Identification of Protein Candidates for Developing Bacterial Ghost Vaccines against
            <i>Brucella</i>

DelVecchio, Vito G.; Alefantis, Tim; Ugalde, Rodolfo A.; Comerci, Diego J.; Marchesini, María Inés; Khan, Akhtar S.; Lubitz, Werner; Mujer, Cesar V.

doi:10.1002/0471973165.ch19

Cited by 7 publications

(5 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To select more suitable candidates for antigen screening, various approaches have been suggested (8,24,34). However, the inability to grow M. haemofelis in vitro and the absence of genomic sequencing for other feline hemoplasmas, including the less virulent "Candidatus Mycoplasma haeamominutum" (10), restricts the use of these methods.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Identification, Bioinformatics Analyses, and Expression of Immunoreactive Antigens of Mycoplasma haemofelis

Messick

Santos

2011

Clin. Vaccine Immunol.

View full text Add to dashboard Cite

Mycoplasma haemofelis infection frequently causes anemia in cats. Despite an intense immune response and/or antibiotic treatment, cats often remain asymptomatic carriers following infection. Our hypothesis is that detection of antibodies to M. haemofelis is a sensitive approach for identifying infected cats, particularly carriers. To date, no immunoassay has been developed. This is due largely to the inability to culture M. haemofelis in vitro; hence, a source of antigen is not readily available. The objective of this study was to identify, express, and purify immunogenic proteins of M. haemofelis. To accomplish this, two whole-genomic expression libraries were created in the Lambda ZapII vector and immunoscreened with preimmune plasma, plasma from specific-pathogen-free cats, and pooled acute-and convalescent-phase plasma from experimentally infected cats. The inserts from 21 immunoreactive clones were sequenced, resulting in the identification of 60 genes coding for putative proteins necessary for diverse cellular functions, along with several novel genes of M. haemofelis. Fragments of selected genes based on bioinformatic analyses were PCR amplified, cloned into a high-level protein expression system, and subsequently expressed in Escherichia coli as a His 6 -fusion protein.The recombinant fusion proteins of M. haemofelis were purified and evaluated as an antigen in a Western blot to verify the findings of previous immunoscreening. Together with bioinformatics analyses of individual genes, this approach provided several putative candidate antigens. Five antigens of M. haemofelis were reactive by Western blotting against the immune plasma and negative against nonimmune plasma; these antigens might be useful serologic or even vaccine targets.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by mass spectrometry and microsequencing, is a commonly used method for identifying these proteins (8,18,24,34). However, low and differentially expressed antigens cannot be identified using this technique.…”

mentioning

confidence: 99%

Identification, Bioinformatics Analyses, and Expression of Immunoreactive Antigens of Mycoplasma haemofelis

Messick

Santos

2011

Clin. Vaccine Immunol.

View full text Add to dashboard Cite

show abstract

“…Bacterial cell surface antigens are prime candidates as they represent the initial point of contact between the pathogen and its host [44]. …”

Section: Subunit Vaccines Against Brucellosismentioning

confidence: 99%

A History of the Development ofBrucellaVaccines

Ávila-Calderón

López-Merino

Sriranganathan

et al. 2013

BioMed Research International

132

131

View full text Add to dashboard Cite

Brucellosis is a worldwide zoonosis affecting animal and human health. In the last several decades, much research has been performed to develop safer Brucella vaccines to control the disease mainly in animals. Till now, no effective human vaccine is available. The aim of this paper is to review and discuss the importance of methodologies used to develop Brucella vaccines in pursuing this challenge.

show abstract

“…Furthermore, gene expression profiling used to define Brucella pathogenesis identified the Omps, lipoproteins, stress response proteins, chaperones, flagellar genes, ABC transport proteins, and genes for LPS and fatty acid biosynthesis as key components for cell envelope or OM biogenesis ( Guzman-Verri et al, 2002 ; Lamontagne et al, 2007 ; Manterola et al, 2007 ; Rossetti et al, 2009 ; Viadas et al, 2009 ). Immunogenic proteins (particularly Omp25, Omp31, Omp2b) found on the bacterial cell surface are superior vaccine candidates as they establish the initial contact site between the pathogen and the host ( Delvecchio et al, 2006 ). Ultimately the knowledge gained on pathogen-associated factors (with a particular interest to Brucella Omps and lipoproteins) have been used for the application of these factors as antigens in vaccine formulation and studies have confirmed their ability to confer protective immunity against Brucella and other intracellular pathogens ( Fu et al, 2012 ).…”

Section: Brucella Pathogenesis and Outer Membrane Biogenesismentioning

confidence: 99%

Analyzing the molecular mechanism of lipoprotein localization in Brucella

et al. 2015

View full text Add to dashboard Cite

Bacterial lipoproteins possess diverse structure and functionality, ranging from bacterial physiology to pathogenic processes. As such many lipoproteins, originating from Brucella are exploited as potential vaccines to countermeasure brucellosis infection in the host. These membrane proteins are translocated from the cytoplasm to the cell membrane where they are anchored peripherally by a multifaceted targeting mechanism. Although much research has focused on the identification and classification of Brucella lipoproteins and their potential use as vaccine candidates for the treatment of Brucellosis, the underlying route for the translocation of these lipoproteins to the outer surface of the Brucella (and other pathogens) outer membrane (OM) remains mostly unknown. This is partly due to the complexity of the organism and evasive tactics used to escape the host immune system, the variation in biological structure and activity of lipoproteins, combined with the complex nature of the translocation machinery. The biosynthetic pathway of Brucella lipoproteins involves a distinct secretion system aiding translocation from the cytoplasm, where they are modified by lipidation, sorted by the lipoprotein localization machinery pathway and thereafter equipped for export to the OM. Surface localized lipoproteins in Brucella may employ a lipoprotein flippase or the β-barrel assembly complex for translocation. This review provides an overview of the characterized Brucella OM proteins that form part of the OM, including a handful of other characterized bacterial lipoproteins and their mechanisms of translocation. Lipoprotein localization pathways in gram negative bacteria will be used as a model to identify gaps in Brucella lipoprotein localization and infer a potential pathway. Of particular interest are the dual topology lipoproteins identified in Escherichia coli and Haemophilus influenza. The localization and topology of these lipoproteins from other gram negative bacteria are well characterized and may be useful to infer a solution to better understand the translocation process in Brucella.

show abstract

Identification of Protein Candidates for Developing Bacterial Ghost Vaccines against Brucella

Cited by 7 publications

References 36 publications

Identification, Bioinformatics Analyses, and Expression of Immunoreactive Antigens of Mycoplasma haemofelis

Identification, Bioinformatics Analyses, and Expression of Immunoreactive Antigens of Mycoplasma haemofelis

A History of the Development ofBrucellaVaccines

Analyzing the molecular mechanism of lipoprotein localization in Brucella

Contact Info

Product

Resources

About