ANTI-IDIOTYPIC ANTIBODIES IN DIAGNOSIS OF OPISTHORCHIASIS
Main Article Content
Authors
A.K. Bulashev
S. Seifullin Kazakh Agro Technical University, 2 Altynsarina str. Astana, 010011, Kazakhstan
Sh.S. Serikova
S. Seifullin Kazakh Agro Technical University, 2 Altynsarina str. Astana, 010011, Kazakhstan
S.Z. Eskendirova
National Center for Biotechnology, 13/1 Valikhanov str., Astana, 010000, Kazakhstan
Abstract
The incidence of people with Opisthorchiasis in Kazakhstan is one of the highest among the CIS countries. In the disease control system the main role is given to the early diagnosis. Effectiveness of the existing diagnostic methods for Opisthorchiasis is rather low. Nowadays diagnostic test-systems developed on the basis of ELISA have promising outlook. Most suitable antigen for serological diagnosing Opisthorchiasis is excretory-secretory antigen (ES-AG). Unfortunately, immunoassays based on the use of this antigen are not enough practical because there are great difficulties in obtaining standardized metabolite of parasite. In this regard, the great interest is the using antigen’s "internal image", i.e. anti-idiotypic antibodies (AIAT) against Fab-fragments that are specific to definite epitopes.
The aim of research was to obtain antibodies to idiotypes of monoclonal antibodies (Mab) specific for Opisthorchis felineus ES-AG epitope and to determine the possibility of using anti-idiotypes as an antigen in ELISA for serological diagnosis of Opisthorchiasis.
Mab 4B3D9 against helminth’s antigen have been used as immunogen for obtaining AIAT. Two hybrid strains designated as 3H10A4 and 4H10D8 producing anti-idiotypes with "internal image" of Opisthorhis felineus’ antigen were created by hybridoma technique. Anti-idiotypes produced by hybridomas belonged to IgG1 and IgM, characterized by sufficient affinity with respect to the used immunogen. As expected, in sandwich immunoassay AIAT of both hybridomas, mimicking ES-AG epitope, were recognized by antibodies of dog, infested with parasite. The results of research suggest the possibility of using anti-idiotypes of hybridomas as specific antigen in the development of immunoassays for the diagnosis of Opisthorchiasis.
Keywords
opisthorchiasis, diagnostics, excretory-secretory antigen, ELISA, hybridoma, monoclonal antibodies, anti-idiotypic antibodies
Article Details
References
Abuladze K.I., Demidov N.V., Nepoklonov A.A., Nikol'skiy S.N., Pavlova N.V., Stepanov A.V. Parazitologiya i invazionnyye bolezni sel'skokhozyaystvennykh zhivotnykh [Parasitology and parasitic diseases of farm animals]. M., Agropromizdat, 1990, 464 р.
Petney T.N. and Andrews R.H. The zoonotic, fish-borne liver flukes Clonorchis sinesnsis, Opisthorchis felineus and Opisthorchis viverrini. International Journal for Parasitology, 2013, vol. 43, no. 12-13, рр. 1031-1046. PMID: 23978669.
Thidarut B., Wu Z., Boonjaruspinyo S., Pinlaor S., Nagano I., Takahashi Y., Kaewsamut B., Yongvanit P. Alterations of gene expression of RB pathway in Opisthorchis viverrini infection-induced cholangiocarcinoma. Parasitology Research, 2009, vol. 105, no. 5, рр. 1273-1281.
Wu Z., Thidarut B., Nagano I., Boonjaraspinyo S., Pinlaor S., Pairojkul S., Chamgramol Y., Takahashi Y. Alteration of galectin-1 during tumorigenesis of Opisthorchis viverriniinfection-induced cholangiocarcinoma and its correlation with clinicopathology. Tumor Biology, 2012, vol. 33, no. 4, рр. 1169-1178. PMID: 22373585.
Khoontawad J., Hongsrichan N., Chamgramol Y., Pinlaor P., Wongkham C., Yongvanit Y., Pairojkul P., Khuntikeo N., Roytrakul S., Boonmars T. Increase of exostosin 1 in plasma as a potential biomarker for opisthorchiasis-associated cholangiocarcinoma. Tumor Biology, 2013, vol. 36, рр. 1275-1283.
Ilyinskikh E.N., Novitskiy V.V., Urazova L.N et al. Assessment of the relationship of chronic Opisthorchiasis to Epstein_Barr virus infection as well as some cytogenetical and immunological parameters in two comparable. Eur. J. Epidemiol, 2000, vol. 16, рр. 993-1002.
Mordvinov V.A., Yurlova N.I., Ogorodova L.M., Katokhin A.V. Opisthorchis felineus and Metorchis bilis are the main agents of liver fluke infection of humans in Russia. Parasitology International, 2012, vol. 61, no. 1, рр. 25-31. PMID: 21840415.
Ogorodova L.M., Freidin M.B., Sazonov A.E., Fedorova O.S., Gerbek I.E., Cherevko N.A., Lebedeva N.Yu. A pilot screening of prevalence of atopic states and opisthorchosis and their relationship in people of Tomsk Oblast. Parasitology Research September, 2007, vol. 101, no. 4, рр. 1165-1168. PMID:17549516.
Innovatsionnyy patent 23891 Respublika Kazakhstan, G01N 33/53 A61B 10/00. Sposob prigotovleniya ekskretorno-sekretornogo antigena dlya serologicheskoy diagnostiki opistorkhoza / A.K. Bulashev, S.N.Borovikov, M.A.Kuybagarov i soavt.; zayavitel' i patentoobladatel' AO «Kazakhskiy agrotekhnicheskiy universitet im.S.Seyfullina» [Innovative patent 23891 Republic of Kazakhstan, G01N 33/53 A61B 10/00. The method of preparing excretory-secretory antigen for serological diagnosis of Opisthorchiasis]. no. 68176; zayavl. 11.02.2010; opubl. 15.04.2011, Byull. no. 4, 4 р.
Bulashev A.K., Suranshiyev Z.A., Serikova S.H. Opisthorchis felineus ekskretorly-sekretorlyk antigeníne telímdí monoklonaldy antidenelerdín immundy khimiyalyk sipattamasy. S.Seyfullin atyndaġy Kazakh agrotekhnikalyk universitetínín Ġylym zharshysy [Immunochemical characterization of monoclonal antibodies specific for excretory-secretory antigen of Opisthorchis felineus. J.Science Review of S.Seyfullin Kazakh Agro Technical University]. Astana, 2013, no. 4(79), рр. 3-8.
Razrabotka sposobov immunofermentnoy diagnostiki opistorkhoza cheloveka i zhivotnykh: otchet o NIR (zaklyuchitel'nyy) / AO «Kazakhskiy agrotekhnicheskiy universitet im.S.Seyfullina»: ruk. Bulashev A.K.; ispoln.: Borovikov S.N. [Development of methods for diagnosis of human and animal Opisthorchiasis on the basis of ELISA: Research report (final). "S.Seifullin Kazakh Agro Technical University"]. Astana, 2011, 60 р., no. 0109RK00476, Inv. no. 0211RK00426.
Dykman L.A. Zolotyye nanochastitsy: sintez, svoystva, biomeditsinskoye primeneniye. L.A.Dykman, V.A.Bogatyrev, S.Yu.Shchogolov, N.G.Khlebtsov; Institut biokhimii i fiziologii rasteniy i mikroorganizmov RAN [Gold nanoparticles: synthesis, properties and biomedical applications; Institute of Biochemistry & Physiology of Plants and Microorganisms RAS]. M., Nauka, 2008, рр. 234-256.
Kohler G and Milstin C. Continuous culture of fused cell secreting antibody of predetermined specificity. Nature, 1975, vol. 256, рр. 495-497. PMID:1172191.
Beatty J., Beatty P., Vlahos W. Measurement of monoclonal antibodi affinity by non – copetitive immunoassay. J. Immunol. Meth., 1987, vol. 100, no. 3, рр. 173-179.
Dykman L.A., Sumaroka M.V., Staroverov S. A. et al. Immunogenic Properties of Colloidal Gold. Biology Bulletin of the Russian Academy of Science, 2004, vol. 31, рр. 75-79.
Dykman L.A. and Khlebtsov N. Gold nanoparticles in biomedical applications: recent advances and perspectives. Chem.Soc.Rev., 2012, vol. 41, рр. 2256-2282.