Leishmania kinetoplastid parasites infect millions of people worldwide and have a distinct cellular architecture depending on location in the host or vector and specific pathogenicity functions. An invagination of the cell body membrane at the base of the flagellum, the flagellar pocket (FP), is an iconic kinetoplastid feature, and is central to processes that are critical for Leishmania pathogenicity. The Leishmania FP has a bulbous region posterior to the FP collar and a distal neck region where the FP membrane surrounds the flagellum more closely. The flagellum is attached to one side of the FP neck by the short flagellum attachment zone (FAZ). We addressed whether targeting the FAZ affects FP shape and its function as a platform for host-parasite interactions. Deletion of the FAZ protein, FAZ5, clearly altered FP architecture and had a modest effect in endocytosis but did not compromise cell proliferation in culture. However, FAZ5 deletion had a dramatic impact in vivo: Mutants were unable to develop late-stage infections in sand flies, and parasite burdens in mice were reduced by >97%. Our work demonstrates the importance of the FAZ for FP function and architecture. Moreover, we show that deletion of a single FAZ protein can have a large impact on parasite development and pathogenicity.
- MeSH
- buněčná membrána metabolismus MeSH
- cilie genetika fyziologie ultrastruktura MeSH
- delece genu MeSH
- endocytóza MeSH
- flagella genetika fyziologie ultrastruktura MeSH
- interakce hostitele a parazita MeSH
- Leishmania genetika patogenita fyziologie ultrastruktura MeSH
- mezibuněčné spoje MeSH
- myši MeSH
- protozoální proteiny genetika metabolismus MeSH
- Psychodidae parazitologie MeSH
- virulence genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Parasitic trypanosomatids diverged from free-living kinetoplastid ancestors several hundred million years ago. These parasites are relatively well known, due in part to several unusual cell biological and molecular traits and in part to the significance of a few - pathogenic Leishmania and Trypanosoma species - as aetiological agents of serious neglected tropical diseases. However, the majority of trypanosomatid biodiversity is represented by osmotrophic monoxenous parasites of insects. In two lineages, novymonads and strigomonads, osmotrophic lifestyles are supported by cytoplasmic endosymbionts, providing hosts with macromolecular precursors and vitamins. Here we discuss the two independent origins of endosymbiosis within trypanosomatids and subsequently different evolutionary trajectories that see entrainment vs tolerance of symbiont cell divisions cycles within those of the host. With the potential to inform on the transition to obligate parasitism in the trypanosomatids, interest in the biology and ecology of free-living, phagotrophic kinetoplastids is beginning to enjoy a renaissance. Thus, we take the opportunity to additionally consider the wider relevance of endosymbiosis during kinetoplastid evolution, including the indulged lifestyle and reductive evolution of basal kinetoplastid Perkinsela.
- MeSH
- biodiverzita MeSH
- biologická evoluce * MeSH
- genom protozoální MeSH
- Kinetoplastida genetika MeSH
- Leishmania genetika fyziologie MeSH
- molekulární evoluce MeSH
- symbióza * MeSH
- Trypanosoma genetika fyziologie MeSH
- Trypanosomatina genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
BACKGROUND: Leishmania parasites are transmitted by phlebotomine sand flies and a crucial step in their life-cycle is the binding to the sand fly midgut. Laboratory studies on sand fly competence to Leishmania parasites suggest that the sand flies fall into two groups: several species are termed "specific/restricted" vectors that support the development of one Leishmania species only, while the others belong to so-called "permissive" vectors susceptible to a wide range of Leishmania species. In a previous study we revealed a correlation between specificity vs permissivity of the vector and glycosylation of its midgut proteins. Lutzomyia longipalpis and other four permissive species tested possessed O-linked glycoproteins whereas none were detected in three specific vectors examined. RESULTS: We used a combination of biochemical, molecular and parasitological approaches to characterize biochemical and biological properties of O-linked glycoprotein of Lu. longipalpis. Lectin blotting and mass spectrometry revealed that this molecule with an apparent molecular weight about 45-50 kDa corresponds to a putative 19 kDa protein with unknown function detected in a midgut cDNA library of Lu. longipalpis. We produced a recombinant glycoprotein rLuloG with molecular weight around 45 kDa. Anti-rLuloG antibodies localize the native glycoprotein on epithelial midgut surface of Lu. longipalpis. Although we could not prove involvement of LuloG in Leishmania attachment by blocking the native protein with anti-rLuloG during sand fly infections, we demonstrated strong binding of rLuloG to whole surface of Leishmania promastigotes. CONCLUSIONS: We characterized a novel O-glycoprotein from sand fly Lutzomyia longipalpis. It has mucin-like properties and is localized on the luminal side of the midgut epithelium. Recombinant form of the protein binds to Leishmania parasites in vitro. We propose a role of this molecule in Leishmania attachment to sand fly midgut.
- MeSH
- glykokonjugáty genetika metabolismus MeSH
- hmyz - vektory metabolismus parazitologie MeSH
- hmyzí proteiny genetika metabolismus MeSH
- Leishmania fyziologie MeSH
- muciny genetika metabolismus MeSH
- Psychodidae genetika metabolismus parazitologie MeSH
- trávicí systém metabolismus parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Sand flies remain the only proven vectors of Leishmania spp. but recent implementation of PCR techniques has led to increasing speculation about "alternative vectors", including biting midges. Here, we summarize that PCR has considerable limits for studing the role of bloodsucking arthropods in the epidemiology of leishmaniasis. The Leishmania life cycle in the sand fly includes a complex series of interactions which are in many cases species-specific, the early phase of the infection is, however, non-specific to sand flies. These facts should be considered in detection of Leishmania in ,"alternative" or "new" vectors to avoid mistaken speculation about their vector competence.
Binding of promastigotes to the sand fly midgut epithelium is regarded as an essential part of the Leishmania life cycle in the vector. Among Leishmania surface molecules putatively involved in attachment to the sand fly midgut, two GPI-anchored molecules are the most prominent: lipophosphoglycan (LPG) and promastigote surface protease gp63. In this work, we examined midgut attachment of Leishmania lines mutated in GPI-anchored molecules and compared results from 2 different techniques: in vivo development in sand flies and in vitro competitive binding assays using fluorescently labelled parasites. In combination with previous studies, our data provide additional support for (1) an LPG-independent parasite-binding mechanism of Leishmania major within the midgut of the permissive vector Phlebotomus perniciosus, and provide strong support for (2) the crucial role of L. major LPG in specific vector Phlebotomus papatasi, and (3) a role for Leishmania amazonensis gp63 in Lutzomyia longipalpis midgut binding. Moreover, our results suggest a critical role for GPI-anchored proteins and gp63 in Leishmania mexicana attachment to L. longipalpis midguts, as the wild type (WT) line accounted for over 99% of bound parasites.
- MeSH
- galaktosyltransferasy genetika metabolismus MeSH
- glykokonjugáty genetika metabolismus MeSH
- glykosfingolipidy genetika metabolismus MeSH
- hmyz - vektory parazitologie MeSH
- kompetitivní vazba MeSH
- Leishmania fyziologie MeSH
- lidé MeSH
- metaloendopeptidasy genetika metabolismus MeSH
- mutace MeSH
- Phlebotomus parazitologie MeSH
- protozoální proteiny genetika metabolismus MeSH
- Psychodidae parazitologie MeSH
- stadia vývoje MeSH
- trávicí systém parazitologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The spread of leishmaniasis to areas where it was previously considered nonendemic has been recently found in the New and Old Worlds, and climate changes are suspected as a crucial factor responsible for this spread. Ambient temperature is known to significantly affect the metabolism of sand flies and their developmental times, but little is known about the effect of temperature on the Leishmania life cycle in vectors. This study assesses the effect of temperature on the development of two closely related New World Viannia species, Leishmania braziliensis and Leishmania peruviana, in the permissive vector Lutzomyia longipalpis, and on the development of New and Old World Leishmania infantum in its natural vectors Lu. longipalpis and Phlebotomus perniciosus, respectively. The mountain species L. peruviana developed well in sand fly females kept at 20 degrees C, whereas at 26 degrees C, most infections were lost during the defecation ofbloodmeal remains; this suggests an adaptation to the slower metabolism of sand flies living at lower ambient temperature. On the contrary, L. infantum and L. braziliensis developed well at both temperatures tested; heavy late-stage infections were observed in a majority of sand fly females maintained at 20 degrees C as well 26 degrees C. Frequent fully developed infections of L. infantum and L. braziliensis at 20 degrees C suggest a certain risk of the spread of these two Leishmania species to higher latitudes and altitudes.
- MeSH
- druhová specificita MeSH
- klimatické změny MeSH
- Leishmania braziliensis růst a vývoj fyziologie MeSH
- Leishmania infantum růst a vývoj fyziologie MeSH
- Leishmania růst a vývoj fyziologie MeSH
- Phlebotomus parazitologie MeSH
- Psychodidae parazitologie MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Leishmaniases are vector-borne parasitic diseases with 0.9 - 1.4 million new human cases each year worldwide. In the vectorial part of the life-cycle, Leishmania development is confined to the digestive tract. During the first few days after blood feeding, natural barriers to Leishmania development include secreted proteolytic enzymes, the peritrophic matrix surrounding the ingested blood meal and sand fly immune reactions. As the blood digestion proceeds, parasites need to bind to the midgut epithelium to avoid being excreted with the blood remnant. This binding is strictly stage-dependent as it is a property of nectomonad and leptomonad forms only. While the attachment in specific vectors (P. papatasi, P. duboscqi and P. sergenti) involves lipophosphoglycan (LPG), this Leishmania molecule is not required for parasite attachment in other sand fly species experimentally permissive for various Leishmania. During late-stage infections, large numbers of parasites accumulate in the anterior midgut and produce filamentous proteophosphoglycan creating a gel-like plug physically obstructing the gut. The parasites attached to the stomodeal valve cause damage to the chitin lining and epithelial cells of the valve, interfering with its function and facilitating reflux of parasites from the midgut. Transformation to metacyclic stages highly infective for the vertebrate host is the other prerequisite for effective transmission. Here, we review the current state of knowledge of molecular interactions occurring in all these distinct phases of parasite colonization of the sand fly gut, highlighting recent discoveries in the field.
- MeSH
- gastrointestinální trakt parazitologie MeSH
- hmyz - vektory MeSH
- Leishmania fyziologie MeSH
- Psychodidae parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
BACKGROUND: In Central Asian foci of zoonotic cutaneous leishmaniases, mixed infections of Leishmania turanica and L. major have been found in a reservoir host (the great gerbil, Rhombomys opimus) as well as in the sand fly vector Phlebotomus papatasi, but hybrids between these two Leishmania species have never been reported. In addition, the role of sand fly species other than P. papatasi in L. turanica circulation is not clear. METHODS: In this work we compared the development of L. turanica in three sand fly species belonging to different subgenera. In addition, we studied experimental co-infections of sand flies by both Leishmania species using GFP transfected L. turanica (MRHO/MN/08/BZ18(GFP+)) and RFP transfected L. major (WHOM/IR/-/173-DsRED(RFP+)). The possibility of Leishmania genetic exchange during the vectorial part of the life cycle was studied using flow cytometry combined with immunofluorescent microscopy. RESULTS: Late-stage infections of L. turanica with frequent colonization of the stomodeal valve were observed in the specific vector P. (Phlebotomus) papatasi and in the permissive vector P. (Adlerius) arabicus. On the other hand, in P. sergenti (the specific vector of L. tropica), L. turanica promatigotes were present only until the defecation of bloodmeal remnants. In their natural vector P. papatasi, L. turanica and L. major developed similarly, and the spatiotemporal dynamics of localization in the sand fly gut was the same for both leishmania species. Fluorescence microscopy in combination with FACS analyses did not detect any L. major / L. turanica hybrids in the experimental co-infection of P. papatasi and P. duboscqi. CONCLUSION: Our data provide new insight into the development of different leishmania parasite species during a mixed infection in the sand fly gut. Despite the fact that both Leishmania species developed well in P. papatasi and P. duboscqi and did not outcompete each other, no genetic exchange was found. However, the ability of L. turanica to establish late-stage infections in these specific vectors of L. major suggests that the lipophosphoglycan of this species must be identical or similar to that of L. major.
- MeSH
- barvení a značení metody MeSH
- fluorescenční mikroskopie MeSH
- gastrointestinální trakt parazitologie MeSH
- Leishmania růst a vývoj fyziologie MeSH
- luminescentní proteiny analýza genetika MeSH
- mikrobiální interakce * MeSH
- Phlebotomus parazitologie MeSH
- zelené fluorescenční proteiny analýza genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
