Cloning and expression of a cDNA encoding an elongation factor 1beta/delta protein from Echinococcus granulosus with immunogenic activity.
A cDNA clone (Eg EF-1beta/delta) of Echinococcus granulosus has been isolated by an expression library screened with immunoglobulin (Ig)E of sera from patients with cystic echinococcosis (CE). The Eg EF-1beta/delta was identified on the basis of sequence homology to the subunits beta or delta of the elongation factor-1. The amino acid sequence deduced from this open reading frame is 244 residues long with a predicted molecular mass of 31 kDa. In Southern blot under high stringent condition, Eg EF-1beta/delta hybridized to genomic DNA of E.granulosus at two bands of 4 and 2.5 Kb. In immunoblotting analysis, the Eg EF-1beta/delta protein shows immunological reactivity with sera from CE patients: 51.7% of sera contained IgE, 41.7% IgG and 18.3% IgG4 specific to the recombinant protein. We identify the Eg EF-1beta/delta by immunoblotting with specific monoclonal antibody both in protoscoleces and in sheep hydatid fluid. The higher percentage of humoral immune response against Eg EF-1beta/delta observed in CE patients with calcified cysts than in patients with active cysts indicates the possible release of the protein in the hydatid fluid after protoscoleces degeneration suggesting the possible use of this antigen in the immunosurveillance of CE. Overall, these findings seem to assign to Eg EF-1beta/delta a key role in the allergic disorders and in the complex host-parasite relationship in CE.
Margutti
P
P
Laboratorio di Immunologia, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Roma, Italy.
Ortona
E
E
Vaccari
S
S
Barca
S
S
Riganò
R
R
Teggi
A
A
Muhschlegel
F
F
Frosch
M
M
Siracusano
A
A
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004443:Echinococcosis / Q000097:blood / Q000276:immunology / Q000469:parasitology; D004446:Echinococcus / Q000235:genetics* / Q000254:growth & development / Q000276:immunology* / Q000302:isolation & purification; D015723:Gene Library; D017238:Genes, Helminth; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D012756:Sheep
D000485:Allergens; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D007074:Immunoglobulin G; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D007073:Immunoglobulin E