Intergenerational inheritance of immune traits linked to epigenetic modifications has been demonstrated in plants and invertebrates. Here we provide evidence for transmission of trained immunity across generations to murine progeny that survived a sublethal systemic infection with Candida albicans or a zymosan challenge. The progeny of trained mice exhibited cellular, developmental, transcriptional and epigenetic changes associated with the bone marrow-resident myeloid effector and progenitor cell compartment. Moreover, the progeny of trained mice showed enhanced responsiveness to endotoxin challenge, alongside improved protection against systemic heterologous Escherichia coli and Listeria monocytogenes infections. Sperm DNA of parental male mice intravenously infected with the fungus C. albicans showed DNA methylation differences linked to immune gene loci. These results provide evidence for inheritance of trained immunity in mammals, enhancing protection against infections.
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Data from the RNA-seq, ATAC-seq and sperm DNA methylation (RRBS) experiments have been deposited in the National Center for Biotechnology Information Gene Expression Omnibus (GEO) database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE130327. Raw sequencing data and sample annotations are available from the GEO under accession number GSE130327. The mouse reference genome (mm10, build GRCm38) was accessed from the UCSC genome browser (https://www.ncbi.nlm.nih.gov/assembly/GCF_000001635.20/). Blacklisted genomic regions were downloaded from http://mitra.stanford.edu/kundaje/akundaje/release/blacklists/mm10-mouse/mm10.blacklist.bed.gz. Prebuilt gene sets for GSEA analysis were downloaded from the GO2MSIG (http://www.bioinformatics.org/go2msig/). The ATAC-seq peaks were annotated using annotatePeaks.pl (http://homer.ucsd.edu/homer/ngs/annotation.html), Reference SNPs and indels from the dbSNP were obtained from the NCBI (http://www.ncbi.nlm.nih.gov/SNP). Repeats were downloaded from Repbase (https://www.girinst.org/repbase/). Differentially methylated regions were annotated using the GENCODE gene model GRCm38.p6 release M20 (https://www.gencodegenes.org/mouse/release_M20.html). Data supporting the findings of this study are available within the article and supplementary information (Supplementary Tables 1–6). No restrictions on data availability apply. Correspondence and requests for materials should be addressed to M.G.N. (Mihai.Netea@radboudumc.nl). Source data are provided with this paper.
Beccaloni, G. W. & Smith, V. S. Celebrations for Darwin downplay Wallace’s role. Nature 451, 1050 (2008).
Cubas, P., Vincent, C. & Coen, E. An epigenetic mutation responsible for natural variation in floral symmetry. Nature 401, 157–161 (1999).
Luna, E., Bruce, T. J. A., Roberts, M. R., Flors, V. & Ton, J. Next-generation systemic acquired resistance. Plant Physiol. 158, 844–853 (2012).
Belicard, T., Jareosettasin, P. & Sarkies, P. The piRNA pathway responds to environmental signals to establish intergenerational adaptation to stress. BMC Biol. 16, 103 (2018).
Ferguson-Smith, A. C. & Patti, M.-E. You are what your dad ate. Cell Metab. 13, 115–117 (2011).
Morgan, H. D., Sutherland, H. G., Martin, D. I. & Whitelaw, E. Epigenetic inheritance at the agouti locus in the mouse. Nat. Genet. 23, 314–318 (1999).
Dias, B. G. & Ressler, K. J. Parental olfactory experience influences behavior and neural structure in subsequent generations. Nat. Neurosci. 17, 89–96 (2014).
Rowe, A. H. & Rowe, M. P. Physiological resistance of grasshopper mice (Onychomys spp.) to Arizona bark scorpion (Centruroides exilicauda) venom. Toxicon 52, 597–605 (2008).
Domínguez-Andrés, J. et al. Inflammatory Ly6Chigh monocytes protect against candidiasis through IL-15-driven NK cell/neutrophil activation. Immunity 46, 1059–1072.e4 (2017).
Saeed, S. et al. Epigenetic programming of monocyte-to-macrophage differentiation and trained innate immunity. Science 345, 1251086 (2014).
Klengel, T., Dias, B. G. & Ressler, K. J. Models of intergenerational and transgenerational transmission of risk for psychopathology in mice. Neuropsychopharmacology 41, 219–231 (2016).
Ciarlo, E. et al. Trained immunity confers broad-spectrum protection against bacterial infections. J. Infect. Dis. 222, 1869–1881 (2020).
Mitroulis, I. et al. Modulation of myelopoiesis progenitors is an integral component of trained immunity. Cell 172, 147–161.e12 (2018).
Mebratu, Y. & Tesfaigzi, Y. How ERK1/2 activation controls cell proliferation and cell death: is subcellular localization the answer? Cell Cycle 8, 1168–1175 (2009).
Zhu, F. G., Reich, C. F. & Pisetsky, D. S. The role of the macrophage scavenger receptor in immune stimulation by bacterial DNA and synthetic oligonucleotides. Immunology 103, 226–234 (2001).
Stein, J. V. et al. APRIL modulates B and T cell immunity. J. Clin. Invest. 109, 1587–1598 (2002).
Volkova, O. et al. Development and characterization of domain-specific monoclonal antibodies produced against human SLAMF9. Monoclon. Antib. Immunodiagn. Immunother. 33, 209–214 (2014).
Kaludercic, N. et al. Monoamine oxidase B prompts mitochondrial and cardiac dysfunction in pressure overloaded hearts. Antioxid. Redox Signal. 20, 267–280 (2014).
Manicone, A. M. & McGuire, J. K. Matrix metalloproteinases as modulators of inflammation. Semin. Cell Dev. Biol. 19, 34–41 (2008).
Nohra, R. et al. RGMA and IL21R show association with experimental inflammation and multiple sclerosis. Genes Immun. 11, 279–293 (2010).
Hedl, M., Zheng, S. & Abraham, C. The IL18RAP region disease polymorphism decreases IL-18RAP/IL-18R1/IL-1R1 expression and signaling through innate receptor-initiated pathways. J. Immunol. 192, 5924–5932 (2014).
Klaver, E. J. et al. Trichuris suis soluble products induce Rab7b expression and limit TLR4 responses in human dendritic cells. Genes Immun. 16, 378–387 (2015).
Hrašovec, S., Hauptman, N., Glavač, D., Jelenc, F. & Ravnik-Glavač, M. TMEM25 is a candidate biomarker methylated and down-regulated in colorectal cancer. Dis. Markers 34, 93–104 (2013).
Cramer, S. D., Ferree, P. M., Lin, K., Milliner, D. S. & Holmes, R. P. The gene encoding hydroxypyruvate reductase (GRHPR) is mutated in patients with primary hyperoxaluria type II. Hum. Mol. Genet. 8, 2063–2069 (1999).
Hong, J. Y. et al. Antibody to FcεRIα suppresses immunoglobulin E binding to high-affinity receptor I in allergic inflammation. Yonsei Med. J. 57, 1412–1419 (2016).
Emin, M. et al. Increased internalization of complement inhibitor CD59 may contribute to endothelial inflammation in obstructive sleep apnea. Sci. Transl. Med. 8, 320ra1 (2016).
Dinarello, C. A. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood 117, 3720–3732 (2011).
Yu, H.-B. et al. NFATc2 mediates epigenetic modification of dendritic cell cytokine and chemokine responses to dectin-1 stimulation. Nucleic Acids Res. 43, 836–847 (2015).
Qin, Q., Lee, S. H., Liang, R. & Kalejta, R. F. Insertion of myeloid-active elements into the human cytomegalovirus major immediate early promoter is not sufficient to drive its activation upon infection of undifferentiated myeloid cells. Virology 448, 125–132 (2014).
Lee, S. H. et al. Runx3 inhibits IL-4 production in T cells via physical interaction with NFAT. Biochem. Biophys. Res. Commun. 381, 214–217 (2009).
Morris, V. A., Cummings, C. L., Korb, B., Boaglio, S. & Oehler, V. G. Deregulated KLF4 expression in myeloid leukemias alters cell proliferation and differentiation through microRNA and gene targets. Mol. Cell. Biol. 36, 559–573 (2016).
Siklenka, K. et al. Disruption of histone methylation in developing sperm impairs offspring health transgenerationally. Science 350, aab2006 (2015).
Sun, W. et al. Cold-induced epigenetic programming of the sperm enhances brown adipose tissue activity in the offspring. Nat. Med. 24, 1372–1383 (2018).
Das, A. et al. High-resolution mapping and dynamics of the transcriptome, transcription factors, and transcription co-factor networks in classically and alternatively activated macrophages. Front. Immunol. 9, 22 (2018).
Pundhir, S. et al. Enhancer and transcription factor dynamics during myeloid differentiation reveal an early differentiation block in Cebpa null progenitors. Cell Rep. 23, 2744–2757 (2018).
Eggert, H., Kurtz, J. & Diddens-de Buhr, M. F. Different effects of paternal trans-generational immune priming on survival and immunity in step and genetic offspring. Proc. Biol. Sci. 281, 20142089 (2014).
Hernández López, J., Schuehly, W., Crailsheim, K. & Riessberger-Gallé, U. Trans-generational immune priming in honeybees. Proc. Biol. Sci. 281, 20140454 (2014).
Tidbury, H. J., Pedersen, A. B. & Boots, M. Within and transgenerational immune priming in an insect to a DNA virus. Proc. Biol. Sci. 278, 871–876 (2011).
Yue, F. et al. Maternal transfer of immunity in scallop Chlamys farreri and its trans-generational immune protection to offspring against bacterial challenge. Dev. Comp. Immunol. 41, 569–577 (2013).
Ramanan, D. et al. An immunologic mode of multigenerational transmission governs a gut Treg setpoint. Cell 181, 1276–1290.e13 (2020).
Weber-Stadlbauer, U. et al. Transgenerational transmission and modification of pathological traits induced by prenatal immune activation. Mol. Psychiatry 22, 102–112 (2017).
Gerbault, P. et al. Evolution of lactase persistence: an example of human niche construction. Philos. Trans. R. Soc. Lond. B Biol. Sci. 366, 863–877 (2011).
Maekawa, T. et al. ATF7 mediates TNF-α-induced telomere shortening. Nucleic Acids Res. 46, 4487–4504 (2018).
Norouzitallab, P., Baruah, K., Biswas, P., Vanrompay, D. & Bossier, P. Probing the phenomenon of trained immunity in invertebrates during a transgenerational study, using brine shrimp Artemia as a model system. Sci. Rep. 6, 21166 (2016).
Kazachenka, A. et al. Identification, characterization, and heritability of murine metastable epialleles: implications for non-genetic inheritance. Cell 175, 1259–1271.e13 (2018).
Oey, H., Isbel, L., Hickey, P., Ebaid, B. & Whitelaw, E. Genetic and epigenetic variation among inbred mouse littermates: identification of inter-individual differentially methylated regions. Epigenetics Chromatin 8, 54 (2015).
Nankabirwa, V. et al. Child survival and BCG vaccination: a community based prospective cohort study in Uganda. BMC Public Health 15, 175 (2015).
Kovacs, E. J. et al. Aging and innate immunity in the mouse: impact of intrinsic and extrinsic factors. Trends Immunol. 30, 319–324 (2009).
Roger, T. et al. Macrophage migration inhibitory factor deficiency is associated with impaired killing of gram-negative bacteria by macrophages and increased susceptibility to Klebsiella pneumoniae sepsis. J. Infect. Dis. 207, 331–339 (2013).
M.G.N. was supported by a European Research Council advanced grant (no. 833247) and a Spinoza grant of the Netherlands Organization for Scientific Research. E.J.G.-B. is funded by the Hellenic Institute for the Study of Sepsis. T.R. was supported by the Swiss National Science Foundation (no. 310030_173123) and grants from Fondation Carigest/Promex Stiftung für die Forschung and Fondation de Recherche en Biochimie. J.D.-A. is supported by the Netherlands Organization for Scientific Research (VENI grant no. 09150161910024). G.R. is funded by the Horizon 2020 Marie Skłodowska-Curie Action-European Sepsis Academy-Innovative Training Network (no. 676129). A.S. holds an Emmy Noether fellowship of the Deutsche Forschungsgemeinschaft (DFG) (no. SCHL2116/1-1). M.G.N., A.S. and J.L.S. are funded by the DFG under Germany’s Excellence Strategy EXC2151 390873048. J.W. and K.L. are funded by the DFG within the SFB 1309 (Chemical Biology of Epigenetic Modifications). M.B. is a member of the excellence cluster ImmunoSensation2. We thank K. Nordström for bioinformatics support.
E.J.G.-B. has received honoraria (paid to the University of Athens) from AbbVie, Abbott, Biotest, Brahms, InflaRx, the Medicines Company, MSD and XBiotech. He has received independent educational grants from AbbVie, Abbott, Astellas Pharma, Axis Shield, bioMérieux, InflaRx, the Medicines Company and XBiotech. M.G.N. is a scientific founder of TTxD.
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(a) Fungal burden assessed in F1-control and F1-exposed mice in liver and kidney 3 and 7 days after infection with C. albicans, n = 5 per group. (b) Myeloperoxidase activity assessed in F1-control and F1-exposed mice in liver and kidney 3 and 7 days after infection with C. albicans, n = 5 per group. Statistical significance was calculated by two-tailed Mann Whitney U test. P values are depicted on the figures; ns, not significant.
Bacterial burden assessed in F3-control and F3-exposed mice in liver, kidney and spleen 3 days after i.v. infection with E. coli. n = 5 per group; Mann Whitney U test. P values are depicted on the figures.
Leukocyte subpopulations in blood collected just before (day 0), 2 and 3 days post-infection with L. monocytogenes (2 ×104 CFU i.v.) in F1-control and F1-exposed mice (n = 8). Statistical significance was calculated by two-tailed Mann Whitney U test. P values are depicted on the figures; ns, not significant.
Extended Data Fig. 4 Responses in the female progeny. Trained parents: females; progeny analyzed: F1 females.
(a) Survival of F1-control and F1-exposed mice after infection with L. monocytogenes. Data are presented as a Kaplan-Maier plot with a log rank test used to compare susceptibility between the two groups, n = 16 per group. (b) Percent of initial weight of mice 48 h after infection, n = 16 per group. (c, d) Bacteria in blood collected 48 (c) and 72 h after infection (d), n = 16 per group. P values are depicted on the figures, Mann-Whitney U test, unless otherwise stated. (e) Leukocyte subpopulations in blood collected just before (day 0), 2 and 3 days post-infection with L. monocytogenes in F1-control and F1-exposed mice, n = 16 per group. Statistical significance was calculated by two-tailed Mann Whitney U test. P values are depicted on the figures; ns, not significant.
Extended Data Fig. 5 Responses in the male progeny. Trained parents: females; progeny analyzed: F1 males.
(a) Survival of F1-control and F1-exposed mice after infection with L. monocytogenes. Data are presented as a Kaplan-Maier plot with a log rank test used to compare susceptibility between the two groups. n = 6 for F1-control, 8 for F1-exposed. (b) Percent of initial weight of mice 48 h after infection, n = 6 for F1-control, 8 for F1-exposed (c, d) Bacteria in blood collected 48 (c) and 72 h after infection (d), n = 6 for F1-control, 8 for F1-exposed. P values are depicted on the figures, Mann-Whitney U test, unless otherwise stated. (e) Leukocyte subpopulations in blood collected just before (day 0), 2 and 3 days post-infection with L. monocytogenes in F1-control and F1-exposed mice, n = 6 for F1-control, 8 for F1-exposed. Statistical significance was calculated by two-tailed Mann Whitney U test. P values are depicted on the figures; ns, not significant.
Extended Data Fig. 6 Tlr1-/-, Tlr2-/-, and Tlr6-/- mice are fully trainable by zymosan and fully resistant to infection with L. monocytogenes.
Survival of wild type, Tlr1-/-, Tlr2-/-, and Tlr6-/- mice female mice trained with zymosan before i.v. challenge with 1.1 ×105 CFU L. monocytogenes. Data are presented as a Kaplan-Maier plot with a log rank test used to compare susceptibility between the two groups. Number of mice per group and p value are depicted in the figure.
(a) Manual gating strategy to phenotype BM myeloid lineages and progenitors using flow cytometry. GMP, cMoP and Ly6chigh monocytes (Mono) were sorted for RNA-seq or ATAC-seq. (b) Quantification of cell populations in the progeny of F1-control or F1-exposed mice. F1-control, n = 17, F1-exposed n = 13, Statistical significance was calculated by two-tailed unpaired t-test; ns, not significant.
Extended Data Fig. 8 Infection induces epigenetic changes in bone marrow progenitors of the F1-control and F1-exposed offspring.
(a) General distribution of all identified peaks by ATAC-seq relative to the distance to the closest gene transcription start site (TSS). (b) Annotation of all identified peaks according to the genomic location. UTR, untranslated region. (c) Hierarchical clustering and heatmap of all differentially accessible (DA) ATAC-seq peaks of sorted GMPs from F1-control and F1-exposed offspring. (1460 opening, 1402 closing regions). (d) Heatmap and hierarchical clustering of the subfraction of DA ATAC-seq peaks located within gene promoter regions (176 opening, 177 closing regions). F1-control/F1-exposed, n = 13 per group. See Supplementary Table 6 for the full list of differentially accessible regions.
(a) Methylation density plot of global methylation levels. (b) Genomic annotation of DMRs. (c) Methylation levels at repetitive elements. (d) KEGG pathways enriched for hypomethylated and hypermethylated DMRs. n = 10 per group.
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Katzmarski, N., Domínguez-Andrés, J., Cirovic, B. et al. Transmission of trained immunity and heterologous resistance to infections across generations. Nat Immunol 22, 1382–1390 (2021). https://doi.org/10.1038/s41590-021-01052-7
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