Propagation of Malaria Parasites In Vitro

Siddiqui, Wasim A.; Palmer, Kevin

doi:10.1016/b978-0-12-007901-8.50012-8

Cited by 7 publications

(4 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Because the vector stages that infect the vertebrate host, such as malaria sporozoites or filarial larvae, may possess effective immunogens (Nussenzweig, 1977) new approaches to the in vitro culture of these parasites are urgently needed. Although the mosquito stages of malarial and filarial parasites have been maintained in culture their survival and potential for growth and development have generally been poor (Devaney and Howells, 1979; Siddiqui and Palmer, 1981). In addition, there is no reliable method to obtain viable uncontaminated mosquito tissues or parasites by surface sterilization of pupae or adults, and researchers have resorted to rearing mosquitoes axenically (Boorman, 1967;Rosales-Ronquillo et al, 1973).…”

Section: Embryos Of the Ticks Rhipicephalus Sanguineus And Anocentor mentioning

confidence: 98%

Anopheles stephensi and Toxorhynchites amboinensis: Aseptic Rearing of Mosquito Larvae on Cultured Cells

Munderloh¹,

Kurtti²,

Maramorosch³

1982

The Journal of Parasitology

View full text Add to dashboard Cite

Aseptic larvae of Anopheles stephensi and Toxorhynchites amboinensis were reared on a continuous cell line (RU TAE 12 V) from the mosquito, T. amboinensis, that grew in suspension as multicellular vesicles. Surface-sterilized eggs were hatched in a 24-well plate containing 0.2 ml of Leibovitz's L-15 medium per well and incubated in a humidified atmosphere. Toxorhynchites amboinensis eggs of 36 hr or older were placed singly to assure hatching and avoid cannibalism. Hatching rates were over 80%. All larval instars were maintained in L-15 medium at 28 C with a 12-hr photoperiod. Anopheles stephensi larvae were reared in 25-cm2 tissue culture flasks containing 10 ml of L-15 medium with 30 to 50 first and second instar larvae or 10 third and fourth instar larvae per flask. Toxorhynchites amboinensis larvae remained in the 24-well plate in 1.5 ml of medium through the second instar; third instar larvae were kept in 12-well plates (3 ml of medium per well) and transferred to 25-cm2 flasks (10 ml per flask) when they reached the fourth instar. First and second instar A. stephensi larvae were fed cultured cells once, and third or fourth instar larvae twice a day. Toxorhynchites amboinensis larvae were fed vesicles once during the first 4 days after hatching, and every 1 or 2 days thereafter. Each A. stephensi larva consumed approximately 2 X 10(6) cells, and T. amboinensis larvae 10 times more cells before pupating. Anopheles stephensi pupated after 7 to 8 days and adults emerged during days 9 to 11. Pupation in T. amboinensis began on day 21 after hatching and adults emerged 5 days later. Cell lines isolated from A. stephensi larvae or embryos of the ticks Rhipicephalus sanguineus and Anocentor (Dermacentor) nitens supported only limited growth of A. stephensi larvae. Defibrinated hamster (Mesocricetus auratus) blood, though readily ingested, did not support the growth of A. stephensi whereas larvae reared on blood cells plus T. amboinensis cells showed limited growth.

show abstract

Section: Embryos Of the Ticks Rhipicephalus Sanguineus And Anocentor mentioning

confidence: 98%

Anopheles stephensi and Toxorhynchites amboinensis: Aseptic Rearing of Mosquito Larvae on Cultured Cells

Munderloh¹,

Kurtti²,

Maramorosch³

1982

The Journal of Parasitology

View full text Add to dashboard Cite

show abstract

“…fjlciparun. Uganda-Palo Alto strain, was grown in RPMI-1640 tissue culture medium (GIBCO Laboratories, Grand Island, N.Y.) supplemented with 10% human type O+ serum and 5% hematocrit of human erythrocytes as reported by Siddiqui and Palmer (22). Metabolic labeling of P. falciparum cultures with [3H]isoleucine.…”

Section: Methodsmentioning

confidence: 99%

Plasmodium falciparum: protein antigens identified by analysis of serum samples from vaccinated Aotus monkeys

Kan¹,

Yamaga²,

Kramer³

et al. 1984

Infect Immun

View full text Add to dashboard Cite

Serum samples from Aotus trivirgatus subsp. griseimembra monkeys obtained at different stages of a vaccination experiment were analyzed for total antibody titer to Plasmodium frlciparum and were used for identifying protective antigens of the human malaria parasite. Total malarial antibody titers were higher in serum samples from protected monkeys (vaccinated with antigen in an adjuvant) than in those from unprotected monkeys (vaccinated with either antigen or adjuvant only). Parasite proteins were labeled with [3H]isoleucine, solubilized with nonionic detergent, and reacted with immune Aotus sera. Immunoprecipitates obtained were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Thirteen protein antigen bands in the molecular weight range 73,000 to 180,000 were resolved. Serum samples obtained from protected Aotus monkeys reacted more intensely with these proteins than samples from unprotected monkeys did. Evidence is presented that the protective antigen is not a single, normally nonimmunogenic, protein that is recognized only in protected monkeys. Rather, the present data indicate that a heightened immune response to multiple proteins correlated with in vivo protection to P. falciparum in Aotus monkeys. This finding may have a significant bearing on strategies for the development of a human P. falciparum vaccine.

show abstract

“…The P. falciparum isolates used were Uganda-Palo Alto (FUP; mefloquine resistant) (10), Falciparum Vietnam Oak Knoll (FVO; chloroquine resistant) (29), Indochina (chloroquine resistant) (31), Thailand (chloroquine, quinine, and mefloquine [multidrug] resistant) (32), GHA (sensitive to all tested drugs), and W-2 CDC Indochina III (resistant to chloroquine, quinine, and pyrimethamine but susceptible to mefloquine). Parasites were cultured by standard methods as previously described (30).…”

Section: Methodsmentioning

confidence: 99%

5HT1A Serotonin Receptor Agonists InhibitPlasmodium falciparumby Blocking a MembraneChannel

Locher

Ruben²,

Gut

et al. 2003

Antimicrob Agents Chemother

View full text Add to dashboard Cite

To identify new leads for the treatment of Plasmodium falciparum malaria, we screened a panel of serotonin (5-hydroxytryptamine [5HT]) receptor agonists and antagonists and determined their effects on parasite growth. The 5HT1A receptor agonists 8-hydroxy-N-(di-n-propyl)-aminotetralin (8-OH-DPAT), 2,5-dimethoxy-4-iodoamphetamine, and 2,5-dimethoxy-4-bromophenylethylamine inhibited the growth of P. falciparum in vitro (50% inhibitory concentrations, 0.4, 0.7, and 1.5 M, respectively). In further characterizing the antiparasitic effects of 8-OH-DPAT, we found that this serotonin receptor agonist did not affect the growth of Leishmania infantum, Trypanosoma cruzi, Trypanosoma brucei brucei, or Trichostrongylus colubriformis in vitro and did not demonstrate cytotoxicity against the human lung fibroblast cell line MRC-5. 8-OH-DPAT had similar levels of growth inhibition against several different P. falciparum isolates having distinct chemotherapeutic resistance phenotypes, and its antimalarial effect was additive when it was used in combination with chloroquine against a chloroquine-resistant isolate. In a patch clamp assay, 8-OH-DPAT blocked a P. falciparum surface membrane channel, suggesting that serotonin receptor agonists are a novel class of antimalarials that target a nutrient transport pathway. Since there may be neurological involvement with the use of 8-OH-DPAT and other serotonin receptor agonists in the treatment of falciparum malaria, new lead compounds derived from 8-OH-DPAT will need to be modified to prevent potential neurological side effects. Nevertheless, these results suggest that 8-OH-DPAT is a new lead compound with which to derive novel antimalarial agents and is a useful tool with which to characterize P. falciparum membrane channels.Discovery of drugs for the treatment of malaria is essential because of the widespread resistance of Plasmodium falciparum to chloroquine and other drugs. Drug resistance contributes importantly to the 1 to 2 million deaths caused by malaria every year (25, 37). Natural products from traditional medicinal plants have formed the basis of new synthetic antimalarial analogues with potent activity, and numerous other lead compounds have been identified, including alkaloids, quinones, terpenes, and flavonoids (17,26). During the screening of plant extracts used in traditional Polynesian medicine and other natural products for antiviral and antimicrobial activity (19), we found that some serotonin (5-hydroxytryptamine [5HT]) receptor agonists have antimalarial properties. The activity of these agonists was accompanied by the blocking of a membrane channel on parasitized erythrocytes. Serotonin agonists may therefore be useful for the characterization of the membrane transport properties of the malaria parasite and may provide new lead candidates for the treatment of malaria. MATERIALS AND METHODSGrowth inhibition assays. The P. falciparum isolates used were Uganda-Palo Alto (FUP; mefloquine resistant) (10), Falciparum Vietnam Oak Knoll (FVO; chloroquine resistant) (29),...

show abstract

Propagation of Malaria Parasites In Vitro

Cited by 7 publications

References 64 publications

Anopheles stephensi and Toxorhynchites amboinensis: Aseptic Rearing of Mosquito Larvae on Cultured Cells

Anopheles stephensi and Toxorhynchites amboinensis: Aseptic Rearing of Mosquito Larvae on Cultured Cells

Plasmodium falciparum: protein antigens identified by analysis of serum samples from vaccinated Aotus monkeys

5HT1A Serotonin Receptor Agonists InhibitPlasmodium falciparumby Blocking a MembraneChannel

Contact Info

Product

Resources

About