Targeted Disruption of Nrf2 Causes Regenerative Immune-Mediated Hemolytic Anemia

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 H2O2-induced hemolysis, and erythrocytebound 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 H2O2injection in vivo, suggesting that the decreased glutathione and increased H2O2render 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.