title: Influence of parasite encoded inhibitors of serine peptidases in early infection of macrophages with Leishmania major.
abstract: Ecotin is a potent inhibitor of family S1A serine peptidases, enzymes lacking in the protozoan parasite Leishmania major. Nevertheless, L. major has three ecotin-like genes, termed inhibitor of serine peptidase (ISP). ISP1 is expressed in vector-borne procyclic and metacyclic promastigotes, whereas ISP2 is also expressed in the mammalian amastigote stage. Recombinant ISP2 inhibited neutrophil elastase, trypsin and chymotrypsin with K(i)s between 7.7 and 83 nM. L. major ISP2-ISP3 double null mutants (Deltaisp2/3) were created. These grew normally as promastigotes, but were internalized by macrophages more efficiently than wild-type parasites due to the upregulation of phagocytosis by a mechanism dependent on serine peptidase activity. Deltaisp2/3 promastigotes transformed to amastigotes, but failed to divide for 48 h. Intracellular multiplication of Deltaisp2/3 was similar to wild-type parasites when serine peptidase inhibitors were present, suggesting that defective intracellular growth results from the lack of serine peptidase inhibition during promastigote uptake. Deltaisp2/3 mutants were more infective than wild-type parasites to BALB/c mice at the early stages of infection, but became equivalent as the infection progressed. These data support the hypothesis that ISPs of L. major target host serine peptidases and influence the early stages of infection of the mammalian host.
authors.affiliation: Glasgow Biomedical Research Centre, Wellcome Centre for Molecular Parasitology and Division of Infection and Immunity, Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8TA, UK.
authors.forename: Sylvain C P
authors.initials: SC
authors.lastname: Eschenlauer
authors.forename: Marilia S
authors.initials: MS
authors.lastname: Faria
authors.forename: Lesley S
authors.initials: LS
authors.lastname: Morrison
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bland
authors.forename: Flavia L
authors.initials: FL
authors.lastname: Ribeiro-Gomes
authors.forename: George A
authors.initials: GA
authors.lastname: DosReis
authors.forename: Graham H
authors.initials: GH
authors.lastname: Coombs
authors.forename: Ana Paula C A
authors.initials: AP
authors.lastname: Lima
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Mottram
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D017353:Gene Deletion; D018320:Leishmania major / Q000276:immunology* / Q000472:pathogenicity*; D016773:Leishmaniasis, Cutaneous / Q000276:immunology / Q000469:parasitology; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors; D008264:Macrophages / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010587:Phagocytosis / Q000276:immunology; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
chemical_list: D015800:Protozoan Proteins; D015842:Serine Proteinase Inhibitors; D002918:Chymotrypsin; D019272:Leukocyte Elastase; D014357:Trypsin