RESUMO
Evolutionary relationships among parasites of the subfamily Leishmaniinae, which comprises pathogen agents of leishmaniasis, were inferred based on differential protein expression profiles from mass spectrometry-based quantitative data using the PhyloQuant method. Evolutionary distances following identification and quantification of protein and peptide abundances using Proteome Discoverer and MaxQuant software were estimated for 11 species from six Leishmaniinae genera. Results clustered all dixenous species of the genus Leishmania, subgenera L. (Leishmania), L. (Viannia), and L. (Mundinia), sister to the dixenous species of genera Endotrypanum and Porcisia. Placed basal to the assemblage formed by all these parasites were the species of genera Zelonia, Crithidia, and Leptomonas, so far described as monoxenous of insects although eventually reported from humans. Inferences based on protein expression profiles were congruent with currently established phylogeny using DNA sequences. Our results reinforce PhyloQuant as a valuable approach to infer evolutionary relationships within Leishmaniinae, which is comprised of very tightly related trypanosomatids that are just beginning to be phylogenetically unraveled. In addition to evolutionary history, mapping of species-specific protein expression is paramount to understand differences in infection processes, tissue tropisms, potential to jump from insects to vertebrates including humans, and targets for species-specific diagnostic and drug development.
Assuntos
Leishmania , Filogenia , Trypanosomatina , Leishmania/genética , Leishmania/metabolismo , Leishmania/classificação , Trypanosomatina/genética , Trypanosomatina/metabolismo , Trypanosomatina/classificação , Evolução Molecular , Animais , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Proteômica/métodos , Proteoma/genética , Proteoma/análise , Proteoma/metabolismo , Crithidia/genética , Crithidia/metabolismoRESUMO
We report a refractory and relapsed visceral leishmaniasis case in a male child patient followed from 2016 to 2020, whose clinical isolates from multiple relapses were analyzed at the genome level. To the best of our knowledge, it is the first report that both visceral leishmaniasis and non-ulcerated cutaneous leishmaniasis have concomitantly manifested in the same patient. Importantly, sequence analysis revealed that the patient was co-infected with Leishmania infantum and a Crithidia-related parasite, which was previously found in a fatal case of visceral leishmaniasis from the same endemic region.
Assuntos
Coinfecção , Leishmania infantum , Leishmaniose Cutânea , Leishmaniose Visceral , Criança , Humanos , Masculino , Leishmaniose Visceral/complicações , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/tratamento farmacológico , Leishmania infantum/genética , Brasil/epidemiologia , Coinfecção/diagnóstico , Leishmaniose Cutânea/parasitologia , CrithidiaRESUMO
We studied infection by Trypanosomatidae in bats captured in two areas with different degradation levels in the Atlantic Forest of Rio de Janeiro state: Reserva Ecológica de Guapiaçu (REGUA) and Estação Fiocruz Mata Atlântica (EFMA). Furthermore, we evaluated whether the diversity of trypanosomatids changes according to bat diversity and the different levels of preservation in the region. The results showed no influence of the level of preservation on bat species richness (15 and 14 species, respectively), with similar chiropterofauna and higher abundance of two common fruit-eating bat species in the tropics: Carollia perspicillata and Artibeus lituratus. Of the 181 bat specimens analyzed by LIT/Schneider hemoculture, we detected 24 infected individuals (13%), including one positive Sturnira lilium individual that was also positive by fresh blood examination. Molecular characterization using nested PCR targeting the 18 SSU rRNA-encoding gene fragment showed similar trypanosomatid infection rates in bats from the two areas: 15% in REGUA and 11% in EFMA (p = 0.46). Trypanosoma dionisii was the most frequently detected parasite (54%), followed by T. cruzi DTUs TcI and TcIV and Trypanosoma sp., in Neotropical phyllostomid bats (RNMO63 and RNMO56); mixed infections by T. dionisii/T. cruzi TcIII and T. dionisii/T. cruzi TcI were also observed. The T. cruzi DTUs TcI and TcIV are the genotypes currently involved in cases of acute Chagas disease in Brazil, and T. dionisii was recently found in the heart tissue of an infected child. Surprisingly, we also describe for the first time Crithidia mellificae, a putative monoxenous parasite from insects, infecting a vertebrate host in the Americas. Bats from the Atlantic Forest of Rio de Janeiro state harbor a great diversity of trypanosomatids, maintaining trypanosomatid diversity in this sylvatic environment.
Assuntos
Quirópteros/parasitologia , Crithidia/genética , Crithidia/isolamento & purificação , Trypanosoma/genética , Trypanosoma/isolamento & purificação , Tripanossomíase/veterinária , Animais , Brasil , DNA de Protozoário/genética , Feminino , Florestas , Genótipo , Masculino , Filogenia , Trypanosoma cruzi/genética , Trypanosoma cruzi/isolamento & purificaçãoRESUMO
Trypanosomatids affecting honey bees, Crithidia mellificae and Lotmaria passim, have been poorly studied in South America. We therefore analyzed their presence in Africanized and European honeybees from Uruguay, Argentina and Chile collected between 1990 and 2011 and assessed their association with other bee parasites and pathogens. Crithidia mellificae was not detected while L. passim was wide-spread. This report shows that L. passim has been present in this region at least since 2007 and it infects both Africanized and European honey bees. L. passim infected colonies showed high V. destructor parasitization levels, suggesting an association between them.
Assuntos
Abelhas/parasitologia , Crithidia , Trypanosomatina , Animais , Argentina , Chile , Coinfecção/parasitologia , Crithidia/genética , Crithidia/parasitologia , DNA de Protozoário , DNA Ribossômico , Patologia Molecular , Trypanosomatina/genética , Trypanosomatina/parasitologia , Uruguai , VarroidaeRESUMO
Conservation biology can profit greatly from incorporating a phylogenetic perspective into analyses of patterns and drivers of species extinction risk. We applied such an approach to analyse patterns of bumblebee (Bombus) decline. We assembled a database representing approximately 43% of the circa 260 globally known species, which included species extinction risk assessments following the International Union fo Conservation of Nature Red List categories and criteria, and information on species traits presumably associated with bumblebee decline. We quantified the strength of phylogenetic signal in decline, range size, tongue length and parasite presence. Overall, about one-third of the assessed bumblebees are declining and declining species are not randomly distributed across the Bombus phylogeny. Susceptible species were over-represented in the subgenus Thoracobombus (approx. 64%) and under-represented in the subgenus Pyrobombus (approx. 6%). Phylogenetic logistic regressions revealed that species with small geographical ranges and those in which none of three internal parasites were reported (i.e. Crithidia bombi, Nosema spp. or Locustacarus buchneri) were particularly vulnerable. Bumblebee evolutionary history will be deeply eroded if most species from threatened clades, particularly those stemming from basal nodes, become finally extinct. The habitat of species with restricted distribution should be protected and the importance of pathogen tolerance/resistance as mechanisms to deal with pathogens needs urgent research.
Assuntos
Abelhas/classificação , Abelhas/microbiologia , Extinção Biológica , Filogenia , Animais , Crithidia/patogenicidade , Ecossistema , Incidência , Nosema/patogenicidadeRESUMO
An initial survey in 2009 carried out at a site in northwestern Patagonia region, Argentina, revealed for the first time in South America the presence of the flagellate Crithidia bombi and the neogregarine Apicystis bombi, two pathogens associated with the Palaearctic invasive bumble bee Bombus terrestris. In order to determine the long-term persistence and dynamics of this microparasite complex, four additional collections at the same site (San Carlos de Bariloche) were conducted along the following seven years. Both protists were detected in all collections: prevalence was 2%-21.6% for C. bombi and 1.2%-14% for A. bombi. In addition, the microsporidium Nosema bombi was recorded for the first time in the country in the last two collections, at prevalences of 12.4% and 2.4% and unusually high infection intensities (Average = 6.56 × 107 spores per individual). Due to the exceptional dispersal ability of the exotic B. terrestris, these three multihost pathogens should be considered as potential threats to South American native bumble bees.
Assuntos
Apicomplexa/isolamento & purificação , Abelhas/microbiologia , Abelhas/parasitologia , Crithidia/isolamento & purificação , Nosema/isolamento & purificação , Animais , Argentina , PrevalênciaRESUMO
Tripanosomatídeos (família Trypanosomatidae) são organismos unicelulares distribuídos mundialmente infectando invertebrados e vertebrados e incluem parasitos dos gêneros Trypanosoma e Leishmania, de grande importância em saúde pública. Os morcegos estão entre os hospedeiros mais antigos destes parasitos e constituem o único grupo de mamíferos com capacidade de voo. A grande capacidade de mobilidade, a longevidade, o hábito alimentar bastante eclético na maioria das espécies e a adaptação aos diversos ambientes, inclusive os sinantrópicos, fazem destes animais potenciais reservatórios (e dispersores) de espécies de tripanosomatídeos. O presente trabalho teve como objetivo avaliar as taxas de infecção por tripanosomatídeos em morcegos provenientes de áreas com diferentes níveis de preservação na Mata Atlântica do Rio de Janeiro. Morcegos foram capturados, utilizando redes de neblina, na Reserva Ecológica de Guapiaçu (REGUA), uma área com alto nível de preservação, e na Estação Biológica Fiocruz da Mata Atlântica (EFMA), que possui elevado grau de interferência antrópica. Nos 181 morcegos capturados, foram realizados: (i) exame direto em sangue; (ii) cultura de sangue e fragmentos de pele, baço e fígado; e (iii) diagnóstico molecular frente a infecção por Leishmania sp. utilizando como alvo um fragmento de kDNA destes parasitos.
Nossos resultados mostraram que a REGUA e a EFMA possuem riqueza de espécies de morcegos similar (15 e 14 espécies, respectivamente). As espécies mais comuns em ambas as áreas foram Carollia perspicillata (n=54 e 29), e Artibeus lituratus (n=18 e 16), além de Sturnira lilium, também abundante na REGUA (n=18). Dos 181 morcegos analisados, 24 (13%) foram positivos no hemocultivo (um deles também no exame a fresco), mas nenhuma cultura de outros tecidos foi positiva. O sequenciamento e análise filogenética utilizando o alvo 18S SSU revelou infecção por T. dionisii em 13 indivíduos (54%), sendo dois destes com infecção mista por T. cruzi. Outros 3 morcegos apresentaram infecção simples por T. cruzi e 4 apresentaram sequências similares a outras de tripanosomas de morcegos neotropicais ainda não identificados. Pela primeira vez detectamos a presença de Crithidia mellificae, um parasito monoxeno, infectando um mamífero, o morcego nectarívoro Anoura caudifer. A taxa de infecção por Leishmania em ambas as áreas foi similar (20% e 15%), sendo a infecção diagnosticada predominantemente em pele (14,4%). A fauna de quirópteros e a diversidade dos tripanosomatídeos a eles associados não diferiram significativamente entre as áreas, reforçando a grande capacidade que esses animais têm de adaptar-se aos ambientes com diferentes níveis de antropização. (AU)
Assuntos
Animais , Trypanosoma , Quirópteros , Crithidia , LeishmaniaRESUMO
Bumblebees are highly valued for their pollination services in natural ecosystems as well as for agricultural crops. These precious pollinators are known to be declining worldwide, and one major factor contributing to this decline are infections by parasites. Knowledge about parasites in wild bumblebee populations is thus of paramount importance for conservation purposes. We here report the geographical distribution of Crithidia and Nosema, two common parasites of bumblebees, in a yet poorly investigated country: Mexico. Based on sequence divergence of the Cytochrome b and Glycosomal glyceraldehyde phosphate deshydrogenase (gGPDAH) genes, we discovered the presence of a new Crithidia species, which is mainly distributed in the southern half of the country. It is placed by Bayesian inference as a sister species to C. bombi. We suggest the name Crithidia mexicana for this newly discovered organism. A population of C. expoeki was encountered concentrated on the flanks of the dormant volcanic mountain, Iztaccihuatl, and microsatellite data showed evidence of a bottleneck in this population. This study is the first to provide a large-scale insight into the health status of endemic bumblebees in Mexico, based on a large sample size (n=3,285 bees examined) over a variety of host species and habitats.
Assuntos
Abelhas/parasitologia , Crithidia/fisiologia , Nosema/fisiologia , Animais , Teorema de Bayes , Conservação dos Recursos Naturais , Crithidia/genética , DNA de Protozoário/química , Especificidade de Hospedeiro , Interações Hospedeiro-Parasita , México , Repetições de Microssatélites , Nosema/genética , Filogenia , Densidade DemográficaRESUMO
We investigated whether ELISA using crude antigens from insect and plant trypanosomatids, which are non-pathogenic and easily cultivated in large scale, has the same positivity data as Leishmania (Leishmania) chagasi, the etiological agent of human visceral leishmaniasis (VL) or canine leishmaniasis (CanL), or as Trypanosoma cruzi, the etiological agent of Chagas disease (CD). The antigens from Crithidia fasciculata, Crithidia luciliae, and Leptomonas seymouri showed 100% cross-reactivity with VL and CanL samples, with no statistically titers differences from L. (L.) chagasi, however, 34% (17/50) of VL samples revealed higher titers using the insect trypanosomatids than the homologous antigen. On the other hand, antigens from Strigomonas culicis, Angomonas deanei, and Phytomonas serpens showed low cross-reactivity with VL and CanL samples. The sera from patients with American tegumentary leishmaniasis showed low levels of cross-reactivity with all trypanosomatids investigated, even with L. (L) chagasi, without titers dissimilarity among them. These parasites were also worthless as antigen source for detection of CD cases, which required homologous antigens to reach 100% positivity. This study showed, by ELISA, that crude extract of Crithidia and Leptomonas have epitopes similar to L. (L.) chagasi, which supports the idea of using them as antigens source for the serodiagnosis of visceral leishmaniasis.
Assuntos
Antígenos de Protozoários/imunologia , Crithidia/imunologia , Epitopos/imunologia , Leishmaniose Visceral/diagnóstico , Trypanosoma cruzi/imunologia , Trypanosomatina/imunologia , Animais , Antígenos de Protozoários/química , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Crithidia/química , Reações Cruzadas , Cães , Epitopos/química , Humanos , Soros Imunes/química , Leishmania donovani/química , Leishmania donovani/imunologia , Leishmania mexicana/química , Leishmania mexicana/imunologia , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/parasitologia , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Masculino , Trypanosoma cruzi/química , Trypanosomatina/químicaRESUMO
The Palaearctic Bombus ruderatus (in 1982/1983) and Bombus terrestris (1998) have both been introduced into South America (Chile) for pollination purposes. We here report on the results of sampling campaigns in 2004, and 2010-2012 showing that both species have established and massively expanded their range. Bombus terrestris, in particular, has spread by some 200 km year(-1) and had reached the Atlantic coast in Argentina by the end of 2011. Both species, and especially B. terrestris, are infected by protozoan parasites that seem to spread along with the imported hosts and spillover to native species. Genetic analyses by polymorphic microsatellite loci suggest that the host population of B. terrestris is genetically diverse, as expected from a large invading founder population, and structured through isolation by distance. Genetically, the populations of the trypanosomatid parasite, Crithidia bombi, sampled in 2004 are less diverse, and distinct from the ones sampled later. Current C. bombi populations are highly heterozygous and also structured through isolation by distance correlating with the genetic distances of B. terrestris, suggesting the latter's expansion to be a main structuring factor for the parasite. Remarkably, wherever B. terrestris spreads, the native Bombus dahlbomii disappears although the reasons remain unclear. Our ecological and genetic data suggest a major invasion event that is currently unfolding in southern South America with disastrous consequences for the native bumblebee species.
Assuntos
Distribuição Animal , Abelhas/fisiologia , Abelhas/parasitologia , Crithidia/isolamento & purificação , Espécies Introduzidas , Nosema/isolamento & purificação , Animais , Argentina , Chile , Crithidia/genética , DNA de Protozoário/genética , Interações Hospedeiro-Parasita , Dados de Sequência Molecular , Nosema/genética , Filogenia , Análise de Sequência de DNARESUMO
Some trypanosomatids, such as Angomonas deanei formerly named as Crithidia deanei, present an obligatory intracellular bacterium, which maintains a mutualistic relationship with the host. Phosphatidylcholine (PC) is the major phospholipid in eukaryotes and an essential component of cell membranes playing structural, biochemical, and physiological roles. However, in prokaryotes, PC is present only in those species closely associated with eukaryotes, either in symbiotic or pathogenic interactions. In trypanosomatids, the endosymbiont envelope is composed by a reduced cell wall and by two membrane units that lack sterols and present cardiolipin (CL) and PC as the major phospholipids. In this study, we tested the effects of miltefosine in A. deanei proliferation, as well as, on the ultrastrucuture and phospholipid composition considering that this drug inhibits the CTP-phosphocholine cytidyltransferase (CCT), a key enzyme in the PC biosynthesis. Besides the low effect of miltefosine in cellular proliferation, treated protozoa presented ultrastructural alterations such as plasma membrane shedding and blebbing, mitochondrial swelling, and convolutions of the endosymbiont envelope. The use of (32) Pi as a tracer revealed that the production of PC, CL, and phosphatidylethanolamine decreased while phosphatidylinositol production remained stable. Mitochondrion and symbiont fractions obtained from protozoa treated with miltefosine also presented a decrease in phospholipid production, reinforcing the idea that an intensive metabolic exchange occurs between the host trypanosomatid and structures of symbiotic origin.
Assuntos
Crithidia/efeitos dos fármacos , Crithidia/microbiologia , Fosforilcolina/análogos & derivados , Simbiose , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Colina-Fosfato Citidililtransferase/metabolismo , Crithidia/metabolismo , Crithidia/ultraestrutura , Microscopia Eletrônica de Transmissão , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/ultraestrutura , Fosfatidilcolinas/biossíntese , Isótopos de Fósforo/metabolismo , Fosforilcolina/farmacologiaRESUMO
Crithidia deanei is a trypanosomatid protozoan that harbours a symbiotic bacterium. The partners maintain a mutualistic relationship, thus constituting an excellent model for studying metabolic exchanges between the host and the symbiont, the origin of organelles and cellular evolution. According to molecular analysis, symbionts of different trypanosomatid species share high identity and descend from a common ancestor, a ß-proteobacterium of the genus Bordetella. The endosymbiont is surrounded by two membranes, like Gram-negative bacteria, but its envelope presents special features, since phosphatidylcholine is a major membrane component and the peptidoglycan layer is highly reduced, as described in other obligate intracellular bacteria. Like the process that generated mitochondria and plastids, the endosymbiosis in trypanosomatids depends on pathways that facilitate the intensive metabolic exchanges between the bacterium and the host protozoan. A search of the annotated symbiont genome database identified one sequence with identity to porin-encoding genes of the genus Bordetella. Considering that the symbiont outer membrane has a great accessibility to cytoplasm host factors, it was important to characterize this single porin-like protein using biochemical, molecular, computational and ultrastructural approaches. Antiserum against the recombinant porin-like molecule revealed that it is mainly located in the symbiont envelope. Secondary structure analysis and comparative modelling predicted the protein 3D structure as an 18-domain ß-barrel, which is consistent with porin channels. Electrophysiological measurements showed that the porin displays a slight preference for cations over anions. Taken together, the data presented herein suggest that the C. deanei endosymbiont porin is phylogenetically and structurally similar to those described in Gram-negative bacteria, representing a diffusion channel that might contribute to the exchange of nutrients and metabolic precursors between the symbiont and its host cell.
Assuntos
Bactérias/metabolismo , Proteínas de Bactérias/química , Crithidia/microbiologia , Porinas/química , Simbiose , Sequência de Aminoácidos , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Fenômenos Fisiológicos Bacterianos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Crithidia/fisiologia , Dados de Sequência Molecular , Filogenia , Porinas/genética , Porinas/metabolismo , Alinhamento de SequênciaRESUMO
In trypanosomatids, cell division involves morphological changes and requires coordinated replication and segregation of the nucleus, kinetoplast and flagellum. In endosymbiont-containing trypanosomatids, like Crithidia deanei, this process is more complex, as each daughter cell contains only a single symbiotic bacterium, indicating that the prokaryote must replicate synchronically with the host protozoan. In this study, we used light and electron microscopy combined with three-dimensional reconstruction approaches to observe the endosymbiont shape and division during C. deanei cell cycle. We found that the bacterium replicates before the basal body and kinetoplast segregations and that the nucleus is the last organelle to divide, before cytokinesis. In addition, the endosymbiont is usually found close to the host cell nucleus, presenting different shapes during the protozoan cell cycle. Considering that the endosymbiosis in trypanosomatids is a mutualistic relationship, which resembles organelle acquisition during evolution, these findings establish an excellent model for the understanding of mechanisms related with the establishment of organelles in eukaryotic cells.
Assuntos
Bactérias/citologia , Divisão Celular , Núcleo Celular/microbiologia , Crithidia/citologia , Crithidia/microbiologia , Simbiose , Bactérias/genética , Fenômenos Fisiológicos Bacterianos , Crithidia/fisiologia , Replicação do DNARESUMO
In trypanosomatids, cell division involves morphological changes and requires coordinated replication and segregation of the nucleus, kinetoplast and flagellum. In endosymbiont-containing trypanosomatids, like Crithidia deanei, this process ismore complex, as each daughter cell contains only a single symbiotic bacterium, indicating that the prokaryote must replicate synchronically with the host protozoan. In this study, we used light and electron microscopy combined with three dimensional reconstruction approaches to observe the endosymbiont shape and division during C. deanei cell cycle. We found that the bacterium replicates before the basal body and kinetoplast segregations and that the nucleus is the last organelle to divide, before cytokinesis. In addition, the endosymbiont is usually found close to the host cell nucleus, presenting different shapes during the protozoan cell cycle. Considering that the endosymbiosis in trypanosomatids is a mutualistic relationship, which resembles organelle acquisition during evolution, these findings establish an excellent model for the understanding of mechanisms related with the establishment of organelles in eukaryotic cells.
Assuntos
Células Eucarióticas/metabolismo , Células Eucarióticas/microbiologia , Células Eucarióticas/ultraestrutura , Organelas/classificação , Organelas/metabolismo , Organelas/microbiologia , Organelas/ultraestrutura , Crithidia/classificação , Crithidia/microbiologia , Microscopia Eletrônica/métodosRESUMO
The flagellate Crithidia bombi and the neogregarine Apicystis bombi have been found in individuals of Bombus terrestris, a Palaearctic species of bumble bee commercially reared and shipped worldwide for pollination services. B. terrestris has recently entered into the northwestern Patagonia region of Argentina from Chile, where it was introduced in 1998. Prevalence was 21.6% for C. bombi and 3.6% for A. bombi (n=111). The pathogens were not detected in 441 bumble bees belonging to five of the eight known Argentine native species (Bombus atratus, Bombus morio, Bombus bellicosus, Bombus opifex, Bombus tucumanus) collected elsewhere in the country. Although the absence of natural occurrence of C. bombi and A. bombi in Argentine native bumble bees cannot be ascertained at present due to the limited surveys performed, it is important to report their detection in invasive B. terrestris. The invasion event is relatively recent and the accompanying pathogens are not species specific within the genus Bombus.
Assuntos
Apicomplexa/isolamento & purificação , Abelhas/parasitologia , Crithidia/isolamento & purificação , Animais , Feminino , MasculinoRESUMO
Renal disease is associated with morbidity and mortality in systemic lupus erythematosus (SLE) and anti-dsDNA antibodies with SLE immunopathogenesis. We investigated the dsDNA antibody profile of 84 Brazilian SLE patients, 27 with lupus nephritis. Thirty-six (39.1%) patients had dsDNA IgG antibodies shown in enzyme-linked immunosorbent assay (454.7 +/- 281.1 WHO units/mL), nine presenting renal disease. The following profile of dsDNA antibodies was demonstrated in Crithidia luciliae test: IgA (seven out of 36; 19.4%), IgG (22 out of 36, 66.1%); IgM (nine out of 36, 25.0%), and IgE (four out of 36, 11.1%). Two or three isotypes of dsDNA antibodies were observed in nine (25.0%) patients, while 11 (30.5%) were seronegative in the C. luciliae test. Patients with dsDNA antibodies had lower serum C3 and C4 when compared with SLE individuals without these immunoglobulins (P < 0.01 and P < 0.001, respectively). There was no association between any dsDNA antibody isotype and lupus kidney disease nor was anti-dsDNA IgM antibody associated with absence of nephritis.
Assuntos
Anticorpos Antinucleares/sangue , DNA/imunologia , Lúpus Eritematoso Sistêmico/etnologia , Lúpus Eritematoso Sistêmico/imunologia , Nefrite Lúpica/etnologia , Nefrite Lúpica/imunologia , Adolescente , Adulto , África/etnologia , Idoso , Animais , Anticorpos Antiprotozoários/metabolismo , Brasil , Crithidia/imunologia , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lúpus Eritematoso Sistêmico/sangue , Nefrite Lúpica/sangue , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Crithidia deanei is an insect trypanosomatid that harbors a bacterial endosymbiont in its cytoplasm. In this work, we have demonstrated the influence of the endosymbiont on the interaction of C. deanei with mammalian fibroblasts, also implicating the surface leishmanolysin-like molecules of C. deanei in this process. The wild strain of C. deanei expressed a higher amount (2-fold) of leishmanolysin-like molecules in the parasite surface than the aposymbiotic strain. The treatment of parasites with anti-leishmanolysin antibodies or the fibroblasts with purified leishmanolysin-like molecules from C. deanei significantly reduced the association index. The aposymbiotic strain of C. deanei presented interaction rates about 2- and 3-fold lower with fibroblasts than the endosymbiont-bearing counterpart after 1 and 2h, respectively. However, the association indexes were similar after 3 and 4h of interaction. Additionally, we observed a 2-fold increase in the association index after 24-96 h of parasite-fibroblast interaction when compared to the interaction process performed for 4h, irrespective to the presence of the endosymbiont, suggesting that fibroblasts support multiplication and survival of C. deanei. Both parasite strains were able to induce fibroblast lysis. Interestingly, the wild strain led to a 2-fold increase in fibroblasts death in comparison to the aposymbiotic strain after 48-96 h. We also showed that both wild and aposymbiotic biotinylated live parasites recognized the same receptor in the fibroblast cells.
Assuntos
Bactérias/patogenicidade , Crithidia/microbiologia , Fibroblastos/parasitologia , Metaloendopeptidases/fisiologia , Simbiose , Células 3T3 , Animais , Crithidia/fisiologia , Eletroforese em Gel de Poliacrilamida , Fibroblastos/metabolismo , Citometria de Fluxo , Interações Hospedeiro-Parasita , Camundongos , Receptores de Superfície Celular/análise , Receptores de Superfície Celular/metabolismoRESUMO
The endosymbiont-bearing trypanosomatids present a typical kDNA arrangement, which is not well characterized. In the majority of trypanosomatids, the kinetoplast forms a bar-like structure containing tightly packed kDNA fibers. On the contrary, in trypanosomatids that harbor an endosymbiotic bacterium, the kDNA fibers are disposed in a looser arrangement that fills the kinetoplast matrix. In order to shed light on the kinetoplast structural organization in these protozoa, we used cytochemical and immunocytological approaches. Our results showed that in endosymbiont-containing species, DNA and basic proteins are distributed not only in the kDNA network, but also in the kinetoflagellar zone (KFZ), which corresponds to the region between the kDNA and the inner mitochondrial membrane nearest the flagellum. The presence of DNA in the KFZ is in accordance with the actual model of kDNA replication, whereas the detection of basic proteins in this region may be related to the basic character of the intramitochondrial filaments found in this area, which are part of the complex that connects the kDNA to the basal body. The kinetoplast structural organization of Bodo sp. was also analyzed, since this protozoan lacks the highly ordered kDNA-packaging characteristic of trypanosomatid and represents an evolutionary ancestral of the Trypanosomatidae family.
Assuntos
DNA de Cinetoplasto/ultraestrutura , DNA de Protozoário/ultraestrutura , Conformação de Ácido Nucleico , Simbiose , Trypanosomatina/ultraestrutura , Animais , Crithidia/microbiologia , Crithidia/ultraestrutura , Evolução Molecular , Flagelos/microbiologia , Flagelos/ultraestrutura , Técnica de Congelamento e Réplica , Histocitoquímica , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Membranas Mitocondriais/microbiologia , Membranas Mitocondriais/ultraestrutura , Trypanosomatina/microbiologiaRESUMO
Crithidia deanei, a monoxenic trypanosomatid, presents an endosymbiotic bacterium in its cytoplasm. Both the protozoan and the bacterium maintain intensive metabolic exchange, resulting in an interesting model to study the coevolution of metabolisms. The relevance of l-proline for the growth of C. deanei and its transport into these cells was studied. Both the endosymbiont-containing (wild) and the endosymbiont-free protozoa (aposymbiont or cured) strains, when grown in medium supplemented with l-proline, reached higher cell densities than those grown in unsupplemented media. We biochemically characterized the uptake of l-proline in both the wild (K(m)=0.153+/-0.022 mM, V(max)=0.239+/-0.011 nmol min(-1) per 4 x 10(7) cells) and the aposymbiont strains (K(m)=0.177+/-0.049 mM, V(max)=0.132+/-0.012 nmol min(-1) per 4 x 10(7) cells). These data suggest a single type of proline transporter whose activity is upregulated by the presence of the symbiotic bacterium. Proline transport was further characterized and was found to be insensitive to the extracellular concentration of Na+, but sensitive to K+ and pH. The abolition of proline uptake by respiratory chain inhibitors and valinomycin indicates that the proline transport in C. deanei is dependent on the plasma membrane K+ gradient.
Assuntos
Crithidia/metabolismo , Crithidia/microbiologia , Prolina/metabolismo , Simbiose , Animais , Antimicina A/análogos & derivados , Antimicina A/farmacologia , Bactérias/metabolismo , Meios de Cultura , DNA Bacteriano/análise , Depressão Química , Concentração de Íons de Hidrogênio , Monensin/farmacologia , Potássio/metabolismo , RNA Ribossômico 16S/análise , Rotenona/farmacologia , Sódio/metabolismo , Temperatura , Fatores de Tempo , Regulação para Cima , Valinomicina/farmacologiaRESUMO
Desde a descoberta dos anticorpos anti DNA fita dupla (DNAafd) em pacientes com LES e sua inclusão nos critérios para classificação do LES pelo colégio Americano de Reumatologia, diferentes técnicas laboratoriais vem sendo utilizadas para sua detecção...Assim, estes resultados sugerem a realização de ambas as técnicas, ELISA e imunofluorescência indireta Crithidia lucilae para a pesquisa sorológica de anticorpos anti DNAfd.