Molecular cloning and characterization of heat shock protein 70 from Trichinella spiralis.
A cDNA encoding heat shock protein 70 of Trichinella spiralis (Ts-Hsp70) was identified by immunoscreening the adult T. spiralis cDNA library with rabbit antisera against T. spiralis adult extracts. The open reading frame of Ts-Hsp70 cDNA encoded a 623-amino acid peptide with a predicted molecular weight of 68.7kDa, which shares a high degree of sequence conservation with HSP70s from other parasites. Recombinant Ts-Hsp70 was expressed in Escherichia coli and purified with nickel column chromatography. Western blot analysis showed that recombinant Ts-Hsp70 could be recognized not only by trichinellosis patient sera, but also by T. spiralis-infected sera from rabbits, swine, and mice. Mice vaccinated with recombinant Ts-Hsp70 formulated with Freund's adjuvant exhibited strong humoral immune responses indicated by high titer of IgG antibody and significant muscle larval reduction (37%) after being challenged with T. spiralis larvae. The present results indicate that Ts-Hsp70 is a possible candidate vaccine against T. spiralis infection.
Wang
Shaohua
S
Department of Parasitology, School of Basic Medical Sciences, Capital Medical University, 10 Xitoutiao, You An Men, Beijing, PR China.
Zhu
Xinping
X
Yang
Yaping
Y
Yang
Jing
J
Gu
Yuan
Y
Wei
Junfei
J
Hao
Ran
R
Boireau
P
P
Cui
Shijuan
S
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009132:Muscles / Q000469:parasitology; D016366:Open Reading Frames; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D013552:Swine; D017160:Trichinella spiralis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D014235:Trichinellosis / Q000276:immunology / Q000517:prevention & control; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology
D000909:Antibodies, Helminth; D018840:HSP70 Heat-Shock Proteins; D015801:Helminth Proteins; D011994:Recombinant Proteins; D022223:Vaccines, Subunit