Article | Published:

The transcription factor XBP1 is selectively required for eosinophil differentiation

Nature Immunology volume 16, pages 829837 (2015) | Download Citation


The transcription factor XBP1 has been linked to the development of highly secretory tissues such as plasma cells and Paneth cells, yet its function in granulocyte maturation has remained unknown. Here we discovered an unexpectedly selective and absolute requirement for XBP1 in eosinophil differentiation without an effect on the survival of basophils or neutrophils. Progenitors of myeloid cells and eosinophils selectively activated the endoribonuclease IRE1α and spliced Xbp1 mRNA without inducing parallel endoplasmic reticulum (ER) stress signaling pathways. Without XBP1, nascent eosinophils exhibited massive defects in the post-translational maturation of key granule proteins required for survival, and these unresolvable structural defects fed back to suppress critical aspects of the transcriptional developmental program. Hence, we present evidence that granulocyte subsets can be distinguished by their differential reliance on secretory-pathway homeostasis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


Primary accessions

Gene Expression Omnibus

Referenced accessions

Gene Expression Omnibus


  1. 1.

    et al. IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature 415, 92–96 (2002).

  2. 2.

    , & Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397, 271–274 (1999).

  3. 3.

    , & Translation reinitiation at alternative open reading frames regulates gene expression in an integrated stress response. J. Cell Biol. 167, 27–33 (2004).

  4. 4.

    , & XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response. Mol. Cell. Biol. 23, 7448–7459 (2003).

  5. 5.

    et al. Stress-independent activation of XBP1s and/or ATF6 reveals three functionally diverse ER proteostasis environments. Cell Rep. 3, 1279–1292 (2013).

  6. 6.

    et al. Plasma cell differentiation requires the transcription factor XBP-1. Nature 412, 300–307 (2001).

  7. 7.

    et al. XBP1 links ER stress to intestinal inflammation and confers genetic risk for human inflammatory bowel disease. Cell 134, 743–756 (2008).

  8. 8.

    , , & XBP-1 is required for biogenesis of cellular secretory machinery of exocrine glands. EMBO J. 24, 4368–4380 (2005).

  9. 9.

    , & The transcription factor XBP-1 is essential for the development and survival of dendritic cells. J. Exp. Med. 204, 2267–2275 (2007).

  10. 10.

    , , , & Neutrophil function: from mechanisms to disease. Annu. Rev. Immunol. 30, 459–489 (2012).

  11. 11.

    , , & Changing roles of eosinophils in health and disease. Ann. Allergy Asthma Immunol. 113, 3–8 (2014).

  12. 12.

    , , , & Nonredundant roles of basophils in immunity. Annu. Rev. Immunol. 29, 45–69 (2011).

  13. 13.

    et al. Eosinophils sustain adipose alternatively activated macrophages associated with glucose homeostasis. Science 332, 243–247 (2011).

  14. 14.

    et al. Eosinophils and type 2 cytokine signaling in macrophages orchestrate development of functional beige fat. Cell 157, 1292–1308 (2014).

  15. 15.

    et al. Type 2 innate signals stimulate fibro/adipogenic progenitors to facilitate muscle regeneration. Cell 153, 376–388 (2013).

  16. 16.

    et al. Regulation of carcinogenesis by IL-5 and CCL11: a potential role for eosinophils in tumor immune surveillance. J. Immunol. 178, 4222–4229 (2007).

  17. 17.

    et al. Eosinophils are required for the maintenance of plasma cells in the bone marrow. Nat. Immunol. 12, 151–159 (2011).

  18. 18.

    & Segregation and packaging of granule enzymes in eosinophilic leukocytes. J. Cell Biol. 45, 54–73 (1970).

  19. 19.

    et al. Identification of eosinophil lineage-committed progenitors in the murine bone marrow. J. Exp. Med. 201, 1891–1897 (2005).

  20. 20.

    , & Eosinophils develop in distinct stages and are recruited to peripheral sites by alternatively activated macrophages. J. Leukoc. Biol. 81, 1434–1444 (2007).

  21. 21.

    , , & TLR activation of the transcription factor XBP1 regulates innate immune responses in macrophages. Nat. Immunol. 11, 411–418 (2010).

  22. 22.

    , , , & Plasma cell differentiation initiates a limited ER stress response by specifically suppressing the PERK-dependent branch of the unfolded protein response. Cell Stress Chaperones 15, 281–293 (2010).

  23. 23.

    et al. Homologous recombination into the eosinophil peroxidase locus generates a strain of mice expressing Cre recombinase exclusively in eosinophils. J. Leukoc. Biol. 94, 17–24 (2013).

  24. 24.

    et al. Eosinophil differentiation in the bone marrow is inhibited by T cell-derived IFN-γ. Blood 116, 2559–2569 (2010).

  25. 25.

    et al. Functionally competent eosinophils differentiated ex vivo in high purity from normal mouse bone marrow. J. Immunol. 181, 4004–4009 (2008).

  26. 26.

    et al. The molecular basis for selective inhibition of unconventional mRNA splicing by an IRE1-binding small molecule. Proc. Natl. Acad. Sci. USA 109, E869–E878 (2012).

  27. 27.

    et al. Transgenic mice expressing a B cell growth and differentiation factor gene (interleukin 5) develop eosinophilia and autoantibody production. J. Exp. Med. 173, 429–437 (1991).

  28. 28.

    et al. XBP1 controls diverse cell type- and condition-specific transcriptional regulatory networks. Mol. Cell 27, 53–66 (2007).

  29. 29.

    , & Eosinophil development, regulation of eosinophil-specific genes, and role of eosinophils in the pathogenesis of asthma. Allergy Asthma Immunol. Res. 4, 68–79 (2012).

  30. 30.

    et al. Critical role of Trib1 in differentiation of tissue-resident M2-like macrophages. Nature 495, 524–528 (2013).

  31. 31.

    et al. The order of expression of transcription factors directs hierarchical specification of hematopoietic lineages. Genes Dev. 20, 3010–3021 (2006).

  32. 32.

    , , & IL-5 triggers a cooperative cytokine network that promotes eosinophil precursor maturation. J. Immunol. 193, 4043–4052 (2014).

  33. 33.

    et al. ER-stress-induced transcriptional regulation increases protein synthesis leading to cell death. Nat. Cell Biol. 15, 481–490 (2013).

  34. 34.

    et al. Discovering hematopoietic mechanisms through genome-wide analysis of GATA factor chromatin occupancy. Mol. Cell 36, 667–681 (2009).

  35. 35.

    et al. FoxOs are critical mediators of hematopoietic stem cell resistance to physiologic oxidative stress. Cell 128, 325–339 (2007).

  36. 36.

    et al. XBP1 governs late events in plasma cell differentiation and is not required for antigen-specific memory B cell development. J. Exp. Med. 206, 2151–2159 (2009).

  37. 37.

    & Eosinophil granule proteins: form and function. J. Biol. Chem. 289, 17406–17415 (2014).

  38. 38.

    , & Mechanism of membrane damage mediated by eosinophil major basic protein. Lancet 1, 1380–1381 (1987).

  39. 39.

    , & Acidic precursor revealed in human eosinophil granule major basic protein cDNA. J. Exp. Med. 168, 1493–1498 (1988).

  40. 40.

    et al. Toxicity of eosinophil MBP is repressed by intracellular crystallization and promoted by extracellular aggregation. Mol. Cell 57, 1011–1021 (2015).

  41. 41.

    & Selective staining of eosinophils and their immature precursors in tissue sections and autoradiographs with Congo red. Stain Technol. 56, 323–325 (1981).

  42. 42.

    & The International Mouse Phenotyping Consortium: past and future perspectives on mouse phenotyping. Mammalian Genome 23, 632–640 (2012).

  43. 43.

    et al. Expression of the secondary granule proteins major basic protein 1 (MBP-1) and eosinophil peroxidase (EPX) is required for eosinophilopoiesis in mice. Blood 122, 781–790 (2013).

  44. 44.

    et al. Global quantification of mammalian gene expression control. Nature 473, 337–342 (2011).

  45. 45.

    et al. Plasma cell differentiation and the unfolded protein response intersect at the transcription factor XBP-1. Nat. Immunol. 4, 321–329 (2003).

  46. 46.

    , , & Function of IRE1 alpha in the placenta is essential for placental development and embryonic viability. Proc. Natl. Acad. Sci. USA 106, 16657–16662 (2009).

  47. 47.

    , , & Airway eosinophils: allergic inflammation recruited professional antigen-presenting cells. J. Immunol. 179, 7585–7592 (2007).

  48. 48.

    et al. Dissociation of inositol-requiring enzyme (IRE1α)-mediated c-Jun N-terminal kinase activation from hepatic insulin resistance in conditional X-box-binding protein-1 (XBP1) knock-out mice. J. Biol. Chem. 287, 2558–2567 (2012).

  49. 49.

    , & HTSeq-a Python framework to work with high-throughput sequencing data. Bioinformatics 31, 166–169 (2015).

  50. 50.

    & Differential expression analysis for sequence count data. Genome Biol. 11, R106 (2010).

Download references


We thank L. Cohen-Gould for assistance with electron microscopy; J. McCormick for assistance with cell sorting; the Starr Foundation Tri-Institutional Stem Cell Derivation Laboratory and Flow Cytometry and Microscopy Core facility for technical assistance, the Weill Cornell Epigenomics Core facility for library preparation and RNA sequencing; J. Cubillos-Ruiz and C. Tan for technical assistance; A. Espinosa, S. Hai and members of the Glimcher laboratory for suggestions and critical reading of this manuscript; A. Doyle for conversations; J.J. Lee and N.A. Lee (Mayo Clinic) for eoCRE mice, antibody to PRG2 and antibody to EPX; C. Gerard (Children's Hospital Medical, Boston) for Il5-transgenic BALB/c mice; and T. Iwawaki (Gunma University) for Ern1f/f mice. Supported by the US National Institutes of Health (R01DK082448 to L.H.G., R01HL095699 to L.A.S. and R37AI020241 to P.F.W.).

Author information


  1. Program in Immunology, Harvard Medical School, Boston, Massachusetts, USA.

    • Sarah E Bettigole
  2. Department of Medicine, Weill Cornell Medical College, Cornell University, New York, New York, USA.

    • Sarah E Bettigole
    • , Stanley Adoro
    •  & Laurie H Glimcher
  3. Sandra and Edward Meyer Cancer Center, Weill Cornell Medical College, New York, New York, USA.

    • Sarah E Bettigole
    • , Stanley Adoro
    •  & Laurie H Glimcher
  4. Ansary Stem Cell Institute, Department of Genetic Medicine, and Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York, USA.

    • Raphael Lis
  5. Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical College, New York, New York, USA.

    • Raphael Lis
  6. Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, Cornell University, New York, New York, USA.

    • Ann-Hwee Lee
  7. Department of Medicine, Division of Allergy and Inflammation, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.

    • Lisa A Spencer
    •  & Peter F Weller


  1. Search for Sarah E Bettigole in:

  2. Search for Raphael Lis in:

  3. Search for Stanley Adoro in:

  4. Search for Ann-Hwee Lee in:

  5. Search for Lisa A Spencer in:

  6. Search for Peter F Weller in:

  7. Search for Laurie H Glimcher in:


S.E.B. and L.H.G. designed and analyzed the experiments; S.E.B. conducted experiments and wrote the manuscript; R.L. performed high-resolution immunofluorescence microscopy imaging. S.A. contributed to the design of certain experiments; A.-H.L. provided reagents; and L.A.S., P.F.W. and L.H.G., supervised the research and edited the manuscript.

Competing interests

L.H.G. is on the board of directors of and holds equity in Bristol Myers Squibb Pharmaceutical Company.

Corresponding author

Correspondence to Laurie H Glimcher.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6

Excel files

  1. 1.

    Supplementary Table 1

    Full DESeq transcriptome statistical analysis results comparing Xbp1f/f and Xbp1Vav1 GMPs

  2. 2.

    Supplementary Table 2

    Enriched gene ontology categories and predicted upstream transcriptional regulators from IPA analyses comparing freshly sorted Xbp1f/f GMPs to Xbp1Vav1 GMPs or GATA2-transduced Xbp1f/f GMPs to GATA2-transduced Xbp1Vav1 GMPs 48 hours after infection

  3. 3.

    Supplementary Table 3

    Full DESeq transcriptome statistical analysis results comparing GATA2-transduced Xbp1f/f and Xbp1Vav1 GMPs 48 hours after infection.

  4. 4.

    Supplementary Table 4

    Table of RT-qPCR primers used in this study

About this article

Publication history





Further reading