5 results on '"Bounkeua V"'
Search Results
2. Plasmodium falciparum ookinete expression of plasmepsin VII and plasmepsin X.
- Author
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Li F, Bounkeua V, Pettersen K, and Vinetz JM
- Subjects
- Amino Acid Sequence, Animals, Anopheles immunology, Aspartic Acid Endopeptidases genetics, Aspartic Acid Endopeptidases immunology, Erythrocytes, Humans, Life Cycle Stages, Malaria, Falciparum transmission, Molecular Sequence Data, Plasmodium falciparum enzymology, Plasmodium falciparum genetics, Protozoan Proteins genetics, Protozoan Proteins immunology, Recombinant Proteins genetics, Recombinant Proteins immunology, Aspartic Acid Endopeptidases metabolism, Malaria, Falciparum parasitology, Plasmodium falciparum metabolism, Protozoan Proteins metabolism, Recombinant Proteins metabolism
- Abstract
Background: Plasmodium invasion of the mosquito midgut is a population bottleneck in the parasite lifecycle. Interference with molecular mechanisms by which the ookinete invades the mosquito midgut is one potential approach to developing malaria transmission-blocking strategies. Plasmodium aspartic proteases are one such class of potential targets: plasmepsin IV (known to be present in the asexual stage food vacuole) was previously shown to be involved in Plasmodium gallinaceum infection of the mosquito midgut, and plasmepsins VII and plasmepsin X (not known to be present in the asexual stage food vacuole) are upregulated in Plasmodium falciparum mosquito stages. These (and other) parasite-derived enzymes that play essential roles during ookinete midgut invasion are prime candidates for transmission-blocking vaccines., Methods: Reverse transcriptase PCR (RT-PCR) was used to determine timing of P. falciparum plasmepsin VII (PfPM VII) and plasmepsin X (PfPM X) mRNA transcripts in parasite mosquito midgut stages. Protein expression was confirmed by western immunoblot and immunofluorescence assays (IFA) using anti-peptide monoclonal antibodies (mAbs) against immunogenic regions of PfPM VII and PfPM X. These antibodies were also used in standard membrane feeding assays (SMFA) to determine whether inhibition of these proteases would affect parasite transmission to mosquitoes. The Mann-Whitney U test was used to analyse mosquito transmission assay results., Results: RT-PCR, western immunoblot and immunofluorescence assay confirmed expression of PfPM VII and PfPM X in mosquito stages. Whereas PfPM VII was expressed in zygotes and ookinetes, PfPM X was expressed in gametes, zygotes, and ookinetes. Antibodies against PfPM VII and PfPM X decreased P. falciparum invasion of the mosquito midgut when used at high concentrations, indicating that these proteases play a role in Plasmodium mosquito midgut invasion. Failure to generate genetic knockouts of these genes limited determination of the precise role of these proteases in parasite transmission but suggests that they are essential during the intraerythrocytic life cycle., Conclusions: PfPM VII and PfPM X are present in the mosquito-infective stages of P. falciparum. Standard membrane feeding assays demonstrate that antibodies against these proteins reduce the infectivity of P. falciparum for mosquitoes, suggesting their viability as transmission-blocking vaccine candidates. Further study of the role of these plasmepsins in P. falciparum biology is warranted.
- Published
- 2016
- Full Text
- View/download PDF
3. Lack of molecular correlates of Plasmodium vivax ookinete development.
- Author
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Bounkeua V, Li F, Chuquiyauri R, Abeles SR, McClean CM, Neyra V, Llanos-Cuentas A, Yori PP, and Vinetz JM
- Subjects
- Antimalarials therapeutic use, Cells, Cultured, Humans, Malaria, Vivax drug therapy, Malaria, Vivax parasitology, Plasmodium vivax cytology, Reverse Transcriptase Polymerase Chain Reaction, Plasmodium vivax physiology
- Abstract
Previous studies of Plasmodium vivax transmission to Anopheles spp. mosquitoes have not been able to predict mosquito infectivity on the basis of microscopic or molecular quantification of parasites (total parasites in the sample or total number of gametocytes) in infected blood. Two methods for production of P. vivax ookinete cultures in vitro, with yields of 10(6) macrogametocytes, 10(4) zygotes, and 10(3) ookinetes, respectively, per 10 mL of P. vivax-infected patient blood with approximately 0.01% parasitemia, were used to study P. vivax sexual stage development. The quantity of gametocytes, determined by counting Giemsa-stained blood smears, and quantity and type of gametocyte as determined by quantitative reverse transcriptase-polymerase chain reaction for Pvalpha tubulin II and macrogametocyte-specific pvg377 did not predict ookinete yield. Factors that affect the efficiency of in vitro P. vivax ookinete transformation remain poorly understood.
- Published
- 2011
- Full Text
- View/download PDF
4. In vitro generation of Plasmodium falciparum ookinetes.
- Author
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Bounkeua V, Li F, and Vinetz JM
- Subjects
- Biomarkers, Culture Media, DNA, Protozoan, Flow Cytometry, Humans, Plasmodium falciparum ultrastructure, Protozoan Proteins metabolism, Reproduction, Time Factors, Cell Culture Techniques methods, Plasmodium falciparum physiology
- Abstract
Plasmodium transmission from the human host to the mosquito depends on the ability of gametocytes to differentiate into ookinetes, the invasive form of the parasite that invades and establishes infection in the mosquito midgut. The biology of P. falciparum ookinetes is poorly understood, because sufficient quantities of this stage of this parasite species have not been obtained for detailed study. This report details methods to optimize production of P. falciparum sexual stage parasites, including ookinetes. Flow cytometric sorting was used to separate diploid/tetraploid zygotes and ookinetes from haploid gametetocytes and unfertilized gametes based on DNA content. Consistent production of 10(6)-10(7) P. falciparum ookinetes per 10 mL culture was observed, with ookinete transformation present in 10-40% of all parasite forms. Transmission electron micrographs of cultured parasites confirmed ookinete development.
- Published
- 2010
- Full Text
- View/download PDF
5. Cytopathic killing of peripheral blood CD4(+) T lymphocytes by human immunodeficiency virus type 1 appears necrotic rather than apoptotic and does not require env.
- Author
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Lenardo MJ, Angleman SB, Bounkeua V, Dimas J, Duvall MG, Graubard MB, Hornung F, Selkirk MC, Speirs CK, Trageser C, Orenstein JO, and Bolton DL
- Subjects
- CD4-Positive T-Lymphocytes pathology, Cytopathogenic Effect, Viral, Gene Products, env genetics, Gene Products, nef physiology, HIV-1 genetics, Humans, Jurkat Cells, Mutation, Recombination, Genetic, Vesicular stomatitis Indiana virus genetics, Viral Envelope Proteins genetics, nef Gene Products, Human Immunodeficiency Virus, Apoptosis, CD4-Positive T-Lymphocytes virology, Gene Products, env physiology, HIV-1 physiology, Membrane Glycoproteins, Necrosis
- Abstract
An important unresolved issue of AIDS pathogenesis is the mechanism of human immunodeficiency virus (HIV)-induced CD4(+) T-lymphocyte destruction. We show here that HIV type 1 (HIV-1) exerts a profound cytopathic effect upon peripheral blood CD4(+) T lymphocytes that resembles necrosis rather than apoptosis. Necrotic cytopathology was found with both laboratory-adapted strains and primary isolates of HIV-1. We carefully investigated the role of env, which has been previously implicated in HIV cytopathicity. HIV-1 stocks with equivalent infectivity were prepared from constructs with either an intact or mutated env coding region and pseudotyped with the glycoprotein of vesicular stomatitis virus (VSV-G) so that the HIV envelope was not rate-limiting for infection. Infected Jurkat T cells died whether or not env was intact; however, the expression of env accelerated death significantly. The accelerated death was blocked by protease inhibitors, indicating that it was due to reinfection by newly produced virus in env(+) cultures. Accordingly, we found no disparity in kinetics in CD4(lo) Jurkat cells. In highly infected peripheral blood T cells, profound necrosis occurred equivalently with both env(+) and env(-) stocks of HIV-1. We also found that HIV-1 cytopathicity was undiminished by the absence of nef. However, viral stocks made by complementation or packaging of HIV-1 genomes with the natural protein-coding sequences replaced by the green fluorescent protein were highly infectious but not cytopathic. Thus, env can accelerate cell death chiefly as an entry function, but one or more viral functions other than env or nef is essential for necrosis of CD4(+) T cells induced by HIV-1.
- Published
- 2002
- Full Text
- View/download PDF
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