Abstract
We recently demonstrated that leukocyte Ig-like receptor 4 (LILRB4) expressed by monocytic acute myeloid leukemia (AML) cells mediates T-cell inhibition and leukemia cell infiltration via its intracellular domain. The cytoplasmic domain of LILRB4 contains three immunoreceptor tyrosine-based inhibitory motifs (ITIMs); the tyrosines at positions 360, 412, and 442 are phosphorylation sites. Here, we analyzed how the ITIMs of LILRB4 in AML cells mediate its function. Our in vitro and in vivo data show that Y412 and Y442, but not Y360, of LILRB4 are required for T-cell inhibition, and all three ITIMs are needed for leukemia cell infiltration. We constructed chimeric proteins containing the extracellular domain of LILRB4 and the intracellular domain of LILRB1 and vice versa. The intracellular domain of LILRB4, but not that of LILRB1, mediates T-cell suppression and AML cell migration. Our studies thus defined the unique signaling roles of LILRB4 ITIMs in AML cells.
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31 January 2020
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
References
Kang, X. et al. Inhibitory leukocyte immunoglobulin-like receptors: immune checkpoint proteins and tumor sustaining factors. Cell Cycle 15, 25–40 (2016).
Hirayasu, K. & Arase, H. Functional and genetic diversity of leukocyte immunoglobulin-like receptor and implication for disease associations. J. Hum. Genet. 60, 703–708 (2015).
Trowsdale, J., Jones, D. C., Barrow, A. D. & Traherne, J. A. Surveillance of cell and tissue perturbation by receptors in the LRC. Immunol. Rev. 267, 117–136 (2015).
Daeron, M., Jaeger, S., Du Pasquier, L. & Vivier, E. Immunoreceptor tyrosine-based inhibition motifs: a quest in the past and future. Immunol. Rev. 224, 11–43 (2008).
Takai, T., Nakamura, A. & Endo, S. Role of PIR-B in autoimmune glomerulonephritis. J. Biomed. Biotechnol. 2011, 275302 (2011).
Katz, H. R. Inhibition of inflammatory responses by leukocyte Ig-like receptors. Adv. Immunol. 91, 251–272 (2006).
Carosella, E. D., Rouas-Freiss, N., Roux, D. T., Moreau, P. & LeMaoult, J. HLA-G: an Immune Checkpoint Molecule. Adv. Immunol. 127, 33–144 (2015).
Colovai, A. I. et al. Expression of inhibitory receptor ILT3 on neoplastic B cells is associated with lymphoid tissue involvement in chronic lymphocytic leukemia. Cytom. Part B, Clin. Cytom. 72, 354–362 (2007).
Zurli, V. et al. Ectopic ILT3 controls BCR-dependent activation of Akt in B-cell chronic lymphocytic leukemia. Blood 130, 2006–2017 (2017).
Zheng, J. et al. Inhibitory receptors bind ANGPTLs and support blood stem cells and leukaemia development. Nature 485, 656–660 (2012).
Kang, X. et al. The ITIM-containing receptor LAIR1 is essential for acute myeloid leukaemia development. Nat. Cell Biol. 17, 665–677 (2015).
Deng, M. et al. LILRB4 signalling in leukaemia cells mediates T cell suppression and tumour infiltration. Nature 562, 605–609 (2018).
John, S. et al. A novel anti-LILRB4 CAR-T cell for the treatment of monocytic AML. Mol. Ther. 26, 2487–2495 (2018).
Suciu-Foca, N. et al. Soluble Ig-like transcript 3 inhibits tumor allograft rejection in humanized SCID mice and T cell responses in cancer patients. J. Immunol. 178, 7432–7441 (2007).
Barkal, A. A. et al. Engagement of MHC class I by the inhibitory receptor LILRB1 suppresses macrophages and is a target of cancer immunotherapy. Nat. Immunol. 19, 76–84 (2018).
Chen, Z. et al. Signalling thresholds and negative B-cell selection in acute lymphoblastic leukaemia. Nature 521, 357–361 (2015).
Perna, F. et al. Integrating proteomics and transcriptomics for systematic combinatorial chimeric antigen receptor therapy of AML. Cancer Cell 32, 506–519 e505 (2017).
Cella, M. et al. A novel inhibitory receptor (ILT3) expressed on monocytes, macrophages, and dendritic cells involved in antigen processing. J. Exp. Med. 185, 1743–1751 (1997).
Inui, M. et al. Human CD43+ B cells are closely related not only to memory B cells phenotypically but also to plasmablasts developmentally in healthy individuals. Int Immunol. 27, 345–355 (2015).
Dobrowolska, H., Vlad, G. & Suciu-Foca, N. Expression of inhibitory receptor ILT3 on normal hematopoietic stem cells and leukemic progenitors. J. Cell Sci. Ther. 4, 98 (2013).
Dobrowolska, H. et al. Expression of immune inhibitory receptor ILT3 in acute myeloid leukemia with monocytic differentiation. Cytom. Part B, Clin. Cytom. 84, 21–29 (2013).
Gui, X. et al. Disrupting LILRB4/APOE interaction by an efficacious humanized antibody reverses T-cell suppression and blocks AML development. Cancer Immunol. Res. 7, 1244–1257 (2019).
Baitsch, D. et al. Apolipoprotein E induces antiinflammatory phenotype in macrophages. Arteriosclerosis, Thrombosis, Vasc. Biol. 31, 1160–1168 (2011).
Samaridis, J. & Colonna, M. Cloning of novel immunoglobulin superfamily receptors expressed on human myeloid and lymphoid cells: structural evidence for new stimulatory and inhibitory pathways. Eur. J. Immunol. 27, 660–665 (1997).
Park, M. et al. A dual positive and negative regulation of monocyte activation by leukocyte Ig-like receptor B4 depends on the position of the tyrosine residues in its ITIMs. Innate Immun. 23, 381–391 (2017).
Ishige, M. et al. A case of metastatic spermatic cord tumor from acute myelogenous leukemia. Nihon Hinyokika Gakkai zasshi Jpn. J. Urol. 103, 631–635 (2012).
Straus, D. J. et al. The acute monocytic leukemias: multidisciplinary studies in 45 patients. Medicine 59, 409–425 (1980).
Wang, Y. et al. Identification of a novel nuclear factor-kappaB sequence involved in expression of urokinase-type plasminogen activator receptor. Eur. J. Biochem. 267, 3248–3254 (2000).
Zhang, H., Meng, F., Chu, C. L., Takai, T. & Lowell, C. A. The Src family kinases Hck and Fgr negatively regulate neutrophil and dendritic cell chemokine signaling via PIR-B. Immunity 22, 235–246 (2005).
Bellon, T., Kitzig, F., Sayos, J. & Lopez-Botet, M. Mutational analysis of immunoreceptor tyrosine-based inhibition motifs of the Ig-like transcript 2 (CD85j) leukocyte receptor. J. Immunol. 168, 3351–3359 (2002).
Munitz, A., McBride, M. L., Bernstein, J. S. & Rothenberg, M. E. A dual activation and inhibition role for the paired immunoglobulin-like receptor B in eosinophils. Blood 111, 5694–5703 (2008).
Lu-Kuo, J. M., Joyal, D. M., Austen, K. F. & Katz, H. R. gp49B1 inhibits IgE-initiated mast cell activation through both immunoreceptor tyrosine-based inhibitory motifs, recruitment of src homology 2 domain-containing phosphatase-1, and suppression of early and late calcium mobilization. J. Biol. Chem. 274, 5791–5796 (1999).
Chang, C. C. et al. Tolerization of dendritic cells by T(S) cells: the crucial role of inhibitory receptors ILT3 and ILT4. Nat. Immunol. 3, 237–243 (2002).
Cousens, L., Najafian, N., Martin, W. D. & De Groot, A. S. Tregitope: immunomodulation powerhouse. Hum. Immunol. 75, 1139–1146 (2014).
Xu, Z. et al. ILT3.Fc-CD166 interaction induces inactivation of p70 S6 kinase and inhibits tumor cell growth. J. Immunol. 200, 1207–1219 (2018).
Acknowledgements
We thank the National Cancer Institute (1R01CA172268), the Cancer Prevention and Research Institute of Texas (RP180435), the Robert A. Welch Foundation (I-1834), China Natural Science Foundation (81872332), Shandong Natural Science Foundation (2018GSF118201), and Yantai Science and Technology Development Plan (2018ZHGY070) for generous support.
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ZL.L., M.D., and CC.Z. designed the study and wrote the paper. ZL.L., M.D., FF.H., HY.C., XY.L., and LC.H. performed the mouse experiments. ZL.L., FF.H., CZ.J., and S.S. performed the flow cytometry analysis and western blotting. ZL.L., M.D., HY.C., XY.L., and AH.S. performed the CRISPR-Cas9 experiments and plasmid constructions. ZL.L. and M.D. performed the T-cell coculture and IHC. ZL.L., and M.D. performed the statistical analysis.
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C.C.Z. is a scientific founder of Immune-Onc Therapeutics. C.C.Z. and M.D. hold equity in and have multiple patents licensed to Immune-Onc Therapeutics. The other authors declare no competing interests.
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Li, Z., Deng, M., Huang, F. et al. LILRB4 ITIMs mediate the T cell suppression and infiltration of acute myeloid leukemia cells. Cell Mol Immunol 17, 272–282 (2020). https://doi.org/10.1038/s41423-019-0321-2
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DOI: https://doi.org/10.1038/s41423-019-0321-2
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