Determining specific proteins in somatic and excretory/secretory antigens of adult and microfilariae ofDirofilaria immitis and investigation of glycoprotein occurrence
Abstract
In this study, somatic and secretional proteins of adult and microfilariae of Dirofilaria immitis were separated with SDS-PAGE. Which separated proteins include specific antigens to the disease was determined with immunblotting technique. Antigenic glycoproteins were determined with glycan kits. According to the immunblotting results which were obtained by somatic and excretory/secretory (E/S) antigens of adult form of parasite it has determined that: 69, 20, 18, 16, 14, 12 kDa proteins are commonly specific, in adults (somatic antigen); 20, 18,16 kDa proteins, adult E/S products; 18 and 16 kDa proteins are glycoproteins. According to the immunblotting results which were obtained by somatic and E/S antigens of microfilariae it has determined that 16, 14 and 12 kDa proteins are commonly specific, in microfilariae and E/S products of microfilariae 14 and 16 kDa proteins are glycoproteins. In all forms of parasite generally mannose, sialic acid and galactose were seen except small differences in sugar ingredients of specific antigens. Determining antigenic structured glycoproteins of parasites will provide clarify of relation between host parasite and also will provide contribution to the vaccine studies
Keywords
Dirofilaria immitis excretory/secretory antigen glycoprotein microfilariae specific protein
References
- Anderson RC (2000): Nematode Parasites of Vertebrates: Their Development and Transmission. 2nd Edition, CABI Publishing, New York.
- Akao N, Kondo K, Fujita K (1991): Immunoblot analysis of Dirofilaria immitis recognized by infected humans. Ann Trop Med Parasitol, 85, 455-460.
- Boto WMO, Powers KG, Levy DA (1984): Antigens of Dirofilaria immitis which are immunogenic in the canine hosts: Detection by immunostaining of protein blots with the antibodies of occult dogs. J Immunol, 133, 975-980.
- Burnie JP, Holland, M, Matthews RC ve ark. (1987): Role of immunblotting in the diagnosis of culture negative and enterococcal endocarditis. J Clin Pathol, 40, 1149- 1158.
- Candiano G, Bruschi M, Musante L ve ark. (2004): Blue silver: A very sensitive colloidal Coomassie G-250 staining for proteome analysis. Electrophoresis, 25, 1327-1333.
- Doğanay A, Şahal M (1987): Türkiye’de köpeklerdeki dirofilariasis sorunu ve insan sağlığı yönünden önemi. Ankara Üniv Vet Fak Derg, 34, 277-287.
- Die I, Cummings RD (2010): Glycan gimmickry by parasitic helminths: A strategy for modulating the host immune response. Glycobiology, 20, 2-12.
- Frank GR, Tripp CA, Grieve RB (1996): Molecular cloning of a developmentally regulated protein isolated from excretory-secretory products of larval Dirofilaria immitis. Mol Biochem Parasitol, 75, 231-240.
- Godel C, Kumar S, Koutsovoulos G ve ark. (2012): The genome of the heartworm, Dirofilaria immitis, reveals drug and vaccine targets. FASEB J, 26, 4650-61. Doi: 10.1096/fj.12-205096.
- Kadirpaşaoğlu KA, Bilge FH, Baier RE (1993): Determination of the role of cuticular carbonhydrates in the hemocompatibility of Dirofilaria immitis (Nematoda). J Biomed Mater Res, 27, 207-216.
- Kaneko H, Hayasaki M, Ohishi İ (1990): Antigenic identification of excretory-secrotory products of adult Dirofilaria immitis. Nippon Juigaku Zasshi, 52, 995-1000.
- Kang S, Cummings RD, McCall JW (1993): Characterisation of the N-linked oligosaccarides in glycoproteins synthesized by microfilarie of Dirofilaria immitis. J Parasitol, 79, 815- 828.
- Laemmli K (1970): Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
- Lombard CW (1987): Heartworm Disease. 275-299. In: Bonagura JD. (Ed), Cardiology. 2nd Edition, Churchill Livingstone, New York.
- Lowry OH, Rosebrough NJ, Farr AL ve ark. (1951): Protein measurement with the folin phenol reagent. J Biol Chem, 193, 265-275.
- Oleaga A, Pérez-Sánchez R, Pagés E ve ark. (2009): Identification of immunoreactive proteins from the dog heartworm (Dirofilaria immitis) differentially recognized by the sera from dogs with patent or occult infections. Mol Biochem Parasitol, 166, 134-141.
- Öge H, Öge S, Yıldırım A ve ark. (2005): İmmunblotting analysis of somatic components of Dirofilaria immitis. Parasite, 12, 179-182
- Philipp M, Davis TB (1986): Biochemical and immunologic characterization of a major surface antigen of Dirofilaria immitis infective larvae. J Immunol, 6, 2621-2627.
- Pou-Barreto C, Quispe-Ricalde MA, Morchón R ve ark. (2008): Galectin and aldolase-like molecules are responsible for the spesific IgE response in humans exposed to Dirofilaria immitis. Parasit Immunol, 30, 596-602.
- Sambrook J, Fritsh EF, Manniatis T (1989): Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York.
- Sarnıç H, Alkan M (1986): Köpeklerdeki dirofilariosis olguları ve insan sağlığı yönünden önemi. Türkiye Parazitol Derg, 2, 169-174.
- Scott AL, Diala C, Moraga DA ve ark. (1988): Dirofilaria immitis: Biochemical and immunological characterization of the surface antigens from adult parasites. Exp Parasitol, 67, 307-23.
- Scott AL, İbrahim MS, Tamashiro WK (1990): Surface associated antigens of second, third and fourth stage larvae of Dirofilaria immitis. Acta Trop, 47, 339-359.
- Sbihi Y, Jansen D, Osuna A (1996): Serologic recognition of Hydatid cyst antigens using different purification methods. Diagn Microbiol Infect Dis, 24, 205-211.
- Song KH, Hayasaki M, Cholig C ve ark. (2002): İmmunological responces of dogs experimentaly infected with Dirofilaria immitis. J Vet Sci, 3, 109-114.
- Song KH, Lee SE, Hayasaki M (2003): Seroprevalance of canine dirofilariosis in South Korea. Vet Parasitol, 114, 231-236.
- Soulsby EJL (1982): Helminths, Arthropods and Protozoa of Domesticated Animals. 7th ed. Bailliere Tindal, London.
- Tamashiro WK, Ehrenberg JP, Levy DA ve ark. (1986): Antigenic peptides on the surface of Dirofilaria immitis microfilarie. Mol Biochem Parasitol, 18, 369-376.
- Ünübol Aypak S, Uysal H (2014): Koyun ve farelerde kist hidatik proteinlerinin karşılaştırmalı analizi ve antijenik proteinlerde glikoprotein varlığının değerlendirilmesi. Ankara Üniv Vet Fak Derg, 61, 243-248.
- Varki A Cummings R, Esko J ve ark. (1999): Essentials of Glycobiology. Cold Spring Harbor Laboratory Press. New york.
- Zingales B (1984): Analysis of proteins by dodecyl sulfate- polyacrylamide gel electrophoresis. In: Genes and Antigenes of Parasites. C.M. Marel Ed., Fiocruz, Rio de Janeiro, pp. 357-363.
Dirofilaria immitis’in ergin ve mikrofilerlerinin somatik ve salgısal antijenlerinde spesifik proteinlerin belirlenmesi ve glikoprotein varlığının araştırılması
Abstract
Bu çalışmada, Dirofilaria immitis’in ergin ve mikrofilerlerinin somatik ve E/S proteinleri SDS-PAGE ile ayrıştırıldı. Ayrıştırılan proteinlerden hangilerinin hastalığa spesifik antijenler içerdiği immunblotting yöntemiyle belirlendi. Antijenik glikoproteinler ise glikan kitleriyle belirlendi. Parazitin ergin formunun somatik ve E/S antijenleri ile yapılan immunblotting sonucunda; 69, 20, 18, 16, 14, 12 kDa’luk proteinlerin ortak şekilde spesifik olduğu, erginlerde (somatik antijen); 20, 18, 16 kDa’luk proteinlerin, ergin salgısal ürünlerinde (E/S antijen); 18 ve 16 kDa’luk proteinlerin glikoprotein olduğu belirlendi. Mikrofilerlerinin somatik ve salgısal antijenleri ile yapılan immunblotting sonucunda; 16, 14 ve 12 kDa'luk proteinlerin ortak şekilde spesifik olduğu, bunlardan mikrofilerlerde ve mikrofilerlerin salgısal ürünlerinde 14 ve 16 kDa’luk proteinlerin glikoprotein olduğu belirlendi. Parazitin bütün formlarında, spesifik antijenlerin şeker içeriklerindeki çok küçük farklılıklar dışında genel olarak mannoz, siyalik asit ve galaktoz bulunduğu tespit edildi. Parazitlerin antijenik yapıdaki glikoproteinlerinin belirlenmesi, konak-parazit arasındaki ilişkilerin aydınlatılmasına ve yapılacak olan aşı çalışmalarına katkı sağlayacaktır
References
- Anderson RC (2000): Nematode Parasites of Vertebrates: Their Development and Transmission. 2nd Edition, CABI Publishing, New York.
- Akao N, Kondo K, Fujita K (1991): Immunoblot analysis of Dirofilaria immitis recognized by infected humans. Ann Trop Med Parasitol, 85, 455-460.
- Boto WMO, Powers KG, Levy DA (1984): Antigens of Dirofilaria immitis which are immunogenic in the canine hosts: Detection by immunostaining of protein blots with the antibodies of occult dogs. J Immunol, 133, 975-980.
- Burnie JP, Holland, M, Matthews RC ve ark. (1987): Role of immunblotting in the diagnosis of culture negative and enterococcal endocarditis. J Clin Pathol, 40, 1149- 1158.
- Candiano G, Bruschi M, Musante L ve ark. (2004): Blue silver: A very sensitive colloidal Coomassie G-250 staining for proteome analysis. Electrophoresis, 25, 1327-1333.
- Doğanay A, Şahal M (1987): Türkiye’de köpeklerdeki dirofilariasis sorunu ve insan sağlığı yönünden önemi. Ankara Üniv Vet Fak Derg, 34, 277-287.
- Die I, Cummings RD (2010): Glycan gimmickry by parasitic helminths: A strategy for modulating the host immune response. Glycobiology, 20, 2-12.
- Frank GR, Tripp CA, Grieve RB (1996): Molecular cloning of a developmentally regulated protein isolated from excretory-secretory products of larval Dirofilaria immitis. Mol Biochem Parasitol, 75, 231-240.
- Godel C, Kumar S, Koutsovoulos G ve ark. (2012): The genome of the heartworm, Dirofilaria immitis, reveals drug and vaccine targets. FASEB J, 26, 4650-61. Doi: 10.1096/fj.12-205096.
- Kadirpaşaoğlu KA, Bilge FH, Baier RE (1993): Determination of the role of cuticular carbonhydrates in the hemocompatibility of Dirofilaria immitis (Nematoda). J Biomed Mater Res, 27, 207-216.
- Kaneko H, Hayasaki M, Ohishi İ (1990): Antigenic identification of excretory-secrotory products of adult Dirofilaria immitis. Nippon Juigaku Zasshi, 52, 995-1000.
- Kang S, Cummings RD, McCall JW (1993): Characterisation of the N-linked oligosaccarides in glycoproteins synthesized by microfilarie of Dirofilaria immitis. J Parasitol, 79, 815- 828.
- Laemmli K (1970): Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
- Lombard CW (1987): Heartworm Disease. 275-299. In: Bonagura JD. (Ed), Cardiology. 2nd Edition, Churchill Livingstone, New York.
- Lowry OH, Rosebrough NJ, Farr AL ve ark. (1951): Protein measurement with the folin phenol reagent. J Biol Chem, 193, 265-275.
- Oleaga A, Pérez-Sánchez R, Pagés E ve ark. (2009): Identification of immunoreactive proteins from the dog heartworm (Dirofilaria immitis) differentially recognized by the sera from dogs with patent or occult infections. Mol Biochem Parasitol, 166, 134-141.
- Öge H, Öge S, Yıldırım A ve ark. (2005): İmmunblotting analysis of somatic components of Dirofilaria immitis. Parasite, 12, 179-182
- Philipp M, Davis TB (1986): Biochemical and immunologic characterization of a major surface antigen of Dirofilaria immitis infective larvae. J Immunol, 6, 2621-2627.
- Pou-Barreto C, Quispe-Ricalde MA, Morchón R ve ark. (2008): Galectin and aldolase-like molecules are responsible for the spesific IgE response in humans exposed to Dirofilaria immitis. Parasit Immunol, 30, 596-602.
- Sambrook J, Fritsh EF, Manniatis T (1989): Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York.
- Sarnıç H, Alkan M (1986): Köpeklerdeki dirofilariosis olguları ve insan sağlığı yönünden önemi. Türkiye Parazitol Derg, 2, 169-174.
- Scott AL, Diala C, Moraga DA ve ark. (1988): Dirofilaria immitis: Biochemical and immunological characterization of the surface antigens from adult parasites. Exp Parasitol, 67, 307-23.
- Scott AL, İbrahim MS, Tamashiro WK (1990): Surface associated antigens of second, third and fourth stage larvae of Dirofilaria immitis. Acta Trop, 47, 339-359.
- Sbihi Y, Jansen D, Osuna A (1996): Serologic recognition of Hydatid cyst antigens using different purification methods. Diagn Microbiol Infect Dis, 24, 205-211.
- Song KH, Hayasaki M, Cholig C ve ark. (2002): İmmunological responces of dogs experimentaly infected with Dirofilaria immitis. J Vet Sci, 3, 109-114.
- Song KH, Lee SE, Hayasaki M (2003): Seroprevalance of canine dirofilariosis in South Korea. Vet Parasitol, 114, 231-236.
- Soulsby EJL (1982): Helminths, Arthropods and Protozoa of Domesticated Animals. 7th ed. Bailliere Tindal, London.
- Tamashiro WK, Ehrenberg JP, Levy DA ve ark. (1986): Antigenic peptides on the surface of Dirofilaria immitis microfilarie. Mol Biochem Parasitol, 18, 369-376.
- Ünübol Aypak S, Uysal H (2014): Koyun ve farelerde kist hidatik proteinlerinin karşılaştırmalı analizi ve antijenik proteinlerde glikoprotein varlığının değerlendirilmesi. Ankara Üniv Vet Fak Derg, 61, 243-248.
- Varki A Cummings R, Esko J ve ark. (1999): Essentials of Glycobiology. Cold Spring Harbor Laboratory Press. New york.
- Zingales B (1984): Analysis of proteins by dodecyl sulfate- polyacrylamide gel electrophoresis. In: Genes and Antigenes of Parasites. C.M. Marel Ed., Fiocruz, Rio de Janeiro, pp. 357-363.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA65ER63CV |
| Journal Section | Research Article |
| Authors | |
| Publication Date | March 1, 2017 |
| Published in Issue | Year 2017Volume: 64 Issue: 1 |