PLASMODIUM-BERGHEI - QUANTITATION OF INVITRO EFFECTS OF ANTIMALARIAL-DRUGS ON EXOERYTHROCYTIC DEVELOPMENT BY A RIBOSOMAL-RNA PROBE

被引:12
作者
LI, J
ZHU, JD
APPIAH, A
MCCUTCHAN, TF
LONG, GW
MILHOUS, WK
HOLLINGDALE, MR
机构
[1] BIOMED RES INST, ROCKVILLE, MD 20852 USA
[2] NIAID, PARASIT DIS LAB, BETHESDA, MD 20892 USA
[3] USN, DEPT INFECT DIS, DIV MALARIA, BETHESDA, MD 20814 USA
[4] WALTER REED ARMY MED CTR, DEPT PARASITOL, DIV EXPTL THERAPEUT, WASHINGTON, DC 20307 USA
关键词
PLASMODIUM-BERGHEI; MALARIA; RIBOSOMAL-RNA PROBE; MALARIAL CHEMOTHERAPY; INVITRO DRUG ASSAY; RIBOSOMAL RNA (RIBOSOMAL-RNA); EXOERYTHROCYTIC (EE); 50-PERCENT INHIBITORY CONCENTRATION (IC50); PRIMAQUINE (PQ) PYRIMETHAMINE (PYR); CHLOROQUINE (CQ); STANDARD SALINE CITRATE (SSC);
D O I
10.1016/0014-4894(91)90091-A
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
A stage-specific ribosomal RNA probe has been used to quantitate exoerythrocytic development of Plasmodium berghei in primary cultures of mouse hepatocytes. Parasite rRNA could be detected as soon as 6 hr after sporozoite invasion and was increased during schizogony to a maximum at 48 hr, when mature schizonts were identified by microscopy. As few as 10 exoerythrocytic schizonts could be detected by filter blot hybridization, followed by autoradiography and liquid scintillation counting. By hybridizing the culture rRNA samples with either parasite-specific or universal rRNA probes, the in vitro tissue schizonticidal activity and hepatotoxicity of primaquine, two of its analogues, and pyrimethamine, could be assessed. After a 48-hr exposure of the culture to serial dilutions of each drug, a quantitative relationship was demonstrated between the decrease of the parasite rRNA and the increase of the drug concentrations. No significant parasite-specific rRNA could be detected at the concentration achieving complete inhibition of schizont formation but causing no cytotoxic effects on host hepatocytes. In contrast to microscopic-based assays, this molecular approach provides an objective and quantitative in vitro method for rapid screening and evaluation of tissue schizonticidal anti-malarials. © 1991.
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页码:450 / 458
页数:9
相关论文
共 21 条
[11]  
KLAUNIG JE, 1981, IN VITRO CELL DEV B, V17, P913
[12]   RIBOSOMAL RNA-BASED DIAGNOSIS OF PLASMODIUM-FALCIPARUM MALARIA [J].
LAL, AA ;
CHANGKASIRI, S ;
HOLLINGDALE, MR ;
MCCUTCHAN, TF .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1989, 36 (01) :67-71
[13]  
LANDAU I, 1984, PRIMAQUINE PHARMACOK, P139
[14]   CULTIVATION OF THE EXOERYTHROCYTIC STAGE OF PLASMODIUM-BERGHEI IN PRIMARY CULTURES OF MOUSE HEPATOCYTES AND CONTINUOUS MOUSE-CELL LINES [J].
LONG, GW ;
LEATH, S ;
SCHUMAN, R ;
HOLLINGDALE, MR ;
BALLOU, WR ;
SIM, BKL ;
HOFFMAN, SL .
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY, 1989, 25 (09) :857-862
[15]   PRIMARY SEQUENCES OF 2 SMALL SUBUNIT RIBOSOMAL-RNA GENES FROM PLASMODIUM-FALCIPARUM [J].
MCCUTCHAN, TF ;
DELACRUZ, VF ;
LAL, AA ;
GUNDERSON, JH ;
ELWOOD, HJ ;
SOGIN, ML .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1988, 28 (01) :63-68
[16]  
MILLET P, 1986, Memorias do Instituto Oswaldo Cruz, V81, P135, DOI 10.1590/S0074-02761986000600022
[17]  
ODUOLA AMJ, 1986, 9TH P INT C INF PAR
[18]   DEVELOPMENTAL REGULATION OF STAGE-SPECIFIC RIBOSOME POPULATIONS IN PLASMODIUM [J].
WATERS, AP ;
SYIN, C ;
MCCUTCHAN, TF .
NATURE, 1989, 342 (6248) :438-440
[19]  
WATERS AP, 1989, LANCET, V1, P1343
[20]  
ZHU JD, 1990, J BIOL CHEM, V265, P12740