IgE is an ancient and conserved immunoglobulin isotype with potent immunological function. Nevertheless, the regulation of IgE responses remains an enigma, and evidence of a role for IgE in host defense is limited. Here we report that topical exposure to a common environmental DNA-damaging xenobiotic initiated stress surveillance by γδTCR+ intraepithelial lymphocytes that resulted in class switching to IgE in B cells and the accumulation of autoreactive IgE. High-throughput antibody sequencing revealed that γδ T cells shaped the IgE repertoire by supporting specific variable-diversity-joining (VDJ) rearrangements with unique characteristics of the complementarity-determining region CDRH3. This endogenous IgE response, via the IgE receptor FcεRI, provided protection against epithelial carcinogenesis, and expression of the gene encoding FcεRI in human squamous-cell carcinoma correlated with good disease prognosis. These data indicate a joint role for immunosurveillance by T cells and by B cells in epithelial tissues and suggest that IgE is part of the host defense against epithelial damage and tumor development.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Nature Communications Open Access 17 March 2022
Remodeling of the tumor microenvironment via disrupting Blimp1+ effector Treg activity augments response to anti-PD-1 blockade
Molecular Cancer Open Access 20 November 2021
Experimental & Molecular Medicine Open Access 12 March 2021
Vernersson, M., Aveskogh, M. & Hellman, L. Cloning of IgE from the echidna (Tachyglossus aculeatus) and a comparative analysis of ε chains from all three extant mammalian lineages. Dev. Comp. Immunol. 28, 61–75 (2004).
Harris, N. & Gause, W. C. To B or not to B: B cells and the Th2-type immune response to helminths. Trends Immunol. 32, 80–88 (2011).
Profet, M. The function of allergy: immunological defense against toxins. Q. Rev. Biol. 66, 23–62 (1991).
Marichal, T. et al. A beneficial role for immunoglobulin E in host defense against honeybee venom. Immunity 39, 963–975 (2013).
Palm, N. W. et al. Bee venom phospholipase A2 induces a primary type 2 response that is dependent on the receptor ST2 and confers protective immunity. Immunity 39, 976–985 (2013).
Starkl, P. et al. IgE antibodies, FcεRIα, and IgE-mediated local anaphylaxis can limit snake venom toxicity. J. Allergy Clin. Immunol. 137, 246–257 (2016).
Palm, N. W., Rosenstein, R. K. & Medzhitov, R. Allergic host defences. Nature 484, 465–472 (2012).
Dalessandri, T. & Strid, J. Beneficial autoimmunity at body surfaces - immune surveillance and rapid type 2 immunity regulate tissue homeostasis and cancer. Front. Immunol. 5, 347 (2014).
Strid, J., Hourihane, J., Kimber, I., Callard, R. & Strobel, S. Disruption of the stratum corneum allows potent epicutaneous immunization with protein antigens resulting in a dominant systemic Th2 response. Eur. J. Immunol. 34, 2100–2109 (2004).
Nelde, A. et al. The impact of the route and frequency of antigen exposure on the IgE response in allergy. Int. Arch. Allergy Immunol. 124, 461–469 (2001).
Dalessandri, T., Crawford, G., Hayes, M., Castro Seoane, R. & Strid, J. IL-13 from intraepithelial lymphocytes regulates tissue homeostasis and protects against carcinogenesis in the skin. Nat. Commun. 7, 12080 (2016).
Strid, J., Sobolev, O., Zafirova, B., Polic, B. & Hayday, A. The intraepithelial T cell response to NKG2D-ligands links lymphoid stress surveillance to atopy. Science 334, 1293–1297 (2011).
Strid, J. et al. Acute upregulation of an NKG2D ligand promotes rapid reorganization of a local immune compartment with pleiotropic effects on carcinogenesis. Nat. Immunol. 9, 146–154 (2008).
Kim, K. H., Jahan, S. A., Kabir, E. & Brown, R. J. A review of airborne polycyclic aromatic hydrocarbons (PAHs) and their human health effects. Environ. Int. 60, 71–80 (2013).
Totlandsdal, A. I. et al. Differential effects of the particle core and organic extract of diesel exhaust particles. Toxicol. Lett. 208, 262–268 (2012).
Modi, B. G. et al. Langerhans cells facilitate epithelial DNA damage and squamous cell carcinoma. Science 335, 104–108 (2012).
Gupta, N. T. et al. Change-O: a toolkit for analyzing large-scale B cell immunoglobulin repertoire sequencing data. Bioinformatics 31, 3356–3358 (2015).
Nakai, K., Kidera, A. & Kanehisa, M. Cluster analysis of amino acid indices for prediction of protein structure and function. Protein Eng. 2, 93–100 (1988).
Collins, A. M., Wang, Y., Roskin, K. M., Marquis, C. P. & Jackson, K. J. The mouse antibody heavy chain repertoire is germline-focused and highly variable between inbred strains. Philos. Trans. R. Soc. Lond. B 370, 20140236 (2015).
Rogosch, T. et al. Plasma cells and nonplasma B cells express differing IgE repertoires in allergic sensitization. J. Immunol. 184, 4947–4954 (2010).
Rosner, K. et al. Third complementarity-determining region of mutated VH immunoglobulin genes contains shorter V, D, J, P, and N components than non-mutated genes. Immunology 103, 179–187 (2001).
Luger, E. et al. Somatic diversity of the immunoglobulin repertoire is controlled in an isotype-specific manner. Eur. J. Immunol. 31, 2319–2330 (2001).
Kim, J. et al. Symptoms of atopic dermatitis are influenced by outdoor air pollution. J. Allergy Clin. Immunol. 132, 495–498 (2013).
Morgenstern, V. et al. Atopic diseases, allergic sensitization, and exposure to traffic-related air pollution in children. Am. J. Respir. Crit. Care Med. 177, 1331–1337 (2008).
Hidaka, T. et al. The aryl hydrocarbon receptor AhR links atopic dermatitis and air pollution via induction of the neurotrophic factor artemin. Nat. Immunol. 18, 64–73 (2017).
Marichal, T. et al. DNA released from dying host cells mediates aluminum adjuvant activity. Nat. Med. 17, 996–1002 (2011).
Girardi, M. et al. Regulation of cutaneous malignancy by γδ T cells. Science 294, 605–609 (2001).
Dema, B. et al. Immunoglobulin E plays an immunoregulatory role in lupus. J. Exp. Med. 211, 2159–2168 (2014).
Henault, J. et al. Self-reactive IgE exacerbates interferon responses associated with autoimmunity. Nat. Immunol. 17, 196–203 (2016).
Messingham, K. A., Holahan, H. M. & Fairley, J. A. Unraveling the significance of IgE autoantibodies in organ-specific autoimmunity: lessons learned from bullous pemphigoid. Immunol. Res. 59, 273–278 (2014).
Altrichter, S. et al. Serum IgE autoantibodies target keratinocytes in patients with atopic dermatitis. J. Invest. Dermatol. 128, 2232–2239 (2008).
Ozcan, E., Notarangelo, L. D. & Geha, R. S. Primary immune deficiencies with aberrant IgE production. J. Allergy Clin. Immunol. 122, 1054–1062 (2008).
Vantourout, P. & Hayday, A. Six-of-the-best: unique contributions of γδ T cells to immunology. Nat. Rev. Immunol. 13, 88–100 (2013).
Wen, L. et al. Germinal center formation, immunoglobulin class switching, and autoantibody production driven by “non α/β” T cells. J. Exp. Med. 183, 2271–2282 (1996).
McCoy, K. D. et al. Natural IgE production in the absence of MHC class II cognate help. Immunity 24, 329–339 (2006).
Josephs, D. H. et al. Anti-folate receptor-α IgE but not IgG recruits macrophages to attack tumors via TNFα/MCP-1 signaling. Cancer Res. 77, 1127–1141 (2017).
Josephs, D. H., Spicer, J. F., Karagiannis, P., Gould, H. J. & Karagiannis, S. N. IgE immunotherapy: a novel concept with promise for the treatment of cancer. MAbs 6, 54–72 (2014).
Rogers, H. W. et al. Incidence estimate of nonmelanoma skin cancer in the United States, 2006. Arch. Dermatol. 146, 283–287 (2010).
Goon, P. K., Greenberg, D. C., Igali, L. & Levell, N. J. Squamous cell carcinoma of the skin has more than doubled over the last decade in the UK. Acta Derm. Venereol. 96, 820–821 (2016).
Lippman, S. M. & Hawk, E. T. Cancer prevention: from 1727 to milestones of the past 100 years. Cancer Res. 69, 5269–5284 (2009).
Leonardi-Bee, J., Ellison, T. & Bath-Hextall, F. Smoking and the risk of nonmelanoma skin cancer: systematic review and meta-analysis. Arch. Dermatol. 148, 939–946 (2012).
Van Hemelrijck, M. et al. Immunoglobulin E and cancer: a meta-analysis and a large Swedish cohort study. Cancer Causes Control 21, 1657–1667 (2010).
Sherman, P. W., Holland, E. & Sherman, J. S. Allergies: their role in cancer prevention. Q. Rev. Biol. 83, 339–362 (2008).
Gentles, A. J. et al. The prognostic landscape of genes and infiltrating immune cells across human cancers. Nat. Med. 21, 938–945 (2015).
Strunk, R. C. & Bloomberg, G. R. Omalizumab for asthma. N. Engl. J. Med. 354, 2689–2695 (2006).
Itohara, S. et al. T cell receptor δ gene mutant mice: independent generation of αβ T cells and programmed rearrangements of γδ TCR genes. Cell 72, 337–348 (1993).
Mombaerts, P., Clarke, A. R., Hooper, M. L. & Tonegawa, S. Creation of a large genomic deletion at the T-cell antigen receptor β-subunit locus in mouse embryonic stem cells by gene targeting. Proc. Natl Acad. Sci. USA 88, 3084–3087 (1991).
Kühn, R., Rajewsky, K. & Müller, W. Generation and analysis of interleukin-4 deficient mice. Science 254, 707–710 (1991).
Kaplan, D. H., Jenison, M. C., Saeland, S., Shlomchik, W. D. & Shlomchik, M. J. Epidermal langerhans cell-deficient mice develop enhanced contact hypersensitivity. Immunity 23, 611–620 (2005).
Mallick-Wood, C. A. et al. Conservation of T cell receptor conformation in epidermal γδ cells with disrupted primary Vγ gene usage. Science 279, 1729–1733 (1998).
Hara, H. et al. Development of dendritic epidermal T cells with a skewed diversity of γδ TCRs in Vδ1-deficient mice. J. Immunol. 165, 3695–3705 (2000).
Oettgen, H. C. et al. Active anaphylaxis in IgE-deficient mice. Nature 370, 367–370 (1994).
Dombrowicz, D., Flamand, V., Brigman, K. K., Koller, B. H. & Kinet, J. P. Abolition of anaphylaxis by targeted disruption of the high affinity immunoglobulin E receptor α chain gene. Cell 75, 969–976 (1993).
Feyerabend, T. B. et al. Cre-mediated cell ablation contests mast cell contribution in models of antibody- and T cell-mediated autoimmunity. Immunity 35, 832–844 (2011).
Barnden, M. J., Allison, J., Heath, W. R. & Carbone, F. R. Defective TCR expression in transgenic mice constructed using cDNA-based α- and β-chain genes under the control of heterologous regulatory elements. Immunol. Cell Biol. 76, 34–40 (1998).
Rickert, R. C., Roes, J. & Rajewsky, K. B lymphocyte-specific, Cre-mediated mutagenesis in mice. Nucleic Acids Res. 25, 1317–1318 (1997).
Hollister, K. et al. Insights into the role of Bcl6 in follicular Th cells using a new conditional mutant mouse model. J. Immunol. 191, 3705–3711 (2013).
Workman, P. et al. Guidelines for the welfare and use of animals in cancer research. Br. J. Cancer 102, 1555–1577 (2010).
Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M. & Altman, D. G. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol. 8, e1000412 (2010).
Wu, Y. C., Kipling, D. & Dunn-Walters, D. Assessment of B cell repertoire in humans. Methods Mol. Biol. 1343, 199–218 (2015).
Hill, M. O. Diversity and evenness: a unifying notation and its consequences. Ecology 54, 427–432 (1973).
Stern, J. N. et al. B cells populating the multiple sclerosis brain mature in the draining cervical lymph nodes. Sci. Transl. Med. 6, 248ra107 (2014).
Martin, V. G. et al. Transitional B cells in early human B cell development - time to revisit the paradigm? Front. Immunol. 7, 546 (2016).
We thank B. Norzawani, C. Margreitter, C. Townsend and B. Hunt for computational assistance and advice during antibody sequencing analysis; H.R. Rodewald (German Cancer Research Center) for Cpa3Cre/+ mice; A. Hayday (King’s College London and The Francis Crick Institute) for Tcrg-V5–/–Tcrd-V1−/− mice; the staff of the Imperial Central Biomedical Services for the care of the animals; the LMS/NIHR Imperial Biomedical Research Centre Flow Cytometry Facility for FACS support for flow cytometry; A. Mowat for critical reading of the manuscript; and colleagues for informed advice. This work was supported by the Wellcome Trust (100999/Z/13/Z) and in part by the Cancer Research UK (C21010/A19788) and the NIHR Newcastle Biomedical Research Centre. C.L. was supported by a Wellcome Trust Research Training Fellowship.
The authors declare no competing interests.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Integrated supplementary information
(a) Quantitative RT-PCR analysis of Rae-1 relative to cyclophylin in skin epithelial cells (n=3) and (b-c) FACS analysis of γH2AX, as a measure of ds-DNA breaks, in skin epithelial cells (n=4), at indicated time-points after topical exposure to a single dose of DMBA to the dorsal ear skin of wild-type FVB mice. UT = untreated (n=4-5). (b) Representative FACS plots of γH2AX staining in CD45- epidermal cells. (d) FACS analysis of γH2AX+ CD45- skin epithelial cells from Langerin-DTA mice and non-transgenic littermate controls (NLC) (n=3/group) after a single or repeated topical DMBA treatment, analysed 3 days after last exposure and (e) FACS analysis of humoral immunity in the skin-draining LNs 7 days after the last DMBA exposure. Total LN cells, B220+CD95+GL7+ GC B cells and IgG1+ and IgE+ FSChiCD95+CD138+ PCs were enumerated (n=8/group). (f) ELISA of serum IgE and (g) FACS analysis of humoral immunity as in (e) in wild-type FVB mice exposed to UV light on shaved back skin at 100mJ/cm2 2-3x a week (n=6). Mice were bled after 4 exposures (week 1.5) and again at the end of the experiment (week 3) after 8 exposures. (h) ELISA of serum IgE in wild-type mice exposed to 200nmol DMBA for 5 consecutive days and then left without further exposure. Mice were bled prior to exposure and at indicated time-points after the last DMBA-treatment (n=13). Some mice started to develop tumors around week 6. Statistics by two-tailed Student’s t-test for unpaired data (d and g), one-way ANOVA multiple comparison (a, c, f) and one-way ANOVA with testing for linear trend of IgE increase with time (h); **p<0.01, ***p<0.001 and ****p<0.0001. All data are expressed as mean ± SEM.
Supplementary Figure 2 Antibody levels in wild-type, Igh7-/-, FceR1a-/- and Il4-/- mice following DMBA carcinogenesis.
(a-c) ELISA of serum antibodies in mice subjected to DMBA carcinogenesis by once weekly exposure to DMBA on shaved back skin. Mice were bled and sera collected at the end of the carcinogenesis experiment. Data are expressed as mean antibody amount ± SEM in sera from (a) BALB/c wild-type and Igh7-/- mice (n=13/group), (b) BALB/c wild-type and FceR1a-/- mice (n=7/group) and (c) FVB wild-type and Il4-/- mice (n=9/group). Statistical analysis in (a-c) was determined using two-tailed Student’s t-test for unpaired data; **p<0.01 and ****p<0.0001. nd = not detected. ns = not significant. WT = wild-type.
Supplementary Figure 3 FcεRI-signaling in basophils is sufficient to protect against carcinogenesis and alters the tumor microenvironment.
(a) FACS analysis of CD45hicKit+FcεRI+ skin mast cells and CD45locKit-FcεRI+ skin basophils in wild-type and Igh7-/- mice after twice topically treatment with DMBA compared to naïve mice (wild-type naïve n=6, wild-type DMBA n=7; Igh7-/- naïve n=3, Igh7-/- DMBA n=4). (b) Tumor susceptibility expressed as tumor latency (time to appearance of first tumor), tumor incidence (average number of tumors per mouse) and tumor area (average tumor size per mouse) in Cpa3Cre/+ (n=10) and Cpa3+/+ (wild-type) littermates (n=13) following DMBA-induced carcinogenesis. Data are expressed as mean ± SEM and statistical significance assessed using Log-rank (Mantel-Cox) test for tumor latency and linear regression for tumor incidence and area. ns = not significant. (c-d) FACS analysis of the degranulation marker CD63 and intracellular cytokine staining in splenic CD45+cKit-CD41+FcεRI+ basophils from wild-type and Igh7-/- mice (n=4/group) left unstimulated (-) or stimulated ex vivo with PMA and ionomycin (+). (c) Representative histograms and (d) enumeration of % basophils positive for indicated marker. (e) Quantitative RT-PCR analysis of selected cytokines relative to cyclophylin in tumor tissue and adjacent skin from wild-type and Igh7-/- mice (n=8/group) treated topically with DMBA once weekly and analysed at week 17. Data are expressed as mean ± SEM. Statistics by two-tailed Student’s t-test for unpaired data; **p<0.01 and ***p<0.001. WT = wild-type.
Supplementary Figure 4 FcεRI+ cells in human skin SSCs accumulate at the interface between the stroma and the neoplastic keratinocytes.
(a-e) Representative SSC histology from 5 patients showing FcεRI staining in brown. 5 μm tissue sections were cut from formalin-fixed paraffin-embedded samples, stained against FcεRI and counterstained with Mayer’s Haematoxylin. Slides were imaged at 20x magnification using an Olympus Dotslide microscope and analysed using OlyVIA software. Insets show zoom of outlined areas.
Supplementary Figure 5 Carcinogen-induced antibody responses require TCR signaling, CD40L and LN IL-4 but not GC B cells.
(a-d) FACS analysis of humoral immunity in the skin draining LNs analysed 7 days following twice topical exposure to DMBA on the dorsal ear skin. (a) C57BL/6 Tcrb-/- mice were reconstituted with wild-type (n=6) or OTII TCR-restricted CD4+ αβ T cells (n=5) 1 day prior to DMBA exposure. (b) Wild-type FVB mice were injected with α-CD40L blocking Ab (MRI clone) (n=4) or isotype control Ab (n=3) prior to DMBA exposure and 3 further times during induction of the response. (c) Lethally irradiated FVB wild-type or Il4-/- mice were reconstituted with either wild-type or Il4-/- BM cells and exposed to DMBA 8 weeks after BM transplant (n=4-6/group). (d) C57BL/6 wild-type mice and mice with a heterozygous or homozygous deletion of Bcl-6 in B cells (CD19creBcl-6fl/wt and CD19creBcl-6fl/fl respectively) were exposed to topical DMBA (n=3-5/group). Graphs show number of total LN cells, B220+CD95+GL7+ GC B cells and IgG1+ or IgE+ FSChiCD95+CD138+ PCs presented as mean ± SEM. Statistics by two-tailed Student’s t-test for unpaired data; *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001. ns = not significant. WT = wild-type. UT = untreated.
Supplementary Figure 6 Analysis of the IgG1 and IgE repertoires induced by topical carcinogen exposure on wild-type and Tcrd-/- mice.
(a-f) High-throughput sequencing and heavy-chain repertoire analysis of IgG1 and IgE in sorted B220+CD95+GL7+ GC B cells and FSChiCD95hiCD138+ PCs from skin draining LNs of wild-type and Tcrd-/- mice 7 days after the last of two topical exposures to DMBA (n=6/group). (a) Average frequency of a given clone size in the entire repertoire is shown for wild-type and Tcrd-/- IgG1+ and IgE+ GC B cells and IgG1+ and IgE+ PCs. (b) Proportion of IgE+ PC clones shared with the IgG1+ PC clones and (c) the fraction of IgE+ PC clones shared with the IgE+ GC B cell clones. (d-f) Total IgHV, IgHD and IgHJ family gene usage within the wild-type IgG1+ GC B cells and PCs as well as wild-type IgE+ GC B cells and PCs. Statistics in (c) by two-tailed Student’s t-test for unpaired data; *p<0.05. WT = wild-type.
(a-e) Examples of autoreactive binding to HEp-2 cells of IgE in serum from (a) DMBA-treated wild-type mice (1:25 dilution), (b) DMBA-treated Tcrd-/- mice (1:5 dilution), (c) TPA-treated wild-type mice (1:5 dilution), (d) DMBA-treated Igh7-/- mouse (1:5 dilution) and (e) naïve wild-type mouse (1:5 dilution). (f-g) Examples of autoreactive binding of IgE to clusters of epithelial cells in acutely DMBA damaged epidermis from FceR1a-/- mice exposed to 200nmol DMBA once on the dorsal ear skin and epidermal sheets isolated 24hr later and stained with serum from (f) DMBA-treated wild-type or (g) Igh7-/- mice (1:25 dilution). Arrows point to hair follicles. IgE binding (red) and nuclei (blue). Each image represents an individual mouse.
About this article
Cite this article
Crawford, G., Hayes, M.D., Seoane, R.C. et al. Epithelial damage and tissue γδ T cells promote a unique tumor-protective IgE response. Nat Immunol 19, 859–870 (2018). https://doi.org/10.1038/s41590-018-0161-8
Nature Communications (2022)
Remodeling of the tumor microenvironment via disrupting Blimp1+ effector Treg activity augments response to anti-PD-1 blockade
Molecular Cancer (2021)
Experimental & Molecular Medicine (2021)
AllergoOncology: ultra-low IgE, a potential novel biomarker in cancer—a Position Paper of the European Academy of Allergy and Clinical Immunology (EAACI)
Clinical and Translational Allergy (2020)
Nature Reviews Cancer (2019)