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Protein-Specific Analysis of Humoral Immune Responses in a Clinical Trial for Vaccines against Contagious Bovine Pleuropneumonia
KTH, School of Biotechnology (BIO), Proteomics. (Proteomik)
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2010 (English)In: Clinical and Laboratory Immunology, ISSN 1556-6811, E-ISSN 1556-679X, Vol. 17, no 5, 853-861 p.Article in journal (Refereed) Published
Abstract [en]

Specific humoral immune responses in a clinical trial on cattle for vaccines against contagious bovine pleuropneumonia (CBPP) were investigated. The trial included a subunit vaccine consisting of five recombinant putative variable surface proteins of the infectious agent Mycoplasma mycoides subspecies mycoides small colony type (M. mycoides SC) compared to the currently approved attenuated vaccine strain T1/44 and untreated controls. Humoral immune responses to 65 individual recombinant surface proteins of M. mycoides SC were monitored by a recently developed bead based array assay. Responses to the subunit vaccine components were found to be weak. Animals vaccinated with this vaccine were not protected and had CBPP lesions similar to the untreated controls. In correlating protein-specific humoral responses to T1/44 induced immunity, five proteins associated with a protective immune response were identified,namely LppQ and those of ORFs MSC_0271, MSC_0136, MSC_0079 and MSC_0431. The five proteins may be important candidates in the development of a novel subunit vaccine against CBPP.

Place, publisher, year, edition, pages
2010. Vol. 17, no 5, 853-861 p.
Keyword [en]
subsp mycoides sc, small colony type, lipoprotein lppq, genetic-characterization, cbpp, cattle, eradication, africa, strategies, infection
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-11107DOI: 10.1128/CVI.00019-10ISI: 000277243200023Scopus ID: 2-s2.0-77951988744OAI: oai:DiVA.org:kth-11107DiVA: diva2:235815
Note
Uppdaterad från manuskript 20100719 Tidigare titel: Protein-specific analysis of humoral immune responses in a CBPP vaccine trial QC 20100719Available from: 2009-09-18 Created: 2009-09-18 Last updated: 2011-01-19Bibliographically approved
In thesis
1. Protein based approaches to understand and prevent contagious bovine pleuropneumonia
Open this publication in new window or tab >>Protein based approaches to understand and prevent contagious bovine pleuropneumonia
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Contagious bovine pleuropneumonia (CBPP) is a severe infectious disease caused by Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) and is a vast problem in Africa. Current CBPP prevention is based on attenuated live strain vaccines, but these are limited by factors such as short-term immunity, cold-chain dependence and retained virulence. CBPP can be diagnosed using post-mortem examination, identification of the agent using culture and PCR based methods as well as serological diagnostic methods, but the latter are generally not sensitive enough and there is also demand for an inexpensive, pen side field test.The research presented in this thesis was focused on using recombinantly expressed surface proteins from M. mycoides SC to characterize humoral immune responses to CBPP. Thereby candidate proteins to be used in development of serological diagnostic methods and possibly subunit vaccines could be identified. As a first step, five putative variable surface proteins of M. mycoides SC were expressed and purified from E. coli in Paper I. These proteins were analyzed using immunoblotting techniques and results showed that one protein, MSC_0364, was variably expressed on the surface of M. mycoides SC in vitro. Paper II presents expanded efforts including cloning and expression of 64 recombinant surface proteins and an assay for high throughput analysis of protein-specific IgG, IgA and IgM titers in hundreds of sera using a bead-based screening assay. The assay was evaluated by protein-specific inhibition experiments, comparisons to Western blotting and monitoring of immune responses over time in a study with sera taken from eight animals over 293 days from a previous vaccine trial.Papers III and IV present applications using the recombinant proteins and bead-based screening assay wherein proteins for diagnostic and vaccine development were identified. In Paper III, the assay was used to screen 61 proteins using well-characterized serum samples from cattle with CBPP and healthy controls, resulting in selection of eight proteins suitable for diagnostic use. These proteins were combined and evaluated in a proof-of-concept ELISA with a discriminative power that enabled 96% correct classification of sera from CBPP-affected and CBPP-free bovines. Paper IV reports the results and protein-specific analyses of a vaccine trial using the recombinant putative variable surface proteins presented in Paper I as a subunit vaccine. The vaccine conferred no protection, but a weak vaccine response could not be excluded as the cause of failure. In an effort to identity other protein candidates to be used in a subunit vaccine, protein-specific analysis of humoral immune responses elicited by the currently approved live strain vaccine, T1/44, were investigated. Here, five proteins with high IgG titers associated to immunity were identified: LppQ, MSC_02714, MSC_0136, MSC_0079 and MSC_0431. These proteins may be important in the development of a novel subunit vaccine against CBPP.

Place, publisher, year, edition, pages
Stockholm: KTH, 2009. x, 77 p.
Series
Trita-BIO-Report, ISSN 1654-2312 ; 2009:17
Keyword
CBPP, Mycoplasma mycoides subsp. mycoides small colony type, humoral immune responses, recombinant surface proteins, ELISA, subunit vaccine, suspension bead array
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-11108 (URN)978-91-7415-417-7 (ISBN)
Public defence
2009-10-16, FD5, AlbaNova Universitetscentrum, Roslagstullsbacken 21, Stockholm, 13:00 (English)
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Supervisors
Note
QC 20100719Available from: 2009-09-25 Created: 2009-09-18 Last updated: 2010-07-19Bibliographically approved

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