Immunoblot analysis of the antibody response to ovine lentivirus infections

Houwers, D.J.; Nauta, I M

doi:10.1016/0378-1135(89)90078-3

Cited by 55 publications

(56 citation statements)

References 16 publications

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“…Radioimmunoprecipitation is technically demanding and rarely used. Western blotting is established more widely as a routine standard for confirming ELISAs [12,61,137].…”

Section: Methods Of Detectionmentioning

confidence: 99%

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Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

et al. 2004

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-Small ruminant lentiviruses (SRLV = maedi-visna in sheep and caprine arthritis encephalitis in goats) are distributed throughout most countries of the world, particularly Europe. Laboratories from 16 European countries established collaborations within the framework of a COST (CO-operation in the field of Scientific and Technical Research) action sponsored by the European Union in order to (i) better organize their research programmes on SRLVs and (ii) to coordinate efforts to combat these two diseases. After five years, a consensus conference -the first one in the veterinary medicine field -concluded the work of this network of laboratories by reviewing the present position and discussing three important questions in the field of SRLVs: routes of transmission, consequences of infection and potential role of eradication programmes at either a European or local level, according to the situation in each country or region. This paper brings together existing information regarding these questions and identifies areas for future research.Maedi-visna / caprine arthritis-encephalitis virus / lentivirus / small ruminants / European consensus conference

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“…Radioimmunoprecipitation is technically demanding and rarely used. Western blotting is established more widely as a routine standard for confirming ELISAs [12,61,137].…”

Section: Methods Of Detectionmentioning

confidence: 99%

“…The mean time to seroconversion is shorter in heavily infected flocks than in those with a low seroprevalence. In addition, animals with a low antibody titre may transiently become seronegative [59,61,78].…”

Section: Serological Testsmentioning

confidence: 99%

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

et al. 2004

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“…Just after challenge (week 16) and until the end of the experimental period, groups receiving B7 genes produced both anti--ENV and anti--GAG antibodies, as revealed by WB, whereas the gag-env group had anti--ENV but a very low production of anti--GAG antibodies at weeks 16 and 20 and the control group did not show either anti--ENV or anti--GAG antibodies until week 20. In natural infections, anti--ENV and GAG p25 antibodies are usually produced the earliest [53] although there can be a delay in the production of anti--ENV antibodies [54]. In this work, the delay was restricted to TM--ENV antibodies, likely because some TM epitopes are less exposed.…”

Section: Discussionmentioning

confidence: 86%

Use of B7 costimulatory molecules as adjuvants in a prime-boost vaccination against Visna/Maedi ovine lentivirus

Andrés

Reina

Ciriza

et al. 2009

Vaccine

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RNA transcripts of the B7 family molecule (CD80) are diminished in blood leukocytes from animals clinically affected with Visna/Maedi virus (VMV) infection. This work investigates whether the use of B7 genes enhances immune responses and protection in immunization--challenge approaches. Sheep were primed by particle--mediated epidermal bombardment with VMV gag and env gene recombinant plasmids together with plasmids encoding both CD80 and CD86 or CD80 alone, boosted with gag and env gene recombinant modified vaccinia Ankara virus and challenged intratracheally with VMV. Immunization in the presence of one or both of the B7 genes resulted in CD4+ T cell activation and antibody production (before and after challenge, respectively), but only immunization with CD80 and CD86 genes together, and not CD80 alone, resulted in a reduced number of infected animals and increased early transient cytotoxic T lymphocytes (CTL) responses. Post--mortem analysis showed an immune activation of lymphoid tissue in challenge--target organs in those animals that had received B7 genes compared to unvaccinated animals. Thus, the inclusion of B7 genes helped to enhance early cellular responses and protection (diminished proportion of infected animals) against VMV infection.

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“…complement fixation (Torfason, 1992), indirect fluorescence and virus neutralization (Gudnadottir, 1990), agar gel immunodiffusion test (Winward et al, 1979) and different indirect ELISAs combined with immunoblot (Houwers and Nauta, 1989;Zanoni et al, 1994;Celer et al, 1998). Indirect ELISA test based on whole virus antigen is commercially available.…”

Section: Introductionmentioning

confidence: 99%

“…The major capsid protein p25 is considered as one of the immunodominant antigens in lentivirus infection (Houwers and Nauta, 1989). It is the first antigen recognized by the immune system of infected animals and the production of p25 specific antibodies persists for a long time.…”

Section: Introductionmentioning

confidence: 99%

Purification of Escherichia coli-expressed HIS-tagged Maedi-Visna p25 core antigen by Ni2+-chelate affinity chromatography

Molínková¹

2001

Vet. Med. - Czech

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In this study, recombinant histidine tagged p25 capsid protein of Maedi-Visna virus was developed. Part of the viral genome coding p25 protein was positioned downstream and in frame with a metal binding domain in pRSET-B vector. Recombinant protein was expressed in E. coli cells and soluble fraction of the protein was subsequently purified by Ni 2+ -chelate affinity chromatography. Purified protein was then used as antigen in an indirect ELISA test. One hundred fifty ovine serum samples were screened for antibodies to p25 protein of the virus. Immunoblot with whole virus antigen was used as a gold standard. The total number of positive results in the ELISA was 38 (25.33%). Immunoblot failed to confirm a positive result in 2 (1.33%) of them and these results were therefore considered to be false positive. The number of true positive results in the ELISA was thus 36 (24%). All immunoblot positive samples were also positive by ELISA test. In conclusion, recombinant His-tagged capsid protein showed very high sensitivity and specificity in detecting antibodies to Maedi-Visna virus.

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Immunoblot analysis of the antibody response to ovine lentivirus infections

Cited by 55 publications

References 16 publications

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

Use of B7 costimulatory molecules as adjuvants in a prime-boost vaccination against Visna/Maedi ovine lentivirus

Purification of Escherichia coli-expressed HIS-tagged Maedi-Visna p25 core antigen by Ni2+-chelate affinity chromatography

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