Biochemical detection of E-ADA on Neospora caninum tachyzoites and the effects of a specific enzymatic inhibitor

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Autores

Alexandre A. Tonin Aleksandro S. Da Silva Victor C. Pimentel Daniele Zanini Maria Rosa C. Schetinger Vera M. Morsch Giovana Camillo Fernanda S.F. Vogel Sonia T.A. Lopes

Resumen

Objective. This study aimed to investigate the presence and activity of the ecto adenosine deaminase (E-ADA) enzyme in tachyzoites of Neospora caninum (Nc-1 strain), as well as to assess the activity of a well-known E-ADA inhibitor, the deoxycoformycin. Materials and methods. The parasites were grown in cell culture, being subsequently separated in a pellet of tachyzoites, on which the E-ADA activity was tested using the concentrations 0 (control), 0.2, 0.4 and 0.8 mg mL-1. Results. The E-ADA showed high activity, progressively increasing its activity according to the enhancement of the protein concentration. The test was carried out with different concentrations of deoxycoformycin, showing that it was able to inhibit the E-ADA present on the free form of the parasite. Conclusions. Based on these results we conclude that the E-ADA is present on tachyzoites of N. caninum, and deoxycoformycin is able to inhibit this enzyme. In this sense, knowing the negative impact of N. caninum on reproductive issue in cattle (mainly abortion), might it is an alternative in order to deal with this parasitic infection.

Key words: adenosine deaminase, deoxycoformycin, neosporosis (Source: CAB, MeSH).

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Detalles del artículo

Referencias

1. Dubey JP, Schares G, Ortega-Mora, LM. Epidemiology and control of neosporosis and Neospora caninum. Clin Microbiol Rev 2007; 20(2):323-367. http://dx.doi.org/10.1128/CMR.00031-06

2. Larson RL, Hardin DK, Pierce VL. Economic considerations for diagnostic and control options for Neospora caninum-induced abortions in endemically infected herds of beef cattle. J Am Vet Med Assoc 2004; 224(10):1597-1604.
http://dx.doi.org/10.2460/javma.2004.224.1597

3. Goodswen SJ, Kennedy PJ, Ellis JP. A review of the infection, genetics, and evolution of Neospora caninum: From the past to the present. Infect Genet Evol 2013; 13:133−150.
http://dx.doi.org/10.1016/j.meegid.2012.08.012

4. Weston JF, Heuer C, Williamson NB. Efficacy of a Neospora caninum killed tachyzoite vaccine in preventing abortion and vertical transmission in dairy cattle. Prev Vet Med 2012; 103(2-3):136-44. http://dx.doi.org/10.1016/j.prevetmed.2011.08.010

5. Quinn HE, Miller CMD, Ellis JT. The cell-mediated immune response to Neospora caninum during pregnancy in the mouse is associated with a bias towards production of interleukin-4. Int J Parasitol 2004; 34(6):723-732. http://dx.doi.org/10.1016/j.ijpara.2004.01.007

6. Ellis J, Sinclair D, Morrison D, Al-Qassab S, Springett K, Ivens A. Microarray analyses of mouse responses to infection by Neospora caninum identifies disease associated cellular pathways in the host response. Mol Biochem Parasitol 2010; 174(2):117-127.
http://dx.doi.org/10.1016/j.molbiopara.2010.08.007

7. Reid AJ, Vermont SJ, Cotton JA, Harris D, Hill-Cawthorne GA, Könen-Waisman S, Latham S. M. Comparative Genomics of the Apicomplexan Parasites Toxoplasma gondii and Neospora caninum: Coccidia Differing in Host Range and Transmission Strategy. PLoS Pathog 2012; 8(3):e1002567. http://dx.doi.org/10.1371/journal.ppat.1002567

8. Kato K, Sugi T, Iwanaga T. Roles of Apicomplexan protein kinases at each life cycle stage. Parasitol Int 2012; 61(2):224-234.
http://dx.doi.org/10.1016/j.parint.2011.12.002

9. Desrosiers MD, Cembrola KM, Fakir MJ, Stephens LA, Jama FM, Shameli A, Mehal WZ, Shi Y. Adenosine deamination sustains dendritic cell activation in inflammation. J Immunol 2007;179(3):1884-1892. http://dx.doi.org/10.4049/jimmunol.179.3.1884

10. Dalla-Rosa L, Da Silva AS, Ruchel JB, Gressler LT, Oliveira CB, França RT, Lopes STA, Leal DBR, Monteiro Silvia G. Influence of treatment with 3--deoxyadenosine associated deoxycoformycin on hematological parameters and activity of adenosine deaminase in infected mice with Trypanosoma evansi. Exp Parasitol 2013; 135(2):357-362. http://dx.doi.org/10.1016/j.exppara.2013.07.019

11. Johnson S, Nguyen V, Coder D. Assessment of cell viability. Curr Protoc Cytom 2013; 9(9.2): 10.1002/0471142956.cy0902s64.

12. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72(1):248-254.
http://dx.doi.org/10.1016/0003-2697(76)90527-3

13. Da Silva AS, Pimentel VC, Jaques JA, Wolkmer P, Tavares KC, Lazzarotto CR, Miletti LC, Schetinger MR, Mazzanti CM, Lopes ST, Monteiro SG. Biochemical detection of adenosine deaminase in Trypanosoma evansi. Exp Parasitol 2011; 128(3):298-300.
http://dx.doi.org/10.1016/j.exppara.2011.03.002

14. Giusti G, Galanti B. Methods of Enzymatic Analysis. 3rd ed. Verlag Chemie. Weinheim, Germany, 1984.

15. Asai T, Howe DK, Nakajima K, Nozaki T, Takeuchi T, Sibley LD. Neospora caninum: tachyzoites express a potent type-I nucleoside triphosphate hydrolase. Exp Parasitol 1998; 90(3):277-285.
http://dx.doi.org/10.1006/expr.1998.4346

16. Fox BA, Gigley JP, Bzik DJ. Toxoplasma gondii lacks the enzymes required for de novo arginine biosynthesis and arginine starvation triggers cyst formation. Int J Parasitol 2004; 34(3):323-31. http://dx.doi.org/10.1016/j.ijpara.2003.12.001

17. Chaudhary K, Darling JA, Fohl LM, Sullivan WJ Jr, Donald RG, Pfefferkorn ER, Ullman B, Roos DS. Purine salvage pathways in the apicomplexan parasite Toxoplasma gondii. J Biol Chem 2004; 279(30):31221-31227. http://dx.doi.org/10.1074/jbc.M404232200

18. Rottenberg ME, Masocha W, Ferella M, Petitto-Assis F, Goto H, Kristensson K, McCaffrey R, Wigzell H. Treatment of African trypanosomiasis with cordycepin and adenosine deaminase inhibitors in a mouse model. J Infect Dis 2005; 192(9):1658-1665.
http://dx.doi.org/10.1086/496896

19. Vodnala SK, Ferella M, Lundén-Miguel H, Betha E, van Reet N. Preclinical Assessment of the Treatment of Second-Stage African Trypanosomiasis with Cordycepin and Deoxycoformycin. PLOS Negl Trop Dis 2009; 3(8):e495. doi: 10.1371/journal.pntd.0000495
http://dx.doi.org/10.1371/journal.pntd.0000495

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