Ricin-resistant mutants of Leishmania major which express modified lipophosphoglycan remain infective for mice.
Glycosylation variants of the virulent Leishmania major clone V121 were generated by mutagenesis with N-methyl-N-nitroso-N-nitroguanidine and selected using the galactose-specific lectin Ricinus communis II (RCA II). Three mutants, 4B9, 1D1 and 1C12, which failed to bind RCA II, were found to have an altered expression of lipophosphoglycan (LPG), a molecule implicated in the attachment to host macrophages and survival within the phagolysosome. There were differences in the antigenicity, molecular weight and localization of LPG from mutant parasites as compared to V121. Expression of gp63, a surface molecule also implicated in attachment to macrophages, was unaltered. All 3 mutants caused disease when injected into genetically susceptible BALB/c mice but lesions developed at a much slower rate than those caused by the virulent V121 clone. This slow rate of lesion development did not correlate with promastigotes' ability to invade macrophages in vitro. Karyotype analysis showed that there was a reduction in the size of chromosome band number 2 in all 3 mutants. The differences in LPG and chromosome band 2 were retained by mutant clones following passage through mice, suggesting that these phenotypes are stable. Although the mutant parasites were infective and caused lesions, the changed structure of the LPG appeared to influence the virulence of the parasites.
Cappai
R
R
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Morris
L
L
Aebischer
T
T
Bacic
A
A
Curtis
J M
JM
Kelleher
M
M
McLeod
K S
KS
Moody
S F
SF
Osborn
A H
AH
Handman
E
E
D000371:Agglutination; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004351:Drug Resistance / Q000235:genetics; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006028:Glycosphingolipids / Q000096:biosynthesis* / Q000235:genetics; D007621:Karyotyping; D018320:Leishmania major / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D012276:Ricin / Q000494:pharmacology*; D013154:Spleen / Q000469:parasitology; D014774:Virulence / Q000235:genetics
D006028:Glycosphingolipids; C008290:lipophosphonoglycan; D012276:Ricin