Immune Netw.  2013 Apr;13(2):70-74. 10.4110/in.2013.13.2.70.

Vitamin C Is an Essential Factor on the Anti-viral Immune Responses through the Production of Interferon-alpha/beta at the Initial Stage of Influenza A Virus (H3N2) Infection

Affiliations
  • 1Laboratory of Anti-oxidant Immunology and Vitamin C, Department of Anatomy, Seoul National University College of Medicine, Seoul 110-799, Korea. genius29@snu.ac.kr, kinglee@snu.ac.kr

Abstract

L-ascorbic acid (vitamin C) is one of the well-known anti-viral agents, especially to influenza virus. Since the in vivo anti-viral effect is still controversial, we investigated whether vitamin C could regulate influenza virus infection in vivo by using Gulo (-/-) mice, which cannot synthesize vitamin C like humans. First, we found that vitamin C-insufficient Gulo (-/-) mice expired within 1 week after intranasal inoculation of influenza virus (H3N2/Hongkong). Viral titers in the lung of vitamin C-insufficient Gulo (-/-) mice were definitely increased but production of anti-viral cytokine, interferon (IFN)-alpha/beta, was decreased. On the contrary, the infiltration of inflammatory cells into the lung and production of pro-inflammatory cytokines, tumor necrosis factor (TNF)-alpha and interleukin (IL)-alpha/beta, were increased in the lung. Taken together, vitamin C shows in vivo anti-viral immune responses at the early time of infection, especially against influenza virus, through increased production of IFN-alpha/beta.

Keyword

Vitamin C; Anti-viral immune response; Influenza A virus; Gulo (-/-) mice

MeSH Terms

Animals
Ascorbic Acid
Cytokines
Humans
Influenza A virus
Influenza, Human
Interferons
Interleukins
Lung
Mice
Mustelidae
Orthomyxoviridae
Tumor Necrosis Factor-alpha
Vitamins
Ascorbic Acid
Cytokines
Interferons
Interleukins
Tumor Necrosis Factor-alpha
Vitamins

Figure

  • Figure 1 Increase of mortality of vitamin C-insufficient Gulo (-/-) mice by the infection of influenza A virus. Twenty hemagglutination units (HAU) of influenza A virus (H3N2/1/68/HongKong) was intranasally inoculated into wild type (n=10), vitamin C-sufficient Gulo (-/-) mice (n=10) and vitamin C-insufficient Gulo (-/-) mice (n=10) was as described in Materials and Methods. And then the survival of mice was monitored for 7 days after virus inoculation. (A) Mice without H3N2 infection, (B) Mice with 20 HAU of H3N2 infection.

  • Figure 2 Increase of influenza A virus replication in the lung of vitamin C-insufficient Gulo (-/-) mice. To analyze the effect of vitamin C on the suppression of viral replication in the lung, the lungs were excised from sacrificed mice (n=10 per each group) and total RNA was purified from lung homogenate as described in Materials and Methods. The virus replication in the presence or absence of vitamin C was determined real time RT-PCR by using of specific primers for influenza virus M2 gene and β2m.

  • Figure 3 Defect on the production of IFN-α/β vitamin C-insufficient Gulo (-/-) mice. The levels of IFN-α (A) and IFN-β (B) in BAL fluids from wild type (n=6), vitamin C-sufficient Gulo (-/-) mice (n=6) and vitamin C-insufficient Gulo (-/-) mice (n=6) were measured by ELISA as described in Materials and Methods, after 1 day of influenza A virus infection. Results are representative of three independent experiments and each performed in triplicates. Values are the mean±SD.

  • Figure 4 Increased production of IL-α/β and TNF-α in the lung of vitamin C-insufficient Gulo (-/-) mice by influenza A virus infection. The levels of IL-1α (A), IL-1β (B) and TNF-α (C) in BAL fluids obtained from wild type (n=6), vitamin C-sufficient Gulo (-/-) mice (n=6) and vitamin C-insufficient Gulo (-/-) mice (n=6) were measured by ELISA as described in Materials and Methods. Results are representative of three independent experiments and each performed in triplicates. Values are the mean±SD. (D) After centrifugation of BAL fluids, the numbers of infiltrated immune cells into the lung upon influenza A virus infection of in the presence or absence of vitamin C were counted under microscope.


Cited by  1 articles

Nutrient modulation of viral infection-implications for COVID-19
Hye-Keong Kim, Chan Yoon Park, Sung Nim Han
Nutr Res Pract. 2021;15(Suppl 1):S1-S21.    doi: 10.4162/nrp.2021.15.S1.S1.


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