Author/Authors :
Amani, Jafar Applied Microbiology Research Center - Systems Biology and Poisonings Institute - Baqiyatallah University of Medical Sciences -Tehran, Iran , Fasihi-Ramandi, Mahdi Systems Biology and Poisonings Institute - Baqiyatallah University of Medical Sciences -Tehran, Iran , Behnaz, Nastaran Department of Genetics and Biotechnology - School of Biological Science - Varamin-Pishva Branch - Islamic Azad Universit -, Varamin, Iran , Karizi, Shohreh Zare Department of Genetics and Biotechnology - School of Biological Science - Varamin-Pishva Branch - Islamic Azad Universit -, Varamin, Iran , Nazarian, Shahram Department of Biology - Faculty of Science - Imam Hossain University - Tehran, Iran , Kazemi, Rouhollah , Motamedi, Mohammad Javad
Abstract :
Background: Among all types of malignant diseases, breast cancer has worldwide importance because of the high mortality rate
in women aged fewer than 50 in the developing countries. Identification of immunogenic antigens and the generation of specific
antibodies against cancer cells are the most successful strategies for early detection and effective treatment of breast cancer.
Objectives: In the current study, a chimeric protein consisting of two specific surface antigens, MUC1 and HER2, were used for the
production of chitosan nanoparticles and evaluated as a vaccine candidate.
Methods: The pET-28a expression vector harboring the HER2-MUC1 gene was constructed. Expression of the protein in E. coli BL21
(DE3) was induced using IPTG. The recombinant HER2-MUC1 (HM) protein was purified using a Ni-NTA column and confirmed by
western blotting. Chitosan nanoparticles containing the target protein were prepared and the lymphocytes viability was evaluated
using MTT assay.
Results: The expression of the recombinant protein with molecular weight of 40 kDa was confirmed using SDS-PAGE andWestern
blotting. The electric charge and the size of the nanoparticles were determined and verified by a Zeta Sizer device. The evaluation of
IgG and IgA titration suggested that inducing humoral and mucosalimmuneresponses by administering nanoparticles containing
the chimeric protein. Analysis of cell-mediated immunity showed that the chimeric HM protein could induce specific splenocyte
proliferation in immunized mice.
Conclusions: It seems thatHMnanoparticles can be utilized as a vaccine candidate for inducing the cellular and humoral immune
response against breast cancer.
Keywords :
Nanovaccine , MUC1 , HER2 , Recombinant Antigen , Breast Cancer