Alvarado IA, Sandoval TAH, de Paz VO, Barrón RBL, Hernández BE, Tenorio GVR, Aguilar SA

Language: English/Spanish
References: 26
Page: 165-175
PDF size: 260.34 Kb.

ABSTRACT

An isolated protein fraction of venom of Agkistrodon piscivorus with hemagglutinant property was evaluated in its capacity to inhibit in vitro the bovine parainfl uenza-3 (PI3) virus, either by its capacity to block cell receptors or its adherence to the hemagglutinin viral spicules. Fraction AL27 acting as a receptor block, at a concentration ≤ 1.062 μg/mL, inhibited any damage or destruction of Madin Darby Bovine Kidney (MDBK) cell cultures by the PI-3 virus (titer 10^5.6 TCID_50%), maintaining a cell viability between 69.43 and 84.86%. The adherence capacity of AL27 to viral hemagglutinin spicules of PI3 virus was determined by incubating both reactants 1:1 (1 mL AL27 at concentration ≤ 1.062 μg/mL + 1 mL PI-3 virus titer 10^5.6 DICC_50%) at 37°C for 1 hour, and a probable adherence to the viral hemagglutinin spicules was detected by electron microscope. The inactivation of PI-3 virus by AL27 was deduced by the reduction of the viral titer from 10^5.6 to 10^2.0 TCID_50%. The apparent identity between AL27 and PI-3 virus was determined by recognition of the common protein 21 kDa when performing a Western blot using hyper-immune anti-Agkistrodon piscivorus serum. This outcome shows the possibility of using AL27 fraction as an antiviral agent.

REFERENCES

1. Collins PL, Chanok RM. McIntosh: Parainfl uenza viruses. In: Fields DM. Knipe PM. Howley et al., editors. Fields Virology. 3a ed. Philadelphia: Lippincott-Raven Publishers, 1996:1177-1204.

2. Graeme LW, Bischofberger N, Webster RG. Disarming fl u viruses. Sci Am 1999:56-65.

3. Hayakawa T, Nikawa T, Kishida A, Akashi M, Baba M. Capture of HIV-1 gp120 and virions by lectin-immobilized polystyrene nanospheres: Potential approach toward prevention of HIV-1 infection. Antiviral Res 1995; 27(1-2):49-57.

4. Alvarado IA, De Paz VO, Campos LY, Ramos RL, Aguilar SA. Aislamiento de una Fracción Crotálida con Actividad Hemoaglutinante y Evaluación In Vivo e In Vitro del Probable Daño a Tejidos. Téc Pecu Méx 2003;42(2):247-260.

5. Manjunatha RK, Evans HJ. Effects of Snake Venom Proteins on Blood Platelets. Toxicon 1990;8:1387-1422.

6. Ogilvie ML, Gartner TK. Identifi cation of lectins in snake venoms. J Herpetol 1984;(18):285-290.

7. Gartner TK, Ogilvie ML. Isolation and characterization of three Ca++ dependent ß-galactoside-specifi c lectins from snake venoms. Biochem J 1984;(224):301-307.

8. Sharon N, Lis H. Lectins: cell-agglutinating and sugar specifi c proteins. Science 1972;(177):949-959.

9. Barret T, Inglis SC. Virology, a practical approach In: Mahy BWJ, editors. Practical approach series. Oxford Washington DC: IRL press, 1991:119-150.

10. Schwimmbeck PL, Lohr M, Tishon A. Viral-receptor binding assay In: Rickwood D, Hames B, editors. Animal virus Pathogenesis. A practical Approach. Oxford: IRL University press, 1990:137-148.

11. Bradford MM. A rapid and sensitive method for the quantitation of micrograms quantities of protein utilizing the principles of protein-dye binding. Annal Biochem 1976;(72):248-254.

12. Jenney EW, Wessman SJ, Spinka FL. Microtitration serology methods for bovine virology. Ames Iowa: US department of Agriculture. Animal and Plant Health Inspection Service, 1978.

13. OMS. La Rabia. Técnicas de laboratorio. 3a ed Ginebra Suiza: Organización Mundial de la Salud, 1975:348.

14. VYDAC. The handbook of analysis and purifi cation on peptides and proteins by reversed phase HPLC. 2nd ed. Hesperia CA 92345 USA: VYDAC (the separations group) 17434 Mojave Street, 1995.

15. Mancin AC, Soares AM, Andriao-Escarso SH et al. The analgesin activity of crotamine, a neurotoxin from Crotalus durissus terrifi cus (south American rattlesnake) venom: a biochemical and pharmacological study. Toxicon 1998;36:1927-1937.

16. Hayes PJ, Scoot R, Wheeler J. In vivo production of tumor necrosis factor-α and interleukin-6 in Balb/c mice inoculated intranasally with a high dose of respiratory syncytial virus. J Med Virol 1994;(42):323-329.

17. Martikainen P, Nyman K, Nevalainen TJ. Toxic effects of human pancreatic and snake and bee venom phospholipases A2 on MCF-7 cells in culture. Toxicon 1993;31:835-843.

18. Wayne WD. Bioestadística. 3a ed. México DF: Ed. Limusa, 1989.

19. Dawes CJ. Biological techniques in electron microscopy. Barnes and Noble International Textbook Series. 1973;17(46):131-150.

20. BIO-RAD. Mini-Protean II. Electrophoresis Cell. Instruction Manual. 2000 Alfred Nobel Drive Hercules, California, USA: Life Science group. No date of publication.

21. Instituto Politécnico Nacional. Manual de Prácticas de Laboratorio de Virología. Departamento de Microbiología. Instituto Nacional de Ciencias Biológicas. Instituto Politécnico Nacional ISBN 968-28-2449-9, 1989:62-77.

22. BIO-RAD. Mini-Protean II. Electrotransfer Cell. Instruction Manual. 2000 Alfred Nobel Drive Hercules, California, USA: Life Science group. Sin fecha de publicación.

23. Cotter TG, Lennon SV, Glynn JG, Martin SJ. Cell death via apoptosis and its relationship to growth, development and differentiation of both tumor and normal cells. Anticancer Res 1990;10(5A):1153-1159.

24. Francischetti IMB, Saliou B, Leduc M, Carlini CR, Hatmi M, Randon J et al. Convulxin, a potent platelet-aggregating protein from Crotalus durissus terrifi cus venom, specifi cally binds to platelets. Toxicon 1997;35(8):1217-1228.

25. Hirabayashi J, Kusunoki T, Ken-ichi K. Complete Primary Structure of a Galactose-specifi c Lectin from the Venom of the Rattlesnake Crotalus atrox. J Biol Chem 1991;286(4):2320-2326.

26. Ogilvie ML, Dockter ME, Wenz L, Gartner TK. Isolation and Characterization of Lactose-Binding Lectins from the venoms of the Snake Lachesis muta and Dendroaspis jamesonni. J Biochem 1986;(100):1425-1431.