Identification of Leishmania major cysteine proteinases as targets of the immune response in humans

Rafati, S; Salmanian, AH; Hashemi, K; Schaff, C; Belli, S; Fasel, N

Identification of Leishmania major cysteine proteinases as targets of the immune response in humans

Rafati, S Salmanian, AH Hashemi, K Schaff, C Belli, S

Fasel, N

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Publication Type:: Journal Article
Citation:: Molecular and Biochemical Parasitology, 2001, 113 (1), pp. 35 - 43
Issue Date:: 2001-03-24

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Field	Value	Language
dc.contributor.author	Rafati, S	en_US
dc.contributor.author	Salmanian, AH	en_US
dc.contributor.author	Hashemi, K	en_US
dc.contributor.author	Schaff, C	en_US
dc.contributor.author	Belli, S https://orcid.org/0000-0003-3845-326X	en_US
dc.contributor.author	Fasel, N	en_US
dc.date.issued	2001-03-24	en_US
dc.identifier.citation	Molecular and Biochemical Parasitology, 2001, 113 (1), pp. 35 - 43	en_US
dc.identifier.issn	0166-6851	en_US
dc.identifier.uri	http://hdl.handle.net/10453/3710
dc.description.abstract	In this study, we report the identification of two parasite polypeptides recognized by human sera of patients infected with Leishmania major. Isolation and sequencing of the two genes encoding these polypeptides revealed that one of the genes is similar to the L. major cathepsin L-like gene family CPB, whereas the other gene codes for the L. major homologue of the cysteine proteinase a (CPA) of L. mexicana. By restriction enzyme digestion of genomic DNA, we show that the CPB gene is present in multiple copies in contrast to the cysteine proteinase CPA gene which could be unique. Specific antibodies directed against the mature regions of both types expressed in Escherichia coli were used to analyze the expression of these polypeptides in different stages of the parasite's life cycle. Polypeptides of 27 and 40 kDa in size, corresponding to CPA and CPB respectively, were detected at higher level in amastigotes than in stationary phase promastigotes. Purified recombinant CPs were also used to examine the presence of specific antibodies in sera from either recovered or active cases of cutaneous leishmaniasis patients. Unlike sera from healthy uninfected controls, all the sera reacted with recombinant CPA and CPB. This finding indicates that individuals having recovered from cutaneous leishmaniasis or with clinically apparent disease have humoral responses to cysteine proteinases demonstrating the importance of these proteinases as targets of the immune response and also their potential use for serodiagnosis. © 2001 Elsevier Science B.V.	en_US
dc.relation.ispartof	Molecular and Biochemical Parasitology	en_US
dc.relation.isbasedon	10.1016/S0166-6851(00)00377-7	en_US
dc.subject.classification	Mycology & Parasitology	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mice, Inbred BALB C	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Mice	en_US
dc.subject.mesh	Leishmania major	en_US
dc.subject.mesh	Leishmaniasis, Cutaneous	en_US
dc.subject.mesh	Cathepsins	en_US
dc.subject.mesh	Cysteine Endopeptidases	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Immune Sera	en_US
dc.subject.mesh	Antibodies, Protozoan	en_US
dc.subject.mesh	Antigens, Protozoan	en_US
dc.subject.mesh	Immunoblotting	en_US
dc.subject.mesh	Blotting, Southern	en_US
dc.subject.mesh	Cloning, Molecular	en_US
dc.subject.mesh	Genome, Protozoan	en_US
dc.subject.mesh	Multigene Family	en_US
dc.subject.mesh	Molecular Sequence Data	en_US
dc.title	Identification of Leishmania major cysteine proteinases as targets of the immune response in humans	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	113	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	closed_access
pubs.issue	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	113	en_US

Abstract:

In this study, we report the identification of two parasite polypeptides recognized by human sera of patients infected with Leishmania major. Isolation and sequencing of the two genes encoding these polypeptides revealed that one of the genes is similar to the L. major cathepsin L-like gene family CPB, whereas the other gene codes for the L. major homologue of the cysteine proteinase a (CPA) of L. mexicana. By restriction enzyme digestion of genomic DNA, we show that the CPB gene is present in multiple copies in contrast to the cysteine proteinase CPA gene which could be unique. Specific antibodies directed against the mature regions of both types expressed in Escherichia coli were used to analyze the expression of these polypeptides in different stages of the parasite's life cycle. Polypeptides of 27 and 40 kDa in size, corresponding to CPA and CPB respectively, were detected at higher level in amastigotes than in stationary phase promastigotes. Purified recombinant CPs were also used to examine the presence of specific antibodies in sera from either recovered or active cases of cutaneous leishmaniasis patients. Unlike sera from healthy uninfected controls, all the sera reacted with recombinant CPA and CPB. This finding indicates that individuals having recovered from cutaneous leishmaniasis or with clinically apparent disease have humoral responses to cysteine proteinases demonstrating the importance of these proteinases as targets of the immune response and also their potential use for serodiagnosis. © 2001 Elsevier Science B.V.

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http://hdl.handle.net/10453/3710