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Braitbard, Ori; Glickstein, Hava; Bishara-Shieban, Janette; Pace, Umberto; Stein, Wilfred D.

doi:10.1186/1477-5956-4-12

Cited by 12 publications

(6 citation statements)

References 12 publications

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“…To analyze for epitope specificity, BLAST search, with parameters adjusted for short input sequence, using the epitope sequence as query was performed against all reported human and dog sequences from NCBI non-redundant protein database. The specific parameters were set to 20.000 expect threshold, the word size length was 2, PAM30 without compositional adjustment was used as substitution matrix and low complexity regions filter was switched off (parameters available at http://www.ncbi.nlm.nih.gov/blast/Why.shtml ) [42] – [44] .…”

Section: Methodsmentioning

confidence: 99%

Improving Serodiagnosis of Human and Canine Leishmaniasis with Recombinant Leishmania braziliensis Cathepsin L-like Protein and a Synthetic Peptide Containing Its Linear B-cell Epitope

et al. 2015

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BackgroundThe early and correct diagnosis of human leishmaniasis is essential for disease treatment. Another important step in the control of visceral leishmaniasis is the identification of infected dogs, which are the main domestic reservoir of L. infantum. Recombinant proteins and synthetic peptides based on Leishmania genes have emerged as valuable targets for serodiagnosis due to their increased sensitivity, specificity and potential for standardization. Cathepsin L-like genes are surface antigens that are secreted by amastigotes and have little similarity to host proteins, factors that enable this protein as a good target for serodiagnosis of the leishmaniasis.Methodology/Principal FindingsWe mapped a linear B-cell epitope within the Cathepsin L-like protein from L. braziliensis. A synthetic peptide containing the epitope and the recombinant protein was evaluated for serodiagnosis of human tegumentary and visceral leishmaniasis, as well as canine visceral leishmaniasis.Conclusions/SignificanceThe recombinant protein performed best for human tegumentary and canine visceral leishmaniasis, with 96.30% and 89.33% accuracy, respectively. The synthetic peptide was the best to discriminate human visceral leishmaniasis, with 97.14% specificity, 94.55% sensitivity and 96.00% accuracy. Comparison with T. cruzi-infected humans and dogs suggests that the identified epitope is specific to Leishmania parasites, which minimizes the likelihood of cross-reactions.

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Section: Methodsmentioning

confidence: 99%

Improving Serodiagnosis of Human and Canine Leishmaniasis with Recombinant Leishmania braziliensis Cathepsin L-like Protein and a Synthetic Peptide Containing Its Linear B-cell Epitope

et al. 2015

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“…High protein concentration in the diluted samples could lead to non-specific binding of the antibodies to antigens other than CagA, VacA and NapA ; specifically, the dissolved peptides compete with the peptides bound to receptors for antibodies. Thus, a diluted concentration of antigens should be used to obtain an accurate result [ 54 , 55 ]. However, the high transcript levels of CagA, VacA and NapA did not completely correspond to the protein yields, which could be because that the quantity of mRNA levels does not restrict translation and that post-translation modification can lead to the lysis of proteins [56] .…”

Section: Discussionmentioning

confidence: 99%

Transient recombinant expression of highly immunogenic CagA, VacA and NapA in Nicotiana benthamiana

Barzigar¹,

Haraprasad

Kumar³

et al. 2022

Biotechnology Reports

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Highlights A platform for rapid expression of genes through agro-infiltration is a suitable alternative to the laborious process of generating stable transgenic lines for producing recombinant immunogenic antigens in plants. This study has successfully developed and demonstrated a reproducible transformation protocol for rapid expression of Helicobacter pylori recombinant CagA, VacA and NapA antigens in Nicotiana benthamiana . This is the first study of recombinant expression CagA, VacA and NapA gene of Helicobacter pylori in Nicotiana benthamiana via syringe-assisted A grobacterium infiltration. A syringe infiltration of a four to five weeks old Nicotiana benthamiana plant with Agrobacterium tumefaciens subtype EHA105 was optimal to produce the maximum mRNA levels of CagA, VacA and NapA mRNA in leaf at the third-day post-Agro-infiltration.

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“…Maxisorp Nunc C-bottom strips were coated overnight at 4 0 C with the indicated amounts of synthetic peptides (Fig. 1 ) dissolved in sodium carbonate buffer pH 9.6 [ 26 ]. After blocking, as in western blot experiments, the OT-R and V1aR antibodies were added for 2h at room temperature at the indicated dilution followed by the secondary anti-rabbit antibody conjugated with horseradish peroxidase.…”

Section: Methodsmentioning

confidence: 99%

Proteomic studies of VEGFR2 in human placentas reveal protein associations with preeclampsia, diabetes, gravidity, and labor

Ho,

Chaput,

Sinkey

et al. 2024

Cell Commun Signal

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VEGFR2 (Vascular endothelial growth factor receptor 2) is a central regulator of placental angiogenesis. The study of the VEGFR2 proteome of chorionic villi at term revealed its partners MDMX (Double minute 4 protein) and PICALM (Phosphatidylinositol-binding clathrin assembly protein). Subsequently, the oxytocin receptor (OT-R) and vasopressin V1aR receptor were detected in MDMX and PICALM immunoprecipitations. Immunogold electron microscopy showed VEGFR2 on endothelial cell (EC) nuclei, mitochondria, and Hofbauer cells (HC), tissue-resident macrophages of the placenta. MDMX, PICALM, and V1aR were located on EC plasma membranes, nuclei, and HC nuclei. Unexpectedly, PICALM and OT-R were detected on EC projections into the fetal lumen and OT-R on 20–150 nm clusters therein, prompting the hypothesis that placental exosomes transport OT-R to the fetus and across the blood–brain barrier. Insights on gestational complications were gained by univariable and multivariable regression analyses associating preeclampsia with lower MDMX protein levels in membrane extracts of chorionic villi, and lower MDMX, PICALM, OT-R, and V1aR with spontaneous vaginal deliveries compared to cesarean deliveries before the onset of labor. We found select associations between higher MDMX, PICALM, OT-R protein levels and either gravidity, diabetes, BMI, maternal age, or neonatal weight, and correlations only between PICALM-OT-R (p < 2.7 × 10–8), PICALM-V1aR (p < 0.006), and OT-R-V1aR (p < 0.001). These results offer for exploration new partnerships in metabolic networks, tissue-resident immunity, and labor, notably for HC that predominantly express MDMX.

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Cited by 12 publications

References 12 publications

Improving Serodiagnosis of Human and Canine Leishmaniasis with Recombinant Leishmania braziliensis Cathepsin L-like Protein and a Synthetic Peptide Containing Its Linear B-cell Epitope

Improving Serodiagnosis of Human and Canine Leishmaniasis with Recombinant Leishmania braziliensis Cathepsin L-like Protein and a Synthetic Peptide Containing Its Linear B-cell Epitope

Transient recombinant expression of highly immunogenic CagA, VacA and NapA in Nicotiana benthamiana

Proteomic studies of VEGFR2 in human placentas reveal protein associations with preeclampsia, diabetes, gravidity, and labor

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