inserts [12]. The in vitro expression of RCN-F1/V307 was examined by western blot analyses at 24 h PI in comparison to RCN-F1 and RCN-V307 (Figure 2). Samples were treated identically and to ensure equal loading, the same samples were used for both blots A and B. No differences were detected in the migration between proteins expressed from the RCN-F1/V307 construct and the individual RCN-F1 and RCN-V307 constructs. For the RCN-F1/V307 construct, Y. pestis antigens were detected in the cell pellet and supernatant of Vero-infected cells. Similar levels of V307 expression were observed in the RCN-V307 and RCN-F1/V307 constructs. In contrast, F1 expression was significantly lower in the RCN-F1/V307 than the RCN-F1 construct. Multiple-sized fragments are due to both glycosylated and non-glycosylated forms of the protein being expressed [8].

Bottom Line: Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens.This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis.The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

View Article: PubMed Central - PubMed

Affiliation: National Wildlife Health Center, U.S. Geological Survey, 6006 Schroeder Rd., Madison, WI 53711, USA. trocke@usgs.gov.

ABSTRACT
In previous studies, we demonstrated in mice and prairie dogs that simultaneous administration of two recombinant raccoon poxviruses (rRCN) expressing Yersinia pestis antigens (F1 and V307-a truncated version of the V protein) provided superior protection against plague challenge compared to individual single antigen constructs. To reduce costs of vaccine production and facilitate implementation of a sylvatic plague vaccine (SPV) control program for prairie dogs, a dual antigen construct is more desirable. Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens. This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis. The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

No MeSH data available.


Related in: MedlinePlus