IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu

Araújo, Flábio R.; Costa, C.; Ramos, Carlos A. N.; Farias, Thaís A; Souza, Ingrid I. F.; Melo, Elaine Silva de Pádua; Elisei, Carina; Rosinha, G. M. S.; Soares, Cleber O.; Fragoso, S. P.; Fonseca, Adivaldo Henrique da

doi:10.1590/s0074-02762008000200010

Cited by 24 publications

(24 citation statements)

References 24 publications

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“…In comparison to the genes of the membrane proteins investigated in the present study, EF-Tu takes on significance as a possible candidate vaccine antigen, as it exhibits the highest percent homology among the predicted amino acid sequences in the isolates analyzed and is considered a highly conserved gene. This finding corroborates a study carried out by Araújo et al (2008), who report the conservation of this gene. Moreover, it is capable of stimulating the proliferative response in T lymphocytes, as reported by Lopez et al (2008).…”

Section: Discussionsupporting

confidence: 93%

“…Ramos et al (2007) assessed the transcription pattern of several membrane proteins' genes in A. marginale, including EF-Tu, which, after analysis, was suggested to be a membrane-associated protein (Lopez et al 2005). Araújo et al (2008) and Lopez et al (2008) expressed EF-Tu in E. coli and the recombinant protein was able to induce a proliferative response in T lymphocytes and titers of IgG2 greater than those of IgG1, thereby demonstrating the desired capacity for immunostimulation.…”

Section: Discussionmentioning

confidence: 99%

“…In studies with other TFSS group members (VirB9-2 and VirB10), Araújo et al (2008) and Vidotto et al (2008) suggested conservation among different isolates of A. marginale and demonstrated that VirB9-2 and VirB10 proteins are recognized by antibodies from naturally and experimentally infected cattle. The VirB3 protein in AMBR isolate has the same conservation profile as other components of the TFSS group analyzed by Araújo et al (2008); thus, it is expected to exhibit the same antigen profile, as the conservation of B cell epitopes between and within isolates is likely important for controlling infection. Lopez et al (2007) also found that VirB9-2, VirB10 and VirB9-1 are conserved among isolates.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale

Daniel

Araújo

Almeida

et al. 2010

Mem. Inst. Oswaldo Cruz

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The sequencing of the complete genome of Anaplasma marginale has enabled the identification of several genes that encode membrane proteins, thereby increasing the chances of identifying candidate immunogens. Little is known regarding the genetic variability of genes that encode membrane proteins in A. marginale isolates. The aim of the present study was to determine the degree of conservation of the predicted amino acid sequences of OMP1, OMP4, OMP5, OMP7, OMP8, OMP10, OMP14, OMP15, SODb, OPAG1, OPAG3, VirB3, VirB9-1, PepA, EF-Tu and AM854 proteins in a Brazilian isolate of A. marginale compared to other isolates. Hence, primers were used to amplify these genes: omp1, omp4, omp5, omp7, omp8, omp10, omp14, omp15, sodb, opag1, opag3, virb3, VirB9-1, pepA, ef-tu and am854. After polimerase chain reaction amplification, the products were cloned and sequenced using the Sanger method and the predicted amino acid sequence were multi-aligned using the CLUSTALW and MEGA 4 programs, comparing the predicted sequences between the Brazilian, Saint Maries, Florida and A. marginale centrale isolates. With the exception of outer membrane protein (OMP) 7, all proteins exhibited 92-100% homology to the other A. marginale isolates. However, only OMP1, OMP5, EF-Tu, VirB3, SODb and VirB9-1 were selected as potential immunogens capable of promoting cross-protection between isolates due to the high degree of homology (over 72%) also found with A. (centrale) marginale

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale

Daniel

Araújo

Almeida

et al. 2010

Mem. Inst. Oswaldo Cruz

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“…Antibody specific for VirB9 in dogs infected with Ehrlichia canis was detected (24), and more recently, antibody specific for VirB9-2 and VirB10 was detected in cattle experimentally or naturally infected with A. marginale in Brazil (4,73). However, there is some evidence that type III secretion system (T3SS) proteins can induce protective immunity in several animal models with human pathogens (34,38,42,69).…”

Section: Discussionmentioning

confidence: 99%

Anaplasma marginaleType IV Secretion System Proteins VirB2, VirB7, VirB11, and VirD4 Are Immunogenic Components of a Protective Bacterial Membrane Vaccine

Sutten

Norimine

Beare³

et al. 2010

Infect Immun

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Anaplasma and related Ehrlichia spp. are important tick-borne, Gram-negative bacterial pathogens of livestock and humans that cause acute infection and disease and can persist. Immunization of cattle with an Anaplasma marginale fraction enriched in outer membranes (OM) can provide complete protection against disease and persistent infection. Serological responses of OM vaccinees to the OM proteome previously identified over 20 antigenic proteins, including three type IV secretion system (T4SS) proteins, VirB9-1, VirB9-2, and VirB10. Subsequent studies showed that these three proteins also stimulated CD4؉ T-cell responses in OM vaccinees. The T4SS, composed of a complex of proteins spanning the inner and outer membranes of certain bacteria, is an important virulence factor but is relatively unexplored as a vaccine target. The goal of this study was to determine if additional T4SS proteins are immunogenic for animals immunized with the protective OM fraction of A. marginale. T4SS proteins expressed by in vitro transcription and translation were screened for stimulating proliferation of T cells from OM vaccinees, and immunogenic proteins were expressed as recombinant proteins in Escherichia coli and their immunogenicity was verified. VirB2, a putative VirB7, VirB11, and VirD4 were immunogenic for OM vaccinees expressing several common major histocompatibility complex (MHC) class II haplotypes. VirB2 is encoded by multiple genes that share a conserved central region, and epitope mapping revealed T-cell epitopes in this region. The discovery of novel immunogenic T4SS proteins recognized by outbred individuals with common MHC haplotypes further justifies evaluating the T4SS as a potential vaccine candidate for pathogenic bacteria.

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“…Nevertheless, it may play additional roles. It has recently been demonstrated that EF-Tu is released/secreted from bacterial cells and that it can modulate pathogen-host interactions (19)(20)(21)(22)(23)(24). However, the pathways by which EF-Tu protein might interfere with immune regulation in the host cell remain unknown.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Genomes of Brucella melitensis M28 and the B. melitensis M5-90 Derivative Vaccine Strain Highlights the Translation Elongation Factor Tu Gene tuf2 as an Attenuation-Related Gene

Wang

Qiao

et al. 2013

Infect Immun

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c Brucella melitensis causes brucellosis, a disease affecting sheep, cattle, and sometimes humans. Attenuated B. melitensis strain M5-90, derived from virulent strain M28, is widely used as a live vaccine in ruminants in China. Genetic differences between the strains may cast light on the mechanism of attenuation. We recently reported the complete genomic sequences of M28 and M5-90. Genome organization is highly conserved between these isolates, and also with virulent strains 16 M and ATCC 23457. Analysis revealed 23 open reading frames (ORFs) with consistent differences between M5-90 and the virulent strains. Notably, the tuf2 gene encoding translation elongation factor EF-Tu from M5-90 contained 50 single nucleotide polymorphisms (SNPs) and 9 gaps (indels) compared to tuf2 of M28 or of the other virulent strains. There were no changes in tuf1. To evaluate the potential role of EF-Tu in pathogenesis, tuf1 and tuf2 mutants of M28 and an M5-90 strain harboring wild-type tuf2 were constructed, and their virulence/attenuation was evaluated in vivo. We report that the tuf2 gene plays an important role in the attenuation of M5-90 virulence. Brucella melitensis, the first species in the genus Brucella to be described, causes abortion in goats and sheep and Malta fever in humans. Currently, there are nine recognized species of the bacterium Brucella, based on host preferences and phenotypic differences (1). The six classically recognized species of this facultative intracellular parasite are B. melitensis (sheep and goats), B. abortus (cattle), B. suis (pigs, reindeer, and hares), B. canis (dogs), B. ovis (sheep), and B. neotomae (desert wood rats) (2). In humans, B. melitensis is the most highly pathogenic, and infection typically takes place via the consumption of unpasteurized milk and milk products. Attenuated B. melitensis M5-90, derived from virulent strain M28, is widely used as a live vaccine in sheep, goats, and cattle in China (3, 4). However, such strains retain pathogenicity for humans and to date no effective vaccine has been approved for human use. Because of its epidemic potential, the absence of a human vaccine, and the effectiveness of aerosol infection, this agent is classified as a biosafety level 3 pathogen and has been designated a potential bioterrorism agent (5).The genome of the first Brucella melitensis strain to be sequenced, 16 M, comprises ϳ3.3 Mb of unique sequence shared between two circular chromosomes: the larger chromosome (chromosome I) is ϳ2.1 Mb, whereas chromosome II is ϳ1.2 Mb. Eleven complete genome sequences representing six species of Brucella are currently available, and the genomes of several other Brucella strains/species are in the process of being sequenced (1).The different genomes are very similar in size and in the numbers of genes and proteins. Sequence comparisons have provided firm evidence of horizontal gene transfer between different Brucella species and strains, despite their predominantly intracellular lifestyle, and phylogenomic analysis of orthologous gene famil...

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IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu

Abstract: Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and

Cited by 24 publications

References 24 publications

Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale

Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale

Anaplasma marginaleType IV Secretion System Proteins VirB2, VirB7, VirB11, and VirD4 Are Immunogenic Components of a Protective Bacterial Membrane Vaccine

Comparison of Genomes of Brucella melitensis M28 and the B. melitensis M5-90 Derivative Vaccine Strain Highlights the Translation Elongation Factor Tu Gene tuf2 as an Attenuation-Related Gene

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