Margutti P, Ortona E, Vaccari S, Barca S, Riganò R, Teggi A, Muhschlegel F, Frosch M, Siracusano A
Laboratorio di Immunologia, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Roma, Italy.
Parasite Immunol. 1999 Sep;21(9):485-92. doi: 10.1046/j.1365-3024.1999.00246.x.
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.
通过用囊性棘球蚴病(CE)患者血清中的免疫球蛋白(Ig)E筛选表达文库,已分离出细粒棘球绦虫的一个cDNA克隆(Eg EF-1β/δ)。根据与延伸因子-1的β或δ亚基的序列同源性鉴定出Eg EF-1β/δ。从该开放阅读框推导的氨基酸序列长244个残基,预测分子量为31 kDa。在高严格条件下的Southern印迹中,Eg EF-1β/δ与细粒棘球绦虫的基因组DNA在4和2.5 Kb的两条带处杂交。在免疫印迹分析中,Eg EF-1β/δ蛋白与CE患者的血清表现出免疫反应性:51.7%的血清含有针对重组蛋白的IgE,41.7%含有IgG,18.3%含有IgG4。我们通过用特异性单克隆抗体进行免疫印迹在原头节和绵羊包虫液中鉴定出Eg EF-1β/δ。在钙化囊肿的CE患者中观察到的针对Eg EF-1β/δ的体液免疫反应百分比高于活动性囊肿患者,这表明原头节退变后该蛋白可能在包虫液中释放,提示该抗原在CE免疫监测中的可能用途。总体而言,这些发现似乎表明Eg EF-1β/δ在CE的过敏性疾病和复杂的宿主-寄生虫关系中起关键作用。