IgY Reduces AFB₁-Induced Cytotoxicity, Cellular Dysfunction, and Genotoxicity in Human L-02 Hepatocytes and Swan 71 Trophoblasts

Abstract

Aflatoxin B (AFB) causes hepatotoxic, genotoxic, and immunotoxic effects in a variety of species. Although various neutralizing agents of AFB toxicity have been studied, the egg yolk immunoglobulin (IgY) detoxification of small molecular toxins and the mechanisms underlying such effects have not yet been reported. In this investigation, anti-AFB IgY against AFB was successfully raised, and a competitive indirect enzyme-linked immunosorbent assay was established with a sensitive half-maximal inhibitory concentration (IC 2.4 ng/mL) and dynamic working range (0.13-43.0 ng/mL). The anti-AFB IgY obtained reduced AFB-induced cytotoxicity, cellular dysfunction, and genotoxicity by protecting cells against apoptotic body formation and DNA strand breaks, preventing G2/M phase cell cycle arrest, reducing AFB-DNA adduct and reactive oxygen species production and maintaining cell migration and invasion and the mitochondrial membrane potential. Anti-AFB IgY significantly inhibited the AFB-induced expression of proteins related to antioxidative, pro-apoptotic, and antiapoptotic processes in a strong dose-dependent manner. These experiments demonstrated that the anti-AFB IgY-bound AFB could not enter cells. This is the first time that IgY has been found to reduce the effects of small molecular toxins, which will be beneficial for the development of antibodies as detoxication agents.

In vitro and in vivo evaluation of AFB1 and OTA-toxicity through immunofluorescence and flow cytometry techniques: A systematic review

“…The aberrant ROS production (1.5-fold higher than control) with MMP disturbance has been also observed. Nevertheless, low doses of IgY showed protective effects against oxidative stress, apoptosis and cell cycle arrest associated with AFB1-treatment (Qiu et al, 2018).…”

Egg Yolk Immunoglobulin Supplementation Prevents Rat Liver from Aflatoxin B1-Induced Oxidative Damage and Genotoxicity

“…The artificial antigen (AFB 1 -GA-BSA) was generated as described by the previous study . Hens were immunized with 400 μg of AFB 1 -GA-BSA in PBS with an equal volume of Freund’s complete adjuvant for the first immunization or Freund’s incomplete adjuvant for four booster immunization.…”

“…17 We have previously reported that anti-AFB 1 IgY formed a strong noncovalent complex with AFB 1 by indirect competitive ELISA. 14 To evaluate the ability of anti-AFB 1 IgY to complex with AFB 1 , AFB 1 (2.5 μM) and different concentrations of anti-AFB 1 IgY were mixed in vitro, and the fluorescent intensity was determined by a fluorescent microplate reader change of the AFB 1 fluorescence spectrum from 380 to 550 nm. There was appropriate linear fluorescence at low concentrations of AFB 1 , as shown in Figure S2A.…”

“…The artificial antigen (AFB 1 -GA-BSA) was generated as described by the previous study. 14 Hens were immunized with 400 μg of AFB 1 -GA-BSA in PBS with an equal volume of Freund's complete adjuvant for the first immunization or Freund's incomplete adjuvant for four booster immunization. The anti-AFB 1 IgY from egg yolk was purified by a combination of several purification technologies including water-soluble fraction under acidic conditions, filtration, and ammonium sulfate precipitation method.…”

One‐step deep eutectic solvent strategy for efficient analysis of aflatoxins in edible oils

“…Aflatoxins are a group of toxic secondary metabolites primarily produced by fungi Aspergillus flavas, Aspergillus parasiticus, and other Aspergillus species, among which aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2) are four major natural aflatoxins with high mutagenicity and carcinogenicity. 1 These aflatoxins exist widely in various grain and animal products. [2][3][4] Among foodstuffs, edible oils such as peanut oil, corn oil, and soybean oil are easily contaminated by various mycotoxins, which could enter the human food chain to potentially impair human growth and increase the incidence of liver cancer.…”