136 results on '"Barillas-Mury C"'
Search Results
2. Milk, lactose and ethanol as dietary factors in cataractogenesis in Guatemala. A case controlled study
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Vettorazzi, C., Canales, D., Rosales, F., Barillas-Mury, C., Van Woert, J., Pineda, O., and Solomons, N.W.
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Guatemala -- Research ,Cataract -- Causes of ,Food/cooking/nutrition - Abstract
A Guatemalan study indicated that cataract risk can be enhanced by ethanol consumption and milk intake. However, ethanol consumption was found to have stronger influence on the risk of cataract, than high milk consumption. The study was conducted on 46 adults who had been treated for cataract and on 46 normal adults. Results of the study indicate that dietary modification can reduce the risk of cataract.
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- 1992
3. Plasmodium berghei P47 is essential for ookinete protection from the Anopheles gambiae complement-like response
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Ukegbu, CV, Giogalli, M, Yassine, H, Luis Ramirez, J, Taxiarchi, C, Barillas-Mury, C, Christophides, GK, and Wellcome Trust
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fungi ,parasitic diseases - Abstract
Malaria is a mosquito-borne disease affecting millions of people every year. The rodent parasite Plasmodium berghei has served as a model for human malaria transmission studies and played a pivotal role in dissecting the mosquito immune response against infection. The 6-cysteine protein P47, known to be important for P. berghei female gamete fertility, is shown to serve a different function in Plasmodium falciparum, protecting ookinetes from the mosquito immune response. Here, we investigate the function of P. berghei P47 in Anopheles gambiae mosquito infections. We show that P47 is expressed on the surface of both female gametocytes and ookinetes where it serves distinct functions in promoting gametocyte-to-ookinete development and protecting ookinetes from the mosquito complement-like response, respectively. The latter function is essential, as ookinetes lacking P47 are targeted for killing while traversing the mosquito midgut cells and eliminated upon exposure to hemolymph proteins of the complement-like system. Silencing key factors of the complement-like system restores oocyst development and disease transmission to rodent hosts. Our data establish a dual role of P. berghei P47 in vivo and reinforce the use of this parasite to study the impact of the mosquito immune response on human malaria transmission.
- Published
- 2017
4. Overexpression and altered nucleocytoplasmic distribution of Anopheles ovalbumin-like SRPN10 serpins in Plasmodium-infected midgut cells
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DANIELLI, ALBERTO, Barillas Mury C., Kumar S., Kafatos F. C., Loukeris T. G., Danielli A., Barillas-Mury C., Kumar S., Kafatos F.C., and Loukeris T.G.
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ANOPHELES ,SERPIN ,OOKINETE MIDGUT INVASION ,fungi ,parasitic diseases ,PLASMODIUM ,MOSQUITO-PARASITE INTERACTIONS - Abstract
The design of effective, vector-based malaria transmission blocking strategies relies on a thorough understanding of the molecular and cellular interactions that occur during the parasite sporogonic cycle in the mosquito. During Plasmodium berghei invasion, transcription from the SRPN10 locus, encoding four serine protease inhibitors of the ovalbumin family, is strongly induced in the mosquito midgut. Herein we demonstrate that intense induction as well as redistribution of SRPN10 occurs specifically in the parasite-invaded midgut epithelial cells. Quantitative analysis establishes that in response to epithelial invasion, SRPN10 translocates from the nucleus to the cytoplasm and this is followed by strong SRPN10 overexpression. The invaded cells exhibit signs of apoptosis, suggesting a link between this type of intracellular serpin and epithelial damage. The SRPN10 gene products constitute a novel, robust and cell-autonomous marker of midgut invasion by ookinetes. The SRPN10 dynamics at the subcellular level confirm and further elaborate the 'time bomb' model of P. berghei invasion in both Anopheles stephensi and Anopheles gambiae. In contrast, this syndrome of responses is not elicited by mutant P. berghei ookinetes lacking the major ookinete surface proteins, P28 and P25. Molecular markers with defined expression patterns, in combination with mutant parasite strains, will facilitate dissection of the molecular mechanisms underlying vector competence and development of effective transmission blocking strategies.
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- 2005
5. Gene synthesis, bacterial expression and proton NMR spectroscopic studies of the rat outer mitochondrial membrane cytochrome b5
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Christensen Ka, Wells Ma, Barillas-Mury C, Mario Rivera, Walker Fa, and Little Jw
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biology ,Stereochemistry ,Cytochrome b ,Cytochrome c ,Cytochrome P450 reductase ,Biochemistry ,chemistry.chemical_compound ,chemistry ,Cytochrome C1 ,Coenzyme Q – cytochrome c reductase ,Cytochrome b5 ,biology.protein ,Cytochrome c oxidase ,Heme - Abstract
The gene coding for the water-soluble domain of the outer mitochondrial membrane cytochrome b5 (OM cytochrome b5) from rat liver has been synthetized and expressed in Escherichia coli. The DNA sequence was obtained by back-translating the known amino acid sequence [Lederer, F., Ghrir, R., Guiard, B., Cortial, S., & Ito, A. (1983) Eur. J. Biochem. 132, 95-102]. The recombinant OM cytochrome b5 was characterized by UV-visible, EPR, and 1H NMR spectroscopy. The UV-visible and EPR spectra of the OM cytochrome b5 are almost identical to the ones obtained from the overexpressed rat microsomal cytochrome b5 [Bodman, S. B. V., Schyler, M. A., Jollie, D. R., & Sligar, S. G. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 9443-9447]. The one-dimensional 1H NMR spectrum of the OM cytochrome b5 indicates that the rhombic perturbation of the ferric center is essentially identical to that in the microsomal beef, rabbit, chicken, and rat cytochromes b5. Two-dimensional 1H NMR spectroscopy (NOESY) and one-dimensional NOE difference spectroscopy were used to assign the contact-shifted resonances that correspond to each of the two isomers that result from the rotation of the heme around its alpha-gamma-meso axis. The assignment of the resonances allowed the determination of the heme orientation ratio in the OM cytochrome b5, which was found to be 1.0 +/- 0.1. It is noteworthy that the two cytochromes b5 that have similar populations of the two heme isomers (large heme disorder) originate from the rat liver.
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- 1992
6. The human malaria parasite Pfs47 gene mediates evasion of the mosquito immune system
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Molina-Cruz, A., Garver, L.S., Alabaster, A., Bangiolo, L., Haile, A., Winikor, J., Ortega, C., Schaijk, B.C.L. van, Sauerwein, R.W., Taylor-Salmon, E., Barillas-Mury, C., Molina-Cruz, A., Garver, L.S., Alabaster, A., Bangiolo, L., Haile, A., Winikor, J., Ortega, C., Schaijk, B.C.L. van, Sauerwein, R.W., Taylor-Salmon, E., and Barillas-Mury, C.
- Abstract
Item does not contain fulltext, Plasmodium falciparum transmission by Anopheles gambiae mosquitoes is remarkably efficient, resulting in a very high prevalence of human malaria infection in sub-Saharan Africa. A combination of genetic mapping, linkage group selection, and functional genomics was used to identify Pfs47 as a P. falciparum gene that allows the parasite to infect A. gambiae without activating the mosquito immune system. Disruption of Pfs47 greatly reduced parasite survival in the mosquito, and this phenotype could be reverted by genetic complementation of the parasite or by disruption of the mosquito complement-like system. Pfs47 suppresses midgut nitration responses that are critical to activate the complement-like system. We provide direct experimental evidence that immune evasion mediated by Pfs47 is critical for efficient human malaria transmission by A. gambiae.
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- 2013
7. PrPCWD lymphoid cell targets in early and advanced chronic wasting disease of mule deer
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Sigurdson, C.J., Barillas-Mury, C., Miller, M.W., Oesch, B., van Keulen, L.J.M., Langeveld, J.P.M., Hoover, E.A., Sigurdson, C.J., Barillas-Mury, C., Miller, M.W., Oesch, B., van Keulen, L.J.M., Langeveld, J.P.M., and Hoover, E.A.
- Abstract
Up to 15% of free-ranging mule deer in northeastern Colorado and southeastern Wyoming, USA, are afflicted with a prion disease, or transmissible spongiform encephalopathy (TSE), known as chronic wasting disease (CWD). CWD is similar to a subset of TSEs including scrapie and variant Creutzfeldt¿Jakob disease in which the abnormal prion protein isoform, PrPCWD, accumulates in lymphoid tissue. Experimental scrapie studies have indicated that this early lymphoid phase is an important constituent of prion replication interposed between mucosal entry and central nervous system accumulation. To identify the lymphoid target cells associated with PrPCWD, we used triple-label immunofluorescence and high-resolution confocal microscopy on tonsils from naturally infected deer in advanced disease. We detected PrPCWD primarily extracellularly in association with follicular dendritic and B cell membranes as determined by frequent co-localization with antibodies against membrane bound immunoglobulin and CD21. There was minimal co-localization with cytoplasmic labels for follicular dendritic cells (FDC). This finding could indicate FDC capture of PrPCWD, potentially in association with immunoglobulin or complement, or PrPC conversion on FDC. In addition, scattered tingible body macrophages in the germinal centre contained coarse intracytoplasmic aggregates of PrPCWD, reflecting either phagocytosis of PrPCWD on FDC processes, apoptotic FDC or B cells, or actual PrPCWD replication within tingible body macrophages. To compare lymphoid cell targets in early and advanced disease, we also examined: (i) PrPCWD distribution in lymphoid cells of fawns within 3 months of oral CWD exposure and (ii) tonsil biopsies from preclinical deer with naturally acquired CWD. These studies revealed that the early lymphoid cellular distribution of PrPCWD was similar to that in advanced disease, i.e. in a pattern suggesting FDC association. We conclude that in deer, PrPCWD accumulates primarily extracellular
- Published
- 2002
8. Immune factor Gambif1, a new rel family member from the human malaria vector, Anopheles gambiae.
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Barillas-Mury, C., primary, Charlesworth, A., additional, Gross, I., additional, Richman, A., additional, Hoffmann, J. A., additional, and Kafatos, F. C., additional
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- 1996
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9. Inducible immune factors of the vector mosquito Anopheles gambiae: biochemical purification of a defensin antibacterial peptide and molecular cloning of preprodefensin cDNA
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Richman, A. M., primary, Bulet, P., additional, Hetru, C., additional, Barillas‐Mury, C., additional, Hoffmann, J. A., additional, and Kafatos, F. C., additional
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- 1996
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10. Aedes aegypti midgut early trypsin is post‐transcriptionally regulated by blood feeding
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Noriega, F. G., primary, Pennington, J. E., additional, Barillas‐Mury, C., additional, Wang, X. Y., additional, and Wells, M. A., additional
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- 1996
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11. Cloning and sequencing of the blood meal-induced late trypsin gene from the mosquito Aedes aegypti and characterization of the upstream regulatory region
- Author
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Barillas-Mury, C., primary and Wells, M. A., additional
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- 1993
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12. Test-retest reproducibility of hydrogen breath test for lactose maldigestion in preschool children.
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Barillas-Mury, Carolina, Solomons, Noel W., Barillas-Mury, C, and Solomons, N W
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- 1987
13. Variance in fasting breath hydrogen concentrations in Guatemalan preschool children.
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Barillas-Mury, Carolina, Solomons, Noel W., Barillas-Mury, C, and Solomons, N W
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- 1987
14. Mosquito immune responses and malaria transmission: lessons from insect model systems and implications for vertebrate innate immunity and vaccine development
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Barillas-Mury, C., Wizel, B., and Han, Y. Soo
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- 2000
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15. Early trypsin, a female-specific midgut protease in Aedes aegypti: isolation, amino-terminal sequence determination, and cloning and sequencing of the gene
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Noriega, F. G., Wang, X.-Y., Pennington, J. E., Barillas-Mury, C. V., and Wells, M. A.
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- 1996
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16. Directional gene expression and antisense transcripts in sexual and asexual stages of Plasmodium falciparum
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López-Barragán María J, Lemieux Jacob, Quiñones Mariam, Williamson Kim C, Molina-Cruz Alvaro, Cui Kairong, Barillas-Mury Carolina, Zhao Keji, and Su Xin-zhuan
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Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background It has been shown that nearly a quarter of the initial predicted gene models in the Plasmodium falciparum genome contain errors. Although there have been efforts to obtain complete cDNA sequences to correct the errors, the coverage of cDNA sequences on the predicted genes is still incomplete, and many gene models for those expressed in sexual or mosquito stages have not been validated. Antisense transcripts have widely been reported in P. falciparum; however, the extent and pattern of antisense transcripts in different developmental stages remain largely unknown. Results We have sequenced seven bidirectional libraries from ring, early and late trophozoite, schizont, gametocyte II, gametocyte V, and ookinete, and four strand-specific libraries from late trophozoite, schizont, gametocyte II, and gametocyte V of the 3D7 parasites. Alignment of the cDNA sequences to the 3D7 reference genome revealed stage-specific antisense transcripts and novel intron-exon splicing junctions. Sequencing of strand-specific cDNA libraries suggested that more genes are expressed in one direction in gametocyte than in schizont. Alternatively spliced genes, antisense transcripts, and stage-specific expressed genes were also characterized. Conclusions It is necessary to continue to sequence cDNA from different developmental stages, particularly those of non-erythrocytic stages. The presence of antisense transcripts in some gametocyte and ookinete genes suggests that these antisense RNA may play an important role in gene expression regulation and parasite development. Future gene expression studies should make use of directional cDNA libraries. Antisense transcripts may partly explain the observed discrepancy between levels of mRNA and protein expression.
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- 2011
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17. Mosquito immune responses and compatibility between Plasmodium parasites and anopheline mosquitoes
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Molina-Cruz Alvaro, Povelones Michael, Ndikuyeze Georges, Rodrigues Janneth, Jaramillo-Gutierrez Giovanna, and Barillas-Mury Carolina
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Microbiology ,QR1-502 - Abstract
Abstract Background Functional screens based on dsRNA-mediated gene silencing identified several Anopheles gambiae genes that limit Plasmodium berghei infection. However, some of the genes identified in these screens have no effect on the human malaria parasite Plasmodium falciparum; raising the question of whether different mosquito effector genes mediate anti-parasitic responses to different Plasmodium species. Results Four new An. gambiae (G3) genes were identified that, when silenced, have a different effect on P. berghei (Anka 2.34) and P. falciparum (3D7) infections. Orthologs of these genes, as well as LRIM1 and CTL4, were also silenced in An. stephensi (Nijmegen Sda500) females infected with P. yoelii (17XNL). For five of the six genes tested, silencing had the same effect on infection in the P. falciparum-An. gambiae and P. yoelii-An. stephensi parasite-vector combinations. Although silencing LRIM1 or CTL4 has no effect in An. stephensi females infected with P. yoelii, when An. gambiae is infected with the same parasite, silencing these genes has a dramatic effect. In An. gambiae (G3), TEP1, LRIM1 or LRIM2 silencing reverts lysis and melanization of P. yoelii, while CTL4 silencing enhances melanization. Conclusion There is a broad spectrum of compatibility, the extent to which the mosquito immune system limits infection, between different Plasmodium strains and particular mosquito strains that is mediated by TEP1/LRIM1 activation. The interactions between highly compatible animal models of malaria, such as P. yoelii (17XNL)-An. stephensi (Nijmegen Sda500), is more similar to that of P. falciparum (3D7)-An. gambiae (G3).
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- 2009
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18. Immunity-Related Genes and Gene Families in Anopheles gambiae
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Gareth J Lycett, Andrey Rzhetsky, Kenneth D. Vernick, Christian von Mering, Jiannong Xu, Frank H. Collins, Claudia Blass, Jules A. Hoffmann, Peer Bork, Stefan M. Kanzok, Jean-Marc Reichhart, Elena A. Levashina, Thanasis G. Loukeris, Stephan Meister, Charles Hetru, Ivica Letunic, Ngo Thi Hoa, Fotis C. Kafatos, Stéphanie Blandin, Liangbiao Zheng, Carolina Barillas-Mury, Mike A. Osta, Hans-Michael Müller, Jennifer Volz, Evgeny M. Zdobnov, Luis F. Moita, George Dimopoulos, Kristin Michel, Paul T. Brey, Laurent Troxler, Dina Vlachou, George K. Christophides, Susan M. Paskewitz, Ewan Birney, Alberto Danielli, Christophides G.K., Zdobnov E., Barillas-Mury C., Birney E., Blandin S., Blass C., Brey P.T., Collins F.H., Danielli A., Dimopoulos G., Hetru C., Hoa N.T., Hoffmann J.A., Kanzok S.M., Letunic I., Levashina E.A., Loukeris T.G., Lycett G., Meister S., Michel K., Moita L.F., Muller H.-M., Osta M.A., Paskewitz S.M., Reichhart J.-M., Rzhetsky A., Troxler L., Vernick K.D., Vlachou D., Volz J., Von Mering C., Xu J., Zheng L., Bork P., and Kafatos F.C.
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Plasmodium ,Serpins/metabolism ,Drosophila melanogaster/genetics/immunology/metabolism ,Anopheles gambiae ,Apoptosis ,Genes, Insect ,Peptides/metabolism ,Insect Protein ,Drosophila Proteins ,Enzyme Precursors/metabolism ,Catechol Oxidase/metabolism ,Phylogeny ,Serpin ,ddc:616 ,Genetics ,Enzyme Precursors ,Genome ,Multidisciplinary ,Drosophila Proteins/chemistry/genetics/metabolism ,Effector ,Serine Endopeptidases ,Insect Proteins/chemistry/genetics/metabolism ,Serine Endopeptidase ,Drosophila melanogaster ,Multigene Family ,Peptide ,Plasmodium/immunology/physiology ,Insect Proteins ,Anophele ,Catechol Oxidase ,Signal Transduction ,Bacteria/immunology ,Biology ,Enzyme Precursor ,Serine Endopeptidases/metabolism ,Immune system ,Anopheles ,Animals ,Anopheles/ genetics/ immunology/metabolism/microbiology/parasitology ,Gene family ,Selection, Genetic ,Gene ,Serpins ,Innate immune system ,Bacteria ,Animal ,Alternative splicing ,Apoptosi ,Computational Biology ,biology.organism_classification ,Immunity, Innate ,Protein Structure, Tertiary ,Alternative Splicing ,Gene Expression Regulation ,Drosophila Protein ,Peptides - Abstract
We have identified 242 Anopheles gambiae genes from 18 gene families implicated in innate immunity and have detected marked diversification relative to Drosophila melanogaster . Immune-related gene families involved in recognition, signal modulation, and effector systems show a marked deficit of orthologs and excessive gene expansions, possibly reflecting selection pressures from different pathogens encountered in these insects' very different life-styles. In contrast, the multifunctional Toll signal transduction pathway is substantially conserved, presumably because of counterselection for developmental stability. Representative expression profiles confirm that sequence diversification is accompanied by specific responses to different immune challenges. Alternative RNA splicing may also contribute to expansion of the immune repertoire.
- Published
- 2002
19. Mosquito midgut stem cell cellular defense response limits Plasmodium parasite infection.
- Author
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Barletta AF, Smith JC, Burkart E, Bondarenko S, Sharakhov IV, Criscione F, O'Brochta D, and Barillas-Mury C
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- Animals, Janus Kinases, STAT Transcription Factors, Signal Transduction, Oocysts, Stem Cells, Plasmodium berghei physiology, Parasites, Plasmodium, Malaria parasitology, Anopheles parasitology
- Abstract
A novel cellular response of midgut progenitors (stem cells and enteroblasts) to Plasmodium berghei infection was investigated in Anopheles stephensi. The presence of developing oocysts triggers proliferation of midgut progenitors that is modulated by the Jak/STAT pathway and is proportional to the number of oocysts on individual midguts. The percentage of parasites in direct contact with enteroblasts increases over time, as progenitors proliferate. Silencing components of key signaling pathways through RNA interference (RNAi) that enhance proliferation of progenitor cells significantly decreased oocyst numbers, while limiting proliferation of progenitors increased oocyst survival. Live imaging revealed that enteroblasts interact directly with oocysts and eliminate them. Midgut progenitors sense the presence of Plasmodium oocysts and mount a cellular defense response that involves extensive proliferation and tissue remodeling, followed by oocysts lysis and phagocytosis of parasite remnants by enteroblasts., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
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- 2024
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20. In-depth transcriptomic analysis of Anopheles gambiae hemocytes uncovers novel genes and the oenocytoid developmental lineage.
- Author
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Saha B, McNinch CM, Lu S, Ho MCW, De Carvalho SS, and Barillas-Mury C
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- Animals, Female, Hemocytes, Gene Expression Profiling, Transcriptome, Proteins metabolism, Anopheles genetics, Anopheles metabolism
- Abstract
Background: Hemocytes are immune cells that patrol the mosquito hemocoel and mediate critical cellular defense responses against pathogens. However, despite their importance, a comprehensive transcriptome of these cells was lacking because they constitute a very small fraction of the total cells in the insect, limiting the study of hemocyte differentiation and immune function., Results: In this study, an in-depth hemocyte transcriptome was built by extensive bulk RNA sequencing and assembly of hemocyte RNAs from adult A. gambiae female mosquitoes, based on approximately 2.4 billion short Illumina and about 9.4 million long PacBio high-quality reads that mapped to the A. gambiae PEST genome (P4.14 version). A total of 34,939 transcripts were annotated including 4,020 transcripts from novel genes and 20,008 novel isoforms that result from extensive differential splicing of transcripts from previously annotated genes. Most hemocyte transcripts identified (89.8%) are protein-coding while 10.2% are non-coding RNAs. The number of transcripts identified in the novel hemocyte transcriptome is twice the number in the current annotation of the A. gambiae genome (P4.14 version). Furthermore, we were able to refine the analysis of a previously published single-cell transcriptome (scRNAseq) data set by using the novel hemocyte transcriptome as a reference to re-define the hemocyte clusters and determine the path of hemocyte differentiation. Unsupervised pseudo-temporal ordering using the Tools for Single Cell Analysis software uncovered a novel putative prohemocyte precursor cell type that gives rise to prohemocytes. Pseudo-temporal ordering with the Monocle 3 software, which analyses changes in gene expression during dynamic biological processes, determined that oenocytoids derive from prohemocytes, a cell population that also gives rise to the granulocyte lineage., Conclusion: A high number of mRNA splice variants are expressed in hemocytes, and they may account for the plasticity required to mount efficient responses to many different pathogens. This study highlights the importance of a comprehensive set of reference transcripts to perform robust single-cell transcriptomic data analysis of cells present in low abundance. The detailed annotation of the hemocyte transcriptome will uncover new facets of hemocyte development and function in adult dipterans and is a valuable community resource for future studies on mosquito cellular immunity., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
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- 2024
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21. The heat shock protein Hsc70-3 mediates an anti-apoptotic response critical for Plasmodium evasion of Anopheles gambiae immunity.
- Author
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Alves E Silva TL, Canepa GE, Sweeney B, Hessab Alvarenga P, Zhao M, Vega-Rodríguez J, Molina-Cruz A, and Barillas-Mury C
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- Animals, Humans, Plasmodium falciparum, Heat-Shock Proteins metabolism, Anopheles parasitology, Plasmodium, Malaria
- Abstract
Importance: Malaria transmission by Anopheles gambiae mosquitoes is very effective, in part because the parasite expresses a surface protein called Pfs47 that allows it to evade the mosquito immune system. Here we investigate how this protein changes the response of mosquito midgut epithelial cells to invasion by the parasite. Pfs47 is known to interact with P47Rec, a mosquito midgut receptor. We found that Pf47Rec inhibits caspase-mediated apoptosis by interacting with the Hsc70-3. This disrupts nitration of midgut epithelial cells invaded by the parasite and the release of hemocyte-derived microvesicles, which are critical for effective activation of the mosquito complement system that eliminates the parasite., Competing Interests: The authors declare no conflict of interest.
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- 2023
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22. Leishmania genetic exchange is mediated by IgM natural antibodies.
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Serafim TD, Iniguez E, Barletta ABF, Cecilio P, Doehl JSP, Short M, Lack J, Nair V, Disotuar M, Wilson T, Coutinho-Abreu IV, Meneses C, Andersen J, Alves E Silva TL, Oliveira F, Vega-Rodriguez J, Barillas-Mury C, Ribeiro JMC, Beverley SM, Kamhawi S, and Valenzuela JG
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- Animals, Hybridization, Genetic, Gene Expression Regulation, Glycoside Hydrolases metabolism, Immunoglobulin M immunology, Leishmania genetics, Leishmania immunology, Psychodidae immunology, Psychodidae parasitology, Reproduction genetics, Host-Parasite Interactions genetics, Host-Parasite Interactions immunology
- Abstract
Host factors that mediate Leishmania genetic exchange are not well defined. Here we demonstrate that natural IgM (IgMn)
1-4 antibodies mediate parasite genetic exchange by inducing the transient formation of a spherical parasite clump that promotes parasite fusion and hybrid formation. We establish that IgMn from Leishmania-free animals binds to the surface of Leishmania parasites to induce significant changes in the expression of parasite transcripts and proteins. Leishmania binding to IgMn is partially lost after glycosidase treatment, although parasite surface phosphoglycans, including lipophosphoglycan, are not required for IgMn-induced parasite clumping. Notably, the transient formation of parasite clumps is essential for Leishmania hybridization in vitro. In vivo, we observed a 12-fold increase in hybrid formation in sand flies provided a second blood meal containing IgMn compared with controls. Furthermore, the generation of recombinant progeny from mating hybrids and parental lines were only observed in sand flies provided with IgMn. Both in vitro and in vivo IgM-induced Leishmania crosses resulted in full genome hybrids that show equal patterns of biparental contribution. Leishmania co-option of a host natural antibody to facilitate mating in the insect vector establishes a new paradigm of parasite-host-vector interdependence that contributes to parasite diversity and fitness by promoting genetic exchange., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)- Published
- 2023
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23. Mosquito midgut stem cell cellular defense response limits Plasmodium parasite infection.
- Author
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Barletta AF, Smith JC, Burkart E, Bondarenko S, Sharakhov I, Criscione F, O'Brochta D, and Barillas-Mury C
- Abstract
A novel cellular response of midgut progenitors (stem cells and enteroblasts) to Plasmodium berghei infection was investigated in Anopheles stephensi. The presence of developing oocysts triggers proliferation of midgut progenitors that is modulated by the Jak/STAT pathway, and proportional to the number of oocysts on individual midguts. The percentage of parasites in direct contact with enteroblasts increases over time, as progenitors proliferate. Enhancing proliferation of progenitors significantly decreases oocyst numbers, while limiting proliferation increases oocyst survival. Live imaging revealed that enteroblasts interact directly with oocysts and eliminate them. Midgut progenitors sense the presence of Plasmodium oocysts and mount a cellular defense response that involves extensive proliferation and tissue remodeling, followed by oocysts lysis and phagocytosis of parasite remnants by enteroblasts., Competing Interests: Competing interests The authors declare no competing financial interests.
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- 2023
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24. Author Correction: A mosquito lipoxin/lipocalin complex mediates innate immune priming in Anopheles gambiae.
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Ramirez JL, de Almeida Oliveira G, Calvo E, Dalli J, Colas RA, Serhan CN, Ribeiro JM, and Barillas-Mury C
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- 2023
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25. Extending the range of Plasmodium falciparum transmission blocking antibodies.
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Simons LM, Ferrer P, Gombakomba N, Underwood K, Herrera R, Narum DL, Canepa G, Acquah F, Amoah L, Duffy PE, Barillas-Mury C, Long C, Lee SM, Locke E, Miura K, and Williamson KC
- Subjects
- Humans, Plasmodium falciparum, Antibodies, Blocking, Epitopes, Antibodies, Protozoan, Antibodies, Monoclonal, Protozoan Proteins, Antigens, Protozoan, Malaria, Falciparum, Malaria Vaccines
- Abstract
Recent work demonstrating that asymptomatic carriers of P. falciparum parasites make up a large part of the infectious reservoir highlights the need for an effective malaria vaccine. Given the historical challenges of vaccine development, multiple parasite stages have been targeted, including the sexual stages required for transmission. Using flow cytometry to efficiently screen for P. falciparum gamete/zygote surface reactivity, we identified 82 antibodies that bound live P. falciparum gametes/zygotes. Ten antibodies had significant transmission-reducing activity (TRA) in a standard membrane feeding assay and were subcloned along with 9 nonTRA antibodies as comparators. After subcloning, only eight of the monoclonals obtained have significant TRA. These eight TRA mAbs do not recognize epitopes present in any of the current recombinant transmission-blocking vaccine candidates, Pfs230D1M, Pfs48/45.6C, Pf47 D2 and rPfs25. One TRA mAb immunoprecipitates two surface antigens, Pfs47 and Pfs230, that are expressed by both gametocytes and gametes/zygotes. These two proteins have not previously been reported to associate and the recognition of both by a single TRA mAb suggests the Pfs47/Pfs230 complex is a new vaccine target. In total, Pfs230 was the dominant target antigen, with five of the eight TRA mAbs and 8 of 11 nonTRA gamete/zygote surface reactive mAbs interacting with Pfs230. Of the three remaining TRA mAbs, two recognized non-reduced, parasite-produced Pfs25 and one bound non-reduced, parasite-produced Pfs48/45. None of the TRA mAbs bound protein on an immunoblot of reduced gamete/zygote extract and two TRA mAbs were immunoblot negative, indicating none of the new TRA epitopes are linear. The identification of eight new TRA mAbs that bind epitopes not included in any of the constructs currently under advancement as transmission-blocking vaccine candidates may provide new targets worthy of further study., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)
- Published
- 2023
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26. The Aedes aegypti peritrophic matrix controls arbovirus vector competence through HPx1, a heme-induced peroxidase.
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Talyuli OAC, Oliveira JHM, Bottino-Rojas V, Silveira GO, Alvarenga PH, Barletta ABF, Kantor AM, Paiva-Silva GO, Barillas-Mury C, and Oliveira PL
- Subjects
- Animals, Humans, Reactive Oxygen Species metabolism, Antioxidants metabolism, Peroxidase metabolism, Mosquito Vectors, Heme metabolism, Peroxidases metabolism, Aedes, Arboviruses, Zika Virus metabolism, Zika Virus Infection
- Abstract
Aedes aegypti mosquitoes are the main vectors of arboviruses. The peritrophic matrix (PM) is an extracellular layer that surrounds the blood bolus. It acts as an immune barrier that prevents direct contact of bacteria with midgut epithelial cells during blood digestion. Here, we describe a heme-dependent peroxidase, hereafter referred to as heme peroxidase 1 (HPx1). HPx1 promotes PM assembly and antioxidant ability, modulating vector competence. Mechanistically, the heme presence in a blood meal induces HPx1 transcriptional activation mediated by the E75 transcription factor. HPx1 knockdown increases midgut reactive oxygen species (ROS) production by the DUOX NADPH oxidase. Elevated ROS levels reduce microbiota growth while enhancing epithelial mitosis, a response to tissue damage. However, simultaneous HPx1 and DUOX silencing was not able to rescue bacterial population growth, as explained by increased expression of antimicrobial peptides (AMPs), which occurred only after double knockdown. This result revealed hierarchical activation of ROS and AMPs to control microbiota. HPx1 knockdown produced a 100-fold decrease in Zika and dengue 2 midgut infection, demonstrating the essential role of the mosquito PM in the modulation of arbovirus vector competence. Our data show that the PM connects blood digestion to midgut immunological sensing of the microbiota and viral infections., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)
- Published
- 2023
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27. Role of Pfs47 in the dispersal of ancestral Plasmodium falciparum malaria through adaptation to different anopheline vectors.
- Author
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Molina-Cruz A, Canepa GE, Dwivedi A, Liu W, Raytselis N, Antonio-Nkondjio C, Hahn BH, Silva JC, and Barillas-Mury C
- Subjects
- Animals, Humans, Plasmodium falciparum genetics, Mosquito Vectors parasitology, Gorilla gorilla, Anopheles genetics, Malaria, Malaria, Falciparum parasitology, Plasmodium
- Abstract
Plasmodium falciparum malaria originated when Plasmodium praefalciparum , a gorilla malaria parasite transmitted by African sylvan anopheline mosquitoes, adapted to humans. Pfs47, a protein on the parasite surface mediates P. falciparum evasion of the mosquito immune system by interacting with a midgut receptor and is critical for Plasmodium adaptation to different anopheline species. Genetic analysis of 4,971 Pfs47 gene sequences from different continents revealed that Asia and Papua New Guinea harbor Pfs47 haplotypes more similar to its ortholog in P. praefalciparum at sites that determine vector compatibility, suggesting that ancestral P. falciparum readily adapted to Asian vectors. Consistent with this observation, Pfs47-receptor gene sequences from African sylvan malaria vectors, such as Anopheles moucheti and An. marshallii , were found to share greater similarity with those of Asian vectors than those of vectors of the African An. gambiae complex. Furthermore, experimental infections provide direct evidence that transformed P. falciparum parasites carrying Pfs47 orthologs of P. praefalciparum or P. reichenowi were more effective at evading the immune system of the Asian malaria vector An. dirus than An. gambiae . We propose that high compatibility of ancestral P. falciparum Pfs47 with the receptors of Asian vectors facilitated the early dispersal of human malaria to the Asian continent, without having to first adapt to sub-Saharan vectors of the An. gambiae complex.
- Published
- 2023
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28. Molecular mechanisms of insect immune memory and pathogen transmission.
- Author
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Gomes FM, Silva M, Molina-Cruz A, and Barillas-Mury C
- Subjects
- Animals, Insect Vectors, Host-Pathogen Interactions, Immunologic Memory, Insecta
- Abstract
Competing Interests: The authors have declared that no competing interests exist,
- Published
- 2022
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29. Deceiving and escaping complement - the evasive journey of the malaria parasite.
- Author
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Inklaar MR, Barillas-Mury C, and Jore MM
- Subjects
- Animals, Humans, Plasmodium falciparum, Anopheles parasitology, Malaria prevention & control, Malaria Vaccines, Parasites
- Abstract
During its life cycle, Plasmodium, the malaria parasite, is exposed to the human and mosquito complement systems. Early experiments demonstrated that activation of complement can pose a serious threat to parasites, but recent studies revealed complement-evasion mechanisms important for parasite survival. Blood-stage parasites and gametes recruit regulators to neutralize human complement activation, while ookinetes inhibit mosquito complement by disrupting epithelial nitration in response to midgut invasion. Here we provide an in-depth overview of the evasion mechanisms currently known and speculate on the existence of others not yet identified. Finally, we discuss how these mechanisms could provide novel targets for urgently needed malaria vaccines and therapeutics., Competing Interests: Declaration of interests The authors declare no competing interests., (Published by Elsevier Ltd.)
- Published
- 2022
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30. Understanding pathogen survival and transmission by arthropod vectors to prevent human disease.
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Barillas-Mury C, Ribeiro JMC, and Valenzuela JG
- Subjects
- Animals, Humans, Leishmaniasis prevention & control, Leishmaniasis transmission, Malaria prevention & control, Malaria transmission, Arthropod Vectors microbiology, Arthropod Vectors parasitology, Host-Pathogen Interactions, Salivary Proteins and Peptides metabolism, Vector Borne Diseases prevention & control, Vector Borne Diseases transmission
- Abstract
Many endemic poverty-associated diseases, such as malaria and leishmaniasis, are transmitted by arthropod vectors. Pathogens must interact with specific molecules in the vector gut, the microbiota, and the vector immune system to survive and be transmitted. The vertebrate host, in turn, is infected when the pathogen and vector-derived factors, such as salivary proteins, are delivered into the skin by a vector bite. Here, we review recent progress in our understanding of the biology of pathogen transmission from the human to the vector and back, from the vector to the host. We also highlight recent advances in the biology of vector-borne disease transmission, which have translated into additional strategies to prevent human disease by either reducing vector populations or by disrupting their ability to transmit pathogens.
- Published
- 2022
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31. Hemocyte differentiation to the megacyte lineage enhances mosquito immunity against Plasmodium .
- Author
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Barletta ABF, Saha B, Trisnadi N, Talyuli OAC, Raddi G, and Barillas-Mury C
- Subjects
- Animals, Hemocytes, Humans, Plastics metabolism, Anopheles, Malaria, Plasmodium
- Abstract
Activation of Toll signaling in Anopheles gambiae by silencing Cactus , a suppressor of this pathway, enhances local release of hemocyte-derived microvesicles (HdMv), promoting activation of the mosquito complement-like system, which eliminates Plasmodium ookinetes. We uncovered the mechanism of this immune enhancement. Cactus silencing triggers a Rel1 -mediated differentiation of granulocytes to the megacyte lineage, a new subpopulation of giant cells, resulting in a dramatic increase in the proportion of circulating megacytes. Megacytes are very plastic cells that are massively recruited to the basal midgut surface in response to Plasmodium infection. We show that Toll signaling modulates hemocyte differentiation and that megacyte recruitment to the midgut greatly enhances mosquito immunity against Plasmodium ., Competing Interests: AB, BS, OT, GR, CB No competing interests declared, NT is affiliated with Atropos Therapeutics Inc The author has no financial interests to declare
- Published
- 2022
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32. Malaria parasite evades mosquito immunity by glutaminyl cyclase-mediated posttranslational protein modification.
- Author
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Kolli SK, Molina-Cruz A, Araki T, Geurten FJA, Ramesar J, Chevalley-Maurel S, Kroeze HJ, Bezemer S, de Korne C, Withers R, Raytselis N, El Hebieshy AF, Kim RQ, Child MA, Kakuta S, Hisaeda H, Kobayashi H, Annoura T, Hensbergen PJ, Franke-Fayard BM, Barillas-Mury C, Scheeren FA, and Janse CJ
- Subjects
- Animals, Glutamic Acid metabolism, Glutamine metabolism, Humans, Plasmodium berghei genetics, Plasmodium berghei immunology, Protozoan Proteins immunology, Aminoacyltransferases immunology, Culicidae immunology, Malaria genetics, Malaria immunology, Malaria parasitology, Protein Processing, Post-Translational immunology, Sporozoites immunology
- Abstract
Glutaminyl cyclase (QC) modifies N-terminal glutamine or glutamic acid residues of target proteins into cyclic pyroglutamic acid (pGlu). Here, we report the biochemical and functional analysis of Plasmodium QC. We show that sporozoites of QC-null mutants of rodent and human malaria parasites are recognized by the mosquito immune system and melanized when they reach the hemocoel. Detailed analyses of rodent malaria QC-null mutants showed that sporozoite numbers in salivary glands are reduced in mosquitoes infected with QC-null or QC catalytically dead mutants. This phenotype can be rescued by genetic complementation or by disrupting mosquito melanization or phagocytosis by hemocytes. Mutation of a single QC-target glutamine of the major sporozoite surface protein (circumsporozoite protein; CSP) of the rodent parasite Plasmodium berghei also results in melanization of sporozoites. These findings indicate that QC-mediated posttranslational modification of surface proteins underlies evasion of killing of sporozoites by the mosquito immune system.
- Published
- 2022
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33. Pfs47 as a Malaria Transmission-Blocking Vaccine Target.
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Molina-Cruz A and Barillas-Mury C
- Abstract
Transmission-blocking vaccines (TBVs), pioneered by Richard Carter and others, aim to prevent parasite development in the mosquito vector and are a promising new tool for malaria elimination. Pfs47, recently identified as a TBV target, is a three-domain 6-cysteine protein on the surface of Plasmodium falciparum sexual stages. Pfs47 allows the parasite to evade mosquito immunity and is key for P. falciparum infection of the dominant malaria vectors Anopheles gambiae, Anopheles dirus, and Anopheles albimanus. Antibodies against Pfs47 domain 2 (D2) have significant transmission-blocking activity that prevents Plasmodium ookinete development and is independent of human complement. Strong transmission-blocking activity has been mapped to a region of 52 amino acids in Pfs47 D2. Efforts to optimize the immunogenicity of the Pfs47 D2 antigen with a viral-like particle have been successful, and the efficacy of a P47-based TBV was confirmed in vivo with Pbs47, the orthologue of Pfs47 in the mouse malaria parasite Plasmodium berghei. The current evidence warrants further development and clinical testing of a Pfs47-based TBV.
- Published
- 2022
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34. Protective effects of combining monoclonal antibodies and vaccines against the Plasmodium falciparum circumsporozoite protein.
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Wang LT, Pereira LS, Kiyuka PK, Schön A, Kisalu NK, Vistein R, Dillon M, Bonilla BG, Molina-Cruz A, Barillas-Mury C, Tan J, Idris AH, Francica JR, and Seder RA
- Subjects
- Animals, Antibodies, Protozoan immunology, Humans, Malaria, Falciparum prevention & control, Mice, Sporozoites immunology, Antibodies, Monoclonal immunology, Immunization, Passive methods, Malaria Vaccines immunology, Plasmodium falciparum immunology, Protozoan Proteins immunology
- Abstract
Combinations of monoclonal antibodies (mAbs) against different epitopes on the same antigen synergistically neutralize many viruses. However, there are limited studies assessing whether combining human mAbs against distinct regions of the Plasmodium falciparum (Pf) circumsporozoite protein (CSP) enhances in vivo protection against malaria compared to each mAb alone or whether passive transfer of PfCSP mAbs would improve protection following vaccination against PfCSP. Here, we isolated a panel of human mAbs against the subdominant C-terminal domain of PfCSP (C-CSP) from a volunteer immunized with radiation-attenuated Pf sporozoites. These C-CSP-specific mAbs had limited binding to sporozoites in vitro that was increased by combination with neutralizing human "repeat" mAbs against the NPDP/NVDP/NANP tetrapeptides in the central repeat region of PfCSP. Nevertheless, passive transfer of repeat- and C-CSP-specific mAb combinations did not provide enhanced protection against in vivo sporozoite challenge compared to repeat mAbs alone. Furthermore, combining potent repeat-specific mAbs (CIS43, L9, and 317) that respectively target the three tetrapeptides (NPDP/NVDP/NANP) did not provide additional protection against in vivo sporozoite challenge. However, administration of either CIS43, L9, or 317 (but not C-CSP-specific mAbs) to mice that had been immunized with R21, a PfCSP-based virus-like particle vaccine that induces polyclonal antibodies against the repeat region and C-CSP, provided enhanced protection against sporozoite challenge when compared to vaccine or mAbs alone. Collectively, this study shows that while combining mAbs against the repeat and C-terminal regions of PfCSP provide no additional protection in vivo, repeat mAbs do provide increased protection when combined with vaccine-induced polyclonal antibodies. These data should inform the implementation of PfCSP human mAbs alone or following vaccination to prevent malaria infection., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: R.A.S., J.F., L.T.W., and R.V. have submitted U.S. Provisional Patent Application No. 62/842,590, filed 3 May 2019, describing mAb L9. All other authors have declared that no competing interests exist.
- Published
- 2021
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35. Double peroxidase and histone acetyltransferase AgTip60 maintain innate immune memory in primed mosquitoes.
- Author
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Gomes FM, Tyner MDW, Barletta ABF, Saha B, Yenkoidiok-Douti L, Canepa GE, Molina-Cruz A, and Barillas-Mury C
- Subjects
- Animals, Anopheles immunology, Anopheles metabolism, Culicidae metabolism, Female, Granulocytes metabolism, Hemocytes immunology, Immunity, Innate immunology, Insect Proteins genetics, Insecta metabolism, Lipoxins metabolism, Malaria immunology, Male, Peroxidase metabolism, Plasmodium metabolism, Plasmodium berghei metabolism, Culicidae immunology, Histone Acetyltransferases metabolism, Immunologic Memory immunology
- Abstract
Immune priming in Anopheles gambiae is mediated by the systemic release of a hemocyte differentiation factor (HDF), a complex of lipoxin A
4 bound to Evokin, a lipid carrier. HDF increases the proportion of circulating granulocytes and enhances mosquito cellular immunity. Here, we show that Evokin is present in hemocytes and fat-body cells, and messenger RNA (mRNA) expression increases significantly after immune priming. The double peroxidase (DBLOX) enzyme, present in insects but not in vertebrates, is essential for HDF synthesis. DBLOX is highly expressed in oenocytes in the fat-body tissue, and these cells increase in number in primed mosquitoes. We provide direct evidence that the histone acetyltransferase AgTip60 (AGAP001539) is also essential for a sustained increase in oenocyte numbers, HDF synthesis, and immune priming. We propose that oenocytes may function as a population of cells that are reprogrammed, and orchestrate and maintain a broad, systemic, and long-lasting state of enhanced immune surveillance in primed mosquitoes., Competing Interests: The authors declare no competing interest.- Published
- 2021
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36. A genotyping assay to determine geographic origin and transmission potential of Plasmodium falciparum malaria cases.
- Author
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Molina-Cruz A, Raytselis N, Withers R, Dwivedi A, Crompton PD, Traore B, Carpi G, Silva JC, and Barillas-Mury C
- Subjects
- Geography, Humans, Genotyping Techniques methods, Malaria, Falciparum transmission, Plasmodium falciparum genetics
- Abstract
As countries work towards malaria elimination, it is important to monitor imported cases to prevent reestablishment of local transmission. The Plasmodium falciparum Pfs47 gene has strong geographic population structure, because only those parasites with Pfs47 haplotypes compatible with the mosquito vector species in a given continent are efficiently transmitted. Analysis of 4,971 world-wide Pfs47 sequences identified two SNPs (at 707 and 725 bp) as sufficient to establish the likely continent of origin of P. falciparum isolates. Pfs47 sequences from Africa, Asia, and the New World presented more that 99% frequency of distinct combinations of the SNPs 707 and 725 genotypes. Interestingly, Papua New Guinea Pfs47 sequences have the highest diversity in SNPs 707 and 725. Accurate and reproducible High-Resolution Melting (HRM) assays were developed to genotype Pfs47 SNPs 707 and 725 in laboratory and field samples, to assess the geographic origin and risk of local transmission of imported P. falciparum malaria cases., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)
- Published
- 2021
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37. Functional human IgA targets a conserved site on malaria sporozoites.
- Author
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Tan J, Cho H, Pholcharee T, Pereira LS, Doumbo S, Doumtabe D, Flynn BJ, Schön A, Kanatani S, Aylor SO, Oyen D, Vistein R, Wang L, Dillon M, Skinner J, Peterson M, Li S, Idris AH, Molina-Cruz A, Zhao M, Olano LR, Lee PJ, Roth A, Sinnis P, Barillas-Mury C, Kayentao K, Ongoiba A, Francica JR, Traore B, Wilson IA, Seder RA, and Crompton PD
- Subjects
- Animals, Humans, Mice, Plasmodium falciparum, Protozoan Proteins, Sporozoites, Antibodies, Protozoan immunology, Immunoglobulin A immunology, Malaria immunology
- Abstract
Immunoglobulin (Ig)A antibodies play a critical role in protection against mucosal pathogens. However, the role of serum IgA in immunity to nonmucosal pathogens, such as Plasmodium falciparum , is poorly characterized, despite being the second most abundant isotype in blood after IgG. Here, we investigated the circulating IgA response in humans to P. falciparum sporozoites that are injected into the skin by mosquitoes and migrate to the liver via the bloodstream to initiate malaria infection. We found that circulating IgA was induced in three independent sporozoite-exposed cohorts: individuals living in an endemic region in Mali, malaria-naïve individuals immunized intravenously with three large doses of irradiated sporozoites, and malaria-naïve individuals exposed to a single controlled mosquito bite infection. Mechanistically, we found evidence in an animal model that IgA responses were induced by sporozoites at dermal inoculation sites. From malaria-resistant individuals, we isolated several IgA monoclonal antibodies that reduced liver parasite burden in mice. One antibody, MAD2-6, bound to a conserved epitope in the amino terminus of the P. falciparum circumsporozoite protein, the dominant protein on the sporozoite surface. Crystal structures of this antibody revealed a unique mode of binding whereby two Fabs simultaneously bound either side of the target peptide. This study reveals a role for circulating IgA in malaria and identifies the amino terminus of the circumsporozoite protein as a target of functional antibodies., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2021
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38. Design of Dissolvable Microneedles for Delivery of a Pfs47-Based Malaria Transmission-Blocking Vaccine.
- Author
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Yenkoidiok-Douti L, Barillas-Mury C, and Jewell CM
- Subjects
- Drug Delivery Systems, Humans, Needles, Plasmodium falciparum, Malaria, Malaria Vaccines
- Abstract
The development of effective malaria vaccines remains a global health priority. In addition to an effective vaccine, there is urgent demand for effective delivery technologies that can be easily deployed. The need for effective vaccine delivery tools is particularly pertinent in resource-poor settings where access to healthcare is limited. Microneedles are micron-scale structures that offer distinct advantages for vaccine delivery by efficiently targeting skin-resident immune cells, eliminating injection-associated pain, and improving patient compliance. Here, we developed and characterized a candidate malaria vaccine loaded and deployed using dissolvable microneedle arrays. Of note, a newly indicated human-relevant antigen was employed, Plasmodium falciparum surface protein P47. P47 and a potent toll-like receptor (TLR9) agonist vaccine adjuvant, CpG, were fabricated into microneedles using a gelatin polymer. Protein binding, ELISA, and fluorescence analysis confirmed the molecular structure, and the function of the P47 antigen and CpG was maintained after fabrication, storage, and release from microneedles. In cell culture, the cargo released from the microneedle arrays triggered TLR9 signaling and activated primary dendritic cells at levels similar to native, unincorporated vaccine components. Together, these studies demonstrate the potential of microneedles as an easily deployable strategy for a P47-based malaria vaccine.
- Published
- 2021
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39. Antibody Therapy Goes to Insects: Monoclonal Antibodies Can Block Plasmodium Transmission to Mosquitoes.
- Author
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Coelho CH, Jore MM, Canepa GE, Barillas-Mury C, Bousema T, and Duffy PE
- Subjects
- Animals, Culicidae immunology, Disease Eradication, Humans, Malaria parasitology, Antibodies, Monoclonal immunology, Culicidae parasitology, Life Cycle Stages immunology, Malaria prevention & control, Malaria transmission, Plasmodium falciparum immunology
- Abstract
Malaria eradication is a global priority but requires innovative strategies. Humoral immune responses attack different parasite stages, and antibody-based therapy may prevent malaria infection or transmission. Here, we discuss targets of monoclonal antibodies in mosquito sexual stages of Plasmodium., (Published by Elsevier Ltd.)
- Published
- 2020
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40. Reply to Patella et al. and Lindestam Arlehamn et al.: Complex pandemic dynamics and effect of bacillus Calmette-Guérin (BCG) vaccination on COVID-19 prevalence and mortality.
- Author
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Barillas-Mury C, Escobar LE, and Molina-Cruz A
- Subjects
- BCG Vaccine, Betacoronavirus, COVID-19, Humans, Patella, Prevalence, SARS-CoV-2, Vaccination, Coronavirus, Coronavirus Infections, Pandemics, Pneumonia, Viral
- Abstract
Competing Interests: The authors declare no competing interest.
- Published
- 2020
- Full Text
- View/download PDF
41. A Potent Anti-Malarial Human Monoclonal Antibody Targets Circumsporozoite Protein Minor Repeats and Neutralizes Sporozoites in the Liver.
- Author
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Wang LT, Pereira LS, Flores-Garcia Y, O'Connor J, Flynn BJ, Schön A, Hurlburt NK, Dillon M, Yang ASP, Fabra-García A, Idris AH, Mayer BT, Gerber MW, Gottardo R, Mason RD, Cavett N, Ballard RB, Kisalu NK, Molina-Cruz A, Nelson J, Vistein R, Barillas-Mury C, Amino R, Baker D, King NP, Sauerwein RW, Pancera M, Cockburn IA, Zavala F, Francica JR, and Seder RA
- Subjects
- Adolescent, Adult, Animals, Cell Line, Cell Line, Tumor, Epitopes immunology, Female, HEK293 Cells, Hepatocytes immunology, Hepatocytes parasitology, Humans, Liver immunology, Liver parasitology, Malaria immunology, Malaria parasitology, Malaria Vaccines immunology, Male, Mice, Mice, Inbred C57BL, Middle Aged, Young Adult, Antibodies, Monoclonal immunology, Antibodies, Neutralizing immunology, Antibodies, Protozoan immunology, Antimalarials immunology, Plasmodium falciparum immunology, Protozoan Proteins immunology, Sporozoites immunology
- Abstract
Discovering potent human monoclonal antibodies (mAbs) targeting the Plasmodium falciparum circumsporozoite protein (PfCSP) on sporozoites (SPZ) and elucidating their mechanisms of neutralization will facilitate translation for passive prophylaxis and aid next-generation vaccine development. Here, we isolated a neutralizing human mAb, L9 that preferentially bound NVDP minor repeats of PfCSP with high affinity while cross-reacting with NANP major repeats. L9 was more potent than six published neutralizing human PfCSP mAbs at mediating protection against mosquito bite challenge in mice. Isothermal titration calorimetry and multiphoton microscopy showed that L9 and the other most protective mAbs bound PfCSP with two binding events and mediated protection by killing SPZ in the liver and by preventing their egress from sinusoids and traversal of hepatocytes. This study defines the subdominant PfCSP minor repeats as neutralizing epitopes, identifies an in vitro biophysical correlate of SPZ neutralization, and demonstrates that the liver is an important site for antibodies to prevent malaria., Competing Interests: Declaration of Interests R.A.S., J.R.F., L.T.W., and R.V. have submitted US Provisional Patent Application 62/842,590, filed 3 May 2019, describing mAb L9. All other authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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42. Live In Vivo Imaging of Plasmodium Invasion of the Mosquito Midgut.
- Author
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Trisnadi N and Barillas-Mury C
- Subjects
- Animals, Anopheles anatomy & histology, Apoptosis, Epithelial Cells pathology, Female, Fluorescent Dyes, Mice, Mice, Inbred BALB C, Microscopy, Confocal methods, Optical Imaging methods, Anopheles parasitology, Digestive System parasitology, Epithelial Cells parasitology, Host-Parasite Interactions, Intravital Microscopy methods, Plasmodium berghei physiology
- Abstract
The mosquito midgut is a critical barrier that Plasmodium parasites must overcome to complete their developmental cycle and be transmitted to a new vertebrate host. Previous confocal studies with fixed infected midguts showed that ookinetes traverse midgut epithelial cells and cause irreversible tissue damage. Here, we investigated the spatiotemporal dynamics of ookinete midgut traversal and the response of midgut cells to invasion. A novel mounting strategy was established, suitable fluorescent dye combinations were identified and protocols optimized to label mosquito tissues in vivo , and live imaging protocols using confocal microscopy were developed. Tracking data showed that ookinetes gliding on the midgut surface travel faster and farther than those that remain in the lumen or those that have invaded the epithelium. Image analysis confirmed that parasite invasion and cell traversal occur within a couple of minutes, while caspase activity in damaged cells, indicative of cellular apoptosis, and F-actin cytoskeletal rearrangements in cells extruded into the gut lumen persist for several hours. This temporal difference highlights the importance of hemocyte-mediated cellular immunity and the mosquito complement system to mount a coordinated and effective antiplasmodial response. This novel in vivo imaging protocol allowed us to continuously observe individual ookinetes in live mosquitoes within the gut lumen and during cell traversal and to capture the subsequent cellular responses to invasion in real time for several hours, without loss of tissue integrity. IMPORTANCE Malaria is one of the most devastating parasitic diseases in humans and is transmitted by anopheline mosquitoes. The mosquito midgut is a critical barrier that Plasmodium parasites must overcome to complete their developmental cycle and be transmitted to a new host. Here, we developed a new strategy to visualize Plasmodium ookinetes as they traverse the mosquito midgut and to follow the response of damaged epithelial cells by imaging live mosquitoes. Understanding the spatial and temporal aspects of these interactions is critical when developing novel strategies to disrupt disease transmission., (Copyright © 2020 Trisnadi and Barillas-Mury.)
- Published
- 2020
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43. Mosquito cellular immunity at single-cell resolution.
- Author
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Raddi G, Barletta ABF, Efremova M, Ramirez JL, Cantera R, Teichmann SA, Barillas-Mury C, and Billker O
- Subjects
- Aedes genetics, Animals, Anopheles genetics, Female, Gene Expression Profiling, Gene Knockdown Techniques, Granulocytes immunology, Hemocytes metabolism, Malaria immunology, Malaria parasitology, Mice, Mosquito Vectors genetics, RNA-Seq, Single-Cell Analysis, Aedes immunology, Anopheles immunology, Hemocytes immunology, Immunity, Cellular, Malaria transmission, Mosquito Vectors immunology
- Abstract
Hemocytes limit the capacity of mosquitoes to transmit human pathogens. Here we profile the transcriptomes of 8506 hemocytes of Anopheles gambiae and Aedes aegypti mosquito vectors. Our data reveal the functional diversity of hemocytes, with different subtypes of granulocytes expressing distinct and evolutionarily conserved subsets of effector genes. A previously unidentified cell type in An. gambiae , which we term "megacyte," is defined by a specific transmembrane protein marker (TM7318) and high expression of lipopolysaccharide-induced tumor necrosis factor-α transcription factor 3 (LL3). Knockdown experiments indicate that LL3 mediates hemocyte differentiation during immune priming. We identify and validate two main hemocyte lineages and find evidence of proliferating granulocyte populations. This atlas of medically relevant invertebrate immune cells at single-cell resolution identifies cellular events that underpin mosquito immunity to malaria infection., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2020
- Full Text
- View/download PDF
44. BCG vaccine protection from severe coronavirus disease 2019 (COVID-19).
- Author
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Escobar LE, Molina-Cruz A, and Barillas-Mury C
- Subjects
- Aged, Betacoronavirus drug effects, COVID-19, Coronavirus Infections virology, Humans, Pneumonia, Viral virology, Prognosis, SARS-CoV-2, Survival Rate, Vaccination, Adjuvants, Immunologic administration & dosage, BCG Vaccine administration & dosage, Betacoronavirus immunology, Coronavirus Infections mortality, Coronavirus Infections prevention & control, Pandemics prevention & control, Pneumonia, Viral mortality, Pneumonia, Viral prevention & control
- Abstract
A series of epidemiological explorations has suggested a negative association between national bacillus Calmette-Guérin (BCG) vaccination policy and the prevalence and mortality of coronavirus disease 2019 (COVID-19). However, these comparisons are difficult to validate due to broad differences between countries such as socioeconomic status, demographic structure, rural vs. urban settings, time of arrival of the pandemic, number of diagnostic tests and criteria for testing, and national control strategies to limit the spread of COVID-19. We review evidence for a potential biological basis of BCG cross-protection from severe COVID-19, and refine the epidemiological analysis to mitigate effects of potentially confounding factors (e.g., stage of the COVID-19 epidemic, development, rurality, population density, and age structure). A strong correlation between the BCG index, an estimation of the degree of universal BCG vaccination deployment in a country, and COVID-19 mortality in different socially similar European countries was observed ( r
2 = 0.88; P = 8 × 10-7 ), indicating that every 10% increase in the BCG index was associated with a 10.4% reduction in COVID-19 mortality. Results fail to confirm the null hypothesis of no association between BCG vaccination and COVID-19 mortality, and suggest that BCG could have a protective effect. Nevertheless, the analyses are restricted to coarse-scale signals and should be considered with caution. BCG vaccination clinical trials are required to corroborate the patterns detected here, and to establish causality between BCG vaccination and protection from severe COVID-19. Public health implications of a plausible BCG cross-protection from severe COVID-19 are discussed., Competing Interests: The authors declare no competing interest., (Copyright © 2020 the Author(s). Published by PNAS.)- Published
- 2020
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45. In vivo Characterization of Plasmodium berghei P47 (Pbs47) as a Malaria Transmission-Blocking Vaccine Target.
- Author
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Yenkoidiok-Douti L, Canepa GE, Barletta ABF, and Barillas-Mury C
- Abstract
An effective vaccine to reduce malaria transmission is central to control and ultimately achieve disease eradication. Recently, we demonstrated that antibodies targeting the Plasmodium falciparum surface protein P47 (Pfs47) reduce parasite transmission to Anopheles gambiae mosquitoes. Here, Plasmodium berghei (Pb) was used as a model to assess the in vivo efficacy of a P47-targeted transmission blocking vaccine (Pbs47). Mice were immunized following a prime/boost regimen and infected with P. berghei . The effect of immunization on infectivity to mosquitoes was evaluated by direct feeding on P. berghei -infected mice. The key region in Pbs47 where antibody binding confers protection was mapped, and the immunogenicity of this protective antigen was enhanced by conjugation to a virus-like particle. Passive immunization with 100 and 50 μg/mL of anti-Pbs47 IgG reduced oocyst density by 77 and 67%, respectively. Furthermore, affinity purified Pbs47-specific IgG significantly reduced oocyst density by 88 and 77%, respectively at doses as low as 10 and 1 μg/mL. These studies suggest that P47 is a promising transmission blocking target and show that antibodies to the same specific region in Pfs47 and Pbs47 confer protection., (Copyright © 2020 Yenkoidiok-Douti, Canepa, Barletta and Barillas-Mury.)
- Published
- 2020
- Full Text
- View/download PDF
46. BCG Vaccine Protection from Severe Coronavirus Disease 2019 (COVID19).
- Author
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Escobar LE, Molina-Cruz A, and Barillas-Mury C
- Abstract
A series of epidemiological explorations has suggested a negative association between national BCG vaccination policy and the prevalence and mortality of COVID-19. However, these comparisons are difficult to validate due to broad differences between countries such as socioeconomic status, demographic structure, rural vs. urban settings, time of arrival of the pandemic, number of diagnostic tests and criteria for testing, and national control strategies to limit the spread of COVID-19. We review evidence for a potential biological basis of BCG cross-protection from severe COVID-19, and refine the epidemiological analysis to mitigate effects of potentially confounding factors (e.g., stage of the COVID-19 epidemic, development, rurality, population density, and age structure). A strong correlation between the BCG index, an estimation of the degree of universal BCG vaccination deployment in a country, and COVID-19 mortality in different socially similar European countries was observed (r2 = 0.88; p = 8 X 10-7), indicating that every 10% increase in the BCG index was associated with a 10.4% reduction in COVID-19 mortality. Results fail to confirm the null hypothesis of no-association between BCG vaccination and COVID-19 mortality, and suggest that BCG could have a protective effect. Nevertheless, the analyses are restricted to coarse-scale signals and should be considered with caution. BCG vaccination clinical trials are required to corroborate the patterns detected here, and to establish causality between BCG vaccination and protection from severe COVID-19. Public health implications of a plausible BCG cross-protection from severe COVID-19 are discussed.
- Published
- 2020
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47. Plasmodium falciparum evades immunity of anopheline mosquitoes by interacting with a Pfs47 midgut receptor.
- Author
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Molina-Cruz A, Canepa GE, Alves E Silva TL, Williams AE, Nagyal S, Yenkoidiok-Douti L, Nagata BM, Calvo E, Andersen J, Boulanger MJ, and Barillas-Mury C
- Subjects
- Animals, Anopheles genetics, Host-Parasite Interactions, Immune Evasion, Insect Proteins genetics, Kinetics, Membrane Glycoproteins genetics, Mosquito Vectors genetics, Plasmodium falciparum genetics, Protozoan Proteins genetics, Anopheles immunology, Anopheles parasitology, Insect Proteins immunology, Membrane Glycoproteins immunology, Mosquito Vectors immunology, Mosquito Vectors parasitology, Plasmodium falciparum immunology, Protozoan Proteins immunology
- Abstract
The surface protein Pfs47 allows Plasmodium falciparum parasites to survive and be transmitted by making them "undetectable" to the mosquito immune system. P. falciparum parasites express Pfs47 haplotypes compatible with their sympatric vectors, while those with incompatible haplotypes are eliminated by the mosquito. We proposed that Pfs47 serves as a "key" that mediates immune evasion by interacting with a mosquito receptor "the lock," which differs in evolutionarily divergent anopheline mosquitoes. Recombinant Pfs47 (rPfs47) was used to identify the mosquito Pfs47 receptor protein (P47Rec) using far-Western analysis. rPfs47 bound to a single 31-kDa band and the identity of this protein was determined by mass spectrometry. The mosquito P47Rec has two natterin-like domains and binds to Pfs47 with high affinity (17 to 32 nM). P47Rec is a highly conserved protein with submicrovillar localization in midgut cells. It has structural homology to a cytoskeleton-interacting protein and accumulates at the site of ookinete invasion. Silencing P47Rec expression reduced P. falciparum infection, indicating that the interaction of Pfs47 with the receptor is critical for parasite survival. The binding specificity of P47Rec from distant anophelines ( Anopheles gambiae , Anopheles dirus , and Anopheles albimanus ) with Pfs47-Africa (GB4) and Pfs47-South America (7G8) haplotypes was evaluated, and it is in agreement with the previously documented compatibility between P. falciparum parasites expressing different Pfs47 haplotypes and these three anopheline species. Our findings give further support to the role of Pfs47 in the adaptation of P. falciparum to different vectors., Competing Interests: The authors declare no competing interest.
- Published
- 2020
- Full Text
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48. Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine.
- Author
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Yenkoidiok-Douti L, Williams AE, Canepa GE, Molina-Cruz A, and Barillas-Mury C
- Subjects
- Animals, Bacteriophages genetics, Bacteriophages immunology, Female, Immunization, Secondary, Malaria Vaccines administration & dosage, Malaria Vaccines immunology, Malaria, Falciparum immunology, Male, Mice, Vaccines, Virus-Like Particle immunology, Antibodies, Protozoan metabolism, Malaria, Falciparum prevention & control, Membrane Glycoproteins immunology, Protozoan Proteins immunology, Vaccines, Virus-Like Particle administration & dosage
- Abstract
We recently characterized Pfs47, a protein expressed on the surface of sexual stages and ookinetes of Plasmodium falciparum, as a malaria transmission-blocking vaccine (TBV) target. Mice immunization induced antibodies that conferred strong transmission-reducing activity (TRA) at a concentration of 200 μg/mL. Here, we sought to optimize the Pfs47 vaccine to elicit higher titers of high-affinity antibodies, capable of inducing strong TRA at a lower concentration. We report the development and evaluation of a Pfs47-based virus-like particle (VLP) vaccine generated by conjugating our 58 amino acid Pfs47 antigen to Acinetobacter phage AP205-VLP using the SpyCatcher:SpyTag adaptor system. AP205-Pfs47 complexes (VLP-P47) formed particles of ~22 nm diameter that reacted with polyclonal anti-Pfs47 antibodies, indicating that the antigen was accessible on the surface of the particle. Mice immunized with VLP-P47 followed by a boost with Pfs47 monomer induced significantly higher antibody titers, with higher binding affinity to Pfs47, than mice that received two immunizations with either VLP-P47 (VLP-P47/VLP-P47) or the Pfs47 monomer (P47/P47). Purified IgG from VLP-P47/P47 mice had strong TRA (83-98%) at concentrations as low as 5 μg/mL. These results indicate that conjugating the Pfs47 antigen to AP205-VLP significantly enhanced antigenicity and confirm the potential of Pfs47 as a TBV candidate.
- Published
- 2019
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49. Mosquito Midgut Prostaglandin Release Establishes Systemic Immune Priming.
- Author
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Barletta ABF, Trisnadi N, Ramirez JL, and Barillas-Mury C
- Abstract
Anopheles gambiae mosquitoes that have been infected with Plasmodium mount a more effective immune response to a subsequent infection. Priming is established when Plasmodium invasion of the mosquito midgut allows contact of the gut microbiota with epithelial cells. This event is followed by a systemic release of a hemocyte differentiation factor (HDF) consisting of Lipoxin A4 bound to Evokin, a lipocalin carrier, which increases the proportion of circulating hemocytes. We show that mosquito midgut cells produce and release prostaglandin E2 (PGE2), which attracts hemocytes to the midgut surface and enhances their patrolling activity. Systemic injection of prostaglandins (PGs) recapitulates the priming response and enhances antiplasmodial immunity by triggering HDF production. Although insects lack cyclooxygenases, two heme peroxidases, HPX7 and HPX8, catalyze essential steps in PG biosynthesis in mosquitoes. Mosquito midgut PGE2 release attracts hemocytes and establishes a long-lasting enhanced systemic cellular immune response to Plasmodium infection., (Published by Elsevier Inc.)
- Published
- 2019
- Full Text
- View/download PDF
50. Infection of anopheline mosquitoes with Wolbachia: Implications for malaria control.
- Author
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Gomes FM and Barillas-Mury C
- Subjects
- Animals, Insect Vectors, Malaria metabolism, Malaria virology, Anopheles microbiology, Malaria prevention & control, Wolbachia virology
- Abstract
Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2018
- Full Text
- View/download PDF
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