Inflammasome Assembly is Required for Intracellular Formation of β2-microglobulin Amyloid Fibrils, Leading to IL-1β Secretion

Overview

Journal Int J Immunopathol Pharmacol

Publisher Sage Publications

Specialties Allergy & Immunology
Pathology
Pharmacology

Date 2022 May 26

PMID 35615856

Authors

Naoe Kaneko

Wakako Mori

Mie Kurata

Toshihiro Yamamoto

Tamotsu Zako

Junya Masumoto

Affiliations

Soon will be listed here.

Abstract

Introduction: Dialysis-related amyloidosis (DRA) caused by β2-microgloblin (B2M) fibrils is a serious complication for patients with kidney failure on long-term dialysis. Deposition of B2M amyloid fibrils is thought to be due not only to serum extracellular B2M but also to infiltrating inflammatory cells, which may have an important role in B2M amyloid deposition in osteoarticular tissues in patients with DRA. Here, we asked whether B2M amyloid fibrils activate the inflammasome and contribute to formation and deposition of amyloid fibrils in cells.

Methods: Amyloid formation was confirmed by a thioflavin T (ThT) spectroscopic assay and scanning electron microscopy (SEM). Activation of inflammasomes was assessed by detecting interleukin (IL)-1β in culture supernatants from human embryonic kidney (HEK) 293T cells ectopically expressing inflammasome components. IL-1β secretion was measured by enzyme-linked immunosorbent assay. Expression and co-localization were analyzed by immunohistochemistry and dual immunofluorescence microscopy.

Results: B2M amyloid fibrils interacted directly with NLRP3/Pyrin and to activate the NLRP3/Pyrin inflammasomes, resulting in IL-1β secretion. When HEK293T cells were transfected with inflammasome components NLRP3 or Pyrin, along with ASC, pro-caspase-1, pro-IL-1β, and B2M, ThT fluorescence intensity increased. This was accompanied by IL-1β secretion, which increased in line with the amount of transfected B2M. In this case, morphological glowing of amyloid fibrils was observed by SEM. In the absence of ASC, there was no increase in ThT fluorescence intensity or IL-1β secretion, or any morphological glowing of amyloid fibrils. NLRP3 or Pyrin and B2M were co-localized in a "speck" in HEK293T cells, and co-expressed in infiltrated monocytes/macrophages in the osteoarticular synovial tissues in a patient with DRA.

Conclusion: Taken together, these data suggest that inflammasome assembly is required for the subsequent triggering of intracellular formation of B2M amyloid fibrils, which may contribute to osteoarticular deposition of B2M amyloid fibrils and inflammation in patients with DRA.

Citing Articles

β2-microglobulin alters the profiles of inflammatory cytokines and of matrix metalloprotease in macrophages derived from the osteoarthritic synovium.

Muneshige K, Uchida K, Aikawa J, Iwase D, Takano S, Miyagi M Cent Eur J Immunol. 2023; 47(4):332-338.

PMID: 36817398 PMC: 9901252. DOI: 10.5114/ceji.2022.124092.

References

Schiffl H . Impact of advanced dialysis technology on the prevalence of dialysis-related amyloidosis in long-term maintenance dialysis patients. Hemodial Int. 2013; 18(1):136-41. DOI: 10.1111/hdi.12057. View

Mokuda S, Kanno M, Takasugi K, Okumura C, Ito Y, Masumoto J . Tocilizumab improved clinical symptoms of a patient with systemic tophaceous gout who had symmetric polyarthritis and fever: An alternative treatment by blockade of interleukin-6 signaling. SAGE Open Med Case Rep. 2016; 2:2050313X13519774. PMC: 4857353. DOI: 10.1177/2050313X13519774. View

Shi H, Murray A, Beutler B . Reconstruction of the Mouse Inflammasome System in HEK293T Cells. Bio Protoc. 2017; 6(21). PMC: 5431583. DOI: 10.21769/BioProtoc.1986. View

Zhou W, Kaneko N, Nakagita T, Takeda H, Masumoto J . A comprehensive interaction study provides a potential domain interaction network of human death domain superfamily proteins. Cell Death Differ. 2021; 28(11):2991-3008. PMC: 8564539. DOI: 10.1038/s41418-021-00796-x. View

Zako T, Sakono M, Kobayashi T, Sorgjerd K, Nilsson K, Hammarstrom P . Cell interaction study of amyloid by using luminescent conjugated polythiophene: implication that amyloid cytotoxicity is correlated with prolonged cellular binding. Chembiochem. 2012; 13(3):358-63. DOI: 10.1002/cbic.201100467. View

Morikawa S, Kaneko N, Okumura C, Taguchi H, Kurata M, Yamamoto T . IAPP/amylin deposition, which is correlated with expressions of ASC and IL-1β in β-cells of Langerhans' islets, directly initiates NLRP3 inflammasome activation. Int J Immunopathol Pharmacol. 2018; 32:2058738418788749. PMC: 6050799. DOI: 10.1177/2058738418788749. View

Masumoto J, Taniguchi S, Ayukawa K, Sarvotham H, Kishino T, Niikawa N . ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. J Biol Chem. 1999; 274(48):33835-8. DOI: 10.1074/jbc.274.48.33835. View

Compan V, Lopez-Castejon G . Functional Reconstruction of NLRs in HEK293 Cells. Methods Mol Biol. 2016; 1417:217-21. DOI: 10.1007/978-1-4939-3566-6_15. View

Mori W, Kaneko N, Nakanishi A, Zako T, Masumoto J . Insulin amyloid fibrils interact directly with the NLRP3, resulting in inflammasome activation and pyroptotic cell death. Int J Immunopathol Pharmacol. 2021; 35:20587384211038357. PMC: 8371720. DOI: 10.1177/20587384211038357. View

10.

Iwasaki T, Kaneko N, Ito Y, Takeda H, Sawasaki T, Heike T . Nod2-Nodosome in a Cell-Free System: Implications in Pathogenesis and Drug Discovery for Blau Syndrome and Early-Onset Sarcoidosis. ScientificWorldJournal. 2016; 2016:2597376. PMC: 4926014. DOI: 10.1155/2016/2597376. View

11.

Masters S, Dunne A, Subramanian S, Hull R, Tannahill G, Sharp F . Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1β in type 2 diabetes. Nat Immunol. 2010; 11(10):897-904. PMC: 3103663. DOI: 10.1038/ni.1935. View

12.

Sugiyama R, Agematsu K, Migita K, Nakayama J, Mokuda S, Ogura F . Defect of suppression of inflammasome-independent interleukin-8 secretion from SW982 synovial sarcoma cells by familial Mediterranean fever-derived pyrin mutations. Mol Biol Rep. 2013; 41(1):545-53. DOI: 10.1007/s11033-013-2890-y. View

13.

Hu J, Han J, Li H, Zhang X, Liu L, Chen F . Human Embryonic Kidney 293 Cells: A Vehicle for Biopharmaceutical Manufacturing, Structural Biology, and Electrophysiology. Cells Tissues Organs. 2018; 205(1):1-8. DOI: 10.1159/000485501. View

14.

Naiki H, Higuchi K, Nakakuki K, Takeda T . Kinetic analysis of amyloid fibril polymerization in vitro. Lab Invest. 1991; 65(1):104-10. View

15.

Ather J, Ckless K, Martin R, Foley K, Suratt B, Boyson J . Serum amyloid A activates the NLRP3 inflammasome and promotes Th17 allergic asthma in mice. J Immunol. 2011; 187(1):64-73. PMC: 3119761. DOI: 10.4049/jimmunol.1100500. View

16.

Portales-Castillo I, Yee J, Tanaka H, Fenves A . Beta-2 Microglobulin Amyloidosis: Past, Present, and Future. Kidney360. 2022; 1(12):1447-1455. PMC: 8815520. DOI: 10.34067/KID.0004922020. View

17.

Kihara M, Chatani E, Sakai M, Hasegawa K, Naiki H, Goto Y . Seeding-dependent maturation of beta2-microglobulin amyloid fibrils at neutral pH. J Biol Chem. 2005; 280(12):12012-8. DOI: 10.1074/jbc.M411949200. View

18.

Nakanishi A, Kaneko N, Takeda H, Sawasaki T, Morikawa S, Zhou W . Amyloid β directly interacts with NLRP3 to initiate inflammasome activation: identification of an intrinsic NLRP3 ligand in a cell-free system. Inflamm Regen. 2018; 38:27. PMC: 6231249. DOI: 10.1186/s41232-018-0085-6. View

19.

Yamamoto S, Kazama J, Narita I, Naiki H, Gejyo F . Recent progress in understanding dialysis-related amyloidosis. Bone. 2009; 45 Suppl 1:S39-42. DOI: 10.1016/j.bone.2009.03.655. View

20.

Halle A, Hornung V, Petzold G, Stewart C, Monks B, Reinheckel T . The NALP3 inflammasome is involved in the innate immune response to amyloid-beta. Nat Immunol. 2008; 9(8):857-65. PMC: 3101478. DOI: 10.1038/ni.1636. View