Comparative analysis of glycosylated and nonglycosylated filarial homologues of the 20-kilodalton retinol binding protein from Onchocerca volvulus (Ov20).
Ov20 is a structurally novel 20-kDa retinol binding protein secreted by Onchocerca volvulus. Immunological and biological investigation of this protein has been hampered by the inability to maintain O. volvulus in a laboratory setting. In an effort to find a system more amenable to laboratory investigation, we have cloned, sequenced, and expressed cDNA encoding homologues of Ov20 from two closely related filarial species, Brugia malayi (Bm20) and Acanthocheilonema viteae (Av20). Sequence comparisons have highlighted differences in glycosylation of the homologues. We present here an analysis of mouse immune responses to Ov20, Bm20, and Av20. The results suggest a strong genetic restriction in response to native Bm20 that is overcome when recombinant, nonnative material is used. Reactivity of human filarial sera to the three recombinant proteins confirmed previous specificity studies with Ov20 but highlighted important differences in the reactivity patterns of the O. volvulus and B. malayi homologues that may be due to differences in glycosylation patterns. Ov20 is a dominant antigen in infected individuals, while Bm20 is not. The availability of the B. malayi homologue enabled us to use defined murine reagents and inbred strains for genetic analysis of responsiveness in a way that is not possible for Ov20. However, the close sequence similarity between Ov20 and Av20 suggests that the A. viteae model may be more suited to the investigation of the biological functions of Ov20.
Nirmalan
N
N
Department of Biological Sciences, University of Salford, Salford M5 4WT, United Kingdom.
Cordeiro
N J
NJ
Kläger
S L
SL
Bradley
J E
JE
Allen
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D017178:Brugia malayi / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004153:Dipetalonema / Q000235:genetics / Q000276:immunology; D005368:Filariasis / Q000469:parasitology; D006031:Glycosylation; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007114:Immunization; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D017181:Onchocerca volvulus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D009855:Onchocerciasis / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D014958:Wuchereria bancrofti / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; C109308:Ov20 antigen; D011994:Recombinant Proteins; D012177:Retinol-Binding Proteins