Antibody Response to Human Extracellular HER2 Subdomain Proteins in Mice
Message:
Abstract:
Background
In addition to passive immunotherapy using anti-HER2 monoclonal
antibodies, active immunotherapy via HER2 targeting is an interesting approach to
inducing specific anti-tumor immune responses. We have recently reported the
immunogenicity of HER2 subdomains following DNA immunization and HER2 protein
boosting. In the present study, we evaluated the immunogenicity of different HER2
extracellular subdomains for the induction of anti-HER2 antibody response in BALB/c
mice.
Objective
To investigate and characterize antibody responses to human
recombinant proteins of HER2 extracellular subdomains in immunized mice.
Methods
Four subdomains of HER2 extracellular domain were expressed in E.coli; subsequently,
purified recombinant proteins were intraperitoneally injected in BALB/c mice with
Freund's adjuvant. The anti-HER2 antibody response was detected by ELISA,
immunoblotting and flow cytometry.
Results
All the four HER2 subdomains along with the full extracellular domain (fECD) were able to induce specific anti-HER2 antibodies. Although anti-HER2 subdomains antibodies could not react with eukaryotic recombinant fECD protein by ELISA, they were able to recognize this protein by immunoblotting under both reduced and non-reduced conditions. Furthermore, only the sera of mice immunized with fECD protein could recognize native HER2 on HER2 overexpressing tumor cells (>99%) by flow cytometry. Moreover, fECD immunized mice sera inhibited the proliferation of tumor cells by XTT assay.
Conclusion
The prokaryotic recombinant proteins of HER2 extracellular subdomains are immunogenic, yet the induced specific antibodies do not react with the native HER2 protein due to the paucity of post-translation modifications and /or distortion of the native conformation of isolated HER2 extracellular subdomains which might be potentially effective for induction of cell mediated immune response against HER2.
Language:
English
Published:
Iranian journal of immunology, Volume:14 Issue: 2, Spring 2017
Pages:
99 to 110
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