doi: 10.14202/vetworld.2018.819-823
Share this article on [Facebook] [LinkedIn]
Article history: Received: 17-01-2018, Accepted: 14-05-2018, Published online: 19-06-2018
Corresponding author: Wiwik Misaco Yuniarti
E-mail: wiwikmisaco@yahoo.com
Citation: Lastuti NDR, Yuniarti WM, Hastutiek P, Suwanti LT, Chrismanto D (2018) Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae, Veterinary World, 11(6): 819-823.Aim: Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae.
Materials and Methods: This research was done as follows, identification and collection of Sarcoptes scabiei var. caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 μg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay.
Results: Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters.
Conclusion: Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research.
Keywords: antigenic protein, CD4, CD8, immunoglobulin G, Sarcoptes scabiei var. caprae, toll-like receptor-9.
1. Tarigan, S. (2004) Antibody responses in naive and sensitised goats infested by Sarcoptes scabiei. Ind. J. Anim. Vet. Sci., 9(4): 258-265.
2. Casais, R., Granda, V., Balseiro, A., Cerro, A.D., Dalt, N., Gonzalez, E., Bra, P., Prieto, J.M. and Montoya, M. (2016) Vaccination of rabbits with immunodominant antigens from Sarcoptes scabiei induced high levels of humoral responses and pro-inflammatory cytokines but confers limited protection. Parasit. Vectors, 9: 435-447. [Crossref] [PubMed] [PMC]
3. Alasaad, S., Rossi, L., Heukelbach, J., Perez, J.M., Hamarsheh, O., Otiende, M. and Zhu, X.Q. (2013) The neglected navigating web of the incomprehensibly emerging and re-emerging sarcoptic mite. Infect. Infect Genet. Evol., 17: 253-259. [Crossref] [PubMed]
4. Gu, X., Xie, Y., Wang, S., Peng, X., Lai, S. and Yang, G. (2014) Immune response induced by candidate Sarcoptes scabiei var. cuniculi DNA vaccine encoding paramyosin in mice. Exp. Appl. Acarol., 63: 401-412. [Crossref] [PubMed]
5. Lastuti, N.D.R. (2009) Specific antigenic protein 57.3 kDa of Sarcoptes scabiei var. Caprae as Material Candidate of Scabies Diagnostic Kit for Goat and Toll-like Receptor-Mediated Immune Responses. Doctoral Dissertation. Postgraduate Program. Universitas Airlangga.
6. Lastuti, N.D.R., Rantam, F.A., Hastutiek, P. and Chrismanto, D. (2017) Toll-like receptor (TLRs) play a role in adaptive immunity in rabbits immunized by Sarcoptes scabiei proteins.VMIC Conf. Proc. KnE Life Sci., 1: 1-9.
7. Kammanadiminti, S.J., Mann, B.J., Dutil, L. and Chadee, K. (2003) Regulation of toll-like receptor 2 expression by gal-lectin of Entamoeba histolytica. Faseb. J., 18: 155-157. [Crossref] [PubMed]
8. Arlian, L.G., Morgan, M.S. and Neal, J.S. (2003) Modulation of cytokine expression in human keratinocytes and fibroblasts by extracts of scabies mites. Am. J. Trop. Med. Hyg., 69: 652-656. [PubMed]
9. Clement, M., Ladell, K., Makinde, J.E., Miles, J.J., Edwards, E.S., Dolton, G., Williams, T., Schauenburg, A.J.A., Cole, D.K., Lauder, S.N., Gallimore, A.M., Godkin, A.J., Burrows, S.R., Price, D.A., Sewell, A.K. and Wooldridge, L. (2011) Anti-CD8 antibodies Can trigger CD8+T cell effector function in the absence of TCR engagement and improve peptide MHC-I tetramer staining. J. Immunol., 187: 654-663. [Crossref] [PubMed] [PMC]
10. Pasare, C. and Medzhitov, R. (2005) Control of B cell responses by toll-like receptor. Nature, 438(7066): 364-368. [Crossref] [PubMed]
11. Blander, J.M. and Medzhitov, R. (2006) Toll-dependent selection of microbial antigens for precentation by dendritic cells. Nature, 440(7085): 808-812. [Crossref] [PubMed]
12. Xiao, H., Li, X. and Abbot, D.W. (2008) In: Gregory, W.K., editor. Analysis of TLR Expression, Regulation and Signalling. Chapter 3: Signalling by Toll-Like Receptor. CRC Press, Taylor & Francis Group, Boca. p39-55.
13. McCluskie, M.J. and Krieg, A.M. (2006) Enhancement of infectious disease vaccines through TLR9 dependent recognition of CpG DNA. Curr. Top. Microbiol., 311: 155-178. [Crossref]
14. Schetter, C. and Vollmer, J. (2004) Toll-like receptors involved in response to microbial pathogens: Development of agonists for toll-like receptor 9. Curr. Opin. Drug. Discov. Dev., 7: 204-210. [PubMed]
15. Bhardwaj, N., Gnjatic, S.S. and Sawhney, N.B. (2010) TLR agonists: Are they good adjuvants? Cancer J., 16(4): 382-391. [Crossref] [PubMed] [PMC]
16. Toussi, D.N. and Massari, P.P. (2014) Review: Immune adjuvant effect of molecularly-defined toll-like receptor ligands. Vaccines, 2: 323-353. [Crossref] [PubMed] [PMC]
17. Soulsby, E.J.L. (1986) Helminths, Arthropods and Protozoa of Domesticated Animal. 7thed. The English and Protozoa of Society and Baillire, Tindall, London. p504-506.
18. Reed, R., Holmes, D., Weyers, J. and Jones, A. (2003) Practical Skills in Biomolecular Sciences. 2nd Ed. Pearson, Prentice Hall. Ashford Colour Press Ltd., Gosport. p237-319.
19. Rantam, F.A. (2003) The Method of Immunology. Airlangga University Press, Surabaya. p145-155.
20. Abbas, A.K. and Litchman, A.H. (2005) Cellular and Molecular Immunology. 5th ed. International Edition. Elsevier Saunders Inc., Philadelphia, PA, Pennsylvania. p41-105, 411-432.
21. Bornstein, S., Frossling, J.K., Zakrisson, G. and Morner, T. (2006) Evaluation of a serological test (indirect ELISA) for the diagnosis of sarcoptic mange in red foxes. Vet. Dermatol., 17(6): 411-416. [Crossref] [PubMed]
22. Rodriguez-Cadenas, F., Carbajal-Gonzales, M.T., Fregeneda-Grandes, J.M., Aller-Gancedo, J.M. and Rojo-Vazquez, F.A. (2010) Clinical evaluation and antibody responses in sheep after primary and secondary experimental challenges with the mange mites Sarcoptes scabiei var. ovis. Vet. Immunol. Immunopathol., 133: 109-116. [Crossref]
23. Tarigan, S. and Huntley, J.E. (2005) Failure to protect goats following vaccination with soluble proteins of Sarcoptes scabiei: Evidence for a role for IgE antibody in protection. Vet. Parasitol., 133(1): 101-109. [Crossref] [PubMed]
24. Casais, R., Millan, J., Rosell, J.M., Dalton, K.P. and Prieto, J.M. (2015) Evaluation of an ELISA using recombinant Ss?20?B3 antigen for the serological diagnosis of Sarcoptes scabiei infestation in domestic and wild rabbits. Vet. Parasitol., 214(3-4): 315-321. [Crossref] [PubMed]