Construction and growth properties of bovine herpesvirus type 5 recombinants defective in the glycoprotein E or thymidine kinase gene or both

Brum, Mário Celso Sperotto; Weiblen, Rudi; Flores, Eduardo Furtado; Chowdhury, Shafiqul I.

doi:10.1590/s0100-879x2009007500008

Cited by 15 publications

(35 citation statements)

References 31 publications

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“…Several BoHV-1 genedeleted viruses have been constructed and some have been used in control/eradication programs BoHV-1 infection in Europe and North America (Kaashoek et al 1995, Strube et al 1996. Gene-deleted BoHV-1 and -5 Brazilian isolates have been developed and are under testing for a future use in vaccine formulations (Franco et al 2002, 2007, Brum et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E

et al. 2010

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RESUMO.-[Imunogenicidade de uma cepa inativada do herpesvírus bovino tipo 5 defectiva na timidina quinase e glicoproteína E.] A imunogenicidade de vacina experimental inativada, produzida com uma cepa do herpesvírus bovino tipo 5 defectiva nos genes da timidina quinase e glicoproteína E (BoHV-5 gE/TKΔ) foi avaliada em bovinos e o resultado foi comparado com a resposta induzida pela cepa parental do The immunogenicity of an inactivated, experimental vaccine based on a bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E (BoHV-5 gE/ TKΔ) was evaluated in cattle and the results were compared with a vaccine containing the parental BoHV-5 strain (SV507/99). To formulate the vaccines, each virus (wildtype SV507/99 and BoHV-5 gE/TKΔ) was multiplied in cell culture and inactivated with binary ethyleneimine (BEI). Each vaccine dose contained approximately of 10 7.5 TCID 50 of inactivated virus mixed with an oil-based adjuvant (46:54). Forty calves, 6 to 9-monthsold, were allocated into two groups of 20 animals each and vaccinated twice (days 0 and 22pv) by the subcutaneous route with either vaccine. Serum samples collected at day 0 and at different intervals after vaccination were tested for virus neutralizing (VN) antibodies against the parental virus and against heterologous BoHV-5 and BoHV-1 isolates. The VN assays demonstrated seroconversion to the respective homologous viruses in all vaccinated animals after the second vaccine dose (mean titers of 17.5 for the wildtype vaccine; 24.1 for the recombinant virus). All animals remained reagents up to day 116 pv, yet showing a gradual reduction in VN titers. Animals from both vaccine groups reacted in similar VN titers to different BoHV-1 and BoHV-5 isolates, yet the magnitude of serological response of both groups was higher against BoHV-5 field isolates. Calves vaccinated with the recombinant virus did not develop antibodies to gE as verified by negative results in a gE-specific ELISA, what would allow serological differentiation from naturally infected animals. Taken together, these results indicate that inactivated antigens of BoHV-5 gE/TK recombinant virus induced an adequate serological response against BoHV-5 and BoHV-1 and thus can be used as an alternative, differential vaccine candidate. inativados emulsionado em adjuvante oleoso (46:54). Quarenta bezerros de raças cruzadas, com idade entre seis a nove meses, foram alocados em dois grupos de 20 animais cada e vacinados duas vezes (dia 0 e 22 pv) pela via subcutânea com uma das vacinas. Amostras de soro foram coletadas no dia 0 e a vários intervalos após vacinação para a pesquisa de anticorpos neutralizantes frente ao vírus homólogo ou frente a isolados de BoHV-5 e BoHV-1. Os testes de soroneutralização (SN) demonstraram que todos os animais soroconverteram após a segunda dose da vacina (títulos médios de 17,5 para o grupo SV507/99; 24,1 para o grupo BoHV-5 gE/TKΔ). Todos os animais mantiveram níveis de anticorpos neutralizantes até o dia 116 pv, no entanto foi observada uma redução ...

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Section: Introductionmentioning

confidence: 99%

Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E

et al. 2010

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“…As BoHV-1 and BoHV-5 co-circulate in Brazilian cattle (Silva et al 2007), a commercial vaccine for use in the country would require the inclusion of both viruses. To this end, we constructed a double deletion BoHV-5 gE-and tk-mutant (BoHV-5gEΔTKΔ, Brum et al 2010a). Hence, a gE-specific ELISA -for serological differentiation of vaccinated from naturally infected animals -would be required as a companion test.…”

Section: Discussionmentioning

confidence: 99%

“…Few vaccines contain BoHV-5 antigens in their formulation, none of them based on live virus (E.F. Flores, unpublished). The antigenic similarity and serological cross-reactivity between BoHV-1 and -5 has led to the assumption that vaccines containing either virus would confer protection upon homologous and heterologous virus challenge (Del Medico Zajac et al 2006, Brum et al 2010a, Anziliero et al 2011. Following the trend of North America and Europe, South American countries are now embarking on the development and use of BoHV-1 DIVA (for differentiating infected from vaccinated animals) vaccines (Brum et al 2010a).…”

Section: Introductionmentioning

confidence: 99%

“…The antigenic similarity and serological cross-reactivity between BoHV-1 and -5 has led to the assumption that vaccines containing either virus would confer protection upon homologous and heterologous virus challenge (Del Medico Zajac et al 2006, Brum et al 2010a, Anziliero et al 2011. Following the trend of North America and Europe, South American countries are now embarking on the development and use of BoHV-1 DIVA (for differentiating infected from vaccinated animals) vaccines (Brum et al 2010a). Herpesvirus DIVA vaccines are usually based on viral strains deleted in genes encoding an envelope, non-essential glycoprotein , Ackermann & Engels 2006, Van Drunen Littel-van den Hurk 2006.…”

Section: Introductionmentioning

confidence: 99%

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Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and its use in an ELISA for gE antibodies

et al. 2013

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Pesq. Vet. Bras. 33(1): 41-46, janeiro 2013 41 RESUMO.-[Expressão procariota de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 e uso em ELISA para anticorpos contra a gE.] Este trabalho relata a expressão de uma forma truncada da glicoproteína E (gE) do herpesvírus bovino tipo 1 (BoHV-1) para uso em imunodiagnóstico. Um fragmento de 651 pares de bases (pb) correspondente ao terço amino-terminal (217 aminoácidos) da gE do BoHV-1 -que compartilha uma alta identidade com a gE do BoHV-5 -foi clonada como proteína de fusão com cauda 6x de histidina em um vetor de expres- This article describes the expression of a truncated form of bovine herpesvirus 1 (BoHV-1) glycoprotein E (gE) for use as immunodiagnostic reagent. A 651 nucleotide fragment corresponding to the amino-terminal third (217 amino acids) of BoHV-1 gE -that shares a high identity with the homologous BoHV-5 counterpart -was cloned as a 6×His-tag fusion protein in an Escherichia coli expression vector. A soluble protein of approximately 25 kDa purified from lysates of transformed E. coli was recognized in Western blot (WB) by anti-6xHis-tag and anti-BoHV-1 gE monoclonal antibodies. In addition, the recombinant protein was specifically recognized in WB by antibodies present in the sera of cattle seropositive to BoHV-1 and BoHV-5. An indirect ELISA using the expressed protein as coating antigen performed comparably to a commercial anti-gE ELISA and was able to differentiate serologically calves vaccinated with a gE-deleted BoHV-5 strain from calves infected with BoHV-1. Thus, the truncated gE may be useful for serological tests designed to differentiate BoHV-1/BoHV-5 infected animals from those vaccinated with gE-negative marker vaccines. bovine herpesvirus, vaccine, DIVA, recombinant protein. são em Escherichia coli. Uma proteína solúvel de aproximadamente 25 kDa purificada de lisados de E.coli foi reconhecida em Western blot (WB) por anticorpos monoclonais anti-6xHis-tag e anti-gE. Além disso, a proteína recombinante purificada foi reconhecida em WB por anticorpos presentes no soro de animais soropositivos ao BoHV-1 e BoHV-5. Um ELISA indireto utilizando a proteína recombinante como antígeno apresentou performance comparável a um ELISA gE comercial e foi capaz e diferenciar sorologicamente animais vacinados com uma cepa gE-negativa de BoHV-5 de animais infectados com o BoHV-1. Portanto, a gE truncada pode ser útil em testes sorológicos diferenciais para uso conjunto com vacinas com marcador antigênico gE para o BoHV-1 e BoHV-5.TERMOS DE INDEXAÇÃO: BoHV-5, herpesvírus bovino, vacina diferencial, proteína recombinante.

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“…Although Brazil and most South American countries have not yet embarked on BoHV-1 eradication programs, the use of differential vaccines (still insipient) represents an attractive strategy for herd certification through animal trade and/or export (Brum et al 2009). The safety and attenuation of the vaccine strain SV265gE-for calves and pregnant cows also represent an advantage over other vaccine strains available (Franco et al 2002b;Spilki et al 2005).…”

Section: Discussionmentioning

confidence: 99%

Genital immunization of heifers with a glycoprotein Edeleted, recombinant bovine herpesvirus 1 strain confers protection upon challenge with a virulent isolate

et al. 2010

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ABSTRACT.-Weiss M., Vogel F.S.F., Martins M., Weiblen R., Roehe P.M., Franco A.C. & Flores E.F. 2010. Genital immunization of heifers with a glycoprotein E-deleted, recombinant bovine herpesvirus 1 strain confers protection upon challenge with a virulent isolate. Pesquisa Veterinária Brasileira 30(1):42-50. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: eduardofurtadoflores@gmail.com Venereal infection of seronegative heifers and cows with bovine herpesvirus type 1.2 (BoHV-1.2) frequently results in vulvovaginitis and transient infertility. Parenteral immunization with inactivated or modified live BoHV-1 vaccines often fails in conferring protection upon genital challenge. We herein report an evaluation of the immune response and protection conferred by genital vaccination of heifers with a glycoprotein E-deleted recombinant virus (SV265gE-). A group of six seronegative heifers was vaccinated with SV265gE-(0,2mL containing 10 6.9 TCID 50 ) in the vulva submucosa (group IV); four heifers were vaccinated intramuscularly (group IM, 1mL containing 10 7.6 TCID 50 ) and four heifers remained as non-vaccinated controls. Heifers vaccinated IV developed mild, transient local edema and hyperemia and shed low amounts of virus for a few days after vaccination, yet a sentinel heifer maintained in close contact did not seroconvert. Attempts to reactivate the vaccine virus in two IV vaccinated heifers by intravenous administration of dexamethasone (0.5mg/kg) at day 70 pv failed since no virus shedding, recrudescence of genital signs or seroconversion were observed. At day 70 pv, all vaccinated and control heifers were challenged by genital inoculation of a highly virulent BoHV-1.2 isolate (SV-56/90, 10 7.1 TCID 50 /animal). After challenge, virus shedding was detected in genital secretions of control animals for 8.2 days (8-9); in the IM group for 6.2 days (4-8 days) and during 5.2 days (5-6 days) in the IV group. Control non-vaccinated heifers developed moderate (2/4) or severe (2/4) vulvovaginitis lasting 9 to 13 days (x: 10.7 days). The disease was characterized by vulvar edema, vulvo-vestibular congestion, vesicles progressing to coalescence and erosions, fibrino-necrotic plaques and fibrinopurulent exudate. IM vaccinated heifers developed mild (1/3) or moderate (3/4) genital lesions, lasting 10 to 12 days (x: 10.7 days); and IV vaccinated heifers developed mild and transient vulvovaginitis (3/4) or mild to moderate genital lesions (1/4). In the IV group, the clinical signs lasted 4 to 8 days (x: 5.5 days). Clinical examination of the animals after challenge revealed that vaccination by both routes conferred some degree of protection, yet IV vaccination was clearly more effective in reducing the severity and duration of clinical 2) pode resultar em vulvovaginite e infertilidade temporária. As vacinas atenuadas ou inativadas administradas pela via parenteral freqüente-mente conferem proteção incompleta frente a desafio pela via genital. Este...

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Construction and growth properties of bovine herpesvirus type 5 recombinants defective in the glycoprotein E or thymidine kinase gene or both

Cited by 15 publications

References 31 publications

Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E

Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E

Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and its use in an ELISA for gE antibodies

Genital immunization of heifers with a glycoprotein Edeleted, recombinant bovine herpesvirus 1 strain confers protection upon challenge with a virulent isolate

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