The Non-classical Major Histocompatibility Complex II Protein SLA-DM is Crucial for African Swine Fever Virus Replication

Overview

Journal Sci Rep

Specialty Science

Date 2023 Aug 21

PMID 37604847

Authors

Katrin Pannhorst

Jolene Carlson

Julia E Holper

Finn Grey

John Kenneth Baillie

Dirk Hoper

Elisabeth Wohnke

Kati Franzke

Axel Karger

Walter Fuchs

Thomas C Mettenleiter

Affiliations

Soon will be listed here.

Abstract

African swine fever virus (ASFV) is a lethal animal pathogen that enters its host cells through endocytosis. So far, host factors specifically required for ASFV replication have been barely identified. In this study a genome-wide CRISPR/Cas9 knockout screen in porcine cells indicated that the genes RFXANK, RFXAP, SLA-DMA, SLA-DMB, and CIITA are important for productive ASFV infection. The proteins encoded by these genes belong to the major histocompatibility complex II (MHC II), or swine leucocyte antigen complex II (SLA II). RFXAP and CIITA are MHC II-specific transcription factors, whereas SLA-DMA/B are subunits of the non-classical MHC II molecule SLA-DM. Targeted knockout of either of these genes led to severe replication defects of different ASFV isolates, reflected by substantially reduced plating efficiency, cell-to-cell spread, progeny virus titers and viral DNA replication. Transgene-based reconstitution of SLA-DMA/B fully restored the replication capacity demonstrating that SLA-DM, which resides in late endosomes, plays a crucial role during early steps of ASFV infection.

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References

Chamorro S, Revilla C, Alvarez B, Lopez-Fuertes L, Ezquerra A, Dominguez J . Phenotypic characterization of monocyte subpopulations in the pig. Immunobiology. 2000; 202(1):82-93. DOI: 10.1016/S0171-2985(00)80055-8. View

Lithgow P, Takamatsu H, Werling D, Dixon L, Chapman D . Correlation of cell surface marker expression with African swine fever virus infection. Vet Microbiol. 2014; 168(2-4):413-9. PMC: 3969584. DOI: 10.1016/j.vetmic.2013.12.001. View

Reith W, LeibundGut-Landmann S, Waldburger J . Regulation of MHC class II gene expression by the class II transactivator. Nat Rev Immunol. 2005; 5(10):793-806. DOI: 10.1038/nri1708. View

Hemmink J, Khazalwa E, M Abkallo H, Oduor B, Khayumbi J, Svitek N . Deletion of the CD2v Gene from the Genome of ASFV-Kenya-IX-1033 Partially Reduces Virulence and Induces Protection in Pigs. Viruses. 2022; 14(9). PMC: 9503863. DOI: 10.3390/v14091917. View

Groenen M, Archibald A, Uenishi H, Tuggle C, Takeuchi Y, Rothschild M . Analyses of pig genomes provide insight into porcine demography and evolution. Nature. 2012; 491(7424):393-8. PMC: 3566564. DOI: 10.1038/nature11622. View

Puschnik A, Majzoub K, Ooi Y, Carette J . A CRISPR toolbox to study virus-host interactions. Nat Rev Microbiol. 2017; 15(6):351-364. PMC: 5800792. DOI: 10.1038/nrmicro.2017.29. View

Holper J, Grey F, Kenneth Baillie J, Regan T, Parkinson N, Hoper D . A Genome-Wide CRISPR/Cas9 Screen Reveals the Requirement of Host Sphingomyelin Synthase 1 for Infection with Pseudorabies Virus Mutant gDPass. Viruses. 2021; 13(8). PMC: 8402627. DOI: 10.3390/v13081574. View

Jouvenet N, Monaghan P, Way M, Wileman T . Transport of African swine fever virus from assembly sites to the plasma membrane is dependent on microtubules and conventional kinesin. J Virol. 2004; 78(15):7990-8001. PMC: 446130. DOI: 10.1128/JVI.78.15.7990-8001.2004. View

Portugal R, Martins C, Keil G . Novel approach for the generation of recombinant African swine fever virus from a field isolate using GFP expression and 5-bromo-2'-deoxyuridine selection. J Virol Methods. 2012; 183(1):86-9. DOI: 10.1016/j.jviromet.2012.03.030. View

10.

Alonso C, Borca M, Dixon L, Revilla Y, Rodriguez F, Escribano J . ICTV Virus Taxonomy Profile: Asfarviridae. J Gen Virol. 2018; 99(5):613-614. DOI: 10.1099/jgv.0.001049. View

11.

Thamamongood T, Aebischer A, Wagner V, Chang M, Elling R, Benner C . A Genome-Wide CRISPR-Cas9 Screen Reveals the Requirement of Host Cell Sulfation for Schmallenberg Virus Infection. J Virol. 2020; 94(17). PMC: 7431805. DOI: 10.1128/JVI.00752-20. View

12.

Hernaez B, Guerra M, Salas M, Andres G . African Swine Fever Virus Undergoes Outer Envelope Disruption, Capsid Disassembly and Inner Envelope Fusion before Core Release from Multivesicular Endosomes. PLoS Pathog. 2016; 12(4):e1005595. PMC: 4844166. DOI: 10.1371/journal.ppat.1005595. View

13.

Chapman D, Tcherepanov V, Upton C, Dixon L . Comparison of the genome sequences of non-pathogenic and pathogenic African swine fever virus isolates. J Gen Virol. 2008; 89(Pt 2):397-408. DOI: 10.1099/vir.0.83343-0. View

14.

Cuesta-Geijo M, Garcia-Dorival I, Del Puerto A, Urquiza J, Galindo I, Barrado-Gil L . New insights into the role of endosomal proteins for African swine fever virus infection. PLoS Pathog. 2022; 18(1):e1009784. PMC: 8820605. DOI: 10.1371/journal.ppat.1009784. View

15.

Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T . Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sci Rep. 2014; 4:5400. PMC: 4066266. DOI: 10.1038/srep05400. View

16.

Summerfield A, Guzylack-Piriou L, Schaub A, Carrasco C, Tache V, Charley B . Porcine peripheral blood dendritic cells and natural interferon-producing cells. Immunology. 2003; 110(4):440-9. PMC: 1783075. DOI: 10.1111/j.1365-2567.2003.01755.x. View

17.

Steimle V, Otten L, Zufferey M, MACH B . Complementation cloning of an MHC class II transactivator mutated in hereditary MHC class II deficiency (or bare lymphocyte syndrome). Cell. 1993; 75(1):135-46. View

18.

Alejo A, Matamoros T, Guerra M, Andres G . A Proteomic Atlas of the African Swine Fever Virus Particle. J Virol. 2018; 92(23). PMC: 6232493. DOI: 10.1128/JVI.01293-18. View

19.

Han J, Perez J, Chen C, Li Y, Benitez A, Kandasamy M . Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication. Cell Rep. 2018; 23(2):596-607. PMC: 5939577. DOI: 10.1016/j.celrep.2018.03.045. View

20.

Bernardes C, Antonio A, Pedroso de Lima M, Valdeira M . Cholesterol affects African swine fever virus infection. Biochim Biophys Acta. 1998; 1393(1):19-25. DOI: 10.1016/s0005-2760(98)00051-4. View