Plasmodium berghei and Plasmodium chabaudi chabaudi: development of simple in vitro erythrocyte invasion assays

Parasitology ◽  
1992 ◽  
Vol 105 (3) ◽  
pp. 355-362 ◽  
Author(s):  
J. McNally ◽  
S. M. O'donovan ◽  
J. P. Dalton
Keyword(s):  
Plasmodium Berghei ◽  
Plasmodium Chabaudi ◽  
Rodent Malaria ◽  
Erythrocyte Invasion ◽  
Malaria Species ◽  
Invasion Assays

SUMMARYErythrocyte invasion assays are described for two species of rodent malaria, namely Plasmodium berghei and P. c. chabaudi. These invasion assays are simple, are carried out using a candle jar and allow a number of assays to be performed simultaneously. Our results demonstrate that both rodent malaria species show an in vitro preference for reticulocytes although the preference of P. c. chabaudi for these cells is not as marked as that of P. berghei. The details of our invasion assays and our results obtained are discussed.

scholarly journals Dissection-independent production of Plasmodium sporozoites from whole mosquitoes

2021 ◽  
Vol 4 (7) ◽  
pp. e202101094
Author(s):  
Joshua Blight ◽  
Katarzyna A Sala ◽  
Erwan Atcheson ◽  
Holger Kramer ◽  
Aadil El-Turabi ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Plasmodium Berghei ◽  
Malaria Vaccine ◽  
Step Method ◽  
Rodent Malaria ◽  
Research Grade ◽  
Parasite Biology ◽  
Malaria Model

Progress towards a protective vaccine against malaria remains slow. To date, only limited protection has been routinely achieved following immunisation with either whole-parasite (sporozoite) or subunit-based vaccines. One major roadblock to vaccine progress, and to pre-erythrocytic parasite biology in general, is the continued reliance on manual salivary gland dissection for sporozoite isolation from infected mosquitoes. Here, we report development of a multi-step method, based on batch processing of homogenised whole mosquitoes, slurry, and density-gradient filtration, which combined with free-flow electrophoresis rapidly produces a pure, infective sporozoite inoculum. Human-infective Plasmodium falciparum and rodent-infective Plasmodium berghei sporozoites produced in this way are two- to threefold more infective than salivary gland dissection sporozoites in in vitro hepatocyte infection assays. In an in vivo rodent malaria model, the same P. berghei sporozoites confer sterile protection from mosquito-bite challenge when immunisation is delivered intravenously or 60–70% protection when delivered intramuscularly. By improving purity, infectivity, and immunogenicity, this method represents a key advancement in capacity to produce research-grade sporozoites, which should impact delivery of a whole-parasite based malaria vaccine at scale in the future.

Development of irreversible lesions in the brain, heart and kidney following acute and chronic murine malaria infection

Parasitology ◽  
1999 ◽  
Vol 119 (6) ◽  
pp. 543-553 ◽  
Author(s):  
P. N. VUONG ◽  
F. RICHARD ◽  
G. SNOUNOU ◽  
F. COQUELIN ◽  
L. RÉNIA ◽  
...  
Keyword(s):  
Drug Treatment ◽  
Tissue Damage ◽  
Malaria Infection ◽  
Control Measures ◽  
Low Grade ◽  
Plasmodium Chabaudi ◽  
Rodent Malaria ◽  
Pathological Lesions ◽  
Malaria Species ◽  
The Brain

Irreversible pathological lesions were noted in the organs of mice infected with 1 of 3 rodent malaria species: Plasmodium chabaudi chabaudi, P. vinckei petteri and P. yoelii nigeriensis at different times during the course of the primary parasitaemia and long after microscopical clearance of the parasites. Moreover, similar lesions were also obtained when parasite levels were kept below 1%by subcurative drug treatment. The frequency and severity of the lesions correlated with the duration of the infection. Accumulation of tissue damage during chronic low-grade malaria infections has implications for the design of control measures.

scholarly journals Transgenic mice expressing human sickle hemoglobin are partially resistant to rodent malaria

Blood ◽  
1993 ◽  
Vol 81 (1) ◽  
pp. 222-226 ◽  
Author(s):  
HL Shear ◽  
EF Jr Roth ◽  
ME Fabry ◽  
FD Costantini ◽  
A Pachnis ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Experimental Model ◽  
Partial Resistance ◽  
Plasmodium Falciparum Malaria ◽  
Plasmodium Chabaudi ◽  
Rodent Malaria ◽  
Sickle Hemoglobin ◽  
Enhanced Resistance

Abstract The polymorphic frequency of the gene for beta s-globin involved in the generation of sickle trait and sickle cell anemia in the human population is caused by the enhanced resistance of sickle trait individuals to Plasmodium falciparum malaria, as supported by epidemiologic and in vitro studies. However, the mechanism for the protective effect of sickle hemoglobin in vivo has not been fully defined. The generation of transgenic mice expressing high levels of human beta s- and alpha-chains has allowed us to study this phenomenon in vivo in an experimental model. We infected the transgenic beta s mice with two species of rodent malaria and found a diminished and delayed increase in parasitemia as compared with controls. This is in contrast to our previous studies involving the introduction of a beta A transgene, which does not alter the infection. The use of this model allowed us to address the question of the mechanism of protection against malaria in mice expressing sickle hemoglobin. We find that splenectomy of transgenic mice completely reverses the protection against Plasmodium chabaudi adami infection. The results reported have shown a relationship between the presence of the beta s gene product and partial resistance to malaria in an experimental model in vivo and shows that the spleen plays an important role in this protection.

scholarly journals In vitro evaluation of the role of the Duffy blood group in erythrocyte invasion by Plasmodium vivax.

1989 ◽  
Vol 169 (5) ◽  
pp. 1795-1802 ◽  
Author(s):  
J W Barnwell ◽  
M E Nichols ◽  
P Rubinstein
Keyword(s):  
Plasmodium Vivax ◽  
Blood Group ◽  
Enzymatic Modification ◽  
Duffy Blood Group ◽  
Alternate Pathway ◽  
Erythrocyte Invasion ◽  
Intact Molecule ◽  
Malaria Species

A short-term in vitro culture system that allows for significant re-invasion of target erythrocytes by Plasmodium vivax was used to study the role of the Duffy blood group antigen as a ligand for merozoite invasion by this human malaria species. Using human Duffy-positive and -negative erythrocytes, various primate erythrocytes, enzymatic modification of erythrocytes, and mAb that defines a new Duffy determinant (Fy6) we conclude that the erythrocyte glycoprotein carrying Duffy determinants is required as a ligand for the invasion of human erythrocytes by P. vivax merozoites. Blockade of invasion by Fab fragments of the anti-Fy6 mAb equal to that of the intact molecule and the correlation of P. vivax susceptibility with the presence of the Fy6 determinant suggests this epitope or a nearby domain may be an active site on the Duffy glycoprotein. However, as for P. knowlesi, there is evidence that an alternate pathway for P. vivax invasion of simian erythrocytes may exist.

scholarly journals An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi

2020 ◽  
Vol 5 ◽  
pp. 71
Author(s):  
Edward J Marr ◽  
Rachel M Milne ◽  
Burcu Anar ◽  
Gareth Girling ◽  
Frank Schwach ◽  
...  
Keyword(s):  
Marker Genes ◽  
Plasmodium Chabaudi ◽  
Chronic Infections ◽  
Selectable Marker Genes ◽  
Erythrocyte Invasion ◽  
Infection Dynamics ◽  
Marker Free ◽  
Parasite Replication

The rodent parasite Plasmodium chabaudi is an important in vivo model of malaria. The ability to produce chronic infections makes it particularly useful for investigating the development of anti-Plasmodium immunity, as well as features associated with parasite virulence during both the acute and chronic phases of infection. P. chabaudi also undergoes asexual maturation (schizogony) and erythrocyte invasion in culture, so offers an experimentally-amenable in vivo to in vitro model for studying gene function and drug activity during parasite replication. To extend the usefulness of this model, we have further optimised transfection protocols and plasmids for P. chabaudi and generated stable, fluorescent lines that are free from drug-selectable marker genes. These mother-lines show the same infection dynamics as wild-type parasites throughout the lifecycle in mice and mosquitoes; furthermore, their virulence can be increased by serial blood passage and reset by mosquito transmission. We have also adapted the large-insert, linear PlasmoGEM vectors that have revolutionised the scale of experimental genetics in another rodent malaria parasite and used these to generate barcoded P. chabaudi gene-deletion and –tagging vectors for transfection in our fluorescent P. chabaudi mother-lines. This produces a tool-kit of P. chabaudi lines, vectors and transfection approaches that will be of broad utility to the research community.

scholarly journals Transgenic mice expressing human sickle hemoglobin are partially resistant to rodent malaria

Blood ◽  
1993 ◽  
Vol 81 (1) ◽  
pp. 222-226 ◽  
Author(s):  
HL Shear ◽  
EF Jr Roth ◽  
ME Fabry ◽  
FD Costantini ◽  
A Pachnis ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Experimental Model ◽  
Partial Resistance ◽  
Plasmodium Falciparum Malaria ◽  
Plasmodium Chabaudi ◽  
Rodent Malaria ◽  
Sickle Hemoglobin ◽  
Enhanced Resistance

The polymorphic frequency of the gene for beta s-globin involved in the generation of sickle trait and sickle cell anemia in the human population is caused by the enhanced resistance of sickle trait individuals to Plasmodium falciparum malaria, as supported by epidemiologic and in vitro studies. However, the mechanism for the protective effect of sickle hemoglobin in vivo has not been fully defined. The generation of transgenic mice expressing high levels of human beta s- and alpha-chains has allowed us to study this phenomenon in vivo in an experimental model. We infected the transgenic beta s mice with two species of rodent malaria and found a diminished and delayed increase in parasitemia as compared with controls. This is in contrast to our previous studies involving the introduction of a beta A transgene, which does not alter the infection. The use of this model allowed us to address the question of the mechanism of protection against malaria in mice expressing sickle hemoglobin. We find that splenectomy of transgenic mice completely reverses the protection against Plasmodium chabaudi adami infection. The results reported have shown a relationship between the presence of the beta s gene product and partial resistance to malaria in an experimental model in vivo and shows that the spleen plays an important role in this protection.

scholarly journals The Plasmodium berghei RC strain is highly diverged and harbors putatively novel drug resistance variants

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3766
Author(s):  
Warangkhana Songsungthong ◽  
Supasak Kulawonganunchai ◽  
Alisa Wilantho ◽  
Sissades Tongsima ◽  
Pongpisid Koonyosying ◽  
...  
Keyword(s):  
Plasmodium Berghei ◽  
Cross Resistance ◽  
Single Nucleotide Variants ◽  
Single Nucleotide ◽  
Rodent Malaria ◽  
Antimalarial Resistance ◽  
Resistance Trait ◽  
Malaria Species ◽  
Carboxyl Terminal ◽  
Novel Drug

Background The current first line drugs for treating uncomplicated malaria are artemisinin (ART) combination therapies. However, Plasmodium falciparum parasites resistant to ART and partner drugs are spreading, which threatens malaria control efforts. Rodent malaria species are useful models for understanding antimalarial resistance, in particular genetic variants responsible for cross resistance to different compounds. Methods The Plasmodium berghei RC strain (PbRC) is described as resistant to different antimalarials, including chloroquine (CQ) and ART. In an attempt to identify the genetic basis for the antimalarial resistance trait in PbRC, its genome was sequenced and compared with five other previously sequenced P. berghei strains. Results We found that PbRC is eight-fold less sensitive to the ART derivative artesunate than the reference strain PbANKA. The genome of PbRC is markedly different from other strains, and 6,974 single nucleotide variants private to PbRC were identified. Among these PbRC private variants, non-synonymous changes were identified in genes known to modulate antimalarial sensitivity in rodent malaria species, including notably the ubiquitin carboxyl-terminal hydrolase 1 gene. However, no variants were found in some genes with strong evidence of association with ART resistance in P. falciparum such as K13 propeller protein. Discussion The variants identified in PbRC provide insight into P. berghei genome diversity and genetic factors that could modulate CQ and ART resistance in Plasmodium spp.

Sign in / Sign up

Export Citation Format

Share Document