Interferon-induced GTPases [guanylate-binding proteins (GBPs)] play an important role in inflammasome activation and mediate innate resistance to many intracellular pathogens, but little is known about their role in leishmaniasis. We therefore studied expression of Gbp2b/Gbp1 and Gbp5 mRNA in skin, inguinal lymph nodes, spleen, and liver after Leishmania major infection and in uninfected controls. We used two different groups of related mouse strains: BALB/c, STS, and CcS-5, CcS-16, and CcS-20 that carry different combinations of BALB/c and STS genomes, and strains O20, C57BL/10 (B10) and B10.O20, OcB-9, and OcB-43 carrying different combinations of O20 and B10 genomes. The strains were classified on the basis of size and number of infection-induced skin lesions as highly susceptible (BALB/c, CcS-16), susceptible (B10.O20), intermediate (CcS-20), and resistant (STS, O20, B10, OcB-9, OcB-43). Some uninfected strains differed in expression of Gbp2b/Gbp1 and Gbp5, especially of Gbp2b/Gbp1 in skin. Uninfected BALB/c and STS did not differ in their expression, but in CcS-5, CcS-16, and CcS-20, which all carry BALB/c-derived Gbp gene-cluster, expression of Gbp2b/Gbp1 exceeds that of both parents. These data indicate trans-regulation of Gbps. Infection resulted in approximately 10× upregulation of Gbp2b/Gbp1 and Gbp5 mRNAs in organs of both susceptible and resistant strains, which was most pronounced in skin. CcS-20 expressed higher level of Gbp2b/Gbp1 than both parental strains in skin, whereas CcS-16 expressed higher level of Gbp2b/Gbp1 than both parental strains in skin and liver. This indicates a trans-regulation present in infected mice CcS-16 and CcS-20. Immunostaining of skin of five strains revealed in resistant and intermediate strains STS, CcS-5, O20, and CcS-20 tight co-localization of Gbp2b/Gbp1 protein with most L. major parasites, whereas in the highly susceptible strain, BALB/c most parasites did not associate with Gbp2b/Gbp1. In conclusion, expression of Gbp2b/Gbp1 and Gbp5 was increased even in organs of clinically asymptomatic resistant mice. It suggests a hidden inflammation, which might contribute to control of persisting parasites. This is supported by the co-localization of Gbpb2/Gbp1 protein and L. major parasites in skin of resistant and intermediate but not highly susceptible mice.

Department of Molecular and Cellular Biology Roswell Park Cancer Institute Buffalo NY United States

Laboratory of Molecular and Cellular Immunology Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czechia

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Meunier E, Broz P. Interferon-inducible GTPases in cell autonomous and innate immunity. Cell Microbiol (2016) 18:168–80. 10.1111/cmi.12546 PubMed DOI

Pilla-Moffett D, Barber MF, Taylor GA, Coers J. Interferon-inducible GTPases in host resistance, inflammation and disease. J Mol Biol (2016) 428:3495–513. 10.1016/j.jmb.2016.04.032 PubMed DOI PMC

Man SM, Place DE, Kuriakose T, Kanneganti T-D. Interferon-inducible guanylate-binding proteins at the interface of cell-autonomous immunity and inflammasome activation. J Leukoc Biol (2017) 101:143–50. 10.1189/jlb.4MR0516-223R PubMed DOI PMC

Vestal DJ, Jeyaratnam JA. The guanylate-binding proteins: emerging insights into the biochemical properties and functions of this family of large interferon-induced guanosine triphosphatase. J Interferon Cytokine Res (2011) 31:89–97. 10.1089/jir.2010.0102 PubMed DOI PMC

Britzen-Laurent N, Bauer M, Berton V, Fischer N, Syguda A, Reipschläger S, et al. Intracellular trafficking of guanylate-binding proteins is regulated by heterodimerization in a hierarchical manner. PLoS One (2010) 5:e14246. 10.1371/journal.pone.0014246 PubMed DOI PMC

Gupta SL, Rubin BY, Holmes SL. Interferon action: induction of specific proteins in mouse and human cells by homologous interferons. Proc Natl Acad Sci U S A (1979) 76:4817–21. 10.1073/pnas.76.10.4817 PubMed DOI PMC

Cheng YSE, Colonno RJ, Yin FH. Interferon induction of fibroblast proteins with guanylate binding activity. J Biol Chem (1983) 258:7746–50. PubMed

Kim B-H, Chee JD, Bradfield CJ, Park E-S, Kumar P, MacMicking JD. Interferon-induced guanylate-binding proteins in inflammasome activation and host defense. Nat Immunol (2016) 17:481–9. 10.1038/ni.3440 PubMed DOI PMC

Kresse A, Konermann C, Degrandi D, Beuter-Gunia C, Wuerthner J, Pfeffer K, et al. Analyses of murine GBP homology clusters based on in silico, in vitro and in vivo studies. BMC Genomics (2008) 9:158. 10.1186/1471-2164-9-158 PubMed DOI PMC

Praefcke GJK, McMahon HT. The dynamin superfamily: universal membrane tubulation and fission molecules? Nat Rev Mol Cell Biol (2004) 5:133–47. 10.1038/nrm1313 PubMed DOI

Daumke O, Praefcke GJK. Invited review: mechanisms of GTP hydrolysis and conformational transitions in the dynamin superfamily. Biopolymers (2016) 105:580–93. 10.1002/bip.22855 PubMed DOI PMC

Cheng YS, Patterson CE, Staeheli P. Interferon-induced guanylate-binding proteins lack an N(T)KXD consensus motif and bind GMP in addition to GDP and GTP. Mol Cell Biol (1991) 11:4717–25. 10.1128/MCB.11.9.4717.Updated PubMed DOI PMC

Prakash B, Praefcke GJK, Renault L, Wittinghofer A, Herrmann C. Structure of human guanylate-binding protein 1 representing a unique class of GTP-binding proteins. Nature (2000) 403:567–71. 10.1038/35000617 PubMed DOI

Degrandi D, Kravets E, Konermann C, Beuter-Gunia C, Klümpers V, Lahme S, et al. Murine guanylate binding protein 2 (mGBP2) controls PubMed DOI PMC

Broz P, Dixit VM. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol (2016) 16:407–20. 10.1038/nri.2016.58 PubMed DOI

Kravets E, Degrandi D, Ma Q, Peulen T-O, Klümpers V, Felekyan S, et al. Guanylate binding proteins directly attack PubMed DOI PMC

Nordmann A, Wixler L, Boergeling Y, Wixler V, Ludwig S. A new splice variant of the human guanylate-binding protein 3 mediates anti-influenza activity through inhibition of viral transcription and replication. FASEB J (2012) 26:1290–300. 10.1096/fj.11-189886 PubMed DOI

Krapp C, Hotter D, Gawanbacht A, McLaren PJ, Kluge SF, Stürzel CM, et al. Guanylate binding protein (GBP) 5 is an interferon-inducible inhibitor of HIV-1 infectivity. Cell Host Microbe (2016) 19:504–14. 10.1016/j.chom.2016.02.019 PubMed DOI

Anderson SL, Carton JM, Lou J, Xing L, Rubin BY. Interferon-induced guanylate binding protein-1 (GBP-1) mediates an antiviral effect against vesicular stomatitis virus and encephalomyocarditis virus. Virology (1999) 256:8–14. 10.1006/viro.1999.9614 PubMed DOI

Carter CC, Gorbacheva VY, Vestal DJ. Inhibition of VSV and EMCV replication by the interferon-induced GTPase, mGBP-2: differential requirement for wild-type GTP binding domain. Arch Virol (2005) 150:1213–20. 10.1007/s00705-004-0489-2 PubMed DOI

Rupper AC, Cardelli JA. Induction of guanylate binding protein 5 by gamma interferon increases susceptibility to PubMed DOI PMC

Tietzel I, El-Haibi C, Carabeo RA. Human guanylate binding proteins potentiate the anti- PubMed DOI PMC

Kim B-H, Shenoy AR, Kumar P, Das R, Tiwari S, MacMicking JD. A family of IFN-inducible 65-kD GTPases protects against bacterial infection. Science (2011) 332:717–21. 10.1126/science.1201711 PubMed DOI

Meunier E, Dick MS, Dreier RF, Schürmann N, Kenzelmann Broz D, Warming S, et al. Caspase-11 activation requires lysis of pathogen-containing vacuoles by IFN-induced GTPases. Nature (2014) 509:366–70. 10.1038/nature13157 PubMed DOI

Yamamoto M, Okuyama M, Ma JS, Kimura T, Kamiyama N, Saiga H, et al. A cluster of interferon-γ-inducible p65 GTPases plays a critical role in host defense against PubMed DOI

Lipoldova M, Demant P. Genetic susceptibility to infectious disease: lessons from mouse models of leishmaniasis. Nat Rev Genet (2006) 7:294–305. 10.1038/nrg1832 PubMed DOI

Dedet JP. Current status of epidemiology of leishmaniases. In: Farrell JP, editor. Leishmania Series: World Class Parasites, Vol. 4 London: Kluwer Academic Press; (2002). p. 1–10.

Lainson R, Shaw JJ. Evolution, classification and geographical distribution. In: Peters W, Killick-Kendrick R, editors. The Leishmaniases in Biology and Medicine. London: Academic Press; (1987). p. 1–20.

Frank B, Marcu A, de Oliveira Almeida Petersen AL, Weber H, Stigloher C, Mottram JC, et al. Autophagic digestion of PubMed DOI PMC

Jayakumar A, Donovan MJ, Tripathi V, Ramalho-Ortigao M, McDowell MA. PubMed DOI PMC

LifeMap Sciences, Inc. (2017). Available from: https://discovery.lifemapsc.com/gene-expression-signals/high-throughput/microarray-analysis-of-skin-lesions-from-leishmaniasis-patients/cotaneous-leishmaniasis-patients-skin-vs-normal-control-subjects-skin

Demant P, Hart AA. Recombinant congenic strains – a new tool for analyzing genetic traits determined by more than one gene. Immunogenetics (1986) 24:416–22. 10.1007/BF00377961 PubMed DOI

Stassen AP, Groot PC, Eppig JT, Demant P. Genetic composition of the recombinant congenic strains. Mamm Genome (1996) 7:55–8. 10.1007/s003359900013 PubMed DOI

Lipoldová M, Svobodová M, Havelková H, Krulová M, Badalová J, Nohýnková E, et al. Mouse genetic model for clinical and immunological heterogeneity of leishmaniasis. Immunogenetics (2002) 54:174–83. 10.1007/s00251-002-0439-7 PubMed DOI

Grekov I, Svobodová M, Nohýnková E, Lipoldová M. Preparation of highly infective PubMed DOI

Lipoldová M, Svobodová M, Krulová M, Havelková H, Badalová J, Nohýnková E, et al. Susceptibility to PubMed DOI

Kobets T, Badalová J, Grekov I, Havelková H, Svobodová M, Lipoldová M. PubMed DOI

Šíma M, Havelková H, Quan L, Svobodová M, Jarošíková T, Vojtíšková J, et al. Genetic control of resistance to PubMed DOI PMC

Staeheli P, Prochazka M, Steigmeier PA, Haller O. Genetic control of interferon action: mouse strain distribution and inheritance of an induced protein with guanylate-binding property. Virology (1984) 137:135–42. 10.1016/0042-6822(84)90016-3 PubMed DOI

Shockley KR, Churchill GA. Gene expression analysis of mouse chromosome substitution strains. Mamm Genome (2006) 17:598–614. 10.1007/s00335-005-0176-y PubMed DOI

Havelková H, Badalová J, Svobodová M, Vojtíšková J, Kurey I, Vladimirov V, et al. Genetics of susceptibility to leishmaniasis in mice: four novel loci and functional heterogeneity of gene effects. Genes Immun (2006) 7:220–33. 10.1038/sj.gene.6364290 PubMed DOI

Sohrabi Y, Havelková H, Kobets T, Šíma M, Volkova V, Grekov I, et al. Mapping the genes for susceptibility and response to PubMed DOI PMC

Stenger S, Donhauser N, Thuring H, Rollinghof M, Bogdan C. Reactivation of latent leishmaniasis by inhibition of inducible nitric oxide synthase. J Exp Med (1996) 183:1501–14. 10.1084/jem.183.4.1501 PubMed DOI PMC

Kobets T, Havelková H, Grekov I, Volkova V, Vojtíšková J, Slapničková M, et al. Genetics of host response to PubMed DOI PMC

Mandell MA, Beverley SM. Continual renewal and replication of persistent PubMed DOI PMC

Blos M, Schleicher U, Soares Rocha FJ, Meißner U, Röllinghoff M, Bogdan C. Organ-specific and stage-dependent control of PubMed DOI

Hammon M, Herrmann M, Bleiziffer O, Pryymachuk G, Andreoli L, Munoz LE, et al. Role of guanylate binding protein-1 in vascular defects associated with chronic inflammatory diseases. J Cell Mol Med (2011) 15:1582–92. 10.1111/j.1582-4934.2010.01146.x PubMed DOI PMC

Goo YH, Son SH, Yechoor VK, Paul A. Transcriptional profiling of foam cells reveals induction of guanylate-binding proteins following western diet acceleration of atherosclerosis in the absence of global changes in inflammation. J Am Heart Assoc (2016) 5. 10.1161/JAHA.115.002663 PubMed DOI PMC

Naschberger E, Croner RS, Merkel S, Dimmler A, Tripal P, Amann KU, et al. Angiostatic immune reaction in colorectal carcinoma: impact on survival and perspectives for antiangiogenic therapy. Int J Cancer (2008) 123:2120–9. 10.1002/ijc.23764 PubMed DOI

Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One (2012) 7:e35671. 10.1371/journal.pone.0035671 PubMed DOI PMC

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