1 2 A nematode immunomodulator suppresses grass pollen-specific allergic 3 responses by controlling excessive Th2 inflammation 4 Emilia Daniłowicz-Luebert a Q1 , Svenja Steinfelder b , Anja A. Kühl c , Gennadiy Drozdenko d , Richard Lucius a , 5 Margitta Worm d , Eckard Hamelmann e , Susanne Hartmann a,b,⇑ 6 a Department of Molecular Parasitology, Institute of Biology, Humboldt University of Berlin, Germany 7 b Department of Veterinary Medicine, Institute of Immunology, Free University Berlin, Germany 8 c Department of Gastroenterology, Infectiology and Rheumatology/Research Center ImmunoSciences (RCIS), University Hospital Charité, Berlin, Germany 9 d Department of Dermatology and Allergy, University Hospital Charité, Berlin, Germany 10 e University Children’s Hospital, Ruhr University Bochum, Germany 11 12 14 article info 15 Article history: 16 Received 4 September 2012 17 Received in revised form 18 October 2012 18 Accepted 24 October 2012 19 Available online xxxx 20 Keywords: 21 Helminth 22 Filarial cystatin 23 AvCystatin 24 Av17 25 Allergy 26 Phl p 5b 27 Timothy grass pollen 28 29 abstract 30 Helminth parasites modulate the immune system by complex mechanisms to ensure persistence in the 31 host. Released immunomodulatory parasite components lead to a beneficial environment for the parasite 32 by targeting different host cells and in parallel to a modulation of unrelated inflammatory responses in 33 the host, such as allergy. The aim of this study was to investigate the effect of the potent helminth immu- 34 nomodulator, filarial cystatin, in a murine model of airway inflammation and hyperreactivity induced by 35 a clinically relevant aeroallergen (timothy grass (Phleum pratense) pollen) and on the function of periph- 36 eral blood mononuclear cells (PBMCs) from timothy grass pollen allergic patients. BALB/c mice were sys- 37 temically sensitised with a recombinant major allergen of timothy grass pollen (rPhl p 5b) and then 38 challenged with timothy grass pollen extract (GPE) via the airways. Filarial cystatin was applied i.p. dur- 39 ing the sensitization phase. Airway hyperresponsiveness to methacholine challenges, inflammation of 40 airways, inflammatory cell recruitment, cytokine production and lung histopathology were investigated. 41 In a translational approach, PBMCs from allergic subjects and healthy controls were treated in vitro with 42 cystatin prior to stimulation with GPE. Administration of filarial cystatin suppressed rPhl p 5b-induced 43 allergen-specific Th2-responses and airway inflammation, inhibited local recruitment of eosinophils, 44 reduced levels of allergen-specific IgE and down-regulated IL-5 and IL-13 in the bronchoalveolar lavage 45 (BAL). Ex vivo restimulation with cystatin of spleen cells from cystatin-treated mice induced the produc- 46 tion of IL-10, while cystatin inhibited allergen-specific IL-5 and IL-13 levels. Human PBMCs from timothy 47 grass pollen allergic patients displayed a shift towards a Th1 response after treatment with cystatin. 48 These results show that filarial cystatin ameliorates allergic inflammation and disease in a clinically rel- 49 evant model of allergy. This data indicate that filarial cystatin has a modulatory effect on grass pollen- 50 specific responses warranting further investigation of potential preventive and therapeutic options in 51 the treatment of allergies. 52 Ó 2012 Published by Elsevier Ltd. on behalf of Australian Society for Parasitology Inc. 53 54 55 1. Introduction 56 Helminth infections have been almost completely eradicated in 57 western countries over the past 50 years while the prevalence of 58 allergic diseases has dramatically increased. One possible explana- 59 tion of these phenomena is the hygiene hypothesis (Strachan, 60 1989; Yazdanbakhsh et al., 2002), which states that improved pub- 61 lic health, use of antibiotics, medications and vaccines that reduce 62 the occurrence of viral, bacterial or helminth infections early in life 63 in return contribute to a higher susceptibility of immune dysregu- 64 lation and a higher prevalence of allergic and autoimmune disor- 65 ders. Moreover, epidemiological studies observed lower levels of 66 immunopathological diseases such as Th1-related autoimmune 67 diseases or aberrant Th2-related diseases e.g. asthma or allergic 68 rhinitis in helminth-infected populations (Fallon and Mangan, 69 2007; Patel et al., 2008; Cooke, 2009; Smits et al., 2010). 70 Many studies in animals and humans have shown that helminth 71 infections primarily induce a Th2 immune response involving the 72 cytokines, IL-4, IL-5, IL-9, IL-10 and IL-13, and immunoglobulin 73 IgE, whereas the immune system is subsequently suppressed dur- 74 ing the chronic phase of the infection (Maizels and Yazdanbakhsh, 75 2003). To down-modulate host protective Th2-type immune 76 responses, helminths induce different regulatory cell types that 0020-7519/$36.00 Ó 2012 Published by Elsevier Ltd. on behalf of Australian Society for Parasitology Inc. http://dx.doi.org/10.1016/j.ijpara.2012.10.014 ⇑ Corresponding author. Address: Free University Berlin, Department of Veteri- nary Medicine, Institute of Immunology, Philippstr. 13, 10115 Berlin, Germany. Tel.: +49 30 2093 6450; fax: +49 30 2093 6051. E-mail address: susanne.hartmann@fu-berlin.de (S. Hartmann). International Journal for Parasitology xxx (2012) xxx–xxx Contents lists available at SciVerse ScienceDirect International Journal for Parasitology journal homepage: www.elsevier.com/locate/ijpara PARA 3473 No. of Pages 10, Model 5G 23 November 2012 Please cite this article in press as: Daniłowicz-Luebert, E., et al. A nematode immunomodulator suppresses grass pollen-specific allergic responses by con- trolling excessive Th2 inflammation. Int. J. Parasitol. (2012), http://dx.doi.org/10.1016/j.ijpara.2012.10.014