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Cloning and expression of DiT33 from Dirofilaria immitis: a specific and early marker of heartworm infection

Published online by Cambridge University Press:  06 April 2009

X. Q. Hong ,
J. Santiago Mejia ,
S. Kumar ,
F. B. Perler  and
C. K. S. Carlow
X. Q. Hong
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
J. Santiago Mejia
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
S. Kumar
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
F. B. Perler
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
C. K. S. Carlow
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
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Summary

Dirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

Keywords

Dirofilaria immitisDiT33Ov33pepsin inhibitordiagnosisheartworm.
Type
Research Article
Information
Parasitology , Volume 112 , Issue 3 , March 1996 , pp. 331 - 338
Copyright
Copyright © Cambridge University Press 1996

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References

REFERENCES

American Heartworm Society (1992). Recommended procedures for the diagnosis and management of heartworm (Dirofilaria immitis) infection. In Proceedings of the Heartworm Symposium '92, (ed. Soll, M. D.), pp. 289–294. Batavia, IL: American Heartworm Society.Google Scholar
Bradley, J. E., Helm, R., Lahaise, M. & Maizels, R. M. (1991). cDNA clones of Onchocerca volvulus low molecular weight antigens provide immunologically specific diagnostic probes. Molecular and Biochemical Parasitology 46, 219–28.Google Scholar
Chandrashekar, R., Curtis, K. C., Ramzy, R. M., Liftis, F., Li, B. M. & Weil, G. J. (1994). Molecular cloning of Brugia malayi antigens for diagnosis of lymphatic filariasis. Molecular and Biochemical Parasitology 64, 261–71.CrossRefGoogle ScholarPubMed
Chandrashekar, R., Masood, K., Alvarez, R. M., Ogunrinade, A. F., Lujan, R., Richards, F. O. Jr., & Weil, G. J. (1991). Molecular cloning and characterization of recombinant parasite antigens for immunodiagnosis of Onchocerciasis. Journal of Clinical Investigation 88, 1460–6.Google Scholar
Courtney, C. H. (1986). Comparison of tests for immunodiagnosis of canine dirofilariasis. In Proceedings of the Heartworm Symposium '86. (ed. Otto, G. F.), pp. 77–82. Washington, DC: American Heartworm Society.Google Scholar
Courtney, C. H., Zeng, Q. Y. & Bean, E. S. (1988). Sensitivity and specificity of the DiroCHEK heartworm antigen test for immunodiagnosis of canine dirofilariasis and a comparison with other immunodiagnostic tests. Journal of the American Animal Hospital Association 24, 27–32.Google Scholar
Dissanayake, S., Xu, M., Nkenfou, C. & Piessens, W. F. (1993). Molecular cloning and serological characterization of a Brugia malayi pepsin inhibitor homolog. Molecular and Biochemical Parasitology 62, 143–6.CrossRefGoogle ScholarPubMed
Dzimianski, M. T. & McCall, J. W. (1986). Evaluation of adult antigen diagnostic test kits using well defined dog sera from laboratory and field trails. In Proceedings of the Heartworm Symposium '86 (ed. Otto, G. F.), pp. 83–86. Washington, DC: American Heartworm Society.Google Scholar
Falls, R. K. & Platt, T. R. (1982). Survey for heartworm, Dirofilaria immitis and Dipetalonema reconditum (Nematoda: Filarioidea) in dogs from Virginia and North Carolina. American Journal of Veterinary Research 43, 738–9.Google ScholarPubMed
Felsenstein, J. (1993). PHYLIP: Phytogeny Inference Package. Department of Genetics, University of Washington, Seattle, Washington.Google Scholar
Grandea, A. G. III, Tuyen, L. K., Asikin, N., Davis, T. B., Philipp, M., Cohen, C. & McReynolds, L. A. (1989). A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin. Molecular and Biochemical Parasitology 35, 31–41.CrossRefGoogle Scholar
Higgens, D. G., Bleasby, A. J. & Fuchs, T. (1992). Clustal V: Improved software for multiple sequence alignment. Computer Application in the Biosciences 8, 189–91.Google Scholar
Hong, X. Q., Mejia, J. S., Nkenfou, C., Perler, F. B. & Carlow, C. K. S. (1995). A promising recombinant antigen for the early detection of heartworm infection. In Proceedings of the Heartworm Symposium '95, (ed. Soil, M. D.). Batavia, IL.: American Heartworm Society. (In press).Google Scholar
Kornfield, R. & Kornfield, S. (1985). Assembly of asparagine-linked oligosaccharides. Annual Reviews in Biochemistry 54, 631–64.Google Scholar
Kumari, S., Lillibridge, C. D., Bakeer, M., Lowrie, R. C. Jr., Jayaraman, K. & Philipp, M. T. (1995). Brugia malayi: The diagnostic potential of recombinant excretory/secretory antigens. Experimental Parasitology 79, 489–505.CrossRefGoogle Scholar
Lobos, E., Weiss, N., Karam, M., Taylor, H. R., Ottesen, E. A. & Nutman, T. B. (1991). An immunogenic Onchocerca volvulus antigen: a specific and early marker of infection. Science 251, 1603–5.Google Scholar
Lucius, R., Erondu, N., Kern, A. & Donelson, J. (1988 b). Molecular cloning of an immunodominant antigen of Onchocerca volvulus. Journal of Experimental Medicine 168, 1199–204.Google Scholar
Lucius, R., Kern, A., Seeber, F., Pogonka, T., Willenbucher, J., Taylor, H. R., Finder, M., Ghalib, H. W., Schulz-Key, H. & Soboslay, P. (1992). Specific and sensitive IgG4 immunodiagnosis of onchocerciasis with a recombinant 33 kD Onchocerca volvulus protein (Ov33). Tropical Medicine and Parasitology 43, 139–45.Google ScholarPubMed
Lucius, R., Schulz-Key, H., Buttner, D. W., Kern, A., Kaltmann, B., Prod'Hon, J., Seeber, F., Walter, R. D., Saxena, K. C. & Diesfeld, H. (1988 a). Characterization of an immunodominant Onchocerca volvulus antigen with patient sera and a monoclonal antibody. Journal of Experimental Medicine 167, 1505–10.Google Scholar
Lustigman, S., Brotman, B., Huima, T. & Prince, A. M. (1991). Characterization of an Onchocerca volvulus cDNA clone encoding a genus-specific antigen present in infective larvae and adult worms. Molecular and Biochemical Parasitology 45, 65–76.CrossRefGoogle ScholarPubMed
Martzen, M. R., McMullen, B. A., Smith, N. E., Fujikawa, K. & Peanasky, R. J. (1990). Primary structure of the major pepsin inhibitor from the intestinal parasitic nematode Ascaris suum. Biochemistry 29, 7366–72.Google Scholar
McCall, J. W., Supakorndej, N., McTier, T. L., Dzimianski, M. T. & Ricketts, R. P. (1993). Commercial antigen test kits detection infections with single female worm but not those with numerous male worms only. Proceedings of the American Association of Veterinary Parasitologists, 38th Annual Meeting, p. 36.Google Scholar
Mejia, J. S., Nkenfou, C., Southworth, M. W., Perler, F. B. & Carlow, C. K. S. (1994). Expression of an Onchocerca volvulus Ov33 homolog in Dirofilaria immitis: potential in immunodiagnosis of heartworm infection. Parasite Immunology 16, 297–303.CrossRefGoogle Scholar
Nkenfou, C. (1993). Molecular cloning of genes coding antigens specific for Onchocerca volvulus: evaluation of expressed proteins for use in the diagnosis of onchocerciasis. Ph.D. thesis, University of Yaounde, Cameroon.Google Scholar
Ozaki, L. S., Mattei, D., Jendoubi, M., Driohle, P., Blisnick, T., Guillotee, M., Puijalon, O. & Da Silva, L. P. (1986). Plaque antibody selection: rapid positive to sera raised against Plasmodium falciparum antigens. Journal of Immunological Methods 89, 213–19.CrossRefGoogle ScholarPubMed
Ramachandran, C. P. (1993). Improved immunodiagnostic tests to monitor onchocerciasis control programmes – a multicenter effort. Parasitology Today 9, 76–9.Google Scholar
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd Edn. Cold Spring Harbor, NY, USA: Cold Spring Harbour Laboratory Press.Google Scholar
Takacs, A. M., Denker, J. A., Perrine, K. G., Maroney, P. A. & Nilsen, T. (1988). A 22 nucleotide spliced leader sequence in the human parasitic nematode Brugia malayi is identical to the trans-spliced leader exon in Caenorhabditis elegans. Proceedings of the National Academy of Sciences, USA 85, 7932–6.CrossRefGoogle Scholar
Von Heijne, G. (1983). Patterns of amino acids near signal-sequence cleavage sites. European Journal of Biochemistry 133, 17–21.Google Scholar
Watson, A. J. L., Porges, W. L. & Testoni, F. J. (1973). A survey of canine filariasis in Sydney. Australian Veterinary Journal 49, 31–4.CrossRefGoogle ScholarPubMed
Weil, G. J., Malane, M. S., Powers, K. G. & Blair, L. S. (1985). Monoclonal antibodies to parasite antigens found in the serum of Dirofilaria immitis-infected dogs. Journal of Immunology 134, 1185–91.Google Scholar
Willenbucher, J., Hofle, W. & Lucius, R. (1993). The filarial antigens Av33/Ov33–3 show striking similarities to the major pepsin inhibitor from Ascaris suum. Molecular and Biochemical Parasitology 57, 349–51.CrossRefGoogle Scholar
Wong, M. M. & Fuller, S. A. (1986). Excretory secretory products (ESP) as antigens in ELISA for serodiagnosis of canine dirofilariasis. In Proceedings of the Heartworm Symposium '86. (ed. Otto, G. F.), pp. 99–105. Washington, DC: American Heartworm Society.Google Scholar
Wong, M. M. & Thomford, J. W. (1991). Serodiagnosis of prepatent dirofilariasis: problems and significance. Journal of the American Animal Hospital Association 27, 33–8.Google Scholar