A library of recombinant Babesia microti cell surface and secreted proteins for diagnostics discovery and reverse vaccinology

Elton, Catherine M.; Rodriguez, Marilis; Mamoun, Choukri Ben; Lobo, Cheryl A.; Wright, Gavin J.

doi:10.1016/j.ijpara.2018.10.003

Cited by 31 publications

(27 citation statements)

References 52 publications

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“…Amino acid sequence alignment analysis revealed that 03g04550 was similar to the secreted antigens from Plasmodium falciparum and Babesia bovis (unpublished data). This result was consistent with previous reports from Elton et al (2019).…”

Section: Anti-bmsa1 Antiserum Inhibits Merozoite Invasionsupporting

confidence: 94%

Surface Antigen 1 Is a Crucial Secreted Protein That Mediates Babesia microti Invasion Into Host Cells

Guo

et al. 2020

Front. Microbiol.

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Babesia microti, a tick-borne intraerythrocytic zoonotic protozoan, causes most of human babesiosis in the world, and patients usually experience intermittent fever, fatigue, and chills, followed by a combination of additional symptoms and even death in severe cases. Unfortunately, there is no curable drug or effective vaccine available, and the mechanism of related virulence factors in invasion to host cells during the merozoite stage is unclear. Here, we evaluated a secreted protein annotated as B. microti surface antigen 1 (BmSA1) and identified from in vitro culture supernatant by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). BmSA1 fragment was expressed in Escherichia coli to prepare polyclonal antiserum. Western blot analysis revealed the existence of BmSA1 in the lysate of the parasites and the hemolysate of infected red blood cells (iRBCs). Laser confocal microscopy confirmed BmSA1 as a secreted protein with diffuse distribution around the parasites in red blood cells (RBCs). The adhesion capacity of BmSA1 against the host RBCs was tested by RBC binding assays using the recombinant BmSA1 protein (rBmSA1), which was shown to specifically bind to host RBCs. Further in vitro antiserum-neutralization test demonstrated that the growth of parasites could be significantly inhibited by the anti-BmSA1 antiserum. These results indicate that BmSA1 is a crucial factor for B. microti invasion into host RBCs with an important role in host-parasite interactions during the merozoite stage and has the potential use as a vaccine candidate due to its high secretion amount.

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Section: Anti-bmsa1 Antiserum Inhibits Merozoite Invasionsupporting

confidence: 94%

Surface Antigen 1 Is a Crucial Secreted Protein That Mediates Babesia microti Invasion Into Host Cells

Guo

et al. 2020

Front. Microbiol.

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“…www.nature.com/scientificreports www.nature.com/scientificreports/ subsequent boosters, respectively. Sera were harvested from tail bleeds of immunised mice 8 days after the final immunisation, with antibody titres determined using ELISA as previously described 66 . Briefly, a fixed concentration of the biotinylated control protein was immobilised on 96-well streptavidin plates (Thermo Scientific) and incubated with serial dilutions of the corresponding sera prepared in HBS/0.1% (v/v) Tween 20 (HBST), containing 2% (w/v) BSA.…”

Section: Methodsmentioning

confidence: 99%

Establishment, optimisation and quantitation of a bioluminescent murine infection model of visceral leishmaniasis for systematic vaccine screening

Ong

Clare

Roberts

et al. 2020

Sci Rep

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Visceral leishmaniasis is an infectious parasitic disease caused by the protozoan parasites Leishmania donovani and Leishmania infantum. The drugs currently used to treat visceral leishmaniasis suffer from toxicity and the emergence of parasite resistance, and so a better solution would be the development of an effective subunit vaccine; however, no approved vaccine currently exists. The comparative testing of a large number of vaccine candidates requires a quantitative and reproducible experimental murine infection model, but the parameters that influence infection pathology have not been systematically determined. to address this, we have established an infection model using a transgenic luciferaseexpressing L. donovani parasite and longitudinally quantified the infections using in vivo bioluminescent imaging within individual mice. We examined the effects of varying the infection route, the site of adjuvant formulation administration, and standardised the parasite preparation and dose. We observed that the increase in parasite load within the liver during the first few weeks of infection was directly proportional to the parasite number in the initial inoculum. finally, we show that immunity can be induced in pre-exposed animals that have resolved an initial infection. this murine infection model provides a platform for systematic subunit vaccine testing against visceral leishmaniasis.Protozoa of the genus Leishmania are obligate intracellular parasites which cause the disease leishmaniasis. The parasite is transmitted by the bite of an infected female phlebotomine sand fly during a blood meal, and every year there are an estimated ~1 million new infections resulting in ~65,000 deaths 1 . Although the disease burden disproportionately affects poor people living in tropical and developing countries, the expanding geographic range of vector distribution caused by climate and environmental changes is an emerging threat outside these regions 2-4 . Outbreaks and re-emergence of leishmaniasis are also linked to other factors including political and socioeconomic upheavals 5-7 . Within the human host, Leishmania spp. cause a spectrum of species-specific clinical manifestations known as cutaneous, mucocutaneous or visceral leishmaniases, and it is the visceral form that is considered to be the most severe, and is ultimately fatal if left untreated. Symptomatic visceral leishmaniasis (VL), results from infections of L. donovani and L. infantum, and while representing only ~10% of all symptomatic leishmanial infections, they are responsible for nearly all leishmaniasis-attributed fatalities 1 . The current front-line drug treatments for VL: liposomal amphotericin B, miltefosine, paromomycin and antimonials, are far from ideal, with major concerns surrounding general toxicity or increased treatment failure 8,9 . While improved drugs are being developed 10,11 , the deployment of an effective vaccine would be an important disease control tool, but to date, no effective vaccine has been licensed for human leishmaniasis.The e...

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“…Furthermore, structures such as the Maurer’s clefts and the tubulovesicular network, previously shown to be present in the cytoplasm of Plasmodium falciparum –infected erythrocytes, are not found in B. microti –infected erythrocytes (Marti et al, 2005; Lanzer et al, 2006; Pelle et al, 2015; Cornillot et al, 2016; de Koning-Ward et al, 2016; Sherling & van Ooij, 2016). Proteomic, immunoproteomic, and genomic efforts used to search for major antigens of the parasite, which could be used as vaccines or diagnostic biomarkers, identified several antigens of B. microti that trigger a strong host immune response in mice and humans (Molestina et al, 2002; Cornillot et al, 2016; Silva et al, 2016; Elton et al, 2018). Among these, BmGPI12 (BmSA1) has been shown to trigger the strongest IgM and IgG responses and has been successfully used as a diagnostic marker of active B. microti infection (Thekkiniath et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Evidence for vesicle-mediated antigen export by the human pathogen Babesia microti

et al. 2019

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The apicomplexan parasite Babesia microti is the primary agent of human babesiosis, a malaria-like illness and potentially fatal tick-borne disease. Unlike its close relatives, the agents of human malaria, B. microti develops within human and mouse red blood cells in the absence of a parasitophorous vacuole, and its secreted antigens lack trafficking motifs found in malarial secreted antigens. Here, we show that after invasion of erythrocytes, B. microti undergoes a major morphogenic change during which it produces an interlacement of vesicles (IOV); the IOV system extends from the plasma membrane of the parasite into the cytoplasm of the host erythrocyte. We developed antibodies against two immunodominant antigens of the parasite and used them in cell fractionation studies and fluorescence and immunoelectron microscopy analyses to monitor the mode of secretion of B. microti antigens. These analyses demonstrate that the IOV system serves as a major export mechanism for important antigens of B. microti and represents a novel mechanism for delivery of parasite effectors into the host by this apicomplexan parasite.

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A library of recombinant Babesia microti cell surface and secreted proteins for diagnostics discovery and reverse vaccinology

Abstract: Graphical abstract

Cited by 31 publications

References 52 publications

Surface Antigen 1 Is a Crucial Secreted Protein That Mediates Babesia microti Invasion Into Host Cells

Surface Antigen 1 Is a Crucial Secreted Protein That Mediates Babesia microti Invasion Into Host Cells

Establishment, optimisation and quantitation of a bioluminescent murine infection model of visceral leishmaniasis for systematic vaccine screening

Evidence for vesicle-mediated antigen export by the human pathogen Babesia microti

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