Ascorbic Acid and α-Tocopherol in the Inactivated SARS-CoV-2 Vaccine Formulation: Induction of the Th1 Pattern in Aged Mice

Viral Immunol. 2024 Sep;37(7):355-370. doi: 10.1089/vim.2024.0023. Epub 2024 Aug 30.

Abstract

Aging is physiologically associated with a decline in the function of the immune system and subsequent susceptibility to infections. Interferon-gamma (IFN-γ), a key element in the activation of cellular immunity, plays an important role in defense against virus infections. Decreased levels of IFN-γ in the elderly may explain their increased risk for viral infectious diseases such as COVID-19. There is accumulating evidence that ascorbic acid (vitamin C [VitC]) and α-tocopherol together help improve the function of the immune system in the elderly, control infections, and decrease the treatment duration. A SARS-CoV-2 strain was isolated from a patient and then cultured in the Vero cell line. The isolated and propagated virus was then inactivated using formalin and purified by the column chromatography. The inactivated SARS-CoV-2 was formulated in the Alum adjuvant combined with VitC or α-tocopherol and/or both of them. The vaccines were injected twice to young and aged C57BL/6 mice. Two weeks later, IFN-γ, IL-4, and IL-2 cytokines were assessed using ELISA Kits. Specific IgG and IgG1/IgG2a were assessed by an in-house ELISA. In addition, the expression of PD1 and TERT genes in the spleen tissue of the mice was measured using real-time PCR. IL-4 and IFN-γ cytokines showed a significant increase in both aged and young mice compared with the Alum-based vaccine. In addition, our results exhibited a significant decrease and increase in specific total IgG and the IgG2a/IgG1 ratio, respectively. Furthermore, the vaccine formulated in α-tocopherol + VitC led to decreased PD1 and increased TERT gene expression levels. In conclusion, our results demonstrated that α-tocopherol + VitC formulated in the inactivated SARS-CoV-2 vaccine led to a shift toward Th1, which may be due to their effect on the physiology of cells, especially aged ones and changing their phenotype toward young cells.

Keywords: adjuvant; alpha-tocopherol; ascorbic acid; elderly; inactivated SARS-CoV-2 vaccine.

MeSH terms

  • Adjuvants, Immunologic / administration & dosage
  • Adjuvants, Immunologic / pharmacology
  • Aging / immunology
  • Animals
  • Antibodies, Viral / blood
  • Ascorbic Acid* / administration & dosage
  • Ascorbic Acid* / pharmacology
  • COVID-19 Vaccines* / administration & dosage
  • COVID-19 Vaccines* / immunology
  • COVID-19* / immunology
  • COVID-19* / prevention & control
  • Chlorocebus aethiops
  • Cytokines / metabolism
  • Female
  • Humans
  • Immunoglobulin G / blood
  • Interferon-gamma / metabolism
  • Mice
  • Mice, Inbred C57BL*
  • Programmed Cell Death 1 Receptor
  • SARS-CoV-2* / immunology
  • Th1 Cells* / immunology
  • Vaccines, Inactivated* / administration & dosage
  • Vaccines, Inactivated* / immunology
  • Vero Cells
  • alpha-Tocopherol* / administration & dosage
  • alpha-Tocopherol* / pharmacology

Substances

  • alpha-Tocopherol
  • Vaccines, Inactivated
  • Ascorbic Acid
  • COVID-19 Vaccines
  • Adjuvants, Immunologic
  • Antibodies, Viral
  • Immunoglobulin G
  • Interferon-gamma
  • Cytokines
  • Programmed Cell Death 1 Receptor