{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T19:51:47Z","timestamp":1772221907012,"version":"3.50.1"},"reference-count":37,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,6,10]],"date-time":"2025-06-10T00:00:00Z","timestamp":1749513600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Agencia estatal de investigaci\u00f3n (Ministerio de Ciencia, Innovaci\u00f3n y universidades, Spain)","award":["AGL2015-68477-P"],"award-info":[{"award-number":["AGL2015-68477-P"]}]},{"name":"Agencia estatal de investigaci\u00f3n (Ministerio de Ciencia, Innovaci\u00f3n y universidades, Spain)","award":["AEI\/10.13039\/501100011033"],"award-info":[{"award-number":["AEI\/10.13039\/501100011033"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Compounds in sand fly saliva elicit specific immune responses that may play a role in the establishment of canine Leishmania infection. Although canine antibodies to anti-sand fly saliva antigens have been extensively studied, little is known about cellular immune responses against Phlebotomus perniciosus salivary proteins. This study aimed to explore humoral and T-cell-mediated immunity against P. perniciosus salivary proteins in dogs (n = 85) from Mallorca (Spain), a leishmaniosis-endemic area, and find correlations with demographic (age, sex, and breed) and parasite-specific immunological parameters. Anti-sand fly saliva IgG was examined using a P. perniciosus whole salivary gland homogenate (SGH) ELISA and recombinant salivary protein rSP03B ELISA. Interferon gamma (IFN-\u03b3) release whole blood assays with L. infantum soluble antigen (LSA), SGH, and rSP03B were also performed. Positive correlations were found between IgG levels in the SGH and rSP03B tests and between concentrations of SGH IFN-\u03b3 and rSP03B IFN-\u03b3. While concentrations of SGH IFN-\u03b3 and rSP03B IFN-\u03b3 were low and produced only by a minority of dogs (less than 20%), high levels and frequencies of LSA IFN-\u03b3 as well as anti-saliva IgG for SGH and rSP03B were detected in a majority of dogs (61% and 75%, respectively). LSA IFN-\u03b3 levels were positively correlated with age and Leishmania-specific antibodies. In conclusion, dogs from a leishmaniosis-endemic area presented high humoral immunity against P. perniciosus salivary proteins, but their cellular immunity to these proteins was low and less frequent.<\/jats:p>","DOI":"10.3390\/pathogens14060576","type":"journal-article","created":{"date-parts":[[2025,6,10]],"date-time":"2025-06-10T06:48:56Z","timestamp":1749538136000},"page":"576","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Humoral and T-Cell-Mediated Immunity Against Phlebotomus perniciosus Salivary Proteins in Dogs from a Leishmaniosis-Endemic Area"],"prefix":"10.3390","volume":"14","author":[{"given":"N\u00faria","family":"Balsells-Aguilar","sequence":"first","affiliation":[{"name":"Departament de Medicina i Cirurgia Animals, Facultat de Veterin\u00e0ria, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9070-5992","authenticated-orcid":false,"given":"Maria Magdalena","family":"Alcover","sequence":"additional","affiliation":[{"name":"Secci\u00f3 de Parasitologia, Departament de Biologia, Salut i Medi Ambient, Facultat de Farm\u00e0cia i Ci\u00e8ncies de l\u2019Alimentaci\u00f3, Universitat de Barcelona, 08028 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1830-4366","authenticated-orcid":false,"given":"Marta","family":"Baxarias","sequence":"additional","affiliation":[{"name":"Departament de Medicina i Cirurgia Animals, Facultat de Veterin\u00e0ria, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alejandra","family":"\u00c1lvarez-Fern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Departament de Medicina i Cirurgia Animals, Facultat de Veterin\u00e0ria, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lourdes","family":"Alarc\u00f3n","sequence":"additional","affiliation":[{"name":"Departament de Medicina i Cirurgia Animals, Facultat de Veterin\u00e0ria, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Petra","family":"Sumova","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 12800 Prague, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1790-1123","authenticated-orcid":false,"given":"Petr","family":"Volf","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 12800 Prague, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8479-4896","authenticated-orcid":false,"given":"Laia","family":"Solano-Gallego","sequence":"additional","affiliation":[{"name":"Departament de Medicina i Cirurgia Animals, Facultat de Veterin\u00e0ria, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e0004770","DOI":"10.1371\/journal.pntd.0004349","article-title":"A historical overview of the classification, evolution, and dispersion of Leishmania parasites and sandflies","volume":"10","author":"Akhoundi","year":"2016","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e0009982","DOI":"10.1371\/journal.pntd.0009982","article-title":"Molecular identification of two newly identified human pathogens causing leishmaniasis using PCR-based methods on the 3\u2032 untranslated region of the heat shock protein 70 (type I) gene","volume":"15","author":"Jariyapan","year":"2021","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1186\/s13071-024-06282-w","article-title":"Canine leishmaniasis in the Americas: Etiology, distribution, and clinical and zoonotic importance","volume":"17","year":"2024","journal-title":"Parasit. Vectors"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e0004458","DOI":"10.1371\/journal.pntd.0004458","article-title":"Seasonal dynamics of Phlebotomine sand fly species proven vectors of mediterranean leishmaniasis caused by Leishmania infantum","volume":"10","author":"Alten","year":"2016","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1186\/s13071-020-04081-7","article-title":"Latest trends in Leishmania infantum infection in dogs in Spain, Part I: Mapped seroprevalence and sand fly distributions","volume":"13","author":"Montoya","year":"2020","journal-title":"Parasit. Vectors"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"105749","DOI":"10.1016\/j.actatropica.2020.105749","article-title":"Seasonal dynamics of phlebotomine sand flies and autochthonous transmission of Leishmania infantum in high-altitude ecosystems in southern Spain","volume":"213","author":"Abattouy","year":"2021","journal-title":"Acta Trop."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e0005600","DOI":"10.1371\/journal.pntd.0005600","article-title":"Insights into the sand fly saliva: Blood-feeding and immune interactions between sand flies, hosts, and Leishmania","volume":"11","author":"Lestinova","year":"2017","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"161","DOI":"10.14411\/fp.2006.022","article-title":"Sand fly saliva: Effects on host immune response and Leishmania transmission","volume":"53","author":"Volf","year":"2006","journal-title":"Folia Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Morales-Yuste, M., Mart\u00edn-S\u00e1nchez, J., and Corpas-Lopez, V. (2022). Canine leishmaniasis: Update on epidemiology, diagnosis, treatment, and prevention. Vet. Sci., 9.","DOI":"10.3390\/vetsci9080387"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.prevetmed.2018.10.015","article-title":"Seroprevalence of canine Leishmania infantum infection in the Mediterranean region and identification of risk factors: The example of North-Eastern and Pyrenean areas of Spain","volume":"162","author":"Velez","year":"2019","journal-title":"Prev. Vet. Med."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Cosma, C., Maia, C., Khan, N., Infantino, M., and Del Riccio, M. (2024). Leishmaniasis in humans and animals: A One Health approach for surveillance, prevention and control in a changing world. Trop. Med. Infect. Dis., 9.","DOI":"10.3390\/tropicalmed9110258"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103293","DOI":"10.1016\/j.meegid.2017.10.005","article-title":"Canine susceptibility to visceral leishmaniasis: A systematic review upon genetic aspects, considering breed factors and immunological concepts","volume":"74","author":"Furtado","year":"2019","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","first-page":"101908","article-title":"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","volume":"90\u201391","author":"Llobat","year":"2022","journal-title":"Comp. Immunol. Microbiol. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1186\/s13071-022-05504-3","article-title":"Immunological and genomic characterization of Ibizan Hound dogs in an endemic Leishmania infantum region","volume":"15","author":"Llobat","year":"2022","journal-title":"Parasit. Vectors"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1186\/s13071-020-3992-8","article-title":"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","volume":"13","author":"Burnham","year":"2020","journal-title":"Parasit. Vectors"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.rvsc.2020.02.004","article-title":"The balancing act: Immunology of leishmaniosis","volume":"130","author":"Toepp","year":"2020","journal-title":"Res. Vet. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1675","DOI":"10.1007\/s11259-023-10123-z","article-title":"Genetic haplotypes associated with immune response to Leishmania infantum infection in dogs","volume":"47","author":"Llobat","year":"2023","journal-title":"Vet. Res. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1111\/mve.12192","article-title":"The recombinant protein rSP03B is a valid antigen for screening dog exposure to Phlebotomus perniciosus across foci of canine leishmaniasis","volume":"31","author":"Kostalova","year":"2017","journal-title":"Med. Vet. Entomol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1186\/s13071-020-3993-7","article-title":"Monitoring Leishmania infection and exposure to Phlebotomus perniciosus using minimal and non-invasive canine samples","volume":"13","author":"Maia","year":"2020","journal-title":"Parasit. Vectors"},{"key":"ref_20","unstructured":"Ordeix, L. (2018). The Spectrum of Cutaneous Manifestations in Canine Leishmaniosis: Insights into Diagnosis and Immune Responses. [Ph.D. Thesis, Universitat Aut\u00f2noma de Barcelona]."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1186\/s13071-016-1598-y","article-title":"Leishmania infantum-specific production of IFN-\u03b3 and IL-10 in stimulated blood from dogs with clinical leishmaniosis","volume":"9","author":"Ordeix","year":"2016","journal-title":"Parasit. Vectors"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1186\/s13071-016-1519-0","article-title":"Early reduction of Leishmania infantum-specific antibodies and blood parasitemia during treatment in dogs with moderate or severe disease","volume":"9","author":"Ordeix","year":"2016","journal-title":"Parasit. Vectors"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0304-4017(99)00084-9","article-title":"Serological and parasitological follow-up in dogs experimentally infected with Leishmania infantum and treated with meglumine antimoniate","volume":"84","author":"Riera","year":"1999","journal-title":"Vet. Parasitol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e0007832","DOI":"10.1371\/journal.pntd.0007832","article-title":"Field study of the improved rapid sand fly exposure test in areas endemic for canine leishmaniasis","volume":"13","author":"Willen","year":"2019","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"839932","DOI":"10.3389\/fcimb.2022.839932","article-title":"Some good and some bad: Sand fly salivary proteins in the control of leishmaniasis and in autoimmunity","volume":"25","author":"Aoki","year":"2022","journal-title":"Front. Cell Infect. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7990","DOI":"10.1038\/s41598-020-64820-9","article-title":"Immunity to vector saliva is compromised by short sand fly seasons in endemic regions with temperate climates","volume":"10","author":"Oliveira","year":"2020","journal-title":"Sci Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e1000441","DOI":"10.1371\/journal.ppat.1000441","article-title":"Sand fly salivary proteins induce strong cellular immunity in a natural reservoir of visceral leishmaniasis with adverse consequences for Leishmania","volume":"5","author":"Collin","year":"2009","journal-title":"PLoS Pathog."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e0005905","DOI":"10.1371\/journal.pntd.0005905","article-title":"Human cellular and humoral immune responses to Phlebotomus papatasi salivary gland antigens in endemic areas differing in prevalence of Leishmania major infection","volume":"11","author":"Naouar","year":"2017","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1186\/s13071-018-3123-y","article-title":"Seasonal dynamics of canine antibody response to Phlebotomus perniciosus saliva in an endemic area of Leishmania infantum","volume":"11","author":"Velez","year":"2018","journal-title":"Parasit. Vectors"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s13071-017-2587-5","article-title":"Antibody response to sand fly saliva is a marker of transmission intensity but not disease progression in dogs naturally infected with Leishmania infantum","volume":"11","author":"Quinnell","year":"2018","journal-title":"Parasit. Vectors"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e0006803","DOI":"10.1371\/journal.pntd.0006803","article-title":"Age trends in asymptomatic and symptomatic Leishmania donovani infection in the Indian subcontinent: A review and analysis of data from diagnostic and epidemiological studies","volume":"12","author":"Chapman","year":"2018","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e0009531","DOI":"10.1371\/journal.pntd.0009531","article-title":"A review of the leishmanin skin test: A neglected test for a neglected disease","volume":"15","author":"Paulini","year":"2021","journal-title":"PLoS Neglected Trop. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e0197430","DOI":"10.1371\/journal.pone.0197430","article-title":"Visceral leishmaniasis in selected communities of Hamar and Banna-Tsamai districts in Lower Omo Valley, South West Ethiopia: Sero-epidemological and Leishmanin Skin Test Surveys","volume":"13","author":"Bekele","year":"2018","journal-title":"PLoS ONE"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"110350","DOI":"10.1016\/j.vetpar.2024.110350","article-title":"Updating the epidemiology of canine leishmaniosis in the United Kingdom through the use of electronic health data","volume":"333","author":"Checa","year":"2025","journal-title":"Vet. Parasitol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Almeida, M., Maia, C., Crist\u00f3v\u00e3o, J.M., Morgado, C., Barbosa, I., Ibars, R.F., Campino, L., Gon\u00e7alves, L., and Cortes, S. (2022). Seroprevalence and risk factors associated with Leishmania infection in dogs from Portugal. Microorganisms, 10.","DOI":"10.3390\/microorganisms10112262"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Lopes, R., Garc\u00eas, A., Silva, A., Brilhante-Sim\u00f5es, P., Martins, \u00c2., Duarte, E.L., Coelho, A.C., and Cardoso, L. (2024). Distribution of and relationships between epidemiological and clinicopathological parameters in canine leishmaniosis: A retrospective study of 15 years (2009\u20132023). Pathogens, 13.","DOI":"10.3390\/pathogens13080635"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"121","DOI":"10.3389\/fimmu.2012.00121","article-title":"Biomarkers for exposure to sand flies bites as tools to aid control of leishmaniasis","volume":"3","author":"Andrade","year":"2012","journal-title":"Front. Immunol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/6\/576\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:49:21Z","timestamp":1760032161000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/6\/576"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,10]]},"references-count":37,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["pathogens14060576"],"URL":"https:\/\/doi.org\/10.3390\/pathogens14060576","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,10]]}}}