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    Vitamin C in the formulation of inactivated COVID-19 vaccine candidate ‎induced Th1 pattern in the aged mice.‎

  • Nika Asefi ‎,1 Parviz Pakzad ‎,2 Akbar Khorasani ‎,3 Mehdi Mahdavi ‎,4,*
    1. Department of Microbiology, Faculty of Basic Sciences, North Tehran Branch, Islamic Azad ‎University, Tehran, Iran‎
    2. Department of Microbiology, Faculty of Basic Sciences, North Tehran Branch, Islamic Azad ‎University, Tehran, Iran‎
    3. ‎2.‎ Department of FMD vaccine production, Razi Vaccine & Serum Research Institute, ‎Agricultural Research, Education & Extension Organization‏ ‏‎(AREEO) Karaj, Iran ‎
    4. Recombinant Vaccine Research Center, Tehran University of Medical Sciences, Tehran, Iranو Immunotherapy Group, The Institute of Pharmaceutical Science (TIPS), Tehran University of ‎Medical Science, Tehran, Iran


  • Introduction: The morality rate of COVID-19 elevated in elderly people by 22%. Moreover, according to the ‎reports, 90% and 80% of death in patients aged ≥60 years and ≥70 years have recorded. Aging is ‎physiologically accompanied by a reduction in the function of the immune system, which there is ‎a relationship between alteration in the immune system and susceptibility to infections.‎ IFN-γ has a crucial role in the defense against viruses. Since the production of IFN-γ declines in ‎older adults. It may explain the reason of the susceptibility of the elderly to viral infectious ‎diseases such as COVID-19. Studies show the potency of vitamin C in the improvement of ‎immune responses of aged people. Here, we formulated inactivated COVID-19 vaccine in ‎vitamin C and the potency of vaccine assessed in the aged and young mice.‎
  • Methods: A SARS-CoV-2 strain was isolated from a patient and propagated in Vero cell line. The virus ‎was inactivated using formalin and purified using column chromatography method and then ‎quantified by Bradford. In order to formulate in Alum-based vaccine, 2µg of inactivated ‎COVID-19 virus in PBS buffer was admixed with 200µg of Alum hydroxide adjuvant (for each ‎dose of vaccine formulation) and shaked at 100 RPM for 60 minutes at room temperature and ‎allowed that the viral particles to adsorb on the surface of Alum gel. And then vitamin C at ‎concentration of 1 and 10mg/dose was added to the vaccine formulation. Young (6-8-week-old, ‎N=32) and aged (16-20-month-old, N=32) male C57bl/6 mice were purchased from Pasteur ‎Institute of Iran and grouped as below;‎ Group 1: Aged mice immunized with inactivated COVID-19-Alum.‎ Group 2: Aged mice immunized with inactivated COVID-19-Alum with1 mg vitamin C.‎ Group 3: Aged mice immunized with inactivated COVID-19-Alum with10 mg vitamin C.‎ Group 4: Aged mice immunized with PBS.‎ Group 5: Young mice immunized with inactivated COVID-19-Alum.‎ Group 6: Young mice immunized with inactivated COVID-19-Alum with1mg vitamin C.‎ Group 7: Young mice immunized with inactivated COVID-19-Alum with10mg vitamin C.‎ Group 8: Young mice immunized with PBS.‎ Experimental mice were immunized with 2µg of inactivated COVID-19 formulated vaccines ‎subcutaneously on days 0 and 14 and two weeks after last shot IFN-γ and IL-4 cytokine were ‎assessed with ELISA and specific IgG titer, IgG1/IgG2a isotypes determined with ELISA. ‎
  • Results: The results of IFN-γ and IL-4 cytokines in experimental groups demonstrated that injection of ‎COVID-19 vaccine formulated with vitamin C in young and aged mice shows a significant ‎increase as compared with the control and mere vaccine groups (P< 0.0401). In addition, specific ‎total IgG and IgG1/IgG2a level improved in young and aged mice.‎
  • Conclusion: It seems that vitamin C is useful in the improvement of specific immune responses against ‎COVID-19 vaccine in young and aged people. ‎
  • Keywords: COVID-19 vaccine, vitamin C, Adjuvant, aging