Expansion of immature, nucleated red blood cells by transient low-dose methotrexate immune tolerance induction in mice

Clin Exp Immunol. 2021 Mar;203(3):409-423. doi: 10.1111/cei.13552. Epub 2020 Dec 16.

Abstract

Biological treatments such as enzyme-replacement therapies (ERT) can generate anti-drug antibodies (ADA), which may reduce drug efficacy and impact patient safety and consequently led to research to mitigate ADA responses. Transient low-dose methotrexate (TLD-MTX) as a prophylactic ITI regimen, when administered concurrently with ERT, induces long-lived reduction of ADA to recombinant human alglucosidase alfa (rhGAA) in mice. In current clinical practice, a prophylactic ITI protocol that includes TLD-MTX, rituximab and intravenous immunoglobulin (optional), successfully induced lasting control of ADA to rhGAA in high-risk, cross-reactive immunological material (CRIM)-negative infantile-onset Pompe disease (IOPD) patients. More recently, evaluation of TLD-MTX demonstrated benefit in CRIM-positive IOPD patients. To more clearly understand the mechanism for the effectiveness of TLD-MTX, non-targeted transcriptional and proteomic screens were conducted and revealed up-regulation of erythropoiesis signatures. Confirmatory studies showed transiently larger spleens by weight, increased spleen cellularity and that following an initial reduction of mature red blood cells (RBCs) in the bone marrow and blood, a significant expansion of Ter-119+ CD71+ immature RBCs was observed in spleen and blood of mice. Histology sections revealed increased nucleated cells, including hematopoietic precursors, in the splenic red pulp of these mice. This study demonstrated that TLD-MTX induced a transient reduction of mature RBCs in the blood and immature RBCs in the bone marrow followed by significant enrichment of immature, nucleated RBCs in the spleen and blood during the time of immune tolerance induction, which suggested modulation of erythropoiesis may be associated with the induction of immune tolerance to rhGAA.

Keywords: erythropoiesis; tolerance induction; transient methotrexate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics
  • Cell Differentiation / immunology
  • Cell Proliferation / drug effects*
  • Dose-Response Relationship, Drug
  • Erythroblasts / cytology
  • Erythroblasts / drug effects*
  • Erythroblasts / metabolism
  • Erythrocytes / drug effects
  • Erythrocytes / immunology
  • Erythrocytes / metabolism
  • Erythropoiesis / drug effects
  • Erythropoiesis / genetics
  • Erythropoiesis / immunology
  • Female
  • Gene Expression Profiling / methods
  • Humans
  • Immune Tolerance / drug effects*
  • Immune Tolerance / genetics
  • Immune Tolerance / immunology
  • Immunosuppressive Agents / administration & dosage
  • Immunosuppressive Agents / immunology
  • Methotrexate / administration & dosage*
  • Methotrexate / immunology
  • Mice
  • Mice, Inbred C57BL
  • Proteomics / methods
  • Spleen / drug effects
  • Spleen / immunology
  • Spleen / metabolism
  • alpha-Glucosidases / administration & dosage

Substances

  • Immunosuppressive Agents
  • GAA protein, human
  • alpha-Glucosidases
  • Methotrexate

Grants and funding