Epitopes of human chorionic gonadotropin and their relationship to immunogenicity and cross-reactivity of beta-chain mutants.
OBJECTIVE
Human chrionic gonadotropin (hCG) is a placental glycoprotein hormone, a heterodimeric molecule, consisting of alpha and beta chains. It induces the synthesis of progesterone, which is essential for the maintenance of the fertilized egg. Antibodies directed against hCG can, therefore, prevent pregnancy and serve as a vaccine. hCG belongs to the glycoprotein hormone family and shares the alpha chain with the other members. The beta chain is a hormone-specific subunit that is unique to hCG, but still possesses 85% amino acid homology with the beta chain of luteinizing hormone (LH), which means that prolonged immunization with hCG produces antibodies that cross-react with LH.


METHODS
We have taken an approach involving the mutation of beta hCG to eliminate cross-reactive epitopes without affecting the natural folding of the polypeptide chain and thus the unique beta hCG-specific epitopes.


RESULTS
Several mutants have been constructed that have maintained the binding to hCG-specific monoclonal antibodies (mAbs) but have lost the ability to bind to a panel of LH cross-reactive mAbs. To investigate the immunogenicity of selected mutants, mice were immunized with expression plasmid DNA, containing the gene for wild-type beta hCG and two mutants: mutant 3, with four amino acid substitutions (68 Arg-->Glu; 74 Arg-->Ser; 75 Gly-->His; 79 Val-->His), and mutant 7, with a single amino acid substitution (68 Arg-->Glu).


CONCLUSIONS
Although both mutants were able to elicit antibody responses in at least some animals, the levels were less than those seen with the wild-type beta hCG DNA, and there seems still to be a residual cross-reactivity with LH. Attempts to improve the immunogenicity of the mutants and to further modify the sequence to remove the cross-reactivity are currently underway.
Porakishvili
N
N
University College London Medical School, Immunology Department, United Kingdom.
Jackson
A M
AM
de Souza
J B
JB
Dalla Chiesa
M
M
Roitt
I M
IM
Delves
P J
PJ
Lund
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000235:genetics* / Q000276:immunology*; D003429:Cross Reactions; D004247:DNA / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007986:Luteinizing Hormone / Q000276:immunology; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D017510:Protein Folding
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D000939:Epitopes; D007986:Luteinizing Hormone; D004247:DNA