Vaccination with recombinant paramyosin against the bovine lungworm Dictyocaulus viviparus considerably reduces worm burden and larvae shedding

Particle and Fibre Toxicology

Georg von Samson-Himmelstjerna²  Thomas Schnieder³  Julia Korrell³  Elisabeth Kremmer¹  Sabine Schicht³  Deborah Joekel³  Sandra Buschbaum³  Ronald Kaminsky⁴  Sandra Schorderet Weber⁴  Heinz Sager⁴  Claas Haake³  Christina Strube³ 

[1] Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Molecular Immunology, Munich, Germany;Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Berlin, Germany;Institute for Parasitology, University of Veterinary Medicine Hannover, Hanover, Germany;Novartis Centre de Recherche Santé Animale, St. Aubin, Switzerland

关键词: Hidden antigen;    Paramyosin;    Immunization;    Vaccination;    Recombinant vaccine;    Parasitic bronchitis;    Bovine lungworm;    Dictyocaulus viviparus;   

Others  :  1146802
DOI  :  10.1186/s13071-015-0733-5

 received in 2014-11-24, accepted in 2015-02-12,  发布年份 2015

【 摘 要 】

Background

The lungworm Dictyocaulus viviparus, causing parasitic bronchitis in cattle, induces a temporary protective immunity that prevents clinical disease. A radiation-attenuated larvae based vaccine is commercially available in a few European countries, but has the disadvantages of a live vaccine. As a recombinant subunit vaccine would overcome these disadvantages, the parasite’s muscle protein paramyosin (PMY) was tested as a recombinant vaccine antigen.

Methods

D. viviparus-PMY was recombinantly expressed in Escherichia coli as a glutathione-S-transferase (GST)-fused protein. Emulsified in adjuvant Saponin Quil A, the protein was given intramuscularly into calves. Two independent recombinant PMY (rPMY) vaccination trials with negative control groups (first trial: adjuvant only; second trial: non-fused GST) as well as an additional positive control group in the second trial, using the Bovilis^©Dictol live vaccine to verify vaccination results, were performed. To determine the vaccination success, shedding of larvae as well as worm burden and worm sizes were analyzed. Additionally, ELISA-based determination of development of immunglobulins IgM, IgA, IgE, IgG as well as the subclasses IgG1 and IgG2 was performed. To analyze PMY localization in the bovine lungworm, immunohistochemical staining of adult worms was carried out.

Results

Immunohistochemical staining revealed that PMY is part of the bovine lungworm’s pharyngeal and body wall muscles. Vaccination with rPMY resulted in 47% [geometric mean: 67%] and 57% (geometric mean: 71%) reduction of larvae shedding in the first and second vaccination trial, respectively. Worm burden was reduced by 54% (geometric mean: 86%) and 31% (geometric mean: 68%), respectively, and worms of rPMY-vaccinated cattle were significantly shorter in both trials. Furthermore, ELISAs showed a clear antibody response towards rPMY with exception of IgE for which titers could not be detected. After challenge infection, rPMY antibodies were only exceptionally elevated among study animals indicating PMY to be a hidden antigen.

Conclusions

Even though vaccination with the attenuated live vaccine was with 94% (geometric mean: 95%) reduction in larvae shedding and 93% (geometric mean: 94%) reduction in worm burden superior to rPMY vaccination, results using the latter are promising and show the potential for further development of a recombinant PMY-based vaccine against the bovine lungworm.

【 授权许可】

   
2015 Strube et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Jarrett WF, Jennings FW, Mc IW, Mulligan W, Urquhart GM. Immunological studies on Dictyocaulus viviparus infection; immunity produced by the administration of irradiated larvae. Immunology. 1960; 3:145-51.
• [2]Nara T, Tanabe K, Mahakunkijcharoen Y, Osada Y, Matsumoto N, Kita K et al.. The B cell epitope of paramyosin recognized by a protective monoclonal IgE antibody to Schistosoma japonicum. Vaccine. 1997; 15:79-84.
• [3]Schmitz KA, Hale TJ, Rajan TV, Yates JA. Localization of paramyosin, myosin, and a heat shock protein 70 in larval and adult Brugia malayi. J Parasitol. 1996; 82:367-70.
• [4]Cancela M, Carmona C, Rossi S, Frangione B, Goni F, Berasain P. Purification, characterization, and immunolocalization of paramyosin from the adult stage of Fasciola hepatica. Parasitol Res. 2004; 92:441-8.
• [5]Epstein HF, Miller DM, Ortiz I, Berliner GC. Myosin and paramyosin are organized about a newly identified core structure. J Cell Biol. 1985; 100:904-15.
• [6]Ardizzi JP, Epstein HF. Immunochemical localization of myosin heavy chain isoforms and paramyosin in developmentally and structurally diverse muscle cell types of the nematode Caenorhabditis elegans. J Cell Biol. 1987; 105:2763-70.
• [7]Lanar DE, Pearce EJ, James SL, Sher A. Identification of paramyosin as schistosome antigen recognized by intradermally vaccinated mice. Science. 1986; 234:593-6.
• [8]Laclette JP, Alagon A, Willms K, Torre-Blanco A. Purification of antigen B from Taenia solium cysticerci by affinity to mammalian collagen. J Parasitol. 1990; 76:273-5.
• [9]Laclette JP, Shoemaker CB, Richter D, Arcos L, Pante N, Cohen C et al.. Paramyosin inhibits complement C1. J Immunol. 1992; 148:124-8.
• [10]Kalinna B, McManus DP. An IgG (Fc gamma)-binding protein of Taenia crassiceps (Cestoda) exhibits sequence homology and antigenic similarity with schistosome paramyosin. Parasitology. 1993; 106:289-96.
• [11]Loukas A, Jones MK, King LT, Brindley PJ, McManus DP. Receptor for Fc on the surfaces of schistosomes. Infect Immun. 2001; 69:3646-51.
• [12]Zhang Z, Yang J, Wei J, Yang Y, Chen X, Zhao X et al.. Trichinella spiralis paramyosin binds to C8 and C9 and protects the tissue-dwelling nematode from being attacked by host complement. PLoS Negl Trop Dis. 2011; 5:e1225.
• [13]Strube C, Buschbaum S, von Samson-Himmelstjerna G, Schnieder T. Stage-dependent transcriptional changes and characterization of paramyosin of the bovine lungworm Dictyocaulus viviparus. Parasitol Int. 2009; 58:334-40.
• [14]Zhang DM, Pan WQ, Qian L, Duke M, Shen LH, McManus DP. Investigation of recombinant Schistosoma japonicum paramyosin fragments for immunogenicity and vaccine efficacy in mice. Parasit Immun. 2006; 28:77-84.
• [15]Zhou S, Liu S, Song G, Xu Y, Sun W. Induction of protective immunity in mice against Schistosoma japonicum by nucleic acid vaccine encoding the full-length paramyosin. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999; 17:321-4.
• [16]Abou-Elhakam H, Rabee I, El Deeb S, El Amir A. Protection against Fasciola gigantica using paramyosin antigen as a candidate for vaccine production. Pak J Biol Sci. 2013; 16:1449-58.
• [17]Chen H, Nara T, Zeng X, Satoh M, Wu G, Jiang W et al.. Vaccination of domestic pig with recombinant paramyosin. against Schistosoma japonicum in China. Vaccine. 2000; 18:2142-6.
• [18]Taylor MG, Huggins MC, Shi F, Lin J, Tian E, Ye P et al.. Production and testing of Schistosoma japonicum candidate vaccine antigens in the natural ovine host. Vaccine. 1998; 16:1290-8.
• [19]McManus DP, Wong JYM, Zhou J, Cai C, Zeng Q, Smyth D et al.. Recombinant paramyosin (rec-Sj-97) tested for immunogenicity and vaccine efficacy against Schistosoma japonicum in mice and water buffaloes. Vaccine. 2001; 20:870-8.
• [20]Li BW, Zhang S, Curtis KC, Weil GJ. Immune responses to Brugia malayi paramyosin in rodents after DNA vaccination. Vaccine. 1999; 18:76-81.
• [21]Li BW, Rush A, Zhang SR, Curtis KC, Weil GJ. Antibody responses to Brugia malayi antigens induced by DNA vaccination. Filaria J. 2004; 3:1. BioMed Central Full Text
• [22]Yang J, Yang Y, Gu Y, Li Q, Wei J, Wang S et al.. Identification and characterization of a full-length cDNA encoding paramyosin of Trichinella spiralis. Biochem Biophys Res Commun. 2008; 365:528-33.
• [23]Britton C, Knox DP, Kennedy MW. Superoxide dismutase (SOD) activity of Dictyocaulus viviparus and its inhibition by antibody from infected and vaccinated bovine hosts. Parasitology. 1994; 109:257-63.
• [24]McKeand JB. Vaccine development and diagnostics of Dictyocaulus viviparus. Parasitology. 2000; 120 Suppl:17-23.
• [25]Matthews JB, Davidson AJ, Beynon RJ. The application of mass spectrometry to identify immunogenic components of excretory/secretory products from adult Dictyocaulus viviparus. Parasitology. 2004; 128 Suppl1:43-7.
• [26]McKeand JB, Knox DP, Duncan JL, Kennedy MW. The immunogenicity of the acetylcholinesterases of the cattle lungworm Dictyocaulus viviparus. Int J Parasitol. 1994; 24:501-10.
• [27]Matthews JB, Davidson AJ, Freeman KL, French NP. Immunisation of cattle with recombinant acetylcholinesterase from Dictyocaulus viviparus and with adult worm ES products. Int J Parasitol. 2001; 31:307-17.
• [28]Vercauteren I, Geldhof P, Vercruysse J, Peelaers I, van den Broeck W, Gevaert K et al.. Vaccination with an Ostertagia ostertagi polyprotein allergen protects calves against homologous challenge infection. Infect Immun. 2004; 72:2995-3001.
• [29]Geldhof P, Meyvis Y, Vercruysse J, Claerebout E. Vaccine testing of a recombinant activation-associated secreted protein (ASP1) from Ostertagia ostertagi. Parasite Immunol. 2008; 30:57-60.
• [30]Smith WD, Smith SK, Murray JM. Protection studies with integral membrane fractions of Haemonchus contortus. Parasite Immunol. 1994; 16:231-41.
• [31]Smith TS, Munn EA, Graham M, Tavernor AS, Greenwood CA. Purification and evaluation of the integral membrane protein H11 as a protective antigen against Haemonchus contortus. Int J Parasitol. 1993; 23:271-80.
• [32]Munn EA, Smith TS, Graham M, Greenwood CA, Tavernor AS, Coetzee G. Vaccination of merino lambs against haemonchosis with membrane-associated proteins from the adult parasite. Parasitology. 1993; 106:63-6.
• [33]Munn EA, Smith TS, Graham M, Tavernor AS, Greenwood CA. The potential value of integral membrane proteins in the vaccination of lambs against Haemonchus contortus. Int J Parasitol. 1993; 23:261-9.
• [34]Reszka N, Rijsewijk FA, Zelnik V, Moskwa B, Bienkowska-Szewczyk K. Haemonchus contortus: characterization of the baculovirus expressed form of aminopeptidase H11. Exp Parasitol. 2007; 117:208-13.
• [35]Pillai S, Dermody K, Metcalf B. Immunogenicity of genetically engineered glutathione S-transferase fusion proteins containing a T-cell epitope from diphtheria toxin. Infect Immun. 1995; 63:1535-40.
• [36]Brophy PM, Pritchard DI. Parasitic helminth glutathione S-transferases: an update on their potential as targets for immuno- and chemotherapy. Exp Parasitol. 1994; 79:89-96.
• [37]Gupta S, Bhandari YP, Reddy MV, Harinath BC, Rathaur S. Setaria cervi: immunoprophylactic potential of glutathione-S-transferase against filarial parasite Brugia malayi. Exp Parasitol. 2005; 109:252-5.
• [38]Veerapathran A, Dakshinamoorthy G, Gnanasekar M, Reddy MV, Kalyanasundaram R. Evaluation of Wuchereria bancrofti GST as a vaccine candidate for lymphatic filariasis. PLoS Negl Trop Dis. 2009; 3:e457.
• [39]Schlein Y, Lewis CT. Lesions in haematophagous flies after feeding on rabbits immunized with fly tissues. Physiol Entomol. 1976; 1:55-9.
• [40]Schlein Y, Sira DT, Jacobson RL. The passage of serum immunoglobulins through the gut of Sarcophaga falculata, Pand. Ann Trop Med Parasitol. 1976; 70:227-30.
• [41]Jarrett WF, Jennings FW, McIntyre WI, Mulligan W. The natural history of parasitic bronchitis with notes on prophylaxis and treatment. Vet Rec. 1957; 69:1329-39.
• [42]Mach JP, Pahud JJ. Secretory IgA, a major immunoglobulin in most bovine external secretions. J Immunol. 1971; 106:552-63.
• [43]Curtain CC, Clark BL, Dufty JH. The origins of the immunoglobulins in the mucous secretions of cattle. Clin Exp Immunol. 1971; 8:335-44.
• [44]Newby TJ, Bourne FJ. The nature of the local immune system of the bovine small intestine. Immunology. 1976; 31:475-80.
• [45]Hillyer GV. Fasciola antigens as vaccines against fascioliasis and schistosomiasis. J Helminthol. 2005; 79:241-7.
• [46]Chauhan JS, Rao A, Raghava GP. In silico platform for prediction of N-O- and C-glycosites in eukaryotic protein sequences. PloS one. 2013; 8:e67008.
• [47]Kooyman FN, Ploeger HW, Hoglund J, van Putten JP. Differential N-glycan- and protein-directed immune responses in Dictyocaulus viviparus-infected and vaccinated calves. Parasitology. 2007; 134:269-79.
• [48]Barnes EH, Dobson RJ, Barger IA. Worm control and anthelmintic resistance: adventures with a model. Parasitol Today. 1995; 11:56-63.
• [49]Hunter RL. Overview of vaccine adjuvants: present and future. Vaccine. 2002; 20 Suppl3:7-12.