Targeted disruption of Nrf2 causes regenerative immune-mediated hemolytic anemia

Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9751-6. doi: 10.1073/pnas.0403620101. Epub 2004 Jun 21.


A basic leucine zipper transcription factor, NF-E2-related factor 2 (Nrf2), plays a critical role in the cellular defense mechanism by mediating a coordinate up-regulation of antioxidant responsive element-driven detoxification and antioxidant genes. Here, we report that targeted disruption of Nrf2 causes regenerative immune-mediated hemolytic anemia due to increased sequestration of damaged erythrocytes. Splenomegaly and spleen toxicity in Nrf2(-/-) mice raised a possibility of hemolytic anemia and splenic extramedullary hematopoiesis in Nrf2(-/-) mice. In support of this, hematology analysis revealed that Nrf2(-/-) mice suffer from anemia with abnormal red cell morphologies (i.e., Howell-Jolly bodies, acantocytes, and schistocytes). In addition, Nrf2(-/-) erythrocytes were more sensitive to H(2)O(2)-induced hemolysis, and erythrocyte-bound IgG levels were markedly increased in Nrf2(-/-) mice compared with Nrf2(+/+) mice. Because IgG bound to erythrocytes in the presence of oxidative damage in erythrocytes (regardless of Nrf2 genotype), these data support that Nrf2(-/-) erythrocytes have higher levels of damage compared with Nrf2(+/+) cells. Finally, Nrf2(-/-) mice showed increased levels of erythrocyte-bound IgG compared with Nrf2(+/+) mice after H(2)O(2) injection in vivo, suggesting that the decreased glutathione and increased H(2)O(2) render the Nrf2(-/-) mice more susceptible to toxicity. Taken together, these observations indicate that a chronic increase in oxidative stress due to decreased antioxidant capacity sensitizes erythrocytes and causes hemolytic anemia in Nrf2(-/-) mice, suggesting a pivotal role of Nrf2-antioxidant responsive element pathway in the cellular antioxidant defense system.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Anemia, Hemolytic, Autoimmune / genetics*
  • Anemia, Hemolytic, Autoimmune / immunology
  • Anemia, Hemolytic, Autoimmune / metabolism
  • Anemia, Hemolytic, Autoimmune / pathology
  • Animals
  • Antioxidants / metabolism
  • Cell Size / drug effects
  • DNA-Binding Proteins / deficiency*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Erythrocytes / drug effects
  • Erythrocytes / immunology
  • Erythrocytes / metabolism
  • Erythrocytes / pathology
  • Gene Deletion*
  • Gene Expression Regulation
  • Genotype
  • Hemolysis / drug effects
  • Hydrogen Peroxide / pharmacology
  • Immunoglobulin G / immunology
  • Mice
  • Mice, Knockout
  • NF-E2-Related Factor 2
  • Oxidative Stress / drug effects
  • Phenotype
  • Response Elements / genetics
  • Spleen / metabolism
  • Spleen / pathology
  • Splenomegaly / genetics
  • Splenomegaly / pathology
  • Tooth / physiology
  • Trans-Activators / deficiency*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism


  • Antioxidants
  • DNA-Binding Proteins
  • Immunoglobulin G
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, mouse
  • Trans-Activators
  • Hydrogen Peroxide