title: Immunogenicity and contraceptive efficacy of Escherichia coli-expressed recombinant porcine zona pellucida proteins.
abstract: OBJECTIVE
  To overcome availability of the purified native zona pellucida (ZP) glycoproteins for immunocontraception, porcine ZP3, and ZP4 were expressed in E. coli.
  
  
  METHODS
  Purified recombinant proteins were characterized by SDS-PAGE and Western blot, and immunogenicity and contraceptive efficacy determined in FvB/J female mice.
  
  
  RESULTS
  Purified ZP3, ZP3 with promiscuous T-cell epitope of tetanus toxoid, ZP4 and ZP4 incorporating promiscuous T-cell epitope of bovine RNase revealed ~44-, ~49-, ~53-, and ~55-kDa bands by SDS-PAGE and Western blot, respectively. Immunization of female mice with recombinant proteins elicited high antibody titers as well as T-cell responses. Immune sera recognized mouse oocyte ZP and also inhibited in vitro fertilization. Immunized mice showed significant decrease in fertility. Recombinant proteins were able to recall memory antibody response in female mice primed with porcine native ZP.
  
  
  CONCLUSIONS
  Availability of recombinant porcine proteins will be useful in the development of contraceptive vaccine.
authors: Reproductive Cell Biology Laboratory, National Institute of Immunology, New Delhi, India. skgupta@nii.ac.in
authors: Neha
authors: N
authors: Gupta
authors: Kausiki
authors: K
authors: Chakrabarti
authors: Krishna
authors: K
authors: Prakash
authors: Neerja
authors: N
authors: Wadhwa
authors: Tripti
authors: T
authors: Gupta
authors: Satish K
authors: SK
authors: Gupta
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000917:Antibody Formation; D003269:Contraception, Immunologic; D003270:Contraceptive Agents / Q000378:metabolism / Q000494:pharmacology*; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D005260:Female; D005298:Fertility / Q000187:drug effects; D005307:Fertilization in Vitro / Q000187:drug effects; D006801:Humans; D007114:Immunization; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology*; D051379:Mice; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D013552:Swine; D022642:Vaccines, Contraceptive / Q000276:immunology; D015044:Zona Pellucida / Q000276:immunology* / Q000378:metabolism; D000072216:Zona Pellucida Glycoproteins
chemical_list: D000906:Antibodies; D003270:Contraceptive Agents; D004527:Egg Proteins; D029968:Escherichia coli Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D022642:Vaccines, Contraceptive; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse
keywords: Contraceptive vaccine; immunogenicity; in vitro fertilization; promiscuous T-cell epitope; recombinant porcine ZP3/ZP4; zona pellucida proteins