Characterization of Plasmodium yoelii monoclonal antibodies directed against stage-specific sporozoite antigens

Charoenvit, Yupin; Leef, Mary F.; Yuan, Leo; Sedegah, Martha; Beaudoin, R. L.

doi:10.1128/iai.55.3.604-608.1987

Cited by 99 publications

(24 citation statements)

References 20 publications

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“…The HSP70 labelling was used to count EEFs. P. falciparum and P. yoelii sporozoites were stained using the anti‐PfCSP mAb E9 (Stuber et al ., 1990) and the anti‐PyCSP mAb NYS1 (Charoenvit et al ., 1987), respectively, using a double labelling procedure as described (Silvie et al ., 2002). For flow cytometric analysis, cells were detached using a non‐enzymatic solution (Invitrogen), washed and stained with saturating concentrations of primary antibody as indicated.…”

Section: Methodsmentioning

confidence: 99%

Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species

et al. 2006

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SummaryPlasmodium sporozoites can enter host cells by two distinct pathways, either through disruption of the plasma membrane followed by parasite transmigration through cells, or by formation of a parasitophorous vacuole (PV) where the parasite further differentiates into a replicative exo-erythrocytic form (EEF). We now provide evidence that following invasion without PV formation, transmigrating Plasmodium falciparum and Plasmodium yoelii sporozoites can partially develop into EEFs inside hepatocarcinoma cell nuclei. We also found that rodent P. yoelii sporozoites can infect both mouse and human hepatocytes, while human P. falciparum sporozoites infect human but not mouse hepatocytes. We have previously reported that the host tetraspanin CD81 is required for PV formation by P. falciparum and P. yoelii sporozoites. Here we show that expression of human CD81 in CD81-knockout mouse hepatocytes is sufficient to confer susceptibility to P. yoelii but not P. falciparum sporozoite infection, showing that the narrow P. falciparum host tropism does not rely on CD81 only. Also, expression of CD81 in a human hepatocarcinoma cell line is sufficient to promote the formation of a PV by P. yoelii but not P. falciparum sporozoites. These results highlight critical differences between P. yoelii and P. falciparum sporozoite infection, and suggest that in addition to CD81, other molecules are specifically required for PV formation during infection by the human malaria parasite.

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Section: Methodsmentioning

confidence: 99%

Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species

et al. 2006

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“…Liver cryosections were treated with 1% NaBH 4 (Sigma) in PBS to reduce autofluorescence. P. yoelii sporozoites were labeled with mouse monoclonal antibody (mAb) NYS1 against the P. yoelii CS protein 46 followed by goat anti-mouse IgG conjugated to FITC (GAM-FITC; Boehringer). The liver tissue was counterstained with 0.1% Evans Blue (Fisher) in PBS.…”

Section: Methodsmentioning

confidence: 99%

Malaria Sporozoites Actively Enter and Pass Through Rat Kupffer Cells Prior to Hepatocyte Invasion

2001

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Malaria sporozoites have to cross the layer of sinusoidal liver cells to reach their initial site of multiplication in the mammalian host, the hepatocytes. To determine the sinusoidal cell type sporozoites use for extravasation, endothelia or Kupffer cells, we quantified sporozoite adhesion to and invasion of sinusoidal cells isolated from rat liver. In vitro invasion assays reveal that Plasmodium berghei and P. yoelii sporozoites attach to and enter Kupffer cells, but not sinusoidal endothelia. Unlike hepatocytes and other nonphagocytic cells, which are invaded in vitro only within the first hour of parasite exposure, the number of intracellular sporozoites in Kupffer cells increases for up to 12 hours. By confocal and electron microscopy, sporozoites are enclosed in a vacuole that does not colocalize with lysosomal markers. Inhibition of phagocytosis with gadolinium chloride has no effect on Kupffer cell invasion, but abolishes phagocytosis of inactivated sporozoites. Malaria-infected anopheline mosquitoes inoculate Plasmodium sporozoites into the avascular dermis and subcutaneous connective tissue of the mammalian host. 1 Within minutes after entering a blood capillary at the mosquito bite site, the sporozoites travel in the bloodstream to the liver where they invade their initial target cells, the hepatocytes. 2 The arrest of the sporozoites in the liver sinusoid is thought to be mediated by the specific binding of the major Plasmodium sporozoite surface proteins, the circumsporozoite (CS) protein and the thrombospondin-related adhesive protein, to the unique heparan sulfate proteoglycans (HSPGs) of the liver. 3-5 For example, recombinant CS protein binds to the HSPGs on the basolateral cell surface of hepatocytes in the space of Disse. This interaction occurs between the conserved regions I and II-plus of the CS protein and highly sulfated, heparin-like oligosaccharides in heparan sulfate. 6 Furthermore, recombinant CS protein is selectively removed from the bloodstream by binding to liver HSPGs, 3,6,7 a clearance mechanism also used by lipoprotein remnants. [8][9][10][11] Since P. berghei sporozoites compete with ␤ very low density lipoprotein for binding to HSPGs, 12 it appears that malaria sporozoites utilize an existing clearance mechanism of the host for targeting the liver sinusoid.After the initial arrest in the liver sinusoid, Plasmodium sporozoites have to cross the sinusoidal cell layer prior to invasion of hepatocytes. The sinusoid is composed of fenestrated endothelial cells and interspersed with Kupffer cells. Kupffer cells are resident macrophages of the liver that are strategically positioned in the sinusoidal lumen and exhibit numerous pseudopodia. To date, the route that sporozoites use for extravasation has remained uncertain. [13][14][15] have proposed that the parasites infect hepatocytes directly by squeezing through the endothelial fenestration, although they exceed the maximum pore size of the sieve plates (0.1 m 16 ) by an order of magnitude. However, this event has never been observ...

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“…P. yoelii sporozoites (17XNL, nonlethal strain) were dissected from Anopheles stephensi salivary glands into medium 199 containing 5% normal mouse serum and were used for the induction of infections. Sporozoites isolated by a discontinuous gradient (11) in medium 199 without serum were used in the preparation of antigen slides for an immunofluorescent antibody test (IFAT) (2).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal antibodies of three different immunoglobulin G isotypes produced by immunization with a synthetic peptide or native protein protect mice against challenge with Plasmodium yoelii sporozoites

et al. 1993

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Passive transfer of monoclonal antibodies (MAbs) against malaria circumsporozoite (CS) proteins protects animals against malaria. Active immunization with synthetic or recombinant peptides induces a level of polyclonal antibodies to sporozoites comparable to those found after passive immunization but does not provide comparable protection. In the Plasmodium yoelii system, synthetic or recombinant peptide-induced antibodies have never been shown to protect. The current studies were designed to determine whether immunogen structure (native protein versus synthetic peptide) or immunoglobulin G (IgG) subclass of antibodies was responsible for the absolute differences between protective, passively transferred MAbs and nonprotective, actively induced polyclonal antibodies. In this study we produced two MAbs, QGP-S1 (IgGl) and QGP-S2 (IgG2b), by immunization with a synthetic peptide based on the P. yoeii CS major repeat, (QGPGAP)4, conjugated to keyhole limpet hemocyanin. These MAbs were compared to NYS1 (IgG3), an anti-CS protein MAb previously produced by immunization with irradiated P. yoelii sporozoites, which recognizes (QGP GAP)2. QGP-S1 and QGP-S2 passively transferred protection. However, when compared with NYS1, there was a hierarchy of protection, NYS1 > QGP-S1 > QGP-S2. There was no correlation between antibody level at challenge as determined by immunofluorescent antibody test against sporozoites or enzyme-linked immunosorbent assay against (QGPGAP)2 or apparent antibody avidity for (QGPGAP)2 by sodium thiocyanate elution assay. The data demonstrate that a synthetic peptide can induce protective antibodies and that a specific antibody subclass is not required for protection. Work to determine whether antibody affinity or fine specificity can explain the hierarchy of protection among the MAbs is under way.

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Characterization of Plasmodium yoelii monoclonal antibodies directed against stage-specific sporozoite antigens

Cited by 99 publications

References 20 publications

Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species

Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species

Malaria Sporozoites Actively Enter and Pass Through Rat Kupffer Cells Prior to Hepatocyte Invasion

Monoclonal antibodies of three different immunoglobulin G isotypes produced by immunization with a synthetic peptide or native protein protect mice against challenge with Plasmodium yoelii sporozoites

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