Abstract
Baylisascaris schroederi is among the most severe intestinal nematodes affecting giant pandas. Developing effective and secure vaccines can be used as a novel strategy for controlling repeated roundworm infection and addressing drug resistance. In our previous study, three recombinant antigens (rBsHP2, rBsGAL, and rBsUP) exhibited promising effects against B. schroederi infection in the mice model. This study extends the findings by formulating four-form cocktail vaccines (GAL+UP, HP2+UP, GAL+HP2, and GAL+HP2+UP) using three B. schroederi recombinant antigens to improve protection in mice further. Additionally, the protective differences after immunizing mice with different doses of cocktail antigens (150 μg, 100 μg, and 50 μg) were analyzed. Administration of rBs(GAL+UP), rBs(HP2+UP), rBs(GAL+HP2), and rBs(GAL+HP2+UP) significantly reduced liver and lung lesions, along with a decrease in L3 larvae by 83.7%, 82.1%, 76.4%, and 75.1%, respectively. These vaccines induced a Th1/Th2 mixed immunity, evidenced by elevated serum antibody levels (IgG, IgG1, IgG2a, IgE, and IgA) and splenocyte cytokines [interferon gamma (IFN-γ), interleukin (IL)-5, and IL-10]. Furthermore, varying cocktail vaccine dosages did not significantly affect protection. The results confirm that a 50 μg rBs(GAL+UP) dosage holds promise as a better candidate vaccine combination against B. schroederi infection, providing a basis for developing the B. schroederi vaccine.
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References
Abraham D, Leon O, Leon S et al (2001) Development of a recombinant antigen vaccine against infection with the filarial worm Onchocerca volvulus. Infect Immun 69(1):262–270. https://doi.org/10.1128/iai.69.1.262-270.2001
Abraham D, Leon O, Schnyder-Candrian S et al (2004) Immunoglobulin E and eosinophil-dependent protective immunity to larval Onchocerca volvulus in mice immunized with irradiated larvae. Infect Immun 72(2):810–817. https://doi.org/10.1128/iai.72.2.810-817.2004
Anand SB, Kodumudi KN, Reddy MV et al (2011) A combination of two Brugia malayi filarial vaccine candidate antigens (BmALT-2 and BmVAH) enhances immune responses and protection in jirds. J Helminthol 85(4):442–452. https://doi.org/10.1017/s0022149x10000799
Ando T, Kitaura J (2021) Tuning IgE: IgE-associating molecules and their effects on IgE-dependent mast cell reactions. Cells 10(7):1697. https://doi.org/10.3390/cells10071697
Arumugam S, Wei J, Liu Z et al (2016) Vaccination of gerbils with Bm-103 and Bm-RAL-2 concurrently or as a fusion protein confers consistent and improved protection against Brugia malayi infection. PLoS Negl Trop Dis 10(4):e0004586. https://doi.org/10.1371/journal.pntd.0004586
Astorgues-Xerri L, Riveiro ME, Tijeras-Raballand A et al (2014) Unraveling galectin-1 as a novel therapeutic target for cancer. Cancer Treat Rev 40(2):307–319. https://doi.org/10.1016/j.ctrv.2013.07.007
Barnes TS, Hinds LA, Jenkins DJ et al (2009) Efficacy of the EG95 hydatid vaccine in a macropodid host, the tammar wallaby. Parasitology 136(4):461–468. https://doi.org/10.1017/s0031182009005526
Bassetto CC, Picharillo M, Newlands GF et al (2014) Attempts to vaccinate ewes and their lambs against natural infection with Haemonchus contortus in a tropical environment. Int J Parasitol 44(14):1049–1054. https://doi.org/10.1016/j.ijpara.2014.07.007
Benavides MV, Souza CJH, Smith WD et al (2021) Evaluation of protection in grazing lambs immunised with different doses of Haemonchus contortus gut membrane glycoproteins in Southern Brazil. Vet Parasitol 290:109360. https://doi.org/10.1016/j.vetpar.2021.109360
Bock CN, Babu S, Breloer M et al (2017) Th2/1 hybrid cells occurring in murine and human Strongyloidiasis share effector functions of Th1 cells. Front Cell Infect Microbiol 7:261. https://doi.org/10.3389/fcimb.2017.00261
Cachat E, Newlands GF, Ekoja SE et al (2010) Attempts to immunize sheep against Haemonchus contortus using a cocktail of recombinant proteases derived from the protective antigen. H-gal-GP. Parasite Immunol 32(6):414–419. https://doi.org/10.1111/j.1365-3024.2010.01208.x
Chauhan N, Banerjee P, Khatri VK et al (2017) Improving the efficacy of a prophylactic vaccine formulation against lymphatic filariasis. Parasitol Res 116(10):2821–2830. https://doi.org/10.1007/s00436-017-5593-9
Chauhan N, Khatri V, Banerjee P et al (2018) Evaluating the vaccine potential of a tetravalent fusion protein (rBmHAXT) vaccine antigen against lymphatic filariasis in a mouse model. Front Immunol 9:1520. https://doi.org/10.3389/fimmu.2018.01520
Dakshinamoorthy G, Kalyanasundaram R (2013) Evaluating the efficacy of rBmHATαc as a multivalent vaccine against lymphatic filariasis in experimental animals and optimizing the adjuvant formulation. Vaccine 32(1):19–25. https://doi.org/10.1016/j.vaccine.2013.10.083
Dakshinamoorthy G, Samykutty AK, Munirathinam G et al (2013) Multivalent fusion protein vaccine for lymphatic filariasis. Vaccine 31(12):1616–1622. https://doi.org/10.1016/j.vaccine.2012.09.055
De Maere V, Vercauteren I, Geldhof P et al (2005) Molecular analysis of astacin-like metalloproteases of Ostertagia ostertagi. Parasitology 130(Pt1):89–98. https://doi.org/10.1017/s0031182004006274
Diemert D, Campbell D, Brelsford J et al (2018) Controlled human hookworm infection: accelerating human hookworm vaccine development. Open Forum. Infect Dis 5(5):ofy083. https://doi.org/10.1093/ofid/ofy083
Ehrens A, Rüdiger N, Heepmann L et al (2021) Eosinophils and neutrophils eliminate migrating strongyloides ratti larvae at the site of infection in the context of extracellular DNA trap formation. Front Immunol 12:715766. https://doi.org/10.3389/fimmu.2021.715766
Falduto GH, Vila CC, Saracino MP et al (2015) Trichinella spiralis: killing of newborn larvae by lung cells. Parasitol Res 114(2):679–685. https://doi.org/10.1007/s00436-014-4233-x
Gazzinelli-Guimarães AC, Gazzinelli-Guimarães PH, Nogueira DS et al (2018) IgG induced by vaccination with Ascaris suum extracts is protective against infection. Front Immunol 9:2535. https://doi.org/10.3389/fimmu.2018.02535
Gazzinelli-Guimarães AC, Nogueira DS, Amorim CCO et al (2021) ASCVac-1, a multi-peptide chimeric vaccine, protects mice against Ascaris suum infection. Front Immunol 12:788185. https://doi.org/10.3389/fimmu.2021.788185
George PJ, Hess JA, Jain S et al (2019) Antibody responses against the vaccine antigens Ov-103 and Ov-RAL-2 are associated with protective immunity to Onchocerca volvulus infection in both mice and humans. PLoS Negl Trop Dis 13(9):e0007730. https://doi.org/10.1371/journal.pntd.0007730
Geurden T, Chartier C, Fanke J et al (2015) Anthelmintic resistance to ivermectin and moxidectin in gastrointestinal nematodes of cattle in Europe. Int J Parasitol Drugs Drug Resist 5(3):163–171. https://doi.org/10.1016/j.ijpddr.2015.08.001
González-Hernández A, Van Coppernolle S, Borloo J et al (2016) Host protective ASP-based vaccine against the parasitic nematode Ostertagia ostertagi triggers NK cell activation and mixed IgG1-IgG2 response. Sci Rep 6:29496. https://doi.org/10.1038/srep29496
He G, Chen S, Wang T et al (2012) Sequence analysis of the Bs-Ag1 gene of Baylisascaris schroederi from the giant panda and an evaluation of the efficacy of a recombinant Baylisascaris schroederi Bs-Ag1 antigen in mice. DNA Cell Biol 31(7):1174–1181. https://doi.org/10.1089/dna.2011.1395
He G, Wang T, Yang G et al (2009) Sequence analysis of Bs-Ag2 gene from Baylisascaris schroederi of giant panda and evaluation of the efficacy of a recombinant Bs-Ag2 antigen in mice. Vaccine 27(22):3007–3011. https://doi.org/10.1016/j.vaccine.2009.02.077
Hernández JN, Hernández A, Stear MJ et al (2016) Potential role for mucosal IgA in modulating Haemonchus contortus adult worm infection in sheep. Vet Parasitol 223:153–158. https://doi.org/10.1016/j.vetpar.2016.04.022
Hess JA, Zhan B, Bonne-Année S et al (2014) Vaccines to combat river blindness: expression, selection and formulation of vaccines against infection with Onchocerca volvulus in a mouse model. Int J Parasitol 44(9):637–646. https://doi.org/10.1016/j.ijpara.2014.04.006
Hess JA, Zhan B, Torigian AR et al (2016) The immunomodulatory role of adjuvants in vaccines formulated with the recombinant antigens Ov-103 and Ov-RAL-2 against Onchocerca volvulus in Mice. PLoS Negl Trop Dis 10(7):e0004797. https://doi.org/10.1371/journal.pntd.0004797
Huang L, Appleton JA (2016) Eosinophils in helminth infection: defenders and dupes. Trends Parasitol 32(10):798–807. https://doi.org/10.1016/j.pt.2016.05.004
Islam MK, Miyoshi T, Tsuji N (2005) Vaccination with recombinant Ascaris suum 24-kilodalton antigen induces a Th1/Th2-mixed type immune response and confers high levels of protection against challenged Ascaris suum lung-stage infection in BALB/c mice. Int J Parasitol 35(9):1023–1030. https://doi.org/10.1016/j.ijpara.2005.03.019
Jaramillo-Hernández DA, Salazar Garcés LF, Pacheco LGC et al (2022) Protective response mediated by immunization with recombinant proteins in a murine model of toxocariasis and canine infection by Toxocara canis. Vaccine 40(6):912–923. https://doi.org/10.1016/j.vaccine.2021.12.052
Jia TW, Melville S, Utzinger J et al (2012) Soil-transmitted helminth reinfection after drug treatment: a systematic review and meta-analysis. PLoS Negl Trop Dis 6(5):e1621. https://doi.org/10.1371/journal.pntd.0001621
Kalyanasundaram R, Balumuri P (2011) Multivalent vaccine formulation with BmVAL-1 and BmALT-2 confer significant protection against challenge infections with Brugia malayi in mice and jirds. Res Rep Trop Med 2:45–56. https://doi.org/10.2147/rrtm.S13679
Kebeta MM, Hine BC, Walkden-Brown SW et al (2021) Protective efficacy of Barbervax® in Merino weaner sheep trickle infected with five doses of Haemonchus contortus infective larvae. Vet Parasitol 292:109386. https://doi.org/10.1016/j.vetpar.2021.109386
Lewis R, Behnke JM, Cassidy JP et al (2007) The migration of Ascaris suum larvae, and the associated pulmonary inflammatory response in susceptible C57BL/6j and resistant CBA/Ca mice. Parasitology 134(Pt9):1301–1314. https://doi.org/10.1017/s0031182007002582
Li C, Li G, Wang Y et al (2019) Preliminary observation on the anthelmintic effect of three benzimidazole drugs on Baylisascaris schroederi. Sichuan. J Zool 38(3):300–304. https://doi.org/10.11984/j.issn.1000-7083.20190034
Li D, He Y, Deng L et al (2015) Anthelmintic effect of ivermectin and pyrantel pamoate on Baylisascaris schroederi in captive giant pandas. Chin Anim Husband. Vet Med 47(6):87–90
Lustigman S, MacDonald AJ, Abraham D (2003) CD4+-dependent immunity to Onchocerca volvulus third-stage larvae in humans and the mouse vaccination model: common ground and distinctions. Int J Parasitol 33(11):1161–1171. https://doi.org/10.1016/s0020-7519(03)00170-x
Luu L, Bah GS, Okah-Nnane NH et al (2022) Co-administration of adjuvanted recombinant Ov-103 and Ov-RAL-2 vaccines confer protection against natural challenge in a bovine Onchocerca ochengi infection model of human Onchocerciasi. Vaccines 10(6):861. https://doi.org/10.3390/vaccines10060861
Makepeace BL, Martin C, Turner JD et al (2012) Granulocytes in helminth infection -- who is calling the shots? Curr Med Chem 19(10):1567–1586. https://doi.org/10.2174/092986712799828337
Matthews J (2011) Clinical forum: bovine lungworm. Livestock 13(6):23–30. https://doi.org/10.1111/j.2044-3870.2008.tb00188.x
Moreau E, Chauvin A (2010) Immunity against helminths: interactions with the host and the intercurrent infections. J Biomed Biotechnol 2010:428593. https://doi.org/10.1155/2010/428593
Muro A, Pérez-Arellano JL (2010) Nitric oxide and respiratory helminthic diseases. J Biomed Biotechnol 2010:958108. https://doi.org/10.1155/2010/958108
Nascimento IP, Leite LC (2012) Recombinant vaccines and the development of new vaccine strategies. Braz J Med Biol Res 45(12):1102–1111. https://doi.org/10.1590/s0100-879x2012007500142
Nisbet AJ, McNeilly TN, Greer AW et al (2016) Protection of ewes against Teladorsagia circumcincta infection in the periparturient period by vaccination with recombinant antigens. Vet Parasitol 228:130–136. https://doi.org/10.1016/j.vetpar.2016.09.002
Nisbet AJ, McNeilly TN, Price DRG et al (2019) The rational simplification of a recombinant cocktail vaccine to control the parasitic nematode Teladorsagia circumcincta. Int J Parasitol 49(3-4):257–265. https://doi.org/10.1016/j.ijpara.2018.10.006
Nisbet AJ, McNeilly TN, Wildblood LA et al (2013) Successful immunization against a parasitic nematode by vaccination with recombinant proteins. Vaccine 31(37):4017–4023. https://doi.org/10.1016/j.vaccine.2013.05.026
Nogueira DS, Gazzinelli-Guimarães PH, Barbosa FS et al (2016) Multiple exposures to Ascaris suum induce tissue injury and mixed Th2/Th17 immune response in mice. PLoS Negl Trop Dis 10(1):e0004382. https://doi.org/10.1371/journal.pntd.0004382
Noon JB, Aroian RV (2017) Recombinant subunit vaccines for soil-transmitted helminths. Parasitology 144(14):1845–1870. https://doi.org/10.1017/s003118201700138x
Prince PR, Madhumathi J, Anugraha G et al (2014) Tandem antioxidant enzymes confer synergistic protective responses in experimental filariasis. J Helminthol 88(4):402–410. https://doi.org/10.1017/s0022149x13000333
Qi X, Yue X, Han Y et al (2018) Characterization of two Trichinella spiralis adult-specific DNase II and their capacity to induce protective immunity. Front Microbiol 9:2504. https://doi.org/10.3389/fmicb.2018.02504
Qin Z, Liu S, Bai M et al (2021) First report of fatal baylisascariasis-induced acute pancreatitis in a giant panda. Parasitol Int 84:102380. https://doi.org/10.1016/j.parint.2021.102380
Roboz GJ, Rafii S (1999) Interleukin-5 and the regulation of eosinophil production. Curr Opin Hematol 6(3):164–168. https://doi.org/10.1097/00062752-199905000-00007
Ruano AL, López-Abán J, Fernández-Soto P et al (2015) Treatment with nitric oxide donors diminishes hyperinfection by Strongyloides venezuelensis in mice treated with dexamethasone. Acta Trop 152:90–95. https://doi.org/10.1016/j.actatropica.2015.08.019
Ryan NM, Hess JA, de Villena FP et al (2021) Onchocerca volvulus bivalent subunit vaccine induces protective immunity in genetically diverse collaborative cross recombinant inbred intercross mice. NPJ Vaccines 6(1):17. https://doi.org/10.1038/s41541-020-00276-2
Sahoo MK, Sisodia BS, Dixit S et al (2009) Immunization with inflammatory proteome of Brugia malayi adult worm induces a Th1/Th2-immune response and confers protection against the filarial infection. Vaccine 27(32):4263–4271. https://doi.org/10.1016/j.vaccine.2009.05.015
Salazar Garcés LF, Santiago LF, Santos SPO et al (2020) Immunogenicity and protection induced by recombinant Toxocara canis proteins in a murine model of toxocariasis. Vaccine 38(30):4762–4772. https://doi.org/10.1016/j.vaccine.2020.04.072
Santano R, Rubio R, Grau-Pujol B et al (2021) Plasmodium falciparum and helminth coinfections increase IgE and parasite-specific IgG responses. Microbiol Spectr 9(3):e0110921. https://doi.org/10.1128/Spectrum.01109-21
Shrivastava N, Singh PK, Nag JK et al (2013) Immunization with a multisubunit vaccine considerably reduces establishment of infective larvae in a rodent model of Brugia malayi. Comp Immunol Microbiol Infect Dis 36(5):507–519. https://doi.org/10.1016/j.cimid.2013.05.003
Slotved H, Eriksen L, Murrell K et al (1997) Comparison of methods for recovery of Ascaris suum larvae from tissues of mice. Int J Parasitol 27(11):1305–1310. https://doi.org/10.1016/s0020-7519(97)00101-x
Sun Y, Li Y, Wu Y et al (2017) Fatty-binding protein and galectin of Baylisascaris schroederi: prokaryotic expression and preliminary evaluation of serodiagnostic potential. PLoS One 12(7):e0182094. https://doi.org/10.1371/journal.pone.0182094
Sutherland IA, Leathwick DM (2011) Anthelmintic resistance in nematode parasites of cattle: a global issue? Trends Parasitol 27(4):176–181. https://doi.org/10.1016/j.pt.2010.11.008
Takatsu K, Kouro T, Nagai Y (2009) Interleukin 5 in the link between the innate and acquired immune response. Adv Immunol 101:191–236. https://doi.org/10.1016/s0065-2776(08)01006-7
Vanam U, Pandey V, Prabhu PR et al (2009) Evaluation of immunoprophylactic efficacy of Brugia malayi transglutaminase (BmTGA) in single and multiple antigen vaccination with BmALT-2 and BmTPX for human lymphatic filariasis. Am J Trop Med Hyg 80(2):319–324. https://doi.org/10.1016/j.actatropica.2008.10.008
Vasta GR (2009) Roles of galectins in infection. Nat Rev Microbiol 7(6):424–438. https://doi.org/10.1038/nrmicro2146
Versteeg L, Wei J, Liu Z et al (2020) Protective immunity elicited by the nematode-conserved As37 recombinant protein against Ascaris suum infection. PLoS Negl Trop Dis 14(2):e0008057. https://doi.org/10.1371/journal.pntd.0008057
Wang T, He G, Yang G et al (2008) Cloning, expression and evaluation of the efficacy of a recombinant Baylisascaris schroederi Bs-Ag3 antigen in mice. Vaccine 26(52):6919–6924. https://doi.org/10.1016/j.vaccine.2008.09.079
Wei F, Swaisgood R, Hu Y et al (2015) Progress in the ecology and conservation of giant pandas. Conserv Biol 29(6):1497–1507. https://doi.org/10.1111/cobi.12582
Wei J, Versteeg L, Liu Z et al (2017) Yeast-expressed recombinant As16 protects mice against Ascaris suum infection through induction of a Th2-skewed immune response. PLoS Negl Trop Dis 11(7):e0005769. https://doi.org/10.1371/journal.pntd.0005769
Wen T, Rothenberg M (2016) The regulatory function of eosinophils. Microbiol Spectr 4(5). https://doi.org/10.1128/microbiolspec.MCHD-0020-2015
Wiśniewski M, Łapiński M, Daniłowicz-Luebert E et al (2016) Vaccination with a cocktail of Ancylostoma ceylanicum recombinant antigens leads to worm burden reduction in hamsters. Acta Parasitol 61(3):556–561. https://doi.org/10.1515/ap-2016-0074
Xiao S, Zhan B, Xue J et al (2008) The evaluation of recombinant hookworm antigens as vaccines in hamsters (Mesocricetus auratus) challenged with human hookworm, Necator americanus. Exp Parasitol 118(1):32–40. https://doi.org/10.1016/j.exppara.2007.05.010
Xie Y, Chen S, Yan Y et al (2013) Potential of recombinant inorganic pyrophosphatase antigen as a new vaccine candidate against Baylisascaris schroederi in mice. Vet Res 44(1):90. https://doi.org/10.1186/1297-9716-44-90
Xie Y, Wang S, Wu S et al (2022) Genome of the giant panda roundworm illuminates its host shift and parasitic adaptation. Genom Proteom Bioinform 20(2):366–381. https://doi.org/10.1016/j.gpb.2021.08.002
Xiong L, Chen L, Chen Y et al (2023) Evaluation of the immunoprotective effects of eight recombinant proteins from Baylisascaris schroederi in mice model. Parasit Vectors 16(1):254. https://doi.org/10.1186/s13071-023-05886-y
Xu J, Gu X, Xie Y et al (2022) A novel cysteine protease inhibitor in Baylisascaris schroederi migratory larvae regulates inflammasome activation through the TLR4-ROS-NLRP3 pathway. Parasit Vectors 15(1):334. https://doi.org/10.1186/s13071-022-05466-6
Zeng J, Zhang XZ, Zhang R et al (2021) Vaccination of mice with recombinant novel aminopeptidase P and cathepsin X alone or in combination induces protective immunity against Trichinella spiralis infection. Acta Trop 224:106125. https://doi.org/10.1016/j.actatropica.2021.106125
Acknowledgements
We are grateful to the workers at Chengdu Research Base of Giant Panda Breeding for their assistance in gathering B. schroederi samples. We would like to thank the native English speaking scientists of Elixigen Company (Huntington Beach, California) for editing our manuscript.
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This work was supported by the Chengdu Research Base of Giant Panda Breeding (grant number CPF2017-24).
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LX participated in conceiving and designing the study, performing the experiments, analyzing the data, and writing the paper. LC participated in the feeding of the experimental animals. YXC contributed to materials/reagents/analysis tools. GYY and RQH participated in developing and designing the study and reviewing the paper. NXS helped with the study design and data review. All authors reviewed and approved the final manuscript.
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The Animal Care and Use Committee of Sichuan Agricultural University reviewed and approved the animal study protocol (No. SYXK 2019-189). All animal procedures adhered to the Guide for the Care and Use of Laboratory Animals (National Research Council, Bethesda, MD, USA) and the Animal Research: Reporting of In Vivo (ARRIVE) Experiments guidelines (http://www.nc3rs.org.uk/arriveguidelines). All applicable institutional and national guidelines for the care and use of animals were followed.
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Xiong, L., Chen, Y., Chen, L. et al. Enhanced protective immunity against Baylisascaris schroederi infection in mice through a multi-antigen cocktail vaccine approach. Parasitol Res 123, 20 (2024). https://doi.org/10.1007/s00436-023-08016-w
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DOI: https://doi.org/10.1007/s00436-023-08016-w