Transcriptomic Analysis of Spleen-derived Macrophages in Response to Lipopolysaccharide Shows Dependency on the MyD88-independent Pathway in Chinese Giant Salamanders (Andrias Davidianus)

Overview

Journal BMC Genomics

Publisher Biomed Central

Specialty Genetics

Date 2024 Oct 28

PMID 39465384

Authors

Jie Deng

Mengdi Han

Jingyu Gong

Hongying Ma

Yinting Hao

Cheng Fang

Han Zhang

Jia Li

Wei Jiang

Affiliations

Soon will be listed here.

Abstract

Background: Gram-negative bacteria are the main bacterial pathogens infecting Chinese giant salamanders (Andrias davidianus; CGS) in captivity and the wild, causing substantial economic losses in the CGS industry. However, the molecular mechanisms underlying pathogenesis following infection remain unclear.

Results: Spleen-derived macrophages from healthy CGS were isolated, cultured, and identified using density gradient centrifugation and immunofluorescence. A macrophage transcriptome database was established 0, 6, and 12 h post lipopolysaccharide stimulation using RNA-sequencing. In the final database 76,743 unigenes and 4,698 differentially expressed genes (DEGs) were functionally annotated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment results showed that DEGs were concentrated in toll-like receptor-nuclear factor kappa B-related immune pathways. Ten DEGs were validated 12 h after lipopolysaccharide (LPS) stimulation. Although the common LPS recognition receptor toll-like receptor 4 was not activated and the key adaptor protein MyD88 showed no significant response, we observed significant up-regulation of the following adaptors: toll/interleukin-1 receptor domain-containing adaptor inducing interferon-β, tumour necrosis factor receptor-associated factor 6, and transforming growth factor-β activated kinase 1, which are located downstream of the non-classical MyD88 pathway.

Conclusions: In contrast to that in other species, macrophage activation in CGS could depend on the non-classical MyD88 pathway in response to bacterial infection. Our study provides insights into the molecular mechanisms regulating CGS antibacterial responses, with implications for disease prevention and understanding immune evolution in amphibians.

References

Yamamoto M, Takeda K . Current views of toll-like receptor signaling pathways. Gastroenterol Res Pract. 2011; 2010:240365. PMC: 3010626. DOI: 10.1155/2010/240365. View

Wang R, Lan C, Benlagha K, Camara N, Miller H, Kubo M . The interaction of innate immune and adaptive immune system. MedComm (2020). 2024; 5(10):e714. PMC: 11401974. DOI: 10.1002/mco2.714. View

Park B, Song D, Kim H, Choi B, Lee H, Lee J . The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. Nature. 2009; 458(7242):1191-5. DOI: 10.1038/nature07830. View

Carpenter S, ONeill L . From periphery to center stage: 50 years of advancements in innate immunity. Cell. 2024; 187(9):2030-2051. PMC: 11060700. DOI: 10.1016/j.cell.2024.03.036. View

Fang X, Li Z, Liu C, Mor G, Liao A . Macrophage memory: Types, mechanisms, and its role in health and disease. Immunology. 2023; 171(1):18-30. DOI: 10.1111/imm.13697. View

Duan T, Du Y, Xing C, Wang H, Wang R . Toll-Like Receptor Signaling and Its Role in Cell-Mediated Immunity. Front Immunol. 2022; 13:812774. PMC: 8927970. DOI: 10.3389/fimmu.2022.812774. View

Hoffman H, Mueller J, Broide D, Wanderer A, Kolodner R . Mutation of a new gene encoding a putative pyrin-like protein causes familial cold autoinflammatory syndrome and Muckle-Wells syndrome. Nat Genet. 2001; 29(3):301-5. PMC: 4322000. DOI: 10.1038/ng756. View

Yan F, Lu J, Zhang B, Yuan Z, Zhao H, Huang S . The Chinese giant salamander exemplifies the hidden extinction of cryptic species. Curr Biol. 2018; 28(10):R590-R592. DOI: 10.1016/j.cub.2018.04.004. View

Martinon F, Burns K, Tschopp J . The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell. 2002; 10(2):417-26. DOI: 10.1016/s1097-2765(02)00599-3. View

10.

Baoprasertkul P, Xu P, Peatman E, Kucuktas H, Liu Z . Divergent Toll-like receptors in catfish (Ictalurus punctatus): TLR5S, TLR20, TLR21. Fish Shellfish Immunol. 2007; 23(6):1218-30. DOI: 10.1016/j.fsi.2007.06.002. View

11.

Wynn T, Chawla A, Pollard J . Macrophage biology in development, homeostasis and disease. Nature. 2013; 496(7446):445-55. PMC: 3725458. DOI: 10.1038/nature12034. View

12.

Pradeu T, Thomma B, Girardin S, Lemaitre B . The conceptual foundations of innate immunity: Taking stock 30 years later. Immunity. 2024; 57(4):613-631. DOI: 10.1016/j.immuni.2024.03.007. View

13.

Liu Y, Li M, Fan S, Lin Y, Lin B, Luo F . A unique feature of Toll/IL-1 receptor domain-containing adaptor protein is partially responsible for lipopolysaccharide insensitivity in zebrafish with a highly conserved function of MyD88. J Immunol. 2010; 185(6):3391-400. DOI: 10.4049/jimmunol.0903147. View

14.

Zaharoff D, Rogers C, Hance K, Schlom J, Greiner J . Chitosan solution enhances both humoral and cell-mediated immune responses to subcutaneous vaccination. Vaccine. 2007; 25(11):2085-94. PMC: 1890043. DOI: 10.1016/j.vaccine.2006.11.034. View

15.

Oshiumi H, Tsujita T, Shida K, Matsumoto M, Ikeo K, Seya T . Prediction of the prototype of the human Toll-like receptor gene family from the pufferfish, Fugu rubripes, genome. Immunogenetics. 2003; 54(11):791-800. DOI: 10.1007/s00251-002-0519-8. View

16.

Zhao X, Chu Q, Cui J, Huo R, Xu T . IRF9 as a negative regulator involved in TRIF-mediated NF-κB pathway in a teleost fish, Miichthys miiuy. Mol Immunol. 2017; 85:123-129. DOI: 10.1016/j.molimm.2017.02.009. View

17.

Cassado A . F4/80 as a Major Macrophage Marker: The Case of the Peritoneum and Spleen. Results Probl Cell Differ. 2017; 62:161-179. DOI: 10.1007/978-3-319-54090-0_7. View

18.

Lu C, Chai J, Murphy R, Che J . Giant salamanders: Farmed yet endangered. Science. 2020; 367(6481):989. DOI: 10.1126/science.abb2375. View

19.

Barton G, Medzhitov R . Toll-like receptor signaling pathways. Science. 2003; 300(5625):1524-5. DOI: 10.1126/science.1085536. View

20.

Meng Z, Zhang X, Guo J, Xiang L, Shao J . Scavenger receptor in fish is a lipopolysaccharide recognition molecule involved in negative regulation of NF-κB activation by competing with TNF receptor-associated factor 2 recruitment into the TNF-α signaling pathway. J Immunol. 2012; 189(8):4024-39. DOI: 10.4049/jimmunol.1201244. View