Home Characterization of immunoreactive proteins of Setaria cervi isolated by preparative polyacrylamide gel electrophoresis
Article
Licensed
Unlicensed Requires Authentication

Characterization of immunoreactive proteins of Setaria cervi isolated by preparative polyacrylamide gel electrophoresis

  • Priyanka Priyadarshi , Piyush Dravid , Inayat Hussain Sheikh , Sunita Saxena , Ashish Tandon , Deep C. Kaushal , Shakir Ali and Nuzhat A. Kaushal EMAIL logo
Published/Copyright: December 28, 2016
Become an author with De Gruyter Brill

Abstract

Filarial parasites are complex mixtures of antigenic proteins and characterization of these antigenic molecules is essential to identify the diagnostically important filaria-specific antigens. In the present study, we have fractionated the somatic extracts from adults of Setaria cervi (bovine filarial parasite) on preparative SDS-polyacrylamide gel electrophoresis and tested the immunoreactivity of the separated gel fractions with polyclonal antibodies against filarial excretory-secretory antigens as well as filarial patients sera. The SDS-PAGE analysis of gel eluted fractions revealed 1 protein band in F-1 fraction, 2 protein bands in F-2 fraction and 2–3 protein bands in all other fractions (F3- F11). Seven gel eluted fractions (F1, F2, F3, F4, F5, F6 and F11) showed high ELISA reactivity with the polyclonal antibody (against excretory-secretory antigen) and four of these fractions (F-1, F-2, F3 and F6) exhibited high ELISA reactivity with antibodies present in filarial patient sera. The reactivities of the gel fractions (F1 and F2), recognized by filarial patients sera, were also tested with the monoclonal antibody (detecting the filarial circulating antigen). The F1 and F2 gel eluted fractions were found to have the target antigen of monoclonal antibody as evident by high reactivity with the monoclonal antibody in ELISA and immunoblotting. The S. cervi gel eluted F1 fraction (containing single antigen) could detect antibodies in filarial patients sera and not in non-filarial sera thereby suggesting its usefulness for specific serodiagnosis of human filariasis.

  1. Conflict of interest:Authors declare no competing interests.

Acknowledgements

The authors would like to thank the Director, CSIR-CDRI and Scientist-in-charge, Parasitology, CSIR-CDRI for their help and encouragement. PP and SS are thankful to ICMR, New Delhi for financial support and two of the authors (PD, IHS) are thankful to CSIR, New Delhi, India. CSIR-CDRI Communication No.: 8552.

References

Alli R., Bhandari Y.P., Reddy M.V., Harinath B.C. 2000. Seroreactivity of purified Brugia malayi microfilarial soluble and excretory-secretory antigens in different clinical presentations of bancroftian filariasis. Indian Journal of Experimental Biology, 38, 791–796Search in Google Scholar

Almeida A.J., Bhopale M.K., Renapurkar D.M. 1990. Evaluation of Setaria cervi antigen in the diagnosis of human filariasis. Folia Parasitologica, 37, 323–329Search in Google Scholar

Bradford M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254. 10.1016/0003-2697(76)90527-3Search in Google Scholar

Cabrera Z., Parkhouse R. 1986. Identification of antigens of Onchocerca volvulus and Onchocerca gibsoni for diagnostic use. Molecular and Biochemical Parasitology, 20, 225–231. 10.1016/0166-6851(86)90102-7Search in Google Scholar

Dissanayake S., Zheng H., Dreyer G., Xu M., Watawana L., Cheng G., Wang S., Morin P., Deng B., Kurniawan L. 1994. Evaluation of a recombinant parasite antigen for the diagnosis of lymphatic filariasis. The American Journal of Tropical Medicine and Hygiene, 50, 727–73410.4269/ajtmh.1994.50.727Search in Google Scholar

Ichimori K., King J.D., Engels D., Yajima A., Mikhailov A., Lammie P., Ottesen E.A. 2014. Global programme to eliminate lymphatic filariasis: the processes underlying programme success. PLoS Neglected Tropical Diseases, 8, e3328. 10.1371/journal.pntd.0003328Search in Google Scholar

Janardhan S., Pandiaraja P., Pandey V., Karande A., Kaliraj P. 2011. Development and characterization of monoclonal antibodies against WbSXP-1 for the detection of circulating filarial antigens. Journal of Helminthology, 85, 1–6. 10.1017/ S0022149X10000118Search in Google Scholar

Kaushal N.A., Hussain R., Ottesen E.A. 1984. Excretory-secretory and somatic antigens in the diagnosis of human filariasis. Clinical and Experimental Immunology, 56, 567–576. http://www.ncbi.nlm.nih.gov/pubmed/6204795Search in Google Scholar

Kaushal N.A., Kaushal D.C., Ghatak S. 1987. Identification of antigenic proteins of Setaria cervi by immunoblotting technique. Immunological Investigations, 16, 139–149. 10.3109/08820138709030571Search in Google Scholar

Kaushal N.A., Kaushal D.C., Ghosh S., Talwar G.P. 1994. Monoclonal antibodies against antigenic epitopes common between Setaria cervi and Brugia malayi. Indian Journal of Experimental Biology, 32, 371–375. http://www.ncbi.nlm.nih.gov/ pubmed/7525468Search in Google Scholar

Kaushal N.A., Srivastava N., Mustafa H., Tandon A., Singh S.K., Kaushal D.C. 2009. Isolation of an antigen fraction from Setaria cervi adults having potential for immunodiagnosis of human filariasis. Immunological Investigations, 38, 749–761. 10.3109/08820130903204093Search in Google Scholar

Laemmli U.K. 1970. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227, 680–685. 10.1038/227680a0Search in Google Scholar

Lowry O.H., Rosebrough N.J., Farr A.L., Randall R.J. 1951. Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry, 193, 265–27510.1016/S0021-9258(19)52451-6Search in Google Scholar

Malhotra A., Kaushal N.A., Ghatak S. 1986. Protein and antigenic composition of adult and microfilarial stages of Setaria cervi. Immunological Investigations, 15, 505–519. 10.3109/0882 0138609026691Search in Google Scholar

Malhotra A., Kaushal N.A., Kaushal D.C., Ghatak S. 1987. Antigenic characterization of excretory-secretory products of Setaria cervi. Tropical Medicine and Parasitology. 38,106–110. http://www.ncbi.nlm.nih.gov/pubmed/3629131Search in Google Scholar

Mohammadian T., Doosti M., Paknejad M., Siavoshi F., Massarrat S. 2010. Preparative SDS-PAGE Electroelution for Rapid Purification of Alkyl Hydroperoxide Reductase from Helicobacter pylori. Iranian Journal of Public Health, 39, 85–91Search in Google Scholar

More S.J., Copeman D.B. 1990. A highly specific and sensitive monoclonal antibody-based ELISA for the detection of circulating antigen in bancroftian filariasis. Tropical Medicine and Parasitology, 41, 403–406. http://www.ncbi.nlm.nih.gov/ pubmed/2075384Search in Google Scholar

Mustafa H., Srivastava N., Kaushal D.C., Kaushal N.A. 1996. Analysis and potential of excretory-secretory antigens of Setaria cervi for immunodiagnosis of human filariasis. Indian Journal of Experimental Biology, 34, 508–512Search in Google Scholar

Ravishankaran R., Shridharan R.N., Vishal L.A., Meenakshisundaram S., Karande A.A., Kaliraj P. 2016. Evaluation of immuno diagnostic assay for the exposure of stage specific filarial infection. Acta Parasitologica, 61, 232–240. 10.1515/ap-2016-0033Search in Google Scholar

Riyong D., Waikagul J., Panasoponkul C., Choochote W., Ito A., Dekumyoy P. 2010. Size and charge antigens of Dirofilaria immitis adult worm for IgG-ELISA diagnosis of bancroftian filariasis. Southeast Asian Journal of Tropical Medicine and Public Health, 41, 285–297Search in Google Scholar

Rubio de Krömer M.T., Krömer M., Lüersen K., Brattig N.W. 1998. Detection of a chemotactic factor for neutrophils in extracts of female Onchocerca volvulus. Acta Tropica, 71, 45–5610.1016/S0001-706X(98)00044-8Search in Google Scholar

Seelert H., Krause F. 2008. Preparative isolation of protein complexes and other bioparticles by elution from polyacrylamide gels. Electrophoresis, 29, 2617–2636. 10.1002/elps.20080 0061Search in Google Scholar

Singh A., Kamal S., Rathaur S. 2010. Filarial selenium glutathione peroxidase: a probable immunodiagnostic marker for lymphatic filariasis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 104, 524–528. 10.1016/j.trstmh. 2010.02.007Search in Google Scholar

Singh V., Kaushal D.C., Rathaur S., Kumar N., Kaushal N.A. 2012. Cloning, overexpression, purification and characterization of Plasmodium knowlesi lactate dehydrogenase. Protein Expression and Purification, 84, 195–203.10.1016/j.pep .2012.05.008Search in Google Scholar

Srivastava S., Srikanth E., Liebau E., Rathaur S. 2010. Identification of Setaria cervi heat shock protein 70 by mass spectrometry and its evaluation as diagnostic marker for lymphatic filariasis. Vaccine, 28, 1429–1436. 10.1016/j.vaccine.2009.06.044Search in Google Scholar

Theodore J.G., Kaliraj P. 1990. Isolation, purification and characterization of surface antigens of the bovine filarial parasite Setaria digitata for the immunodiagnosis of bancroftian filariasis. Journal of Helminthology, 64, 105–11410.1017/S0022149X00012001Search in Google Scholar

Towbin H., Staehelin T., Gordon J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350–435410.1073/pnas.76.9.4350Search in Google Scholar

Voller A., Bidwell D., Huldt G., Engvall E. 1974. A microplate method of enzyme-linked immunosorbent assay and its application to malaria. Bulletin of the World Health Organization, 51, 209–211Search in Google Scholar

WHO 2015. Lymphatic filariasis WHO. URL: http://www.who.int/ entity/ mediacentre/ factsheets/fs102/en/index.htmlSearch in Google Scholar

Wickremanayake M.N., Ekanayake S., Karunanayake E.H. 2001. Wuchereria bancrofti: detection of microfilariae in asymptomatic microfilaremic individuals with Setaria digitata antigens. The Southeast Asian journal of tropical medicine and public health, 32, 230–234Search in Google Scholar

Weil G.J., Lammie P.J., Weiss N. 1997. The ICT Filariasis Test: A rapid-format antigen test for diagnosis of bancroftian filariasis. Parasitology Today, 13, 401–40410.1016/S0169-4758(97)01130-7Search in Google Scholar

Received: 2016-2-14
Revised: 2016-8-22
Accepted: 2016-8-31
Published Online: 2016-12-28
Published in Print: 2017-3-1

© 2017 W. Stefański Institute of Parasitology, PAS

Articles in the same Issue

  1. A taxonomic review of hymenolepidids (Eucestoda, Hymenolepididae) from dormice (Rodentia, Gliridae), with descriptions of two new species
  2. Trichuris suis secrete products that reduce disease severity in a multiple sclerosis model
  3. Characterization of immunoreactive proteins of Setaria cervi isolated by preparative polyacrylamide gel electrophoresis
  4. New records of host-parasite relationships between Coenagrion scitulum (Rambur, 1842) (Odonata) and water mite larvae (Hydrachnidia) in core and edge host populations
  5. A new species of myocoptid mites Myocoptes (Comyocoptes) nesokia sp. nov. (Acariformes: Myocoptidae) parasitizing Nesokia indica (Rodentia: Muridae) in Pakistan
  6. Two new species of digeneans (Lecithasteridae and Haploporidae) of the intertidal blenny Scartichthys viridis (Valenciennes) from the central coast of Chile
  7. Phylogenetic analysis of genetically distinct Enterocytozoon bieneusi infecting renal transplant recipients
  8. Prochristianella clarkeae Beveridge, 1990 (Eutetrarhynchidae): a species complex or a species with intraspecific variation in the distribution of its tegumental microtriches?
  9. The genetic structure of hypoderatid mites (Actinotrichida: Astigmata) parasitizing great cormorant (Phalacrocorax carbo) during host post-breeding dispersal in Milicz, SW Poland
  10. Prevalence of three Oesophagostomum spp. from Tibetan Pigs analyzed by Genetic Markers of nad1, cox3 and ITS1
  11. Anti-Anisakis sp. antibodies in serum of healthy subjects. Relationship with αβ and γδ T cells
  12. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran
  13. Spermatological characteristics of the enigmatic monogenean Dictyocotyle coeliaca Nybelin, 1941 (Monopisthocotylea: Monocotylidae) reveal possible adaptation to endoparasitism
  14. Occurrence of two novel actinospore types (Cnidaria: Myxosporea) in Brazilian fish farms, and the creation of a novel actinospore collective group, Seisactinomyxon
  15. Dictyocaulosis in dairy cows in Brazil: an epidemiological, clinical-pathological and therapeutic approach
  16. Intra and inter species genetic variability of transferrin receptor gene regions in Trypanosoma evansi isolates of different livestock and geographical regions of India
  17. Transcriptional immune response in mesenteric lymph nodes in pigs with different levels of resistance to Ascaris suum
  18. The first successful report of the in vitro life cycle of Chinese Leishmania: the in vitro conversion of Leishmania amastigotes has been raised to 94% by testing 216 culture medium compound
  19. Reduced [14C]-methionine uptake and fecundity in Schistosoma mansoni females treated with recombinant tumor necrosis factor α in vitro
  20. A new feather mite of the genus Neodectes Park and Atyeo 1971 (Acari: Proctophyllodidae) from New Zealand wrens (Passeriformes: Acanthisittidae)
  21. Exploring possibilities for an alternative approach in experimental schistosomiasis mansoni: the peritoneal cavity of mice
  22. Molecular diagnosis of Entamoeba spp. versus microscopy in the Great Cairo
  23. Genetic diversity among Babesia rossi detected in naturally infected dogs in Abeokuta, Nigeria, based on 18S rRNA gene sequences
  24. Morphological and molecular characterization of Cucullanus bourdini Petter et Le Bel, 1992 (Ascaridida: Cucullanidae) from the yellowback fusilier Caesio xanthonota Bleeker (Perciformes: Caesionidae)
  25. Prevalence and molecular characterization of Theileria equi and Babesia caballi in jereed horses in Erzurum, Turkey
  26. Report of fatal mixed infection with Cryptosporidium parvum and Giardia intestinalis in neonatal calves
  27. Case Report
  28. Lethal Angiostrongylus vasorum infection in a Hungarian dog
  29. Research Note
  30. Serosurvey of anti-Toxoplasma gondii antibodies in sport horses from Paraiba state, Northeastern Brazil
  31. Erratum to: Acta Parasitologica 2016, 61(4): Table of contents
  32. Erratum to: Acta Parasitologica 2016, 61(4): Table of contents
  33. Erratum to: Acta Parasitologica (2015), 60: 354
  34. Coccidiosis in European rabbit (Oryctolagus cuniculus algirus) populations in the Iberian Peninsula
Downloaded on 21.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/ap-2017-0003/html
Scroll to top button