Abstract
Leishmaniasis is a zoonotic parasitic disease, and the main reservoir of the parasite is the dog, although recent years have seen an increase in other mammalian species. In the Mediterranean region, where it is an endemic disease, it is caused by the species Leishmania infantum. The Ibizan hound, an autochthonous breed of this region, appears to have a genetic resistance to parasitic infection, whereas other canine breeds, such as the Boxer, are susceptible to infection. These differences are related to the differentiated activation of the immune response, with the Ibizan hound activating the Th1 immune response, whereas the Boxer breed triggers the Th2 immune response. Cytokine levels and genomic haplotypes of several genes involved in the immune response were analysed in twenty-eight Ibizan hound (resistant canine breed model) and twenty-four Boxer (susceptible canine breed) without clinical signs in the Mediterranean region. Cytokine levels were analysed by ELISA commercial kits and haplotypes were studied using CanineHD DNA Analysis BeadChip including 165,480 mapped positions. The results show 126 haplotypes associated with differential immune response in dogs. Specifically, haplotypes in IL12RB1, IL6R, CIITA, THEMIS, NOXA1, HEY2, RAB38, SLC35D2, SLC28A3, RASEF and DAPK1 genes are associated with serum levels of IFN-γ, IL-2, IL-8, and IL-18. These results suggest that the resistance or susceptibility to Leishmania infantum infection could be a consequence of haplotypes in several genes related to immune response. Future studies are needed to elucidate the relationship of these haplotypes with immune response and gene expression regulation.
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References
Adhikari A, Cobb B, Eddington S et al (2020) IFN-γ and CIITA modulate IL-6 expression in skeletal muscle. Cytokine X 2:100023. https://doi.org/10.1016/j.cytox.2020.100023
Ahmed G, Thakur AK, Pushpanjali null, et al (2019) Modulation of the immune response and infection pattern to Leishmania donovani in visceral leishmaniasis due to arsenic exposure: An in vitro study. PLoS One 14:e0210737. https://doi.org/10.1371/journal.pone.0210737
Ahuir-Baraja AE, Ruiz MP, Garijo MM, Llobat L (2021) Feline Leishmaniosis: An Emerging Public Health Problem. Vet Sci 8:173. https://doi.org/10.3390/vetsci8090173
Akhoundi M, Kuhls K, Cannet A et al (2016) A Historical Overview of the Classification, Evolution, and Dispersion of Leishmania Parasites and Sandflies. PLoS Negl Trop Dis 10:e0004349. https://doi.org/10.1371/journal.pntd.0004349
Alvar J, Vélez ID, Bern C et al (2012) Leishmaniasis worldwide and global estimates of its incidence. PLoS One 7:e35671. https://doi.org/10.1371/journal.pone.0035671
Álvarez L, Marín-García P-J, Llobat L (2022) Serum levels and genetic variations of cytokines in two canine breeds (Ibizan hound and boxer) in the Mediterranean region, in terms of Leishmania infantum infection. Comp Immunol Microbiol Infect Dis 90–91:101908. https://doi.org/10.1016/j.cimid.2022.101908
Álvarez L, Marín-García P-J, Llobat L (2022b) Immunological and genomic characterization of Ibizan Hound dogs in an endemic Leishmania infantum region. Parasit Vectors 15:445. https://doi.org/10.1186/s13071-022-05504-3
Álvarez L, Marín-García P-J, Rentero-Garrido P, Llobat L (2022c) Immune and Genomic Analysis of Boxer Dog Breed and Its Relationship with Leishmania infantum Infection. Vet Sci 9:608. https://doi.org/10.3390/vetsci9110608
Athanasiou LV, Katsogiannou EG, Tsokana CN et al (2021) Wild Rabbit Exposure to Leishmania infantum, Toxoplasma gondii, Anaplasma phagocytophilum and Babesia caballi Evidenced by Serum and Aqueous Humor Antibody Detection. Microorganisms 9:2616. https://doi.org/10.3390/microorganisms9122616
Batista LFS, Utsunomiya YT, Silva TBF et al (2016) Genome-Wide Association Study of Cell-Mediated Response in Dogs Naturally Infected by Leishmania infantum. Infect Immun 84:3629–3637. https://doi.org/10.1128/IAI.00486-16
Bodas M, Jain N, Awasthi A et al (2006) Inhibition of IL-2 induced IL-10 production as a principle of phase-specific immunotherapy. J Immunol 177:4636–4643. https://doi.org/10.4049/jimmunol.177.7.4636
Brar HK, Roy G, Kanojia A et al (2022) Chromatin-Remodeling Factor BRG1 Is a Negative Modulator of L donovani in IFNγ Stimulated and Infected THP-1 Cells. Front Cell Infect Microbiol 12:860058. https://doi.org/10.3389/fcimb.2022.860058
Brzostek J, Gautam N, Zhao X et al (2020) T cell receptor and cytokine signal integration in CD8+ T cells is mediated by the protein Themis. Nat Immunol 21:186–198. https://doi.org/10.1038/s41590-019-0570-3
Burnham AC, Ordeix L, Alcover MM et al (2020) Exploring the relationship between susceptibility to canine leishmaniosis and anti-Phlebotomus perniciosus saliva antibodies in Ibizan hounds and dogs of other breeds in Mallorca. Spain Parasit Vectors 13:129. https://doi.org/10.1186/s13071-020-3992-8
Butticè G, Miller J, Wang L, Smith BD (2006) Interferon-gamma induces major histocompatibility class II transactivator (CIITA), which mediates collagen repression and major histocompatibility class II activation by human aortic smooth muscle cells. Circ Res 98:472–479. https://doi.org/10.1161/01.RES.0000204725.46332.97
Carrillo E, Moreno J (2009) Cytokine profiles in canine visceral leishmaniasis. Vet Immunol Immunopathol 128:67–70. https://doi.org/10.1016/j.vetimm.2008.10.310
Chabod M, Pedros C, Lamouroux L et al (2012) A spontaneous mutation of the rat Themis gene leads to impaired function of regulatory T cells linked to inflammatory bowel disease. PLoS Genet 8:e1002461. https://doi.org/10.1371/journal.pgen.1002461
Chakraborty S, Srivastava A, Jha MK et al (2015) Inhibition of CD40-induced N-Ras activation reduces leishmania major infection. J Immunol 194:3852–3860. https://doi.org/10.4049/jimmunol.1401996
Chang CC, Chow CC, Tellier LC et al (2015) Second-generation PLINK: rising to the challenge of larger and richer datasets. Gigascience 4:7. https://doi.org/10.1186/s13742-015-0047-8
Corradin SB, Mauël J (1991) Phagocytosis of Leishmania enhances macrophage activation by IFN-gamma and lipopolysaccharide. J Immunol 146:279–285
Cunha-Júnior EF, Andrade-Neto VV, Lima ML et al (2017) Cyclobenzaprine Raises ROS Levels in Leishmania infantum and Reduces Parasite Burden in Infected Mice. PLoS Negl Trop Dis 11:e0005281. https://doi.org/10.1371/journal.pntd.0005281
Edo M, Marín-García PJ, Llobat L (2021) Is the Prevalence of Leishmania infantum Linked to Breeds in Dogs? Characterization of Seropositive Dogs in Ibiza. Animals (basel) 11:2579. https://doi.org/10.3390/ani11092579
Ejghal R, Hida M, Idrissi ML et al (2014) SLC11A1 polymorphisms and susceptibility to visceral leishmaniasis in Moroccan patients. Acta Trop 140:130–136. https://doi.org/10.1016/j.actatropica.2014.08.013
Escobar TA, Dowich G, Dos Santos TP et al (2019) Assessment of Leishmania infantum infection in equine populations in a canine visceral leishmaniosis transmission area. BMC Vet Res 15:381. https://doi.org/10.1186/s12917-019-2108-1
Giner J, Villanueva-Saz S, Fernández A et al (2022) Detection of Anti-Leishmania infantum Antibodies in Wild European and American Mink (Mustela lutreola and Neovison vison) from Northern Spain, 2014–20. J Wildl Dis 58:198–204. https://doi.org/10.7589/JWD-D-21-00027
Gramiccia M (2011) Recent advances in leishmaniosis in pet animals: epidemiology, diagnostics and anti-vectorial prophylaxis. Vet Parasitol 181:23–30. https://doi.org/10.1016/j.vetpar.2011.04.019
Gurjar D, Kumar Patra S, Bodhale N et al (2022) Leishmania intercepts IFN-γR signaling at multiple levels in macrophages. Cytokine 157:155956. https://doi.org/10.1016/j.cyto.2022.155956
Hondowicz BD, Park AY, Elloso MM, Scott P (2000) Maintenance of IL-12-responsive CD4+ T cells during a Th2 response in Leishmania major-infected mice. Eur J Immunol 30:2007–2014. https://doi.org/10.1002/1521-4141(200007)30:7%3c2007::AID-IMMU2007%3e3.0.CO;2-W
Khattak FA, Akbar NU, Riaz M et al (2021) Novel IL-12Rβ1 deficiency-mediates recurrent cutaneous leishmaniasis. Int J Infect Dis 112:338–345. https://doi.org/10.1016/j.ijid.2021.08.049
Kohno K, Kataoka J, Ohtsuki T et al (1997) IFN-gamma-inducing factor (IGIF) is a costimulatory factor on the activation of Th1 but not Th2 cells and exerts its effect independently of IL-12. J Immunol 158:1541–1550
Lake SL, Lyon H, Tantisira K et al (2003) Estimation and tests of haplotype-environment interaction when linkage phase is ambiguous. Hum Hered 55:56–65. https://doi.org/10.1159/000071811
Liu Y, Cong Y, Niu Y et al (2022) Themis is indispensable for IL-2 and IL-15 signaling in T cells. Sci Signal 15:eab9983. https://doi.org/10.1126/scisignal.abi9983
Maia C, Campino L (2012) Cytokine and Phenotypic Cell Profiles of Leishmania infantum Infection in the Dog. J Trop Med 2012:541571. https://doi.org/10.1155/2012/541571
Maia C, Campino L (2018) Biomarkers Associated With Leishmania infantum Exposure, Infection, and Disease in Dogs. Front Cell Infect Microbiol 8:302. https://doi.org/10.3389/fcimb.2018.00302
Marín-García PJ, Llobat L (2022) Canine Cytokines Profile in an Endemic Region of L infantum: Related Factors. Vet Sci 9:305. https://doi.org/10.3390/vetsci9060305
Martín-Sánchez J, Torres-Medina N, Morillas-Márquez F et al (2021) Role of wild rabbits as reservoirs of leishmaniasis in a non-epidemic Mediterranean hot spot in Spain. Acta Trop 222:106036. https://doi.org/10.1016/j.actatropica.2021.106036
Mendonça VRR, Souza LCL, Garcia GC et al (2014) DDX39B (BAT1), TNF and IL6 gene polymorphisms and association with clinical outcomes of patients with Plasmodium vivax malaria. Malar J 13:278. https://doi.org/10.1186/1475-2875-13-278
Mendoza-Roldan JA, Latrofa MS, Tarallo VD et al (2021) Leishmania spp in Squamata reptiles from the Mediterranean basin. Transbound Emerg Dis. https://doi.org/10.1111/tbed.14438
Moafi M, Rezvan H, Sherkat R et al (2017) Evaluation of IL-12RB1, IL-12B, CXCR-3 and IL-17a expression in cases affected by a non-healing form of cutaneous leishmaniasis: an observational study design. BMJ Open 7:e013006. https://doi.org/10.1136/bmjopen-2016-013006
Morales-Yuste M, Martín-Sánchez J, Corpas-Lopez V (2022) Canine Leishmaniasis: Update on Epidemiology, Diagnosis, Treatment, and Prevention. Vet Sci 9:387. https://doi.org/10.3390/vetsci9080387
Ohkusu K, Yoshimoto T, Takeda K et al (2000) Potentiality of interleukin-18 as a useful reagent for treatment and prevention of Leishmania major infection. Infect Immun 68:2449–2456. https://doi.org/10.1128/IAI.68.5.2449-2456.2000
Okamura H, Tsutsi H, Komatsu T et al (1995) Cloning of a new cytokine that induces IFN-gamma production by T cells. Nature 378:88–91. https://doi.org/10.1038/378088a0
Ortuño M, Nachum-Biala Y, García-Bocanegra I et al (2022) An epidemiological study in wild carnivores from Spanish Mediterranean ecosystems reveals association between Leishmania infantum, Babesia spp. and Hepatozoon spp. infection and new hosts for Hepatozoon martis, Hepatozoon canis and Sarcocystis spp. Transbound Emerg Dis 69:2110–2125. https://doi.org/10.1111/tbed.14199
Parvaneh N, Barlogis V, Alborzi A, et al (2017) Visceral leishmaniasis in two patients with IL-12p40 and IL-12Rβ1 deficiencies. Pediatr Blood Cancer 64:. https://doi.org/10.1002/pbc.26362
Pedros C, Gaud G, Bernard I et al (2015) An Epistatic Interaction between Themis1 and Vav1 Modulates Regulatory T Cell Function and Inflammatory Bowel Disease Development. J Immunol 195:1608–1616. https://doi.org/10.4049/jimmunol.1402562
Petry A, Weitnauer M, Görlach A (2010) Receptor activation of NADPH oxidases. Antioxid Redox Signal 13:467–487. https://doi.org/10.1089/ars.2009.3026
Pradhan S, Schwartz RA, Patil A et al (2022) Treatment options for leishmaniasis. Clin Exp Dermatol 47:516–521. https://doi.org/10.1111/ced.14919
Ramia E, Chiaravalli AM, Bou Nasser Eddine F et al (2019) CIITA-related block of HLA class II expression, upregulation of HLA class I, and heterogeneous expression of immune checkpoints in hepatocarcinomas: implications for new therapeutic approaches. Oncoimmunology 8:1548243. https://doi.org/10.1080/2162402X.2018.1548243
Shweash M, Adrienne McGachy H, Schroeder J et al (2011) Leishmania mexicana promastigotes inhibit macrophage IL-12 production via TLR-4 dependent COX-2, iNOS and arginase-1 expression. Mol Immunol 48:1800–1808. https://doi.org/10.1016/j.molimm.2011.05.013
Singh N, Gedda MR, Tiwari N et al (2017) Solute carrier protein family 11 member 1 (Slc11a1) activation efficiently inhibits Leishmania donovani survival in host macrophages. J Parasit Dis 41:671–677. https://doi.org/10.1007/s12639-016-0864-4
Solano-Gallego L, Llull J, Ramos G et al (2000) The Ibizian hound presents a predominantly cellular immune response against natural Leishmania infection. Vet Parasitol 90:37–45. https://doi.org/10.1016/s0304-4017(00)00223-5
Solano-Gallego L, Miró G, Koutinas A et al (2011) LeishVet guidelines for the practical management of canine leishmaniosis. Parasit Vectors 4:86. https://doi.org/10.1186/1756-3305-4-86
Sortica VA, Cunha MG, Ohnishi MDO et al (2014) Role of IL6, IL12B and VDR gene polymorphisms in Plasmodium vivax malaria severity, parasitemia and gametocytemia levels in an Amazonian Brazilian population. Cytokine 65:42–47. https://doi.org/10.1016/j.cyto.2013.09.014
Sousa-Franco J, Araújo-Mendes E, Silva-Jardim I et al (2006) Infection-induced respiratory burst in BALB/c macrophages kills Leishmania guyanensis amastigotes through apoptosis: possible involvement in resistance to cutaneous leishmaniasis. Microbes Infect 8:390–400. https://doi.org/10.1016/j.micinf.2005.07.007
Torre S, Faucher SP, Fodil N et al (2015) THEMIS is required for pathogenesis of cerebral malaria and protection against pulmonary tuberculosis. Infect Immun 83:759–768. https://doi.org/10.1128/IAI.02586-14
van de Vosse E, Haverkamp MH, Ramirez-Alejo N et al (2013) IL-12Rβ1 deficiency: mutation update and description of the IL12RB1 variation database. Hum Mutat 34:1329–1339. https://doi.org/10.1002/humu.22380
Velez R, Gállego M (2020) Commercially approved vaccines for canine leishmaniosis: a review of available data on their safety and efficacy. Trop Med Int Health 25:540–557. https://doi.org/10.1111/tmi.13382
Vergadi E, Ieronymaki E, Lyroni K et al (2017) Akt Signaling Pathway in Macrophage Activation and M1/M2 Polarization. J Immunol 198:1006–1014. https://doi.org/10.4049/jimmunol.1601515
Wei Z, Li P, He R et al (2021) DAPK1 (death associated protein kinase 1) mediates mTORC1 activation and antiviral activities in CD8+ T cells. Cell Mol Immunol 18:138–149. https://doi.org/10.1038/s41423-019-0293-2
Wujcicka W, Gaj Z, Wilczyński J, Nowakowska D (2015) Contribution of IL6 -174 G>C and IL1B +3954 C>T polymorphisms to congenital infection with Toxoplasma gondii. Eur J Clin Microbiol Infect Dis 34:2287–2294. https://doi.org/10.1007/s10096-015-2481-z
Yang C, Blaize G, Marrocco R et al (2022) THEMIS enhances the magnitude of normal and neuroinflammatory type 1 immune responses by promoting TCR-independent signals. Sci Signal 15:eab15343. https://doi.org/10.1126/scisignal.abl5343 (https://www.who.int/es/news-room/fact-sheets/detail/leishmaniasis)
Acknowledgements
We sincerely thank the “Podenco Ibicenco Association” and “Boxer club España” for allowing their data and samples of Ibizan hound and Boxer dogs. We are grateful to the INCLIVA Precision Medicine Unit for carrying out the genomic analysis and to University Cardenal Herrera-CEU.
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This work was supported by IDOC (IDOC22-05) and PUENTE (PUENTE22-01) projects of University CEU Cardenal Herrera. The funding agency was not involved in the study design, data collection, data analysis or drafting of the manuscript.
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LA, PJMG performed data analysis and edited the manuscript. LL conceived of the research, designed the methods, and wrote the manuscript. All authors read and approved the final manuscript.
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Álvarez, L., Marín-García, PJ. & Llobat, L. Genetic haplotypes associated with immune response to Leishmania infantum infection in dogs. Vet Res Commun 47, 1675–1685 (2023). https://doi.org/10.1007/s11259-023-10123-z
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DOI: https://doi.org/10.1007/s11259-023-10123-z