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A novel NLRP3 homologue lacking the PYD domain activates IL-1β expression and promotes the bacterial elimination in Qihe crucian carp Carassius auratus

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Abstract

Nod-like receptor pyrin domain-containing protein 3 (NLRP3) is a crucial member of NLRs family, which plays an important role in host defense. However, knowledge about inflammation response of NLRP3 in fish was scarce. Herein, a novel NLRP3 homologue (designated as CaNLRP3-like) was identified in Qihe crucian carp Carassius auratus. Full-length cDNA of CaNLRP3-like was composed of 3898 bp, which contained a 3093-bp open reading frame encoding. Structural analysis revealed that CaNLRP3-like lacked the PYD domain at N-terminus. Multiple sequence alignment and phylogenetic analysis revealed that CaNLRP3-like showed the high identity with the fishes in Cyprinidae, distinct from the branch formed by the mammals. Spatial expression analysis revealed that CaNLRP3-like was expressed in all examined tissues, with the highest expression levels in gill. Immune challenge experiments revealed that the expression of CaNLRP3-like was significantly up-regulated in six tissues in cases of challenge with Aeromonas hydrophila, LPS, and polyI:C. The recombined protein of CaNLRP3-like with 3Flag-tag was observed obviously in the cytoplasm. Furthermore, knockdown of CaNLRP3-like reduced the mRNA level of IL-1β, and increased the bacterial loading in liver, spleen, and head kidney. Vice versa, overexpression of CaNLRP3-like increased the mRNA expression of IL-1β, and decreased the bacterial loading in three detected tissues. Taken together, CaNLRP3-like gene, as a NLRP3 homologue, was identified and characterized in C. auratus, and these results revealed its role in regulating IL-1β expression and restricting bacterial infection in vivo, which shed light on the potential of CaNLRP3-like as a therapeutic target upon bacterial infection in teleost fish.

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Data availability

The data that support the findings of this study are available from the corresponding author but restrictions apply to the availability of these data, which were used under license for the current study, and so are not publicly available. Data are however available from the authors upon reasonable request and with permission of participating local authority areas.

References

  • Bilen S, Biswas G, Otsuyama S, Kono T, Sakai M, Hikima J (2014) Inflammatory responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells stimulated with an inflammasome-inducing agent, nigericin. Dev Comp Immunol 46(2):222–230

    Article  CAS  PubMed  Google Scholar 

  • Cassel SL, Joly S, Sutterwala FS (2009) The NLRP3 inflammasome: a sensor of immune danger signals. Semin Immunol 21(4):194–198

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen H, Ding S, Tan J, Yang D, Zhang Y, Liu Q (2020) Characterization of the Japanese flounder NLRP3 inflammasome in restricting Edwardsiella piscicida colonization in vivo. Fish Shellfish Immunol 103:169–180

    Article  CAS  PubMed  Google Scholar 

  • Deng N, Zhao Y, Xu J, Ouyang H, Wu Z, Lai W, Lu Y, Lin H, Zhang Y, Lu D (2022) Molecular characterization and functional study of the NLRP3 inflammasome genes in Tetraodon nigroviridis. Fish Shellfish Immunol 131:570–581

    Article  CAS  PubMed  Google Scholar 

  • Elliott EI, Sutterwala FS (2015) Initiation and perpetuation of NLRP3 inflammasome activation and assembly. Immunol Rev 265(1):35–52

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guarda G, Zenger M, Yazdi AS, Schroder K, Ferrero I, Menu P, Tardivel A, Mattmann C, Tschopp J (2011) Differential expression of NLRP3 among hematopoietic cells. J Immunol 186(4):2529–2534

    Article  CAS  PubMed  Google Scholar 

  • Hatscher L, Amon L, Heger L, Dudziak D (2021) Inflammasomes in dendritic cells: friend or foe? Immunol Lett 234:16–32

    Article  CAS  PubMed  Google Scholar 

  • Heilig R, Broz P (2018) Function and mechanism of the pyrin inflammasome. Eur J Immunol 48(2):230–238

    Article  CAS  PubMed  Google Scholar 

  • He Y, Hara H, Núñez G (2016) Mechanism and regulation of NLRP3 inflammasome activation. Trends Biochem Sci 41(12):1012–1021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Inoue M, Shinohara ML (2013) NLRP3 Inflammasome and MS/EAE. Autoimmun Dis 2013:859145

    Google Scholar 

  • Laing KJ, Purcell MK, Winton JR, Hansen JD (2008) A genomic view of the NOD-like receptor family in teleost fish: identification of a novel NLR subfamily in zebrafish. BMC Evol Biol 8:42

    Article  PubMed  PubMed Central  Google Scholar 

  • Lamkanfi M, Dixit VM (2014) Mechanisms and functions of inflammasomes. Cell 157(5):1013–1022

    Article  CAS  PubMed  Google Scholar 

  • Li S, Chen X, Hao G, Geng X, Zhan W, Sun J (2016) Identification and characterization of a novel NOD-like receptor family CARD domain containing 3 gene in response to extracellular ATP stimulation and its role in regulating LPS-induced innate immune response in Japanese flounder (Paralichthys olivaceus) head kidney macrophages. Fish Shellfish Immunol 50:79–90

    Article  CAS  PubMed  Google Scholar 

  • Li S, Peng W, Li J, Hao G, Geng X, Sun J (2017) Characterization of Japanese flounder (Paralichthys olivaceus) Caspase1 involved in extracellular ATP-mediated immune signaling in fish. Fish Shellfish Immunol 67:536–545

    Article  CAS  PubMed  Google Scholar 

  • Li Z, Guo J, Bi L (2020a) Role of the NLRP3 inflammasome in autoimmune diseases. Biomed Pharmacother 130:110542

    Article  CAS  PubMed  Google Scholar 

  • Li JY, Wang YY, Shao T, Fan DD, Lin AF, Xiang LX, Shao JZ (2020b) The zebrafish NLRP3 inflammasome has functional roles in ASC-dependent interleukin-1β maturation and gasdermin E-mediated pyroptosis. J Biol Chem 295(4):1120–1141

    Article  PubMed  Google Scholar 

  • Liu X, Huang M, Huang X, Guan J, Li X, Xie D, Xu C (2022) NLRP3 is a promising target for regulating high glucose-induced inflammatory response in Megalobrama amblycephala. Aquaculture 555:738220

    Article  CAS  Google Scholar 

  • Lu A, Magupalli VG, Ruan J, Yin Q, Atianand MK, Vos MR, Schröder GF, Fitzgerald KA, Wu H, Egelman EH (2014) Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. Cell 156(6):1193–1206

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lu H, Su H, Liu Y, Yin K, Wang D, Li B, Wang Y, Xing M (2022) NLRP3 inflammasome is involved in the mechanism of the mitigative effect of lycopene on sulfamethoxazole-induced inflammatory damage in grass carp kidneys. Fish Shellfish Immunol 123:348–357

    Article  CAS  PubMed  Google Scholar 

  • Lv Z, Wei Z, Zhang Z, Li C, Shao Y, Zhang W, Zhao X, Li Y, Duan X, Xiong J (2017) Characterization of NLRP3-like gene from Apostichopus japonicus provides new evidence on inflammation response in invertebrates. Fish Shellfish Immunol 68:114–123

    Article  CAS  PubMed  Google Scholar 

  • Morimoto N, Kono T, Sakai M, Hikima JI (2021) Inflammasomes in teleosts: structures and mechanisms that induce pyroptosis during bacterial infection. Int J Mol Sci 22(9):4389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ohto U, Kamitsukasa Y, Ishida H, Zhang Z, Murakami K, Hirama C, Maekawa S, Shimizu T (2022) Structural basis for the oligomerization-mediated regulation of NLRP3 inflammasome activation. P Nati Acad Sci USA 119(11):e2121353119

    Article  CAS  Google Scholar 

  • Rathinam VA, Fitzgerald KA (2016) Inflammasome complexes: emerging mechanisms and effector functions. Cell 165(4):792–800

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ranson N, Veldhuis M, Mitchell B, Fanning S, Cook AL, Kunde D, Eri R (2018) NLRP3-dependent and -independent processing of interleukin (IL)-1β in active ulcerative colitis. Int J Mol Sci 20(1):57

    Article  PubMed  PubMed Central  Google Scholar 

  • Sharma D, Kanneganti TD (2016) The cell biology of inflammasomes: mechanisms of inflammasome activation and regulation. J Cell Biochem 213(6):617–629

    CAS  Google Scholar 

  • Song Y, Dong X, Hu G (2022) Transcriptome analysis of turbot (Scophthalmus maximus) head kidney and liver reveals immune mechanism in response to Vibrio anguillarum infection. J Fish Dis 45(7):1045–1057

    Article  CAS  PubMed  Google Scholar 

  • Sun J, Zhao XL, Pei C, Zhu L, Zhang J, Kong XH (2022) Molecular characterization of NLRC3 and its interaction with other inflammasome components and regulation on the bacterial colonization in Qihe crucian carp Carassius auratus. Fish Shellfish Immunol 131:958–971

    Article  CAS  PubMed  Google Scholar 

  • Sun J, Zhao XL, Pei C, Zhu L, Zhang J, Kong XH (2023) Molecular characteristics and the roles of CaASC and its restriction to Aeromonas hydrophila in Carassius auratus. Fish Shellfish Immunol 132:108452

    Article  CAS  PubMed  Google Scholar 

  • Swanson KV, Deng M, Ting JP (2019) The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol 19(8):477–489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Taechavasonyoo A, Kondo H, Nozaki R, Suzuki Y, Hirono I (2013) Identification of novel interleukin 1 beta family genes in Japanese flounder Paralichthys olivaceus. Fish Shellfish Immunol 34(1):393–396

    Article  CAS  PubMed  Google Scholar 

  • Vojtech LN, Scharping N, Woodson JC, Hansen JD (2012) Roles of inflammatory caspases during processing of zebrafish interleukin-1β in Francisella noatunensis infection. Infect Immun 80(8):2878–2885

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xie J, Hodgkinson JW, Li C, Kovacevic N, Belosevic M (2014) Identification and functional characterization of the goldfish (Carassius auratus L.) high mobility group box 1 (HMGB1) chromatin-binding protein. Dev Comp Immunol 44(1):245–253

    Article  CAS  PubMed  Google Scholar 

  • Zhang T, Du H, Feng S, Wu R, Chen T, Jiang J, Peng Y, Ye C, Fang R (2019) NLRP3/ASC/Caspase-1 axis and serine protease activity are involved in neutrophil IL-1β processing during Streptococcus pneumoniae infection. Biochem Bioph Res Co 513(3):675–680

    Article  CAS  Google Scholar 

  • Zhao S, Chen F, Yin Q, Wang D, Han W, Zhang Y (2020) Reactive oxygen species interact with NLRP3 inflammasomes and are involved in the inflammation of sepsis: from mechanism to treatment of progression. Front Physiol 11:571810

    Article  PubMed  PubMed Central  Google Scholar 

  • Zheng Z, Tang D, Zhao W, Wan Z, Yu M, Huang Z, Li L, Aweya JJ, Zhang Y (2021) NLRP3-like protein negatively regulates the expression of antimicrobial peptides in Penaeus vannamei hemocyates. Fish Shellfish Immun Rep 2:100039

    Article  Google Scholar 

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Funding

This research was supported by the National Natural Science Foundation of China (U2004154).

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Authors and Affiliations

  1. College of Life Science, Henan Normal University, Xinxiang, China

    Juan Sun & Xianghui Kong

  2. Engineering Lab of Henan Province for Aquatic Animal Disease Control, College of Fisheries, Henan Normal University, Xinxiang, 453007, China

    Juan Sun, Xianliang Zhao, Chao Pei, Lei Zhu, Jie Zhang & Xianghui Kong

  3. School of Nursing, Xinxiang Medical University, Xinxiang, China

    Juan Sun

Authors
  1. Juan Sun
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  2. Xianliang Zhao
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  3. Chao Pei
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  4. Lei Zhu
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  5. Jie Zhang
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  6. Xianghui Kong
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Contributions

Juan Sun and Xianghui Kong wrote the manuscript, Xianliang Zhao and Chao Pei performed experiments and data analysis, Lei Zhu interpreted the data, and Xianliang Zhao, Jie Zhang, and Xianghui Kong conceptualized and designed the project.

Corresponding author

Correspondence to Xianghui Kong.

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The study was performed in strict compliance with the recommendations set forth in the Animal Ethics Procedures and Guidelines of the People’s Republic of China. All animal protocols were approved by the Animal Administration and Ethics Committee of Henan Normal University.

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The authors declare no competing interests.

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Sun, J., Zhao, X., Pei, C. et al. A novel NLRP3 homologue lacking the PYD domain activates IL-1β expression and promotes the bacterial elimination in Qihe crucian carp Carassius auratus. Aquacult Int 32, 1135–1150 (2024). https://doi.org/10.1007/s10499-023-01209-1

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  • DOI: https://doi.org/10.1007/s10499-023-01209-1

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