Wuchereria bancrofti: cloning and characterization of heat shock protein 70 from the human lymphatic filarial parasite.
Heat shock protein 70 (HSP70) was identified as an immunodominant antigen by screening a Wuchereria bancrofti (Wb) microfilarial cDNA library with pooled Wb-infected sera, with 28% of the immunopositive clones coding for Wb-HSP70. The deduced amino acid sequence showed greater than 97 and 85% identity with HSP70 from filarial nematodes and humans, respectively. Recombinant HSP70 (74 kDa) and a recombinant protein from the C-terminal portion (43 kDa) also reacted with pooled Wb-infected sera, suggesting that the C-terminal region of HSP70 contains at least one antibody epitope. Brugia malayi L3 larvae showed increasing levels of HSP70 with increasing temperatures. Further, a polyclonal mouse anti-Wb-HSP70 antibody had reactivity to the HSP70 of cattle filarial parasite Settaria digitata and to human HSP70 derived from a Hep-2 cell line. Immune reactivity to Wb-HSP70 was strong, with uninfected non-endemic normal sera showing significantly greater reactions than sera from filaria-infected individuals. Both immunodominant self-HSP70 and HSP70 from other microbial infections may be primary targets for developing autoantibodies naturally.
Ravi
V
V
Centre for Biotechnology, Anna University, Chennai, India.
Kubofcik
J
J
Bandopathyaya
S
S
Geetha
M
M
Narayanan
R B
RB
Nutman
T B
TB
Kaliraj
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003429:Cross Reactions; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015723:Gene Library; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006358:Hot Temperature; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D014958:Wuchereria bancrofti / Q000235:genetics / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D018840:HSP70 Heat-Shock Proteins; D007106:Immune Sera; D016056:Immunodominant Epitopes; D011994:Recombinant Proteins