Developmental changes in the expression of Leishmania chagasi gp63 and heat shock protein in a human macrophage cell line.
The ability of the protozoan Leishmania chagasi to infect a vertebrate host depends on its ability to survive intracellularly in a mammalian macrophage. Novel patterns of gene expression are probably important for conversion from the extracellular promastigote to the obligate intracellular amastigote parasite form. We found that the human macrophage-like cell line U937 provided an in vitro model of phagocytosis of L. chagasi promastigotes and intracellular conversion to amastigotes, allowing examination of parasite protein and RNA expression. The Leishmania surface protease gp63 assumed three isoforms during stage conversion, and a 64-kDa form of gp63 not present in promastigotes became the most prominent form in amastigotes. gp63 RNAs derived from the three different classes of msp genes (mspS, mspL, and mspC) were also differentially expressed. Infectious promastigotes contained mRNAs from mspS and mspC genes, whereas converting parasites expressed only mspL and mspC mRNAs. Sequence analysis of clones from an amastigote cDNA library confirmed the presence of gp63 mRNAs only from mspL and mspC class genes in tissue-derived amastigotes. Finally, 24 h after phagocytosis, there was a transient increase in the level of hsp70 and hsp90 proteins that subsequently decreased to baseline; this increase was not due to heat shock alone. We conclude that a unique pattern of selected L. chagasi proteins and RNAs is induced following phagocytosis by macrophages.
Streit
J A
JA
Department of Medicine, University of Iowa, Iowa City 52242, USA.
Donelson
J E
JE
Agey
M W
MW
Wilson
M E
ME
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017125:Genes, Protozoan; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D018314:Leishmania infantum / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D008264:Macrophages / Q000276:immunology / Q000469:parasitology; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D010587:Phagocytosis; D015800:Protozoan Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D016054:DNA, Protozoan; D006360:Heat-Shock Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D008666:Metalloendopeptidases; C055146:glycoprotein gp63, Leishmania