The Experts below are selected from a list of 2250 Experts worldwide ranked by ideXlab platform
Iliano V Coutinhoabreu - One of the best experts on this subject based on the ideXlab platform.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:Background The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines. Here, we investigated PpSP15 predicted amino acid sequence variability and mRNA profile of P. Papatasi field populations from the Middle East. In addition, predicted MHC class II T-cell epitopes were obtained and compared to areas of amino acid sequence variability within the secreted protein. The analysis of PpSP15 expression from field populations revealed significant intra- and interpopulation variation.. In spite of the variability detected for P. Papatasi populations, common epitopes for MHC class II binding are still present and may potentially be used to boost the response against Le. major infections. Conserved epitopes of PpSP15 could potentially be used in the development of a salivary gland antigen-based vaccine.
-
expression plasticity of Phlebotomus Papatasi salivary gland genes in distinct ecotopes through the sand fly season
BMC Ecology, 2011Co-Authors: Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Gwen Stayback, Mariha Wadsworth, Dilkushi De Alwis Pitts, Mahmoud N AboshehadaAbstract:Sand fly saliva can drive the outcome of Leishmania infection in animal models, and salivary components have been postulated as vaccine candidates against leishmaniasis. In the sand fly Phlebotomus Papatasi, natural sugar-sources modulate the activity of proteins involved in meal digestion, and possibly influence vectorial capacity. However, only a handful of studies have assessed the variability of salivary components in sand flies, focusing on the effects of environmental factors in natural habitats. In order to better understand such interactions, we compared the expression profiles of nine P. Papatasi salivary gland genes of specimens inhabiting different ecological habitats in Egypt and Jordan and throughout the sand fly season in each habitat. The majority of investigated genes were up-regulated in specimens from Swaymeh late in the season, when the availability of sugar sources is reduced due to water deprivation. On the other hand, these genes were not up-regulated in specimens collected from Aswan, an irrigated area less susceptible to drought effects. Expression plasticity of genes involved with vectorial capacity in disease vectors may play an important epidemiological role in the establishment of diseases in natural habitats.
-
targeting the midgut secreted ppchit1 reduces leishmania major development in its natural vector the sand fly Phlebotomus Papatasi
PLOS Neglected Tropical Diseases, 2010Co-Authors: Iliano V Coutinhoabreu, Narinder K Sharma, Maricela Roblesmurguia, Marcelo RamalhoortigaoAbstract:Background During its developmental cycle within the sand fly vector, Leishmania must survive an early proteolytic attack, escape the peritrophic matrix, and then adhere to the midgut epithelia in order to prevent excretion with remnants of the blood meal. These three steps are critical for the establishment of an infection within the vector and are linked to interactions controlling species-specific vector competence. PpChit1 is a midgut-specific chitinase from Phlebotomus Papatasi presumably involved in maturation and degradation of the peritrophic matrix. Sand fly midgut chitinases, such as PpChit1, whether acting independently or in a synergistic manner with Leishmania-secreted chitinase, possibly play a role in the Leishmania escape from the endoperitrophic space. Thus, we predicted that silencing of sand fly chitinase will lead to reduction or elimination of Leishmania within the gut of the sand fly vector.
-
differential expression of salivary gland genes in the female sand fly Phlebotomus Papatasi diptera psychodidae
Journal of Medical Entomology, 2010Co-Authors: Iliano V Coutinhoabreu, Marcelo Ramalhoortigao, Gwen Stayback, Mariha Wadsworth, Mary Ann McdowellAbstract:Saliva from blood-sucking arthropods modulates host homostasis and immunity, making salivary components potential candidates to be used against pathogens transmitted by these biting insects. Functional characterization of salivary molecules is fundamental to gain a better understanding into their roles during blood feeding and to determine under which conditions such molecules are expressed in the insect saliva. In the current study, we investigated the expression profile of 10 salivary genes from the sand fly Phlebotomus Papatasi (Scopoli) (Diptera: Psychodidae), a principal vector of Leishmania major. Our analyses using quantitative polymerase chain reaction were aimed at defining whether diet or senescence influences the expression of P. Papatasi salivary gland-expressed genes in laboratory-reared female sand flies. Our results demonstrate that at least one of the most abundant salivary transcripts, SP44, is consistently modulated by either senescence or diet. In contrast, another abundant transcript, SP32, was expressed without any influence from the diet received or the age of the sand fly. Differential expression of the other eight transcripts was not consistently regulated by either diet or age, suggesting that other factors may have a greater influence on differential expression of these salivary gland proteins.
Marcelo Ramalhoortigao - One of the best experts on this subject based on the ideXlab platform.
-
effects of specific antisera targeting peritrophic matrix associated proteins in the sand fly vector Phlebotomus Papatasi
Acta Tropica, 2016Co-Authors: Juliana Malta, Gustavo Ferreira Martins, Julin Weng, Kenner Morais Fernandes, M L Munford, Marcelo RamalhoortigaoAbstract:In many hematophagous insects, the peritrophic matrix (PM) is formed soon after a blood meal (PBM) to compartmentalize the food bolus. The PM is an important component of vector competence, functioning as a barrier to the development of many pathogens including parasites of the genus Leishmania transmitted by sand flies. PM morphology and permeability are associated with the proteins that are part of the PM scaffolding, including several peritrophins, and chitin fibers. Here, we assessed the effects of specific antisera targeting proteins thought to be an integral part of the PM scaffolding and its process of maturation and degradation. Phlebotomus Papatasi sand flies were fed with red blood cells reconstituted with antisera targeting the chitinase PpChit1, and the peritrophin PpPer2. Sand fly midguts were dissected at different time points and processed for light microscopy (LM), confocal and transmission electron (TEM) microscopies (24, 42-46, 48 and 72h PBM), scanning electron (SEM) (48h PBM) and atomic force (AFM) (30h PBM) microscopies. TEM and WGA-FITC staining indicate PM degradation was significantly delayed following feeding of flies on anti-PpChit1. AFM analysis at 30h PBM point to an increase in roughness' amplitude of the PM of flies that fed on either anti-PpChit1 or anti-PpPer2. Collective, our data suggest that antibodies targeting PM-associated proteins affects the kinetics of PM maturation, delaying its degradation and disruption and are potential targets on transmission-blocking vaccines strategies.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:Background The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines. Here, we investigated PpSP15 predicted amino acid sequence variability and mRNA profile of P. Papatasi field populations from the Middle East. In addition, predicted MHC class II T-cell epitopes were obtained and compared to areas of amino acid sequence variability within the secreted protein. The analysis of PpSP15 expression from field populations revealed significant intra- and interpopulation variation.. In spite of the variability detected for P. Papatasi populations, common epitopes for MHC class II binding are still present and may potentially be used to boost the response against Le. major infections. Conserved epitopes of PpSP15 could potentially be used in the development of a salivary gland antigen-based vaccine.
-
the characterization of the Phlebotomus Papatasi transcriptome
Insect Molecular Biology, 2013Co-Authors: Jenica L Abrudan, Phillip G Lawyer, Marcelo Ramalhoortigao, Shawn T Oneil, Gwen Stayback, Mariha Wadsworth, Megan Bernard, Doug Shoue, Scott J Emrich, Shaden KamhawiAbstract:As important vectors of human disease, phlebotomine sand flies are of global significance to human health, transmitting several emerging and re-emerging infectious diseases. The most devastating of the sand fly transmitted infections are the leishmaniases, causing significant mortality and morbidity in both the Old and New World. Here we present the first global transcriptome analysis of the Old World vector of cutaneous leishmaniasis, Phlebotomus Papatasi (Scopoli) and compare this transcriptome to that of the New World vector of visceral leishmaniasis, Lutzomyia longipalpis. A normalized cDNA library was constructed using pooled mRNA from Phlebotomus Papatasi larvae, pupae, adult males and females fed sugar, blood, or blood infected with Leishmania major. A total of 47 615 generated sequences was cleaned and assembled into 17 120 unique transcripts. Of the assembled sequences, 50% (8837 sequences) were classified using Gene Ontology (GO) terms. This collection of transcripts is comprehensive, as demonstrated by the high number of different GO categories. An in-depth analysis revealed 245 sequences with putative homology to proteins involved in blood and sugar digestion, immune response and peritrophic matrix formation. Twelve of the novel genes, including one trypsin, two peptidoglycan recognition proteins (PGRP) and nine chymotrypsins, have a higher expression level during larval stages. Two novel chymotrypsins and one novel PGRP are abundantly expressed upon blood feeding. This study will greatly improve the available genomic resources for P. Papatasi and will provide essential information for annotation of the full genome.
-
targeting the midgut secreted ppchit1 reduces leishmania major development in its natural vector the sand fly Phlebotomus Papatasi
PLOS Neglected Tropical Diseases, 2010Co-Authors: Iliano V Coutinhoabreu, Narinder K Sharma, Maricela Roblesmurguia, Marcelo RamalhoortigaoAbstract:Background During its developmental cycle within the sand fly vector, Leishmania must survive an early proteolytic attack, escape the peritrophic matrix, and then adhere to the midgut epithelia in order to prevent excretion with remnants of the blood meal. These three steps are critical for the establishment of an infection within the vector and are linked to interactions controlling species-specific vector competence. PpChit1 is a midgut-specific chitinase from Phlebotomus Papatasi presumably involved in maturation and degradation of the peritrophic matrix. Sand fly midgut chitinases, such as PpChit1, whether acting independently or in a synergistic manner with Leishmania-secreted chitinase, possibly play a role in the Leishmania escape from the endoperitrophic space. Thus, we predicted that silencing of sand fly chitinase will lead to reduction or elimination of Leishmania within the gut of the sand fly vector.
Mahmoud N Aboshehada - One of the best experts on this subject based on the ideXlab platform.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:Background The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines.
-
Phlebotomus Papatasi sp15 mrna expression variability and amino acid sequence polymorphisms of field populations
Parasites & Vectors, 2015Co-Authors: Marcelo Ramalhoortigao, Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Valdir De Queiroz Balbino, Carlos Alberto Santiago Figueiredo, Hussan Dayem, Mahmoud N AboshehadaAbstract:The Phlebotomus Papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines. Here, we investigated PpSP15 predicted amino acid sequence variability and mRNA profile of P. Papatasi field populations from the Middle East. In addition, predicted MHC class II T-cell epitopes were obtained and compared to areas of amino acid sequence variability within the secreted protein. The analysis of PpSP15 expression from field populations revealed significant intra- and interpopulation variation.. In spite of the variability detected for P. Papatasi populations, common epitopes for MHC class II binding are still present and may potentially be used to boost the response against Le. major infections. Conserved epitopes of PpSP15 could potentially be used in the development of a salivary gland antigen-based vaccine.
-
expression plasticity of Phlebotomus Papatasi salivary gland genes in distinct ecotopes through the sand fly season
BMC Ecology, 2011Co-Authors: Iliano V Coutinhoabreu, Rami Mukbel, Hanafi A Hanafi, Shabaan S Elhossary, Emadeldin Y Fawaz, Gwen Stayback, Mariha Wadsworth, Dilkushi De Alwis Pitts, Mahmoud N AboshehadaAbstract:Sand fly saliva can drive the outcome of Leishmania infection in animal models, and salivary components have been postulated as vaccine candidates against leishmaniasis. In the sand fly Phlebotomus Papatasi, natural sugar-sources modulate the activity of proteins involved in meal digestion, and possibly influence vectorial capacity. However, only a handful of studies have assessed the variability of salivary components in sand flies, focusing on the effects of environmental factors in natural habitats. In order to better understand such interactions, we compared the expression profiles of nine P. Papatasi salivary gland genes of specimens inhabiting different ecological habitats in Egypt and Jordan and throughout the sand fly season in each habitat. The majority of investigated genes were up-regulated in specimens from Swaymeh late in the season, when the availability of sugar sources is reduced due to water deprivation. On the other hand, these genes were not up-regulated in specimens collected from Aswan, an irrigated area less susceptible to drought effects. Expression plasticity of genes involved with vectorial capacity in disease vectors may play an important epidemiological role in the establishment of diseases in natural habitats.
Omar Hamarsheh - One of the best experts on this subject based on the ideXlab platform.
-
Development of polymorphic EST microsatellite markers for the sand fly, Phlebotomus Papatasi (Diptera: Psychodidae)
Parasites and Vectors, 2018Co-Authors: Omar Hamarsheh, Mohammad Reza Yaghoobi-ershadi, Ahmad Amro, Mehmet Karakuş, Kifaya Azmi, Kaouther Jaouadi, Andreas Krüger, Mohamed Kenawy, Mostafa Dokhan, Ziad AbdeenAbstract:BACKGROUND: Phlebotomus Papatasi is a widely distributed sand fly species in different tropical and sub-tropical regions including the Middle East and North Africa. It is considered an important vector that transmits Leishmania major parasites, the causative agents of cutaneous leishmaniasis. The development of microsatellite markers for this sand fly vector is of high interest to understand its population structure and to monitor its geographic dispersal. RESULTS: Fourteen polymorphic microsatellite markers were developed with simple di-, tri- and tetra-nucleotide repeats. The F statistics calculated for the 14 markers revealed high genetic diversity; expected heterozygosity (He) ranged from 0.407 to 0.767, while observed heterozygosity (Ho) was lower and ranged from 0.083 to 0.514. The number of alleles sampled fall in the range of 9-29. Three out of 14 markers deviated from Hardy-Weinberg expectations, no significant linkage disequilibrium was detected and high values for inbreeding coefficient (FIS) were likely due to inbreeding. CONCLUSIONS: The development of these functional microsatellites enable a high resolution of P. Papatasi populations. It opens a path for researchers to perform multi locus-based population genetic structure analyses, and comparative mapping, a part of the efforts to uncover the population dynamics of this vector, which is an important global strategy for understanding the epidemiology and control of leishmaniasis.
-
development of polymorphic est microsatellite markers for the sand fly Phlebotomus Papatasi diptera psychodidae
Parasites & Vectors, 2018Co-Authors: Omar Hamarsheh, Mohammad Reza Yaghoobiershadi, Ahmad Amro, Mehmet Karakuş, Kifaya Azmi, Kaouther Jaouadi, Andreas Krüger, Mohamed A Kenawy, Mostafa DokhanAbstract:Phlebotomus Papatasi is a widely distributed sand fly species in different tropical and sub-tropical regions including the Middle East and North Africa. It is considered an important vector that transmits Leishmania major parasites, the causative agents of cutaneous leishmaniasis. The development of microsatellite markers for this sand fly vector is of high interest to understand its population structure and to monitor its geographic dispersal. Fourteen polymorphic microsatellite markers were developed with simple di-, tri- and tetra-nucleotide repeats. The F statistics calculated for the 14 markers revealed high genetic diversity; expected heterozygosity (He) ranged from 0.407 to 0.767, while observed heterozygosity (Ho) was lower and ranged from 0.083 to 0.514. The number of alleles sampled fall in the range of 9–29. Three out of 14 markers deviated from Hardy-Weinberg expectations, no significant linkage disequilibrium was detected and high values for inbreeding coefficient (FIS) were likely due to inbreeding. The development of these functional microsatellites enable a high resolution of P. Papatasi populations. It opens a path for researchers to perform multi locus-based population genetic structure analyses, and comparative mapping, a part of the efforts to uncover the population dynamics of this vector, which is an important global strategy for understanding the epidemiology and control of leishmaniasis.
-
characterization of simple sequence repeats ssrs from Phlebotomus Papatasi diptera psychodidae expressed sequence tags ests
Parasites & Vectors, 2011Co-Authors: Omar Hamarsheh, Ahmad AmroAbstract:Background: Phlebotomus Papatasi is a natural vector of Leishmania major, which causes cutaneous leishmaniasis in many countries. Simple sequence repeats (SSRs), or microsatellites, are common in eukaryotic genomes and are short, repeated nucleotide sequence elements arrayed in tandem and flanked by non-repetitive regions. The enrichment methods used previously for finding new microsatellite loci in sand flies remain laborious and time consuming; in silico mining, which includes retrieval and screening of microsatellites from large amounts of sequence data from sequence data bases using microsatellite search tools can yield many new candidate markers. Results: Simple sequence repeats (SSRs) were characterized in P. Papatasi expressed sequence tags (ESTs) derived from a public database, National Center for Biotechnology Information (NCBI). A total of 42,784 sequences were mined, and 1,499 SSRs were identified with a frequency of 3.5% and an average density of 15.55 kb per SSR. Dinucleotide motifs were the most common SSRs, accounting for 67% followed by tri-, tetra-, and penta-nucleotide repeats, accounting for 31.1%, 1.5%, and 0.1%, respectively. The length of microsatellites varied from 5 to 16 repeats. Dinucleotide types; AG and CT have the highest frequency. Dinucleotide SSR-ESTs are relatively biased toward an excess of (AX)n repeats and a low GC base content. Forty primer pairs were designed based on motif lengths for further experimental validation. Conclusion: The first large-scale survey of SSRs derived from P. Papatasi is presented; dinucleotide SSRs identified are more frequent than other types. EST data mining is an effective strategy to identify functional microsatellites in P. Papatasi.
-
Distribution of Leishmania major zymodemes in relation to populations of Phlebotomus Papatasi sand flies
Parasites & Vectors, 2011Co-Authors: Omar HamarshehAbstract:Phlebotomus Papatasi (Scopoli) (Diptera: Psychodidae) is the main vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the causative agent of zoonotic cutaneous leishmaniasis in the Old World. Multilocus enzyme electrophoresis (MLEE) was extensively used to type different L. major stocks allover the world. Multilocus microsatellite typing (MLMT) has been recently used to investigate P. Papatasi sand flies at population and subpopulation levels. In this article, the association between geographical distribution of L. major zymodemes and the distribution of populations and subpopulations of L. major vector; P. Papatasi are discussed.
-
molecular markers for Phlebotomus Papatasi diptera psychodidae and their usefulness for population genetic analysis
Transactions of The Royal Society of Tropical Medicine and Hygiene, 2009Co-Authors: Omar Hamarsheh, W. Presber, Amer Aljawabreh, Ziad Abdeen, Ahmad Amro, Gabriele SchonianAbstract:Three molecular typing tools: multilocus microsatellite typing, cytochrome b sequence analysis and internal transcribed spacer 2 (ITS2) sequence analysis, were evaluated for their usefulness in inferring the population structure of Phlebotomus Papatasi sand flies. ITS2 sequence analysis did not prove suitable for inferring phylogenetic and population genetic relationships across P. Papatasi sand flies. Microsatellite markers showed high resolution in differentiating globally distributed P. Papatasi populations, whereas cytochrome b sequence analysis provided insight into the relationships between closely related populations from the Mediterranean. Population structure, differentiation and demographic history among P. Papatasi are important for understanding patterns of dispersal in this species and for planning appropriate control measures.
Shaden Kamhawi - One of the best experts on this subject based on the ideXlab platform.
-
Phlebotomus Papatasi yellow related and apyrase salivary proteins are candidates for vaccination against human cutaneous leishmaniasis
Journal of Investigative Dermatology, 2017Co-Authors: Aymen Tlili, Shaden Kamhawi, Soumaya Marzouki, Maha Abdeladhim, Fabiano Oliveira, Emna Chabaane, Wafa Kammounrebai, Rahma Sakkouhi, Nabil Belhadj Hmida, Hechmi LouzirAbstract:Nowadays, there is no available vaccine for human leishmaniasis. Animal experiments demonstrate that pre-exposure to sand fly saliva confers protection against leishmaniasis. Our preceding work in humans indicates that Phlebotomus Papatasi saliva induces the production of IL-10 by CD8+ T lymphocytes. The neutralization of IL-10 enhanced the activation of a T-cell CD4+ population-producing IFN-γ. Herein, we used a biochemical and functional genomics approach to identify the sand fly salivary components that are responsible for the activation of the T helper type 1 immune response in humans, therefore constituting potential vaccine candidates against leishmaniasis. Fractionated P. Papatasi salivary extracts were first tested on T lymphocytes of immune donors. We confirmed that the CD4+ lymphocytes proliferate and produce IFN-γ in response to stimulation with the proteins of molecular weight >30 kDa. Peripheral blood mononuclear cells from immune donors were transfected with plasmids coding for the most abundant proteins from the P. Papatasi salivary gland cDNA library. Our result showed that the "yellow related proteins," PPTSP42 and PPTSP44, and "apyrase," PPTSP36, are the proteins responsible for the aforementioned cellular immune response and IFN-γ production. Strikingly, PPTSP44 triggered the highest level of lymphocyte proliferation and IFN-γ production. Multiplex cytokine analysis confirmed the T helper type 1-polarized response induced by these proteins. Importantly, recombinant PPTSP44 validated the results observed with the DNA plasmid, further supporting that PPTSP44 constitutes a promising vaccine candidate against human leishmaniasis.
-
Validation of Recombinant Salivary Protein PpSP32 as a Suitable Marker of Human Exposure to Phlebotomus Papatasi, the Vector of Leishmania major in Tunisia
PLoS Neglected Tropical Diseases, 2015Co-Authors: Soumaya Marzouki, Maha Abdeladhim, Hamide Aslan, Wafa Kammoun-rebai, Jihène Bettaieb, Saoussen Hadj Kacem, Rania Abdelkader, Sami Gritli, Jomâa Chemkhi, Shaden KamhawiAbstract:Background During a blood meal, female sand flies, vectors of Leishmania parasites, inject saliva into the host skin. Sand fly saliva is composed of a large variety of components that exert different pharmacological activities facilitating the acquisition of blood by the insect. Importantly, proteins present in saliva are able to elicit the production of specific anti-saliva antibodies, which can be used as markers for exposure to vector bites. Serological tests using total sand fly salivary gland extracts are challenging due to the difficulty of obtaining reproducible salivary gland preparations. Previously, we demonstrated that PpSP32 is the immunodominant salivary antigen in humans exposed to Phlebotomus Papatasi bites and established that humans exposed to P. perniciosus bites do not recognize it. Methodology/Principal Findings Herein, we have validated, in a large cohort of 522 individuals, the use of the Phlebotomus Papatasi recombinant salivary protein PpSP32 (rPpSP32) as an alternative method for testing exposure to the bite of this sand fly. We also demonstrated that screening for total anti-rPpSP32 IgG antibodies is sufficient, being comparable in efficacy to the screening for IgG2, IgG4 and IgE antibodies against rPpSP32. Additionally, sera obtained from dogs immunized with saliva of P. perniciosus, a sympatric and widely distributed sand fly in Tunisia, did not recognize rPpSP32 demonstrating its suitability as a marker of exposure to P. Papatasi saliva. Conclusions/Significance Our data indicate that rPpSP32 constitutes a useful epidemiological tool to monitor the spatial distribution of P. Papatasi in a particular region, to direct control measures against zoonotic cutaneous leishmaniasis, to assess the efficiency of vector control interventions and perhaps to assess the risk of contracting the disease.
-
Seasonal and Physiological Variations of Phlebotomus Papatasi Salivary Gland Antigens in Central Iran
Tehran University of Medical Sciences, 2015Co-Authors: Nasibeh Hosseini-vasoukolaei, Jesus G Valenzuela, Shaden Kamhawi, Ali Khamesipour, Ahmad-reza Mahmoudi, Mohammad Hossein Arandian, Mohammad Reza Yaghoobi-ershadi, H. Mirhendi, Shaghayegh Emami, Zahra SaeidiAbstract:Background: Sand fly saliva helps parasite establishment and induce immune responses in vertebrate hosts. In the current study, we investigated the modulation of Phlebotomus Papatasi salivary gland antigen expression by seasonal and biological factors.Methods: Sand flies were grouped according to physiological stages such as unfed, fed, semi-gravid, gravid, parous, nulliparous, infected or non-infected with Leishmania major and based on the season in which they were collected. Salivary gland antigens (SGAs) were analyzed using SDS-PAGE and the antibody response against SGAs in Rhombomys opimus was determined by ELISA and Western blot.Results: The highest protein content was found in the salivary glands of unfed sand flies. The saliva content was higher in parous compared to nulliparous, in summer compared to spring, and in Leishmania-infected compared to non-infected flies. The salivary gland lysate (SGL) electrophoretic pattern variations were observed among sand flies with various physiological stages particularly from 4–9 protein bands of 14–70 kDa. The SGL of unfed and gravid flies had extra protein bands compared to fed and semi-gravid sand flies. There was missing protein bands in SGL of parous compared to nulliparous; and in summer compared to spring collected flies. Rhombomys opimus serum reacted strongly with an antigenic band of around 28 kDa in the SGL of all sand fly groups.Conclusion: Certain biological and environmental characteristics of wild populations of vector sand flies affect the protein content and antigenicity of saliva. This might have an important implication in the design of vector-based vaccines.
-
Leishmania major Survival in Selective Phlebotomus Papatasi Sand Fly Vector Requires a Specific SCG- Encoded Lipophosphoglycan Galactosylation Pattern
2013Co-Authors: Deborah E. Dobson, Salvatore J. Turco, Shaden Kamhawi, Phillip Lawyer, Stephen M. Beverley, David L SacksAbstract:Phlebotomine sand flies that transmit the protozoan parasite Leishmania differ greatly in their ability to support different parasite species or strains in the laboratory: while some show considerable selectivity, others are more permissive. In ‘‘selective’ ’ sand flies, Leishmania binding and survival in the fly midgut typically depends upon the abundant promastigote surface adhesin lipophosphoglycan (LPG), which exhibits species- and strain-specific modifications of the dominant phosphoglycan (PG) repeat units. For the ‘‘selective’ ’ fly Phlebotomus Papatasi PpapJ, side chain galactosyl-modifications (scGal) of PG repeats play key roles in parasite binding. We probed the specificity and properties of this scGal-LPG PAMP (Pathogen Associated Molecular Pattern) through studies of natural isolates exhibiting a wide range of galactosylation patterns, and of a panel of isogenic L. major engineered to express similar scGal-LPG diversity by transfection of SCGencoded b1,3-galactosyltransferases with different activities. Surprisingly, both ‘poly-scGal ’ and ‘null-scGal ’ lines survived poorly relative to PpapJ-sympatric L. major FV1 and other ‘mono-scGal ’ lines. However, survival of all lines was equivalent in P. duboscqi, which naturally transmit L. major strains bearing ‘null-scGal’-LPG PAMPs. We then asked whether scGal-LPGmediated interactions were sufficient for PpapJ midgut survival by engineering Leishmania donovani, which normally express unsubstituted LPG, to express a ‘PpapJ-optimal ’ scGal-LPG PAMP. Unexpectedly, these ‘‘L. major FV1-cloaked’ ’ L. donovani-SCG lines remained unable to survive within PpapJ flies. These studies establish that midgut survival of L. major i
-
the characterization of the Phlebotomus Papatasi transcriptome
Insect Molecular Biology, 2013Co-Authors: Jenica L Abrudan, Phillip G Lawyer, Marcelo Ramalhoortigao, Shawn T Oneil, Gwen Stayback, Mariha Wadsworth, Megan Bernard, Doug Shoue, Scott J Emrich, Shaden KamhawiAbstract:As important vectors of human disease, phlebotomine sand flies are of global significance to human health, transmitting several emerging and re-emerging infectious diseases. The most devastating of the sand fly transmitted infections are the leishmaniases, causing significant mortality and morbidity in both the Old and New World. Here we present the first global transcriptome analysis of the Old World vector of cutaneous leishmaniasis, Phlebotomus Papatasi (Scopoli) and compare this transcriptome to that of the New World vector of visceral leishmaniasis, Lutzomyia longipalpis. A normalized cDNA library was constructed using pooled mRNA from Phlebotomus Papatasi larvae, pupae, adult males and females fed sugar, blood, or blood infected with Leishmania major. A total of 47 615 generated sequences was cleaned and assembled into 17 120 unique transcripts. Of the assembled sequences, 50% (8837 sequences) were classified using Gene Ontology (GO) terms. This collection of transcripts is comprehensive, as demonstrated by the high number of different GO categories. An in-depth analysis revealed 245 sequences with putative homology to proteins involved in blood and sugar digestion, immune response and peritrophic matrix formation. Twelve of the novel genes, including one trypsin, two peptidoglycan recognition proteins (PGRP) and nine chymotrypsins, have a higher expression level during larval stages. Two novel chymotrypsins and one novel PGRP are abundantly expressed upon blood feeding. This study will greatly improve the available genomic resources for P. Papatasi and will provide essential information for annotation of the full genome.