Immunoproteomic analysis of Lawsonia intracellularis identifies candidate neutralizing antibody targets for use in subunit vaccine development

Publication date: Available online 19 July 2019Source: Veterinary MicrobiologyAuthor(s): Milan Obradovic, J. Alex Pasternak, Siew Hon Ng, Brenda Allan, Robert Brownlie, Heather L. WilsonAbstractLawsonia intracellularis is an obligate intracellular microorganism and the causative agent of porcine proliferative enteropathy. Due to its obligate intracellular nature, characterization of antigens and proteins involved in host-pathogen interaction and immune recognition have been difficult to achieve using conventional microbiological techniques. In this work, we used 2-dimensional gel electrophoresis coupled with Western-immunoblotting, mass spectrometry and bioinformatics to identify bacterial proteins that interact in vitro with pig intestinal cells (IPEC-1), have immunogenic properties and the potential to be used as subunit vaccine antigens. We detected eleven immunogenic bacterial proteins from which fliC (LI0710), LI1153 (annotated by NCBI as Putative protein N), and LI0649 (annotated as autotransporter) were predicted to be expressed on the outer membrane while LI0169 (oppA; annotated as ABC dipeptide transport system)was predicted to be periplasmic with a transmembrane domain forming a central pore through the plasma membrane.. Genes coding for these four proteins were cloned and expressed in Escherichia coli and the corresponding recombinant proteins were purified using affinity chromatography. Porcine hyperimmune serum against whole Lawsonia lysate established that all f...
Source: Veterinary Microbiology - Category: Veterinary Research Source Type: research

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Heat-labile toxin (LT) is a well-characterized powerful enterotoxin produced by enterotoxigenic Escherichia coli (ETEC). This toxin is known to contribute to diarrhea in young children in developing countries, international travelers, as well as many different species of young animals. Interestingly, it has also been revealed that LT is involved in other activities in addition to its role in enterotoxicity. Recent studies have indicated that LT toxin enhances enteric pathogen adherence and subsequent intestinal colonization. LT has also been shown to act as a powerful adjuvant capable of upregulating vaccine antigenicity; ...
Source: Frontiers in cellular and infection microbiology - Category: Microbiology Source Type: research
Zuñiga Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus causing high morbidity and mortality in porcine herds worldwide. Although both inactivated and live attenuated vaccines have been extensively used, the emergence of highly virulent strains and the recurrent outbreaks even in vaccinated farms highlight the need of effective vaccines. Engineering of genetically defined live attenuated vaccines is a rational approach for novel vaccine development. In this line, we engineered an attenuated virus based on the transmissible gastroenteritis virus (TGEV) genome, expressing a chimeric spike prote...
Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research
Viruses, Vol. 11, Pages 682: Recombinant Chimeric Transmissible Gastroenteritis Virus (TGEV)—Porcine Epidemic Diarrhea Virus (PEDV) Virus Provides Protection against Virulent PEDV Viruses doi: 10.3390/v11080682 Authors: Alejandro Pascual-Iglesias Carlos M. Sanchez Zoltan Penzes Isabel Sola Luis Enjuanes Sonia Zuñiga Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus causing high morbidity and mortality in porcine herds worldwide. Although both inactivated and live attenuated vaccines have been extensively used, the emergence of highly virulent strains and the recurrent outbreaks...
Source: Viruses - Category: Virology Authors: Tags: Article Source Type: research
Publication date: Available online 23 July 2019Source: Veterinary MicrobiologyAuthor(s): Haoju Wang, Kang Liang, Qingke Kong, Qing LiuAbstractAvian pathogenic Escherichia coli (APEC) typically causes colibacillosis and is a major concern for the poultry industry and public health. As a vaccine platform, the outer membrane vesicles (OMVs) derived from various gram-negative bacteria and even some gram-positive bacteria have been reported to be immunogenic in laboratories or upon commercial usage worldwide. Here, we purified OMVs from APEC serotype O78 strain by ultracentrifugation and gradient isolation. By SDS-PAGE and LC-M...
Source: Veterinary Microbiology - Category: Veterinary Research Source Type: research
Abstract C2 toxin produced from Clostridium botulinum serotypes C and D has a potential role in many pathophysiological mechanisms in birds and animals. It has encompassed an ADP ribosyltransferase subunit (C2I) and a translocation/binding subunit (C2II). In the present study, we intended to produce C2I mutant proteins as recombinant subunit vaccines by using glutathione-S-transferase-gene fusion system. The mutants of this study were previously evaluated from their evolutionary imprints and suggested as suitable candidates for subunit vaccines. A synthetic C2 gene was inserted in a pGEX-2T vector, cloned and expr...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
eição FR Abstract Clostridium perfringens type A is the causative agent of gas gangrene and gastroenteric ("yellow lamb disease") disease in ruminants, with C. perfringens alpha toxin (CPA) being the main virulence factor in the pathogenesis of these illnesses. In the present study, we have developed recombinant Escherichia coli bacterins expressing rCPA and used it to vaccinate rabbits and sheep. Doses of up to 200 μg of rCPA used for inoculation, induced 13,82 IU.mL-1 of neutralizing antitoxin in rabbits, which is three times higher than that recommended by the USDA (4 IU.mL-1). In she...
Source: Anaerobe - Category: Microbiology Authors: Tags: Anaerobe Source Type: research
Authors: Martiñón S, Cisneros A, Villicaña S, Hernández-Miramontes R, Mixcoha E, Calderón-Vargas P Abstract Adjuvants are a diverse family of substances whose main objective is to increase the strength, quality, and duration of the immune response caused by vaccines. The most commonly used adjuvants are aluminum-based, oil-water emulsion, and bacterial-origin adjuvants. In this paper, we will discuss how the election of adjuvants is important for the adjuvant-mediated induction of immunity for different types of vaccines. Aluminum-based adjuvants are the most commonly used, the sa...
Source: Journal of Immunology Research - Category: Allergy & Immunology Tags: J Immunol Res Source Type: research
AbstractAn unusual group A rotavirus (RVA) strain MAR/ma31/2011/G8P[14] was detected for the first time in Morocco in a stool sample from hospitalized child aged 18  months suffering from acute gastroenteritis and fever in 2011. Complete genome sequencing of the ma31 strain was done using the capillary sequencing technology. The analysis revealed the G8-P[14]-I2-R2-C2-M2-A11-N2-T6-E2-H3 constellation and the backbone genes: I2-R2-C2-M2-A11-N2-T6-E2-H3 are comm only found in RVA strains from artiodactyls such as cattle. The constellation was shared with another Italian zoonotic G8P[14] strains (BA01 and BA02), two Hung...
Source: Virus Genes - Category: Genetics & Stem Cells Source Type: research
Publication date: Available online 4 June 2019Source: Research in Veterinary ScienceAuthor(s): Reza Hoseini Shahidi, Gholamreza Hashemi Tabar, Mohammad Reza Bassami, Abdollah Jamshidi, Hesam DehghaniAbstractAn Escherichia coli (E. coli) O2:K1 bacterial ghost was produced by controlled expression of bacteriophage PhiX 174 lysis gene E. Temperature controlled expression of this gene caused tunnels and holes in the cell wall of E. coli O2:K1 bacterium, leading to loss of cytoplasmic contents. Formation of E. coli O2:K1 ghost was confirmed by scanning electron microscopy and determination of colony forming units. To evaluate t...
Source: Research in Veterinary Science - Category: Veterinary Research Source Type: research
CONCLUSIONS: This is the first report of the design and study of a rMEP vaccine against A. baumannii. The results indicate that the rMEP is a promising vaccine candidate for the control of infections caused by A. baumannii. PMID: 31203576 [PubMed - in process]
Source: Swiss Medical Weekly - Category: General Medicine Authors: Tags: Swiss Med Wkly Source Type: research
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