Leishmania (Viannia) braziliensis metacyclic promastigotes purified using Bauhinia purpurea lectin are complement resistant and highly infective for macrophages in vitro and hamsters in vivo

Pinto-da-Silva, Lúcia Helena; Camurate, Marcelle; Costa, Karina A.; Oliveira, Sandra Maria de; Cunha-e-Silva, Narcisa L.; Saraiva, Elvira M.

doi:10.1016/s0020-7519(02)00137-6

Cited by 48 publications

(53 citation statements)

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“…Consequently, for these species, the identification and characterization of metacyclic stage promastigotes have required the establishment of techniques with which to enrich for metacyclic promastigotes. Many of these are lectin-based enrichment techniques that capitalize on characteristics of surface glycosylation that vary between metacyclic and non-metacyclic promastigote stages (Saraiva et al, 1986; Pinto-da-Silva et al, 2002), while others use alternative procedures, including density gradient centrifugation (Spath and Beverley, 2001). The need for such enrichment techniques in L. chagasi are greatly reduced, given that studies of axenic cultured L. chagasi have determined that metacyclic promastigotes comprise a relatively large proportion of the promastigote forms found within cultures at stationary growth phase (Dahlin-Laborde et al, 2005; Yao et al, 2008; Lei et al, 2010).…”

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confidence: 99%

Population Changes in Leishmania chagasi Promastigote Developmental Stages Due to Serial Passage

Lei

Romine

Beetham

2010

Journal of Parasitology

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Leishmania chagasi causes visceral leishmaniasis, a potentially fatal disease of humans. Within the sand fly vector, L. chagasi replicates as promastigotes which undergo complex changes in morphology as they progress from early stage procyclic promastigotes, to intermediate stage leptomonad and nectomonad promastigotes, and ultimately to terminal stage metacyclic promastigotes that are highly infective to vertebrates. This developmental progression is largely recapitulated in vitro using axenic promastigote cultures that have been passaged only a few times. Within a single passage (which takes about a week), axenic cultures progress from logarithmic to stationary growth phases; parasites within those growth phases progress from stages that do not have metacyclic cell properties to ones that do. Interestingly, repeated serial passage of promastigote cultures will result in cell populations that exhibit perturbations in developmental progression, in expression levels of surface macromolecules (major surface protease, MSP, and promastigote surface antigen, PSA), and in virulence properties, including resistance to serum lysis. Experiments were performed to determine whether there exists a direct relationship between promastigote developmental form and perturbations associated with repeated serial passage. Passage 2 to passage 4 L. chagasi cultures at stationary growth phase were predominately (>85%) comprised of metacyclic promastigotes and exhibited high resistance to serum lysis and high levels of MSP and PSA. Serial passaging 8, or more, times resulted in a stationary phase population that was largely (>85%) comprised of nectomonad promastigotes, almost completely devoid (<2%) of metacyclic promastigotes, and that exhibited low resistance to serum lysis and low levels of MSP and PSA. The study suggests that the loss of particular cell properties seen in cells from serially passaged cultures is principally due to a dramatic reduction in the proportion of metacyclic promastigotes. Additionally, the study suggests that serially passaged cultures may be a highly enriched source of nectomonad-stage promastigotes, a stage that has largely been characterized only in mixtures containing other promastigote forms.

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Population Changes in Leishmania chagasi Promastigote Developmental Stages Due to Serial Passage

Lei

Romine

Beetham

2010

Journal of Parasitology

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“…9-O-AcSA is absent in normal erythrocytes, but detection with a lectin agglutination test showed that it can be used as an important biomarker reflecting indirectly the parasite presence (Sharma et al 1998). Metacyclogenesis in an axenic culture of Leishmania braziliensis, which is the causative agent of mucocutaneous leismaniasis in the New World, was characterized by negative agglutination with BPA lectin (Pinto-da-Silva et al 2002).…”

Section: Phaseolus Vulgarismentioning

confidence: 99%

Lectinomics I. Relevance of exogenous plant lectins in biomedical diagnostics

et al. 2009

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This review focuses on utilization of plant lectins as medical diagnostic reagents and tools. The lectin-related diagnostic is aimed at detection of several diseases connected to alteration of the glycosylation profiles of cells and at identification of microbial and viral agents in clinical microbiology. Certain lectins, proposed for or used as diagnostic tools could even recognize those cellular determinants, which are not detected by available antibodies. Broad information is presented on the lectinomics field, illustrating that lectin diagnostics might become practical alternative to antibody-based diagnostic products. In addition, the rising trend of lectin utilization in biomedical diagnostics might initiate a development of innovative methods based on better analytical technologies. Lectin microarray, a rapid and simple methodology, can be viewed as an example for such initiative. This technology could provide simple and efficient screening tools for analysis of glycosylation patterns in biological samples (cellular extracts, tissues and the whole cells), allowing thus personalized detection of changes associated with carbohydrate-related diseases.

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“…Ingested amastigotes quickly differentiate into procyclic stage promastigotes that attach to the midgut wall and, over a period of 1 to several weeks, replicate and progress through a series of developmental stages, ultimately yielding the metacyclic promastigote stage. Metacyclic promastigotes are characterized by their lack of replication, infectivity to mammals, and resistance to lysis by the complement component of serum, while the other promastigote forms are sensitive to lysis by complement (Sacks and Perkins, 1984; Franke et al, 1985; Pinto-da-Silva et al, 2002; Rogers et al, 2002). A naive vertebrate host becomes infected when metacyclic promastigotes are inoculated into the vertebrate by a blood feeding female sandfly that bears a mature infection.…”

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“…promastigotes to CML during development can also be observed during development in vitro culture (Franke et al, 1985; Joshi et al, 1998; Noronha et al, 1998; Pinto-da-Silva et al, 2002). In a specific example of this phenomenon with L. chagasi , we showed that upon exposure to normal human serum (NHS), cultures of promastigotes are CML-sensitive when in the logarithmic growth phase but become CML-resistant upon transition to the stationary growth phase (Lincoln et al, 2004; Dahlin-Laborde et al, 2005, 2008).…”

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Differential Surface Deposition of Complement Proteins on Logarithmic and Stationary PhaseLeishmania chagasiPromastigotes

Ramer‐Tait¹,

Lei²,

Bellaire³

et al. 2012

Journal of Parasitology

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Previous works demonstrated that various species of Leishmania promastigotes exhibit differential sensitivity to complement-mediated lysis (CML) during development. Upon exposure to normal human serum (NHS), cultures of Leishmania chagasi promastigotes recently isolated from infected hamsters (fewer than 5 in vitro passages) are CML-sensitive when in the logarithmic growth phase but become CML-resistant upon transition to the stationary culture phase. Visualization by light and electron microscopy revealed dramatic morphological differences between promastigotes from the 2 culture phases following exposure to NHS. Flow cytometric analysis demonstrated that surface deposition of the complement components C3, C5, and C9 correlated inversely with promastigote CML-resistance. The highest levels of complement protein surface accumulation were observed for logarithmic phase promastigotes, while stationary phase promastigotes adsorbed the least amount of complement proteins. Additionally, fluorescence microscopy revealed that C3 and C5 localized in a fairly uniform pattern to the plasma membrane of promastigotes from logarithmic phase cultures, while the staining of promastigotes from stationary phase cultures was indistinguishable from background. By Western blot analysis, high levels of the complement proteins C3, C5, and C9 were detected in the total lysates of NHS-exposed logarithmic phase L. chagasi promastigotes, relative to NHS-exposed stationary phase promastigotes; this finding indicates that the low levels of C3 and C5 seen on the surface of stationary phase promastigotes were not due to protein uptake/internalization. Together, these data demonstrate the differential deposition of complement proteins on the surfaces of logarithmic and stationary phase L. chagasi promastigotes. The data support a model wherein stationary phase L. chagasi promastigotes resist CML by limiting the deposition of C3 and its derivatives, which, in turn, limit surface levels of complement proteins (including C5 and C9) that form the lytic membrane attack complex.

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Leishmania (Viannia) braziliensis metacyclic promastigotes purified using Bauhinia purpurea lectin are complement resistant and highly infective for macrophages in vitro and hamsters in vivo

Cited by 48 publications

References 30 publications

Population Changes in Leishmania chagasi Promastigote Developmental Stages Due to Serial Passage

Population Changes in Leishmania chagasi Promastigote Developmental Stages Due to Serial Passage

Lectinomics I. Relevance of exogenous plant lectins in biomedical diagnostics

Differential Surface Deposition of Complement Proteins on Logarithmic and Stationary PhaseLeishmania chagasiPromastigotes

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