Expression and immune recognition of Brugia malayi VAL-1, a homologue of vespid venom allergens and Ancylostoma secreted proteins.
Several important nematode parasites have been found to express members of a gene family variously termed as venom allergen antigen homologue (vah) or Ancylostoma secreted protein (asp). In some cases these products are secreted by infective larval stages and have been suggested to be effective vaccine immunogens. We isolated the corresponding gene from the human filarial nematode, Brugia malayi, by first searching the expressed sequence tag (EST) dataset generated by the Filarial Genome Project and then using gene-specific nondegenerate primers matching the selected gene for PCR, from B. malayi cDNA libraries. We report here the full-length gene sequence, which we have designated as Bm-val-1, for vah/asp-like. The corresponding protein (Bm-VAL-1) contains 232 amino acids in a single homology unit, unlike products from some other species in which there is a tandem repeat. A putative signal sequence is present at the 5' end and there are two potential N-glycosylation sites. Murine antibodies to recombinant Bm-VAL-1 react with a 28 kDa protein in L3 extracts and recombinant Bm-VAL-1 is recognised by murine T cells primed with soluble L3 proteins. Of 82 ESTs corresponding to Bm-val-1, 72 are recorded from the infective larval (L3) stage. However, PCR on the first-strand cDNA from later mammalian stages revealed some expression at most subsequent time points. Over 95% (20/21) of microfilaraemic human filariasis patients are seropositive for antibodies to Bm-VAL-1, with particularly high levels of IgG3 and IgG4 isotypes. The IgG4 subclass may indicate stimulation by adult and/or microfilarial-derived immunogens. The association of Bm-VAL-1 with the infective stage and its recognition by humans exposed to filariasis suggests that further evaluation of this antigen as a vaccine candidate should be performed.
Murray
J
J
Ashworth Laboratories, Institute of Cell, Animal and Population Biology, University of Edinburgh, West Mains Road, EH9 3JT, Edinburgh, UK.
Gregory
W F
WF
Gomez-Escobar
N
N
Atmadja
A K
AK
Maizels
R M
RM
D000328:Adult; D000485:Allergens / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017178:Brugia malayi / Q000235:genetics / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism; D005368:Filariasis / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control; D005849:Gerbillinae; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013601:T-Lymphocytes / Q000276:immunology; D014611:Vaccination; D014612:Vaccines / Q000276:immunology; D014862:Wasp Venoms / Q000235:genetics*
C099548:ASP protein, Ancylostoma caninum; D000485:Allergens; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D014612:Vaccines; D014862:Wasp Venoms; C446035:venom allergen antigen-like protein 1, Brugia malayi