Cloning, characterization and serological evaluation of K9 and K26: two related hydrophilic antigens of Leishmania chagasi.
We report here the molecular cloning and characterization of two related hydrophilic antigens of Leishmania chagasi. These two antigens have predicted molecular weights of approximately 9 and 26 kDa and detect antibodies in sera of patients with kala-azar (k). Thus, to maintain consistency with nomenclature of the previously described 39 kDa diagnostic antigen of L. chagasi (k39 [1]), these antigens are being referred to as k9 and k26. A significant difference between k9 and k26 is the presence of 11 copies of a 14 amino acid repeat in the open reading frame of k26. The region flanking the repeats of k26 shares a 69% identity with the open reading frame of k9. The recombinant proteins encoded by both antigens are very hydrophilic and show aberrant migration on SDS PAGE. Results of Southern blot analysis reveal that k9 and k26 are conserved to varying degrees among various Leishmania species. Interestingly, the repeat region of k26 is specific to L. chagasi and L. donovani while the flanking region is conserved among several other species. Transcript levels of k26 are significantly upregulated in the amastigote stage of the parasite. Our results show that recombinant K26 is specific in detecting antibodies in infection sera from visceral leishmaniasis (VL) patients. Thus rK26 may complement rK39 in a more accurate diagnosis of VL in the old and the new world.
Bhatia
A
A
Corixa Corporation, Seattle, WA 98104, USA.
Daifalla
N S
NS
Jen
S
S
Badaro
R
R
Reed
S G
SG
Skeiky
Y A
YA
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D017125:Genes, Protozoan; D006801:Humans; D007891:Leishmania / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D018320:Leishmania major / Q000235:genetics / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D017422:Sequence Analysis, DNA
D000953:Antigens, Protozoan; C400962:K26 antigen, Leishmania chagasi; C400961:K9 protein, Leishmania; D011994:Recombinant Proteins