Inducible resistance to oxidant stress in the protozoan Leishmania chagasi.
Leishmania sp. protozoa are introduced into a mammalian skin by a sandfly vector, whereupon they encounter increased temperature and toxic oxidants generated during phagocytosis. We studied the effects of 37 degrees C "heat shock" or sublethal menadione, which generates superoxide and hydrogen peroxide, on Leishmania chagasi virulence. Both heat and menadione caused parasites to become more resistant to H(2)O(2)-mediated toxicity. Peroxide resistance was also induced as promastigotes developed in culture from logarithmic to their virulent stationary phase form. Peroxide resistance was not associated with an increase in reduced thiols (trypanothione and glutathione) or increased activity of ornithine decarboxylase, which is rate-limiting in trypanothione synthesis. Membrane lipophosphoglycan increased in size as parasites developed to stationary phase but not after environmental exposures. Instead, parasites underwent a heat shock response upon exposure to heat or sublethal menadione, detected by increased levels of HSP70. Transfection of promastigotes with L. chagasi HSP70 caused a heat-inducible increase in resistance to peroxide, implying it is involved in antioxidant defense. We conclude that leishmania have redundant mechanisms for resisting toxic oxidants. Some are induced during developmental change and others are induced in response to environmental stress.
Veterans Affairs Medical Center, the Departments of Internal Medicine and Microbiology, and the Interdisciplinary Immunology Program, University of Iowa, Iowa City, Iowa 52242, USA.
M A
MA
Miller
S E
SE
McGowan
K R
KR
Gantt
M
M
Champion
S L
SL
Novick
K A
KA
Andersen
C J
CJ
Bacchi
N
N
Yarlett
B E
BE
Britigan
M E
ME
Wilson
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D006028:Glycosphingolipids / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D006861:Hydrogen Peroxide / Q000633:toxicity*; D018314:Leishmania infantum / Q000187:drug effects* / Q000378:metabolism; D009955:Ornithine Decarboxylase / Q000096:biosynthesis; D018384:Oxidative Stress; D013438:Sulfhydryl Compounds / Q000378:metabolism
D000975:Antioxidants; D006028:Glycosphingolipids; D018840:HSP70 Heat-Shock Proteins; D013438:Sulfhydryl Compounds; C008290:lipophosphonoglycan; D006861:Hydrogen Peroxide; D009955:Ornithine Decarboxylase