The Effect of Medium in Properties and Activity of Acid Phosphatase (ACP) in the Infective Form of Leishmania Major Promastigotes

Document Type : Original Article (s)

Authors

1 Department of Parasitology and Mycology, Alzahra Hospital, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: The wet cutaneus leishmaniasis caused by Leishmania major infection is one of the common skin diseases in many parts of the world. Reports indicate that some properties of Leishmania, such as time of infection, is affected of medium. Respecting the use of artificial medium in the majority of the studies on Leismania, and the role of acid phosphatase (ACP) in the pathogenesis of promastigotes, in this research, we studied the effects of medium in properties of acid phosphatase and time of infecting in Leishmania major promastigotes.Methods: In this cross-sectional study, to culture promastigotes, Leishmania major (MRHO/IR/75/ER) from previously infected Balb/c mice was transferred to NNN (Novy-MacNeal-Nicolle) and RPMI1640 (Roswell Park memorial institute) medium. Growth curve was generated and stationary stages were divided. Frozen promastigotes of each medium were homogenized using sodium acetate and Triton-X-100 and acid phosphatase was measured via calorimetric assay.Findings: Stationary parasites were collected in NNN and RPMI-1640 medium in seventh and fifth day, respectively. The rate of acid phosphatase activity was determined as 1.02±0.08 in NNN and 1.8 ± 0.01 in RPMI-1640. Also, Km (Michaelis–Menten) and Vmax (Maximum velocity) of this enzyme was 105.26 ± 1.11 µM and 98.37 ± 1.48 µM/min/mg protein in NNN and 106.39 ± 1.14 µM and 98.04 ± 0.96 µM/min/mg protein in RPMI1640.Conclusion: Despite the infecting time in promastigotes, rate of agglutination, rate of kinase synthesis and virulent are different in two mediums. It seems that there are no significant differences in properties of acid phosphatase in stationary promastigotes in the two mediums.

Keywords

References

  1. do Monte-Neto RL, Coelho AC, Raymond F, Legare D, Corbeil J, Melo MN, et al. Gene expression profiling and molecular characterization of antimony resistance in Leishmania amazonensis. PLoS Negl Trop Dis 2011; 5(5): e1167.
  2. Bern C, Maguire JH, Alvar J. Complexities of assessing the disease burden attributable to leishmaniasis. PLoS Negl Trop Dis 2008; 2(10): e313.
  3. Wilson R, Bates MD, Dostalova A, Jecna L, Dillon RJ, Volf P, et al. Stage-specific adhesion of Leishmania promastigotes to sand fly midguts assessed using an improved comparative binding assay. PLoS Negl Trop Dis 2010; 4(9).
  4. Rogers ME, Chance ML, Bates PA. The role of promastigote secretory gel in the origin and transmission of the infective stage of Leishmania mexicana by the sandfly Lutzomyia longipalpis. Parasitology 2002; 124(Pt 5): 495-507.
  5. Bates PA, Hermes I, Dwyer DM. Leishmania donovani: immunochemical localization and secretory mechanism of soluble acid phosphatase. Exp Parasitol 1989; 68(3): 335-46.
  6. Fernandes AC, Soares DC, Saraiva EM, Meyer-Fernandes JR, Souto-Padron T. Different secreted phosphatase activities in Leishmania amazonensis. FEMS Microbiol Lett 2013; 340(2): 117-28.
  7. Dutra PM, Dias FA, Rodrigues CO, Romeiro A, Attias M, De SW, et al. Platelet-activating factor modulates a secreted phosphatase activity of the trypanosomatid parasite Herpetomonas muscarum muscarum. Curr Microbiol 2001; 43(4): 288-92.
  8. Baghaei M, Mesripour M. Characterization of acid phosphatase in the promastigotes of three isolates of leishmania major . Iran J Med Sci 2003; 28(1): 1-8.
  9. el-On J, Sneier R, Elias E. Leishmania major: bacterial contamination of cutaneous lesions in experimental animals. Isr J Med Sci 1992; 28(12): 847-51.
  10. Soleimanifard S, Arjmand R, Hejazi SH. Investigation and comparison of Leishmania major promastigote and amastigote protein content by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Sci J Hamadan Univ Med Sci 2013; 20 (1): 1-8. [In Persian].
  11. Dey T, Afrin F, Anam K, Ali N. Infectivity and virulence of Leishmania donovani promastigotes: a role for media, source, and strain of parasite. J Eukaryot Microbiol 2002; 49(4): 270-4.
  12. Sarkari B, Chance M, Hommel M. Antigenuria in visceral leishmaniasis: detection and partial characterisation of a carbohydrate antigen. Acta Trop 2002; 82(3): 339-48.
  13. Rezazadeh MF, Shakeri R, Kaboudanian AS, Tahghighi A, Foroumadi A. In vitro immunobiological studies of novel 5-(5-nitrofuran-2-yl)-1, 3, 4-thiadiazoles with piperazinyl-linked benzamidine substituents against Leishmania major. Iran J Allergy Asthma Immunol 2013; 12(4): 368-76.
  14. Sant'anna MR, Nascimento A, Alexander B, Dilger E, Cavalcante RR, Diaz-Albiter HM, et al. Chicken blood provides a suitable meal for the sand fly Lutzomyia longipalpis and does not inhibit Leishmania development in the gut. Parasit Vectors 2010; 3(1): 3.
  15. Schlein Y, Jacobson RL. Haemoglobin inhibits the development of infective promastigotes and chitinase secretion in Leishmania major cultures. Parasitology 1994; 109 (Pt 1): 23-8.
  16. Sacks DL, Modi G, Rowton E, Spath G, Epstein L, Turco SJ, et al. The role of phosphoglycans in Leishmania-sand fly interactions. Proc Natl Acad Sci U S A 2000; 97(1): 406-11.
  17. Kbaier-Hachemi H, Guerbouj S, Turki-Mannoubi L, Kaabi B, Guizani I. In vitro growth kinetics, differentiation and morphological characterisation of Tunisian Leishmania infantum parasites. Trans R Soc Trop Med Hyg 2012; 106(1): 20-5.
  18. Sysoev VV. A comparison of the growth of Leishmania major, L. turanica and L. gerbilli on NNN medium. Med Parazitol (Mosk) 1995; (1): 13-7. [In Russian].
Journal of Isfahan Medical School
Volume 32, Issue 295 - Serial Number 295
May and June 2014
Pages 1166-1174

Files

History

  • Receive Date: 29 October 2013
  • Revise Date: 01 May 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics