Schistosoma mansoni hexokinase: cDNA cloning and immunogenicity studies.
DNA encoding a Schistosoma mansoni hexokinase (SHEX) was amplified from cDNA by the polymerase chain reaction using opposing oligonucleotide primers designed to hybridize with two short segments of hexokinase coding sequences that are well-conserved through evolution. The resulting DNA fragment was then used as a probe to identify a full-length hexokinase cDNA clone. SHEX cDNA encodes a 50-kDa protein that is approximately 46% homologous to rat hexokinase, 40% to rat glucokinase, and 34% to yeast hexokinase A. SHEX coding DNA was expressed within Escherichia coli cells and the 50-kDa recombinant product (rSHEX) was partially purified. Mice repeatedly immunized with rSHEX produced antibodies which recognize rSHEX but this offered no significant protection against subsequent cercarial challenge. On Western blots, rSHEX is weakly recognized by antisera against rat brain hexokinase but not by sera from three strains of mice experimentally infected with S. mansoni parasites or from numerous human schistosomiasis patients. Thus, unlike other reported S. mansoni glycolytic enzymes, hexokinase appears to be poorly immunogenic during schistosome infection and of limited potential as a vaccine candidate.
Shoemaker
C B
CB
Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115.
Reynolds
S R
SR
Wei
G
G
Tielens
A G
AG
Harn
D A
DA
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006593:Hexokinase / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D012555:Schistosomiasis mansoni / Q000276:immunology / Q000517:prevention & control; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014611:Vaccination
D000909:Antibodies, Helminth; D017931:DNA Primers; D018509:DNA, Helminth; D007106:Immune Sera; D012333:RNA, Messenger; D011994:Recombinant Proteins; D006593:Hexokinase