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Licensed Unlicensed Requires Authentication Published by De Gruyter December 28, 2016

Transcriptional immune response in mesenteric lymph nodes in pigs with different levels of resistance to Ascaris suum

Per Skallerup, Peter Nejsum, Susanna Cirera, Kerstin Skovgaard, Christian B. Pipper, Merete Fredholm, Claus B. Jørgensen and Stig M. Thamsborg
From the journal Acta Parasitologica


A single nucleotide polymorphism on chromosome 4 (SNP TXNIP) has been reported to be associated with roundworm (Ascaris suum) burden in pigs. The objective of the present study was to analyse the immune response to A. suum mounted by pigs with genotype AA (n = 24) and AB (n = 23) at the TXNIP locus. The pigs were repeatedly infected with A. suum from eight weeks of age until necropsy eight weeks later. An uninfected control group (AA; n = 5 and AB; n = 5) was also included. At post mortem, we collected mesenteric lymph nodes and measured the expression of 28 selected immune-related genes. Recordings of worm burdens confirmed our previous results that pigs of the AA genotype were more resistant to infection than AB pigs. We estimated the genotype difference in relative expression levels in infected and uninfected animals. No significant change in expression levels between the two genotypes due to infection was observed for any of the genes, although IL-13 approached significance (P = 0.08; Punadjusted = 0.003). Furthermore, statistical analysis testing for the effect of infection separately in each genotype showed significant up-regulation of IL-13 (P<0.05) and CCL17 (P<0.05) following A. suum infection in the ‘resistant’ AA genotype and not in the ‘susceptible’ AB genotype. Pigs of genotype AB had higher expression of the high-affinity IgG receptor (FCGR1A) than AA pigs in both infected and non-infected animals (P = 1.85*10-11).


The authors wish to thank all those who contributed to the research project which forms the basis of this paper: Anne Strandsby, Tina Neergaard Mahler and Karin Tarp provided excellent technical assistance. We would also like to acknowledge Dries Masure for providing input to the selection of genes. P.S. is supported by a University of Copenhagen (Denmark) Ph.D. grant. This work was supported by a Danish Agency for Science, Technology and Innovation grant awarded to P.N.


Adugna S., Kebede Y., Moges F., Tiruneh M. 2007. Efficacy of mebendazole and albendazole for Ascaris lumbricoides and hookworm infections in an area with long time exposure for antihelminthes, Northwest Ethiopia. Ethiopian Medical Journal, 45, 301–306Search in Google Scholar

Albers G.A., Gray G.D., Piper L.R., Barker J.S., Le Jambre L.F., Barger I.A. 1987. The genetics of resistance and resilience to Haemonchus contortus infection in young Merino sheep. International Journal for Parasitology, 17, 1355–136310.1016/0020-7519(87)90103-2Search in Google Scholar

Andersen C.L., Jensen J.L., Orntoft T.F. 2004. Normalization of realtime quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Research, 64, 5245–525010.1158/0008-5472.CAN-04-0496Search in Google Scholar

Andronicos N., Hunt P., Windon R. 2010. Expression of genes in gastrointestinal and lymphatic tissues during parasite infection in sheep genetically resistant or susceptible to Trichostrongylus colubriformis and Haemonchus contortus. International Journal for Parasitology, 40, 417–429. 10.1016/j.ijpara. 2009.09.007Search in Google Scholar

Anthony R.M., Rutitzky L.I., Urban J.F., Jr., Stadecker M.J., Gause W.C. 2007. Protective immune mechanisms in helminth infection. Nature Reviews Immunology, 7, 975–98710.1038/nri2199Search in Google Scholar

Anthony R.M., Urban J.F., Jr, Alem F., Hamed H.A., Rozo C.T., Boucher J.L., et al. 2006. Memory TH2 cells induce alternatively activated macrophages to mediate protection against nematode parasites. Nature Medicine, 12, 955-96010.1038/nm1451Search in Google Scholar

Araujo R.N., Padilha T., Zarlenga D., Sonstegard T., Connor E.E., Van Tassel C., et al. 2009. Use of a candidate gene array to delineate gene expression patterns in cattle selected for resistance or susceptibility to intestinal nematodes. Veterinary Parasitology, 162, 106–115. 10.1016/j.vetpar.2008.12.017Search in Google Scholar

Barger I.A. 1993. Influence of sex and reproductive status on susceptibility of ruminants to nematode parasitism. International Journal for Parasitology, 23, 463–46910.1016/0020-7519(93)90034-VSearch in Google Scholar

Burke M.L., McGarvey L., McSorley H.J., Bielefeldt-Ohmann H., McManus D.P., Gobert G.N. 2011. Migrating Schistosoma japonicum schistosomula induce an innate immune response and wound healing in the murine lung. Molecular Immunology, 49, 191–200. 10.1016/j.molimm.2011.08.014Search in Google Scholar PubMed

Bustin S.A., Benes V., Garson J.A., Hellemans J., Huggett J., Kubista M., et al. 2009. The MIQE guidelines: Minimum information for publication of quantitative real-time PCR 10.1373/clinchem.2008.11279710.1373/clinchem.2008.112797Search in Google Scholar PubMed

Cooper P.J., Figuieredo C.A. 2013. Immunology of Ascaris and immunomodulation. In: (Ed. C. V. Holland) Ascaris: The neglected parasite. Elsevier, London, 3-1910.1016/B978-0-12-396978-1.00001-XSearch in Google Scholar

Dawson H., Solano-Aguilar G., Beal M., Beshah E., Vangimalla V., Jones E., et al. 2009. Localized Th1-, Th2, T regulatory cell, and inflammation-associated hepatic and pulmonary immune responses in Ascaris suum-infected swine are increased by retinoic acid. Infection and Immunity, 77, 2576–2587. 10.1128/IAI.00827-07Search in Google Scholar PubMed PubMed Central

Dawson H.D., Beshah E., Nishi S., Solano-Aguilar G., Morimoto M., Zhao A., et al. 2005. Localized multigene expression patterns support an evolving Th1/Th2-like paradigm in response to infections with Toxoplasma gondii and Ascaris suum. Infection and Immunity, 73, 1116–112810.1128/IAI.73.2.1116-1128.2005Search in Google Scholar PubMed PubMed Central

Else K.J., Hultner L., Grencis R.K. 1992. Cellular immune responses to the murine nematode parasite Trichuris muris. II. Differential induction of TH-cell subsets in resistant versus susceptible mice. Immunology, 75, 232–237Search in Google Scholar

Geiger S.M., Jardim-Botelho A., Williams W., Alexander N., Diemert D.J., Bethony J.M. 2013. Serum CCL11 (eotaxin-1) and CCL17 (TARC) are serological indicators of multiple helminth infections and are driven by Schistosoma mansoni infection in humans. Tropical Medicine and International Health, 18, 750–760. 10.1111/tmi.12095Search in Google Scholar PubMed

Gossner A., Wilkie H., Joshi A., Hopkins J. 2013. Exploring the abomasal lymph node transcriptome for genes associated with resistance to the sheep nematode Teladorsagia circum-cincta. Veterinary Research, 44, 68. 10.1186/1297-9716-44-68Search in Google Scholar PubMed PubMed Central

Gossner A.G., Venturina V.M., Shaw D.J., Pemberton J.M., Hopkins, J. 2012. Relationship between susceptibility of blackface sheep to Teladorsagia circumcincta infection and an inflammatory mucosal T cell response. Veterinary Research, 43, 26. 10.1186/129797164326Search in Google Scholar

Groenen M.A., Archibald A.L., Uenishi H., Tuggle C.K., Takeuchi Y., Rothschild M.F., et al. 2012. Analyses of pig genomes provide insight into porcine demography and evolution. Nature 491, 393–398. 10.1038/nature11622Search in Google Scholar PubMed PubMed Central

Gutiérrez-Gil B., Perez J., Alvarez L., Martinez-Valladares M., de la Fuente L.F., Bayon Y., et al. 2009. Quantitative trait loci for resistance to trichostrongylid infection in Spanish Churra sheep. Genetics Selection Evolution, 41, 46. 10.1186/1297-9686-41-46Search in Google Scholar PubMed PubMed Central

Hanotte O., Ronin Y., Agaba M., Nilsson P., Gelhaus A., Horstmann R., et al. 2003. Mapping of quantitative trait loci controlling trypanotolerance in a cross of tolerant West African N’Dama and susceptible East African Boran cattle. Proceedings of the National Academy of Sciences U. S. A, 100, 7443–744810.1073/pnas.1232392100Search in Google Scholar PubMed PubMed Central

Hartl D., Lee C.G., Da Silva C.A., Chupp G.L., Elias J.A. 2009. Novel biomarkers in asthma: Chemokines and chitinase-like proteins. Current Opinion in Allergy and Clinical Immunology, 9, 60–66. 10.1097/ACI.0b013e32831f8ee0Search in Google Scholar PubMed

Hassan M., Good B., Hanrahan J.P., Campion D., Sayers G., Mulcahy G., Sweeney T. 2011a. The dynamic influence of the DRB1*1101 allele on the resistance of sheep to experimental Teladorsagia circumcincta infection. Veterinary Research, 42, 46. 10.1186/1297-9716-42-46Search in Google Scholar PubMed PubMed Central

Hassan M., Hanrahan J.P., Good B., Mulcahy G., Sweeney T. 2011b. A differential interplay between the expression of Th1/Th2/Treg related cytokine genes in Teladorsagia circumcincta infected DRB1*1101 carrier lambs. Veterinary Research, 42, 45. 10.1186/1297-9716-42-45Search in Google Scholar PubMed PubMed Central

Hindorff L.A., Sethupathy P., Junkins H.A., Ramos E.M., Mehta J.P., Collins F.S., Manolio T.A. 2009. Potential etiologic and functional implications of genome-wide association loci for human diseases and traits. Proceedings of the National Academy of Sciences U. S. A, 106, 9362–9367. 10.1073/pnas. 0903103106Search in Google Scholar

Hothorn T., Bretz F., Westfall P. 2008. Simultaneous inference in general parametric models. Biometrical Journal, 50, 346–363. 10.1002/bimj.200810425Search in Google Scholar PubMed

Ingham A., Reverter A., Windon R., Hunt P., Menzies M. 2008. Gastrointestinal nematode challenge induces some conserved gene expression changes in the gut mucosa of genetically resistant sheep. International Journal for Parasitology, 38, 431–44210.1016/j.ijpara.2007.07.012Search in Google Scholar PubMed

Keiser J., Utzinger J. 2008. Efficacy of current drugs against soiltransmitted helminth infections: Systematic review and metaanalysis. Journal of the American Medical Association, 299, 1937–1948. 10.1001/jama.299.16.1937Search in Google Scholar PubMed

LaPorte S.L., Juo Z.S., Vaclavikova J., Colf L.A., Qi X., Heller N.M., Keegan A.D., Garcia K.C. 2008. Molecular and structural basis of cytokine receptor pleiotropy in the interleukin-4/13 system. Cell, 132, 259–272. 10.1016/j.cell.2007.12.030Search in Google Scholar PubMed PubMed Central

Masure D., Vlaminck J., Wang T., Chiers K., Van den Broeck W., Vercruysse J., Geldhof P. 2013. A role for eosinophils in the intestinal immunity against infective Ascaris suum larvae. PLoS Neglected Tropical Diseases, 7, e213810.1371/journal.pntd.0002138Search in Google Scholar PubMed PubMed Central

Matika O., Pong-Wong R., Woolliams J.A., Bishop S.C. 2011. Confirmation of two quantitative trait loci regions for nematode resistance in commercial British terminal sire breeds. Animal, 5, 1149–1156. 10.1017/S175173111100022XSearch in Google Scholar PubMed

Nejsum P., Roepstorff A., Jorgensen C.B., Fredholm M., Göring H.H., Anderson T.J., Thamsborg S.M. 2009. High heritability for Ascaris and Trichuris infection levels in pigs. Heredity, 102, 357–364. 10.1038/hdy.2008.131Search in Google Scholar PubMed

Noyes H., Brass A., Obara I., Anderson S., Archibald A.L., Bradley D.G., et al. 2011. Genetic and expression analysis of cattle identifies candidate genes in pathways responding to Trypanosoma congolense infection. Proceedings of the National Academy of Sciences U. S. A, 108, 9304-9309. 10.1073/ pnas.1013486108Search in Google Scholar

Oksanen A., Eriksen L., Roepstorff A., Ilsoe B., Nansen P., Lind P. 1990. Embryonation and infectivity of Ascaris suum eggs. A comparison of eggs collected from worm uteri with eggs isolated from pig faeces. Acta Veterinaria Scandinavica, 31, 393–39810.1186/BF03547520Search in Google Scholar

Pernthaner A., Cole S.A., Morrison L., Hein W.R. 2005. Increased expression of interleukin-5 (IL-5), IL-13, and tumor necrosis factor alpha genes in intestinal lymph cells of sheep selected for enhanced resistance to nematodes during infection with Trichostrongylus colubriformis. Infection and Immunity, 73, 2175–218310.1128/IAI.73.4.2175-2183.2005Search in Google Scholar PubMed PubMed Central

R Core Team 2013. R: A language and environment for statistical computing. R foundation for statistical computing, Vienna, AustriaSearch in Google Scholar

Reinemeyer C.R. 2012. Anthelmintic resistance in non-strongylid parasites of horses. Veterinary Parasitology, 185, 9–15. 10.1016/j.vetpar.2011.10.009Search in Google Scholar PubMed

Roepstorff A., Eriksen L., Slotved H.C., Nansen, P. 1997. Experimental Ascaris suum infection in the pig: Worm population kinetics following single inoculations with three doses of infective eggs. Parasitology, 115, 443–45210.1017/S0031182097001480Search in Google Scholar PubMed

Rozen S., Skaletsky H. 2000. Primer3 on the WWW for general users and for biologist programmers. Methods in Molecular Biology, 132, 365–38610.1385/1-59259-192-2:365Search in Google Scholar

Schaschl H., Aitman T.J., Vyse T.J. 2009. Copy number variation in the human genome and its implication in autoimmunity. Clinical and Experimental Immunology, 156, 12–16. 10.1111/j.1365-2249.2008.03865.xSearch in Google Scholar PubMed PubMed Central

Skallerup P., Nejsum P., Jorgensen C.B., Göring H.H., Karlskov-Mortensen P., Archibald A.L., et al. 2012. Detection of a quantitative trait locus associated with resistance to Ascaris suum infection in pigs. International Journal for Parasitology, 42, 383–391. 10.1016/j.ijpara.2012.02.010Search in Google Scholar PubMed

Skallerup P., Thamsborg S.M., Jorgensen C.B., Enemark H.L., Yoshida A., Göring H.H., et al. 2014. Functional study of a genetic marker allele associated with resistance to Ascaris suum in pigs. Parasitology, 141, 777–787. 10.1017/ S0031182013002175Search in Google Scholar

Skovgaard K., Cirera S., Vasby D., Podolska A., Breum S.O., Durrwald R., et al. 2013. Expression of innate immune genes, proteins and microRNAs in lung tissue of pigs infected experimentally with influenza virus (H1N2). Innate Immunity, 19, 531–544. 10.1177/1753425912473668Search in Google Scholar PubMed

Slotved H.C., Barnes E.H., Eriksen L., Roepstorff A., Nansen P., Bjorn H. 1997. Use of an agar-gel technique for large scale application to recover Ascaris suum larvae from intestinal contents of pigs. Acta Veterinaria Scandinavica, 38, 207–212.10.1186/BF03548483Search in Google Scholar

Steenhard N.R., Jungersen G., Kokotovic B., Beshah E., Dawson H.D., Urban J.F., Jr, et al. 2009. Ascaris suum infection negatively affects the response to a Mycoplasma hyopneumoniae vaccination and subsequent challenge infection in pigs. Vaccine 27, 5161–5169. 10.1016/j.vaccine.2009.05.075Search in Google Scholar PubMed

Taylor M.D., van der Werf N., Maizels R.M. 2012. T cells in helminth infection: The regulators and the regulated. Trends in Immunology, 33, 181–189. 10.1016/ in Google Scholar PubMed

Thamsborg S.M., Nejsum P., Mejer H. 2013. Impact of Ascaris suum in livestock. In: (Ed. C. V. Holland) Ascaris: The neglected parasite. Elsevier, London, 363–38110.1016/B978-0-12-396978-1.00014-8Search in Google Scholar

van der Poel C.E., Spaapen R.M., van de Winkel J.G., Leusen J.H. 2011. Functional characteristics of the high affinity IgG receptor, FcyRI. Journal of Immunology, 186, 2699–2704. 10.4049/jimmunol.1003526Search in Google Scholar PubMed

Vlaminck J., Geldhof P. 2013. Diagnosis and control of ascariasis in pigs. In: (Ed. C. V. Holland) Ascaris: The neglected parasite. Elsevier, London, 395–42510.1016/B978-0-12-396978-1.00016-1Search in Google Scholar

Wakelin D. 1975. Genetic control of immune responses to parasites: Immunity to Trichuris muris in inbred and random-bred strains of mice. Parasitology 71, 51–6010.1017/S0031182000053142Search in Google Scholar

Williams-Blangero S., VandeBerg J.L., Subedi J., Aivaliotis M.J., Rai D.R., Upadhayay R.P., et al. 2002. Genes on chromosomes 1 and 13 have significant effects on Ascaris infection. Proceedings of the National Academy of Sciences U. S. A, 99, 5533–553810.1073/pnas.082115999Search in Google Scholar PubMed PubMed Central

Williams-Blangero S., VandeBerg J.L., Subedi J., Jha B., Corrêa-Oliveira R., Blangero J. 2008. Localization of multiple quantitative trait loci influencing susceptibility to infection with Ascaris lumbricoides. The Journal of Infectious Diseases, 197, 66–71. 10.1086/524060Search in Google Scholar PubMed

Zaros L.G., Bricarello P.A., Amarante A.F., Rocha R.A., Kooyman F.N., De Vries E., Coutinho L.L. 2010. Cytokine gene expression in response to Haemonchus placei infections in Nelore cattle. Veterinary Parasitology, 171, 68–73. 10.1016/j.vetpar.2010.03.020Search in Google Scholar PubMed

Zhu J., Yamane H., Paul W.E. 2010. Differentiation of effector CD4 T cell populations. Annual Review of Immunology, 28, 445–489. 10.1146/annurev-immunol-030409-101212Search in Google Scholar PubMed PubMed Central

Received: 2015-12-18
Revised: 2016-9-19
Accepted: 2016-10-14
Published Online: 2016-12-28
Published in Print: 2017-3-1

© 2017 W. Stefañski Institute of Parasitology, PAS