Plasmodium yoelii and Plasmodium berghei: Isolation of infected erythrocytes from blood by colloidal silica gradient centrifugation

Tosta, Carlos Eduardo; Sedegah, Martha; Henderson, D. C.; Wedderburn, Nina

doi:10.1016/0014-4894(80)90003-x

Cited by 51 publications

(24 citation statements)

References 10 publications

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“…For large scale culture and growth inhibition assay with human serum, 5 mg/ml AlbuMax (Invitrogen) was used in place of 10% human serum. For synchronization, schizont-rich parasites were purified by 63% (v/v) Percoll (Amersham Biosciences) density centrifugation (32) and incubated within 4 h in fresh medium with 3% erythrocyte prior to 5% sorbitol treatment.…”

Section: Methodsmentioning

confidence: 99%

Serine Repeat Antigen (SERA5) Is Predominantly Expressed among the SERA Multigene Family of Plasmodium falciparum, and the Acquired Antibody Titers Correlate with Serum Inhibition of the Parasite Growth

Aoki

Itagaki

et al. 2002

Journal of Biological Chemistry

100

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The Plasmodium falciparum serine repeat antigen (SERA) is one of the blood stage malaria vaccine candidates. The malaria genome project has revealed that SERA is a member of the SERA multigene family consisting of eight SERA homologues clustered on chromosome 2 and one SERA homologue on chromosome 9. Northern blotting and real time quantitative reverse transcription-PCR with five independent parasite strains, including three allelic representative forms of the SERA gene, have shown that all of the SERA homologues are transcribed most actively at trophozoite and schizont stages and that SERA5 (SERA/SERP) is transcribed predominantly among the family. Polyclonal antibodies were raised against recombinant proteins representing the Nterminal portions of four significantly transcribed SERA homologues (SERA3 to -6) in the center of the cluster on chromosome 2. Using these antibodies, indirect immunofluorescence microscopy detected the expression of SE-RA3 to -6, with similar localization, in all trophozoite-and schizont-infected erythrocytes. We have examined 40 sera from Ugandan adults for their antibody reactivity and found that enzyme-linked immunosorbent assay titer against SERA5 N-terminal domain, but not against other SERA proteins, is positively correlated with the inhibition of in vitro parasite growth by individual sera. Our data confirm the usefulness of the N-terminal domain of SERA5 as a promising malaria candidate vaccine.

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Section: Methodsmentioning

confidence: 99%

Serine Repeat Antigen (SERA5) Is Predominantly Expressed among the SERA Multigene Family of Plasmodium falciparum, and the Acquired Antibody Titers Correlate with Serum Inhibition of the Parasite Growth

Aoki

Itagaki

et al. 2002

Journal of Biological Chemistry

100

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“…Each wash consisted of resuspending the erythrocytes in 20 volumes of the medium and centrifugation with removal of the supernatant fluid. Parasitized and non-parasitized red blood cells were separated using percoll density gradient centrifugation according to Tosta et al (1980). After separation, parasitized erythrocytes were lysed and membranes were prepared essentially according to Konigk and Mirtsch (1977).…”

Section: Animals and Infectionmentioning

confidence: 99%

ERYTHROCYTE MEMBRANE‐BOUND ENZYMES IN MASTOMYS NATALENSIS DURING PLASMODIUM BERGHEI INFECTION

Khare

Saxena

Sen

et al. 1984

Aust J Exp Biol Med

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Summary The pattern of activity of certain membrane‐associated enzymes was followed in the erythrocytes of Plasmodium berghei‐infected Mastomys natalensis. Parasitized erythrocytes were separated from non‐parasitized populations by percoll‐density gradient centrifugation. The activity of adenylate cyclase was markedly increased while those of ATPase, acid phosphatase, β‐glucuronidase and N‐acetyl‐β‐D‐glucosaminidase were considerably decreased in the membrane preparations of parasitized erythrocytes as compared to normal erythrocytes. There was a decrease in the activity of ATPase and an increase of adenylate cyclase in the membrane preparations of non‐parasitized erythrocytes. However, other enzymes did not alter to a significant extent in non‐parasitized erythrocytes. Chloroquine (in vitro) stimulated adenylate cyclase, Na+ K+‐ATPase and Ca++Mg++‐ATPase while acetylcholinesterase was significantly inhibited.

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“…However, the data suggested that fluorescence was proportional to DNA content, and that more than 99% of infected cells could be identified by DNA-specific fluorescence and separated from uninfected cells by sorting. This degree of separation cannot be obtained by any other available technique (2,22,24).…”

mentioning

confidence: 97%

“…Methods exist for removal of mammalian leukocytes (WBC) (3,6) and platelets (18) from infected blood to a contamination level of 1% or less. It is not, however, generally posssible to obtain more than a fair separation of the RBC population into its three components-parasitized RBC (pRBC), uninfected immature RBC (imRBC), and uninfected mature RBC (mRBC) by current techniques (2,22,24). Further, in most systems it is not possible to do more than enrich for the pRBC containing the growing stages of the parasite (2,11,14,16,17,24).…”

mentioning

confidence: 99%

“…It is not, however, generally posssible to obtain more than a fair separation of the RBC population into its three components-parasitized RBC (pRBC), uninfected immature RBC (imRBC), and uninfected mature RBC (mRBC) by current techniques (2,22,24). Further, in most systems it is not possible to do more than enrich for the pRBC containing the growing stages of the parasite (2,11,14,16,17,24). The necessity either of achieving better purification or of monitoring cellular heterogeneity and using proper controls in biochemical studies has been stressed (4).…”

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confidence: 99%

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Analysis of malaria parasite‐infected blood by flow cytometry

1983

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The use of flow cytometry in the quantitative analysis of blood from mice infected with Plasmodium vinckei has been studied. Several fluorescent dyes responsive to cell membrane potential were screened and one dye, 3,3'-dimethyloxacarbocyanine (I)ioc1(3)), was chosen for further study. Mature red blood cells (mRBC), immature RBC (imRBC), and parasitized RBC (pRBC) could be recognized and counted in the flow cytometer. When infected blood was separated on a Percoll gradient and fractions analyzed by flow cytometry using DiOC1(3), distinct populations of pRBC were recognized, the frequency of which varied with density. These subpopulations could not be correlated with distinct morphologic stages but varied with the size or age of the growing parasite. Methods combining the use of DiOC1(3) with a DNA specificdye, Hoechst 33342, are discussed as an approach to more complete analysis of the blood of malaria-infected animals.

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Plasmodium yoelii and Plasmodium berghei: Isolation of infected erythrocytes from blood by colloidal silica gradient centrifugation

Cited by 51 publications

References 10 publications

Serine Repeat Antigen (SERA5) Is Predominantly Expressed among the SERA Multigene Family of Plasmodium falciparum, and the Acquired Antibody Titers Correlate with Serum Inhibition of the Parasite Growth

Serine Repeat Antigen (SERA5) Is Predominantly Expressed among the SERA Multigene Family of Plasmodium falciparum, and the Acquired Antibody Titers Correlate with Serum Inhibition of the Parasite Growth

ERYTHROCYTE MEMBRANE‐BOUND ENZYMES IN MASTOMYS NATALENSIS DURING PLASMODIUM BERGHEI INFECTION

Analysis of malaria parasite‐infected blood by flow cytometry

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