Биохимические и иммунологические свойства экскреторно-секреторного антигена Opisthorchis felineus
Ключевые слова:
иммунология, Opisthorchis felineus, мариты, экскреторно-секреторный антиген, электрофорез, иммуногенность.Аннотация
Основным компонентом при постановке серологических реакций является антиген, который должен обладать необходимыми биохимическими, иммунологическими свойствами, высокой специфичностью и активностью. В статье приводятся данные по получению экскреторно-секреторного антигена возбудителя Opisthorchis felineus. Приведена схема получения антигенного продукта, основанная на культивировании марит описторха in vitro в искусственной среде RPMI-1640 и дальнейшей очисткой методом центрифугирования. Сравнительные эксперименты по детекции белковых компонентов антигена, показывают преимущество метода окраски оксидом серебра над стандартным методом, основанном на использовании раствора Кумасси. Учитывая данный факт, автором было доказано, что в состав экскреторно-секреторного антигена входит до 21 белковой фракции. В статье приводятся данные по изучению активности, специфичности и иммуногенности экскреторно-секреторного антигена. Использование антигена в непрямом варианте ИФА с сыворотками зараженных собак, показало его высокую активность и возможность применения для выявления специфических антител против возбудителя описторхоза. Кроме этого, использование указанного антигена для иммунизации лабораторных мышей и сирийских хомяков показало высокую иммунологическую активность антигена и способность вызывать иммунный ответ в организме лабораторных животных.
Библиографические ссылки
Литература
1. Wongratanacheewin S., Sermswan R.W., Sirisinha S. Immunology and molecular biology of Opisthorchis viverrini infection // Acta Trop. – 2003. – Vol. 88(3). – P. 195-207. DOI:10.1016/j.actatropica.2003.02.002
2. Sun T., Gibson J.B. Antigens of Clonorchis sinensis in experimental and human infections // Am. J. Trop. Med. Hyg. – 1969. – Vol. 18. – P. 241-252.
3. Sun T., Gibson J.B. Metabolic products of adult Clonorchis sinensis: their composition and antigenic potential // J. Helminthol. – 1969. – Vol. 43. P. 395-402.
4. Kojima S., Yokogawa M., Tada T. Production and properties of reaginic antibodies in rabbits infected with Clonorchis sinensis or Schistosoma japonicum // Exp. Parasitol. – 1974. – Vol. 35. – P. 141-149.
5. Chen C.Y., Hsieh W.C., Shih H.H., Chen S.N. Immunodiagnosis of clonorchiasis by enzyme-linked immunosorbent assay // Southeast Asian J. Trop. Med. Publ. Health. – 1988. – Vol. 19. – P. 117-121.
6. Klebanovskaia I.A. Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. I. A method of isolating erythrocyte samples and the results of their diagnostic use in experimental opisthorchiasis in golden hamsters // Med. Parazitol. (Mosk). – 1981. – Vol. 50. – P. 20-23.
7. Klebanovskaia I.A. Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. 3. An economic method of obtaining a dried, erythrocyte opisthorchid antigenic diagnostic agent // Med. Parazitol. (Mosk). – 1985. – Vol. 54. – P. 25-27.
8. Glupov V.V., Khokhlova N.I., Khvoshchevskaia M.F., Vodianitskaia S.N., Iurlova, N.I. The use of immunoblotting for studying Opisthorchis felineus (Rivolta, 1884) antigens // Med. Parazitol. (Mosk). – 1997. – Vol. 66. – P. 17-19.
9. Kotelkin A.T., Razumov I.A., Pokrovskii I.V., Loktev V.B. A comparative study of the somatic, excretory-secretory and egg antigens of Opisthorchis felineus // Med. Parazitol. (Mosk). – 1997. – Vol. 66. – P. 12-16.
10. Meniavtseva T.A., Ratner G.M., Struchkova S.V., Kolmakova M.V., Stepanova T.F., Lepekhin A.V., Maier V.A., Postnikova T.F. Immunoenzyme analysis in the diagnosis of opisthorchiasis. I. The development of an immunoenzyme method for determining IgM antibodies to the Opisthorchis antigen // Med. Parazitol. (Mosk). – 1996. – Vol. 65. – P. 41-43.
11. Wongratanacheewin S., Sirisinha S. Analysis of Opisthorchis viverrini antigens: physicochemical characterization and antigen localization // Southeast Asian J. Trop. Med. Publ. Health. – 1987. – Vol. 18. – P. 511-520.
12. Wongratanacheewin S., Chawengkirttikul R., Bunnag D., Sirisinha S. Analysis of Opisthorchis viverrini antigens by immunoprecipitation and polyacrylamide gel electrophoresis // Parasitology. – 1988. – Vol. 96. – P. 119-128.
13. Боровиков С.Н., Куйбагаров М.А., Сураншиев Ж.А., Баешева Д.А., Атыгаева С.К., Халикова А.С. Получение и изучение иммунохимических свойств антигенов описторхов // Биотехнология. Теория и практика. – 2010. – №4. – С. 70-74.
14. Lvova M.N., Tangkawattana S., Balthaisong S., Katokhin A.V., Mordvinov V.A., Sripa B. Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke // Parasitol Int. – 2012. – Vol. 61(1). – P. 167-172. DOI: 10.1016/j.parint.2011.08.005.
15. Фридлянская И.И. Получение поликлональных антител // Методы культивирования клеток: Сб. науч. трудов, Л.: Наука. – 1988. – С. 194-205.
16. Киян В.С., Токпан С.С., Байболин Ж.К., Тетерин А.В., Мертасов А.Г. Гематологические показатели собак при искусственном заражении описторхозом // Вестник ПГУ. Серия Химико-биологическая, Павлодар. – 2015. – №3. – С. 12-20.
References
1. Wongratanacheewin S, Sermswan RW, Sirisinha S (2003) Immunology and molecular biology of Opisthorchis viverrini infection, Acta Trop, 88:195-207. DOI:10.1016/j.actatropica.2003.02.002
2. Sun T, Gibson JB (1969) Antigens of Clonorchis sinensis in experimental and human infections, Am. J. Trop. Med. Hyg., 18:241-252.
3. Sun T, Gibson JB (1969) Metabolic products of adult Clonorchis sinensis: their composition and antigenic potential, J. Helminthol., 43:395-402.
4. Kojima S, Yokogawa M, Tada T (1974) Production and properties of reaginic antibodies in rabbits infected with Clonorchis sinensis or Schistosoma japonicum, Exp. Parasitol., 35:141-149.
5. Chen CY, Hsieh WC, Shih HH, Chen SN (1988) Immunodiagnosis of clonorchiasis by enzyme-linked immunosorbent assay, Southeast Asian J. Trop. Med. Publ. Health., 19:117-121.
6. Klebanovskaia I.A. (1981) Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. I. A method of isolating erythrocyte samples and the results of their diagnostic use in experimental opisthorchiasis in golden hamsters, Med. Parazitol. (Mosk)., 50:20-23.
7. Klebanovskaia IA (1985) Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. 3. An economic method of obtaining a dried, erythrocyte opisthorchid antigenic diagnostic agent, Med. Parazitol. (Mosk)., 54:25-27.
8. Glupov VV, Khokhlova NI, Khvoshchevskaia MF, Vodianitskaia SN, Iurlova NI (1997) The use of immunoblotting for studying Opisthorchis felineus (Rivolta, 1884) antigens, Med. Parazitol. (Mosk)., 66:17-19.
9. Kotelkin AT, Razumov IA, Pokrovskii IV, Loktev VB (1997) A comparative study of the somatic, excretory-secretory and egg antigens of Opisthorchis felineus, Med. Parazitol. (Mosk)., 66:12-16.
10. Meniavtseva TA, Ratner GM, Struchkova SV, Kolmakova MV, Stepanova TF, Lepekhin AV, Maier VA, Postnikova TF (1996) Immunoenzyme analysis in the diagnosis of opisthorchiasis. I. The development of an immunoenzyme method for determining IgM antibodies to the Opisthorchis antigen, Med. Parazitol. (Mosk)., 65:41-43.
11. Wongratanacheewin S, Sirisinha S (1987) Analysis of Opisthorchis viverrini antigens: physicochemical characterization and antigen localization, Southeast Asian J. Trop. Med. Publ. Health., 18:511-520.
12. Wongratanacheewin S, Chawengkirttikul R, Bunnag D, Sirisinha S (1988) Analysis of Opisthorchis viverrini antigens by immunoprecipitation and polyacrylamide gel electrophoresis, Parasitology, 96:119-128.
13. Borovikov SN, Kuibagarov MA, Suranshiev ZhA, Baesheva DA, Atygaeva SK, Khalikova AS (2010) Preparation and study immunochemical properties of the opistorchis antigens [Poluchenie i izuchenie immunokhimicheskikh svoistv antigenov opistorkhov], Biothechnology. Theory and practice, 4:70-74. (In Russian)
14. Lvova MN, Tangkawattana S, Balthaisong S, Katokhin AV, Mordvinov VA, Spira B (2012) Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke, Parasitol Int., 61:167-172. DOI: 10.1016/j.parint.2011.08.005.
15. Fridlianskaia II (1988) Preparation the polyclonal antibody. Cell culture techniques [Poluchenie poliklonal'nykh antitel. Metody kul'tivirovaniia kletok], Bullet of scientific papers, Nauka, Leningrad, 194-205. (In Russian)
16. Kiyan VS, Tokpan SS, Baibolin ZhK, Teterin AV, Mertasov AG (2015) Hematologic indicators of the dogs under artificial infection by opisthorchiasis [Gematologicheskie pokazateli sobak pri iskusstvennom zarazhenii opistorkhozom], Bulletin of PSU. A series of chemical and biological, Pavlodar, 3:12-20. (In Russian)
1. Wongratanacheewin S., Sermswan R.W., Sirisinha S. Immunology and molecular biology of Opisthorchis viverrini infection // Acta Trop. – 2003. – Vol. 88(3). – P. 195-207. DOI:10.1016/j.actatropica.2003.02.002
2. Sun T., Gibson J.B. Antigens of Clonorchis sinensis in experimental and human infections // Am. J. Trop. Med. Hyg. – 1969. – Vol. 18. – P. 241-252.
3. Sun T., Gibson J.B. Metabolic products of adult Clonorchis sinensis: their composition and antigenic potential // J. Helminthol. – 1969. – Vol. 43. P. 395-402.
4. Kojima S., Yokogawa M., Tada T. Production and properties of reaginic antibodies in rabbits infected with Clonorchis sinensis or Schistosoma japonicum // Exp. Parasitol. – 1974. – Vol. 35. – P. 141-149.
5. Chen C.Y., Hsieh W.C., Shih H.H., Chen S.N. Immunodiagnosis of clonorchiasis by enzyme-linked immunosorbent assay // Southeast Asian J. Trop. Med. Publ. Health. – 1988. – Vol. 19. – P. 117-121.
6. Klebanovskaia I.A. Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. I. A method of isolating erythrocyte samples and the results of their diagnostic use in experimental opisthorchiasis in golden hamsters // Med. Parazitol. (Mosk). – 1981. – Vol. 50. – P. 20-23.
7. Klebanovskaia I.A. Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. 3. An economic method of obtaining a dried, erythrocyte opisthorchid antigenic diagnostic agent // Med. Parazitol. (Mosk). – 1985. – Vol. 54. – P. 25-27.
8. Glupov V.V., Khokhlova N.I., Khvoshchevskaia M.F., Vodianitskaia S.N., Iurlova, N.I. The use of immunoblotting for studying Opisthorchis felineus (Rivolta, 1884) antigens // Med. Parazitol. (Mosk). – 1997. – Vol. 66. – P. 17-19.
9. Kotelkin A.T., Razumov I.A., Pokrovskii I.V., Loktev V.B. A comparative study of the somatic, excretory-secretory and egg antigens of Opisthorchis felineus // Med. Parazitol. (Mosk). – 1997. – Vol. 66. – P. 12-16.
10. Meniavtseva T.A., Ratner G.M., Struchkova S.V., Kolmakova M.V., Stepanova T.F., Lepekhin A.V., Maier V.A., Postnikova T.F. Immunoenzyme analysis in the diagnosis of opisthorchiasis. I. The development of an immunoenzyme method for determining IgM antibodies to the Opisthorchis antigen // Med. Parazitol. (Mosk). – 1996. – Vol. 65. – P. 41-43.
11. Wongratanacheewin S., Sirisinha S. Analysis of Opisthorchis viverrini antigens: physicochemical characterization and antigen localization // Southeast Asian J. Trop. Med. Publ. Health. – 1987. – Vol. 18. – P. 511-520.
12. Wongratanacheewin S., Chawengkirttikul R., Bunnag D., Sirisinha S. Analysis of Opisthorchis viverrini antigens by immunoprecipitation and polyacrylamide gel electrophoresis // Parasitology. – 1988. – Vol. 96. – P. 119-128.
13. Боровиков С.Н., Куйбагаров М.А., Сураншиев Ж.А., Баешева Д.А., Атыгаева С.К., Халикова А.С. Получение и изучение иммунохимических свойств антигенов описторхов // Биотехнология. Теория и практика. – 2010. – №4. – С. 70-74.
14. Lvova M.N., Tangkawattana S., Balthaisong S., Katokhin A.V., Mordvinov V.A., Sripa B. Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke // Parasitol Int. – 2012. – Vol. 61(1). – P. 167-172. DOI: 10.1016/j.parint.2011.08.005.
15. Фридлянская И.И. Получение поликлональных антител // Методы культивирования клеток: Сб. науч. трудов, Л.: Наука. – 1988. – С. 194-205.
16. Киян В.С., Токпан С.С., Байболин Ж.К., Тетерин А.В., Мертасов А.Г. Гематологические показатели собак при искусственном заражении описторхозом // Вестник ПГУ. Серия Химико-биологическая, Павлодар. – 2015. – №3. – С. 12-20.
References
1. Wongratanacheewin S, Sermswan RW, Sirisinha S (2003) Immunology and molecular biology of Opisthorchis viverrini infection, Acta Trop, 88:195-207. DOI:10.1016/j.actatropica.2003.02.002
2. Sun T, Gibson JB (1969) Antigens of Clonorchis sinensis in experimental and human infections, Am. J. Trop. Med. Hyg., 18:241-252.
3. Sun T, Gibson JB (1969) Metabolic products of adult Clonorchis sinensis: their composition and antigenic potential, J. Helminthol., 43:395-402.
4. Kojima S, Yokogawa M, Tada T (1974) Production and properties of reaginic antibodies in rabbits infected with Clonorchis sinensis or Schistosoma japonicum, Exp. Parasitol., 35:141-149.
5. Chen CY, Hsieh WC, Shih HH, Chen SN (1988) Immunodiagnosis of clonorchiasis by enzyme-linked immunosorbent assay, Southeast Asian J. Trop. Med. Publ. Health., 19:117-121.
6. Klebanovskaia I.A. (1981) Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. I. A method of isolating erythrocyte samples and the results of their diagnostic use in experimental opisthorchiasis in golden hamsters, Med. Parazitol. (Mosk)., 50:20-23.
7. Klebanovskaia IA (1985) Indirect hemagglutination reaction in the diagnosis of the early phase of opisthorchiasis. 3. An economic method of obtaining a dried, erythrocyte opisthorchid antigenic diagnostic agent, Med. Parazitol. (Mosk)., 54:25-27.
8. Glupov VV, Khokhlova NI, Khvoshchevskaia MF, Vodianitskaia SN, Iurlova NI (1997) The use of immunoblotting for studying Opisthorchis felineus (Rivolta, 1884) antigens, Med. Parazitol. (Mosk)., 66:17-19.
9. Kotelkin AT, Razumov IA, Pokrovskii IV, Loktev VB (1997) A comparative study of the somatic, excretory-secretory and egg antigens of Opisthorchis felineus, Med. Parazitol. (Mosk)., 66:12-16.
10. Meniavtseva TA, Ratner GM, Struchkova SV, Kolmakova MV, Stepanova TF, Lepekhin AV, Maier VA, Postnikova TF (1996) Immunoenzyme analysis in the diagnosis of opisthorchiasis. I. The development of an immunoenzyme method for determining IgM antibodies to the Opisthorchis antigen, Med. Parazitol. (Mosk)., 65:41-43.
11. Wongratanacheewin S, Sirisinha S (1987) Analysis of Opisthorchis viverrini antigens: physicochemical characterization and antigen localization, Southeast Asian J. Trop. Med. Publ. Health., 18:511-520.
12. Wongratanacheewin S, Chawengkirttikul R, Bunnag D, Sirisinha S (1988) Analysis of Opisthorchis viverrini antigens by immunoprecipitation and polyacrylamide gel electrophoresis, Parasitology, 96:119-128.
13. Borovikov SN, Kuibagarov MA, Suranshiev ZhA, Baesheva DA, Atygaeva SK, Khalikova AS (2010) Preparation and study immunochemical properties of the opistorchis antigens [Poluchenie i izuchenie immunokhimicheskikh svoistv antigenov opistorkhov], Biothechnology. Theory and practice, 4:70-74. (In Russian)
14. Lvova MN, Tangkawattana S, Balthaisong S, Katokhin AV, Mordvinov VA, Spira B (2012) Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke, Parasitol Int., 61:167-172. DOI: 10.1016/j.parint.2011.08.005.
15. Fridlianskaia II (1988) Preparation the polyclonal antibody. Cell culture techniques [Poluchenie poliklonal'nykh antitel. Metody kul'tivirovaniia kletok], Bullet of scientific papers, Nauka, Leningrad, 194-205. (In Russian)
16. Kiyan VS, Tokpan SS, Baibolin ZhK, Teterin AV, Mertasov AG (2015) Hematologic indicators of the dogs under artificial infection by opisthorchiasis [Gematologicheskie pokazateli sobak pri iskusstvennom zarazhenii opistorkhozom], Bulletin of PSU. A series of chemical and biological, Pavlodar, 3:12-20. (In Russian)
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