TY - JOUR
T1 - Donor natural killer cells trigger production of β-2-microglobulin to enhance post–bone marrow transplant immunity
AU - Ruggeri, Loredana
AU - Urbani, Elena
AU - Chiasserini, Davide
AU - Susta, Federica
AU - Orvietani, Pier Luigi
AU - Burchielli, Emanuela
AU - Ciardelli, Sara
AU - Sola, Rosaria
AU - Bruscoli, Stefano
AU - Cardinale, Antonella
AU - Pierini, Antonio
AU - Piersma, Sander R.
AU - Pasquino, Stefano
AU - Locatelli, Franco
AU - Ramarli, Dunia
AU - Velardi, Enrico
AU - Binaglia, Luciano
AU - Jimenez, Connie R.
AU - Holländer, Georg A.
AU - Velardi, Andrea
N1 - Funding Information:
This work was supported by grants from the Italian Association for Cancer Research (AIRC), the Swiss National Science Foundation , the Leukemia and Lymphoma Society , and the Italian Ministry for Further Education. Vrije Universiteit Medical Center-Cancer Center Amsterdam is acknowledged for support of the mass spectrometry infrastructure. E.V. was supported by grants from the Amy Strelzer Manasevit Research Program and AIRC.
Funding Information:
The authors thank Temistocle Ragni (Ospedale Santa Maria della Misericordia, Perugia) and Adriano Carotti (Ospedale Bambino Gesù, Roma) for thymic tissue samples obtained during corrective cardiovascular surgery. The authors also thank Massimo F. Martelli, Professor Emeritus, University of Perugia, for critical review of the manuscript and helpful advice. This work was supported by grants from the Italian Association for Cancer Research (AIRC), the Swiss National Science Foundation, the Leukemia and Lymphoma Society, and the Italian Ministry for Further Education. Vrije Universiteit Medical Center-Cancer Center Amsterdam is acknowledged for support of the mass spectrometry infrastructure. E.V. was supported by grants from the Amy Strelzer Manasevit Research Program and AIRC. Contribution: L.R. designed and performed experiments and wrote paper; E.U. S.C. and R. S. performed murine transplant experiments and human cell cultures; F.S. and P.L.O. performed biochemical analyses; E.B. performed murine immune reconstitution analyses; D.C. designed and performed proteomics analyses and wrote paper; S.R.P. performed proteomics analyses; S.B. performed human RNA silencing experiments; D.R. contributed to human thymocyte culture experiments; S.P. as heart surgeon at the Ospedale Santa Maria della Misericordia, Perugia, provided human thymic tissue samples obtained during corrective cardiovascular surgery; L.B. conceived, designed, and supervised biochemical analyses and critically revised the manuscript; C.R.J. supervised proteomics analyses and critically revised the manuscript; G.A.H. designed experiments, provided constructive suggestions, and wrote paper; E.V. and A.C performed GSEA and revised the manuscript; A.P. and F.L. revised the manuscript; and A.V. conceived and supervised the project, designed experiments, and wrote the paper.
Publisher Copyright:
© 2022 The American Society of Hematology
PY - 2022/12/1
Y1 - 2022/12/1
N2 - Allogeneic hematopoietic transplantation is a powerful treatment for hematologic malignancies. Posttransplant immune incompetence exposes patients to disease relapse and infections. We previously demonstrated that donor alloreactive natural killer (NK) cells ablate recipient hematopoietic targets, including leukemia. Here, in murine models, we show that infusion of donor alloreactive NK cells triggers recipient dendritic cells (DCs) to synthesize β-2-microglobulin (B2M) that elicits the release of c-KIT ligand and interleukin-7 that greatly accelerate posttransplant immune reconstitution. An identical chain of events was reproduced by infusing supernatants of alloreactive NK/DC cocultures. Similarly, human alloreactive NK cells triggered human DCs to synthesize B2M that induced interleukin-7 production by thymic epithelial cells and thereby supported thymocyte cellularity in vitro. Chromatography fractionation of murine and human alloreactive NK/DC coculture supernatants identified a protein with molecular weight and isoelectric point of B2M, and mass spectrometry identified amino acid sequences specific of B2M. Anti-B2M antibody depletion of NK/DC coculture supernatants abrogated their immune-rebuilding effect. B2M knock-out mice were unable to undergo accelerated immune reconstitution, but infusion of (wild-type) NK/DC coculture supernatants restored their ability to undergo accelerated immune reconstitution. Similarly, silencing the B2M gene in human DCs, before coculture with alloreactive NK cells, prevented the increase in thymocyte cellularity in vitro. Finally, human recombinant B2M increased thymocyte cellularity in a thymic epithelial cells/thymocyte culture system. Our studies uncover a novel therapeutic principle for treating posttransplant immune incompetence and suggest that, upon its translation to the clinic, patients may benefit from adoptive transfer of large numbers of cytokine-activated, ex vivo–expanded donor alloreactive NK cells.
AB - Allogeneic hematopoietic transplantation is a powerful treatment for hematologic malignancies. Posttransplant immune incompetence exposes patients to disease relapse and infections. We previously demonstrated that donor alloreactive natural killer (NK) cells ablate recipient hematopoietic targets, including leukemia. Here, in murine models, we show that infusion of donor alloreactive NK cells triggers recipient dendritic cells (DCs) to synthesize β-2-microglobulin (B2M) that elicits the release of c-KIT ligand and interleukin-7 that greatly accelerate posttransplant immune reconstitution. An identical chain of events was reproduced by infusing supernatants of alloreactive NK/DC cocultures. Similarly, human alloreactive NK cells triggered human DCs to synthesize B2M that induced interleukin-7 production by thymic epithelial cells and thereby supported thymocyte cellularity in vitro. Chromatography fractionation of murine and human alloreactive NK/DC coculture supernatants identified a protein with molecular weight and isoelectric point of B2M, and mass spectrometry identified amino acid sequences specific of B2M. Anti-B2M antibody depletion of NK/DC coculture supernatants abrogated their immune-rebuilding effect. B2M knock-out mice were unable to undergo accelerated immune reconstitution, but infusion of (wild-type) NK/DC coculture supernatants restored their ability to undergo accelerated immune reconstitution. Similarly, silencing the B2M gene in human DCs, before coculture with alloreactive NK cells, prevented the increase in thymocyte cellularity in vitro. Finally, human recombinant B2M increased thymocyte cellularity in a thymic epithelial cells/thymocyte culture system. Our studies uncover a novel therapeutic principle for treating posttransplant immune incompetence and suggest that, upon its translation to the clinic, patients may benefit from adoptive transfer of large numbers of cytokine-activated, ex vivo–expanded donor alloreactive NK cells.
UR - https://www.scopus.com/pages/publications/85142447957
U2 - 10.1182/blood.2021015297
DO - 10.1182/blood.2021015297
M3 - Article
C2 - 35984965
SN - 0006-4971
VL - 140
SP - 2323
EP - 2334
JO - Blood
JF - Blood
IS - 22
ER -
SDG 3 Good Health and Well-being