Volume 7, Issue 3 (7-2019)                   JoMMID 2019, 7(3): 72-78 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Kazemirad E, Reisi Nafchi H, Latifi A, Raoofian R, Mohebali M, Hajjaran H. Comparison of Cysteine Protease B Gene Expression between Clinical Isolates of Leishmania tropica, Leishmania major and Leishmania infantum. JoMMID 2019; 7 (3) :72-78
URL: http://jommid.pasteur.ac.ir/article-1-221-en.html
Comparison of Cysteine Protease B Gene Expression between Clinical Isolates of Leishmania tropica, Leishmania major and Leishmania infantum
Elham Kazemirad , Hossien Reisi Nafchi , Alireza Latifi , Reza Raoofian , Mehdi Mohebali , Homa Hajjaran *
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Abstract:   (5032 Views)
Introduction: Leishmania cysteine protease B (CPB) is a parasite virulence factor that plays a vital role in host-parasitic interactions. Regarding the importance of the CPB gene, we used a quantitative real-time RT-PCR to investigate the expression of CPB in the Iranian isolates of different Leishmania species. Methods: In this study, 36 clinical samples were collected, out of which 29 belonged to cutaneous leishmaniasis (CL), 3 to viscerotropic leishmaniasis (VTL), and 5 to visceral leishmaniasis (VL) patients. Among CL isolates, 8 were Leishmania major, and 21 were Leishmania tropica, including 3 isolates from the lupoid type. After RNA extraction and cDNA synthesis, gene expression analysis was performed by qPCR. Results: Our results showed the highest expression of CPB in isolates of Leishmania infantum, followed by L. major and L. tropica. Among L. tropica isolates, in the lupoid forms, the mean expression of CPB was ≈6.4 times higher than that of non-lupoid isolates. In L. major isolates, a significant difference was observed between the level of gene expression and the duration of the infection. The expression level of CPB was associated with the severity of the infection in L. infantum isolates. Conclusion: The CPB gene expressed in all Leishmania species. The highest expression was in L. infantum species and the least expression in L. tropica. The transcript level of CPB increased in L. major isolates derived from patients with a higher number and duration of ulcers; however, further studies on more clinical samples are needed to explore our findings. 
Keywords: Cysteine protease B (CPB), Real-time RT- PCR, Leishmania species, Iran
Full-Text [PDF 602 kb]   (778 Downloads)    
Type of Study: Original article | Subject: Microbial pathogenesis
Received: 2019/10/5 | Accepted: 2019/12/17 | Published: 2020/03/9
References
1. Murray HW, Berman JD, Davies CR, Saravia NG. Advances in leishmaniasis. Lancet 2005; 366 (9496): 1561-77. [DOI:10.1016/S0140-6736(05)67629-5]
2. Magill AJ, Grögl M, Gasser RA Jr, Sun W, Oster CN. Visceral infection caused by Leishmania tropica in veterans of Operation Desert Storm. N Engl J Med. 1993; 328 (19): 1383-7. [DOI:10.1056/NEJM199305133281904]
3. Hajjaran H, Mohebali M, Teimouri A, Oshaghi MA, Mirjalali H, Kazemi-Rad E, et al. Identification and phylogenetic relationship of Iranian Strains of various Leishmania species isolated from cutaneous and visceral cases of Leishmaniasis based on N acetyl glucosamine-1-phosphate transferase gene. Infect Genet Evol. 2014; 26, 203-212. [DOI:10.1016/j.meegid.2014.05.026]
4. Matlashewski G. Leishmania infection and virulence. Med Microbial Immunol. 2001; 190 (1-2): 37-42. [DOI:10.1007/s004300100076]
5. Silva-Almeida M, Pereira BA, Ribeiro-Guimarães ML, Alves CR. Proteinases as virulence factors in Leishmania spp. infection in mammals. Parasit Vectors. 2012; 7; 5: 160. [DOI:10.1186/1756-3305-5-160]
6. Verma S, Dixit R, Pandey KC. Cysteine Proteases: Modes of Activation and Future Prospects as Pharmacological Targets. Front Pharmacol. 2016; 7: 107. [DOI:10.3389/fphar.2016.00107]
7. Klemba M, Goldberg DE. Biological roles of proteases in parasitic protozoa. Annu Rev Biochem. 2002; 71: 275-305. [DOI:10.1146/annurev.biochem.71.090501.145453]
8. Sajid M, McKerrow JH. Cysteine proteases of parasitic organisms. Mol Biochem Parasitol. 2002; 120: 1-21. [DOI:10.1016/S0166-6851(01)00438-8]
9. Rafati S, Fasel N, Masina S. Leishmania cysteine proteinases: from gene to subunit vaccine. Curr Genom. 2003; 4, 109-121. [DOI:10.2174/1389202033490439]
10. Mottram JC, Brooks DR, Coombs GH. Roles of cysteine proteinases of trypanosomes and Leishmania in host-parasite interactions. Curr Opin Microbiol. 1998; 1 (4): 455-60. [DOI:10.1016/S1369-5274(98)80065-9]
11. Poot H, Denise DC, Herrmann JC, Mottram GH, Coombs AN. Vermeulen, Virulence and protective potential of several Cysteine peptidase knockout strains of Leishmania infantum in hamsters, in: Poot J. Utrecht (Ed.), Experimental Challenge Models for Canine Leishmaniasis in Hamsters and Dogs, Optimization and Application in Vaccine Research, vol. 2006, Utrecht University press, Netherlands, 2006; 93e107.
12. Bañuls AL, Hide M, Prugnolle F. Leishmania and the leishmaniases: a parasite genetic update and advances in taxonomy, epidemiology and pathogenicity in humans. Adv Parasitol. 2007; 64: 1-109. [DOI:10.1016/S0065-308X(06)64001-3]
13. Khamesipour A, Rafati S, Davoudi N, Maboudi F, Modabber F. Leishmaniasis vaccine candidates for development: a global overview. Indian J Med Res. 2006; 123: 423-38.
14. Rafati S, Kariminia A, Seye Eslami S, Narimani M, Taheri T, Lebbatard M. Recombinant cysteine proteinase-based vaccine against Leishmania major in BALB/c: the role partial protection relies on interferon gamma CD8+ T lymphocyte activation. Vaccine. 2002; 20: 2439-47. [DOI:10.1016/S0264-410X(02)00189-5]
15. Rafati S, Salmanian AH, Taheri T, Vafa M, Fasel N. A protective cocktail vaccine against murine cutaneous leishmaniasis with DNA encoding cysteine proteinases of Leishmania major. Vaccine. 2001; 14; 19 (25-26): 3369-75. [DOI:10.1016/S0264-410X(01)00081-0]
16. Rafati S, Nakhaee A, Tahere T, Ghashghaii A, Salmanian AH, Jimenez M, et al. Expression of cysteine proteinase type I and II of Leishmania infantum and their recognition by sera during canine and human visceral leishmaniasis. Exp Parasitol. 2003; 103: 143-51. [DOI:10.1016/S0014-4894(03)00097-3]
17. Nakhaee A, Taheri T, Taghikhani M, Mohebali M, Salmanian AH, Fasel N, et al. Humoral and cellular immune responses against Type I cysteine proteinase of Leishmania infantum are higher in asymptomatic than symptomatic dogs selected from a naturally infected population. Vet Parasitol. 2004; 119: 107-123. [DOI:10.1016/j.vetpar.2003.11.013]
18. Coombs GH. Proteinases of Leishmania mexicana and other flagellate protozoa. Parasitology. 1982; 84: 149-155. [DOI:10.1017/S003118200005174X]
19. Hajjaran H, Mohebali M, Mamishi S, Vasigheh F, Oshaghi MA, Naddaf SR, et al. Molecular identification and polymorphism determination of cutaneous and visceral leishmaniasis agents isolated from human and animal hosts in Iran. Biomed Res Int. 2013; 2013: 789326. [DOI:10.1155/2013/789326]
20. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, Hajjaran H, Hadighi R, Khamesipour A, et al. Overexpression of ubiquitin and amino acid permease genes in association with antimony resistance in Leishmania tropica field isolates. Korean J Parasitol. 2013; 51: 413-419. [DOI:10.3347/kjp.2013.51.4.413]
21. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res. 2002; 1; 30 (9): e36. [DOI:10.1093/nar/30.9.e36]
22. Kramer S. "Developmental regulation of gene expression in the absence of transcriptional control: the case of kinetoplastids." Mol Biochem Parasitol. 2012; 181 (2): 61-72. [DOI:10.1016/j.molbiopara.2011.10.002]
23. Nafchi HR, Kazemi-Rad E, Mohebali M, Raoofian R, Ahmadpour NB, Oshaghi MA, et al. Expression analysis of viscerotropic leishmaniasis gene in Leishmania species by real-time RT-PCR. Acta Parasitol. 2016; 61 (1): 93-7. [DOI:10.1515/ap-2016-0011]
24. Silva-Almeida M, Pereira BA, Ribeiro-Guimarães ML, Alves CR. Proteinases as virulence factors in Leishmania spp. infection in mammals. Parasit Vectors. 2012; 5 (1): 1-10. [DOI:10.1186/1756-3305-5-160]
25. Pereira BAS, Britto C, Alves CR. Expression of infection-related genes in parasites and host during murine experimental infection with Leishmania Leishmania amazonensis. Microb Pathog. 2012; 52 (2): 101-8. [DOI:10.1016/j.micpath.2011.11.004]
26. Mottram JC, Coombs GH, Alexander J. Cysteine peptidases as virulence factors of Leishmania. Curr Opin Microbiol. 2004; 7 (4): 357-81. [DOI:10.1016/j.mib.2004.06.010]
27. Kariyawasam R, Mukkala AN, Lau R, Valencia BM, Llanos-Cuentas A, Boggild AK. Virulence factor RNA transcript expression in the Leishmania Viannia subgenus: influence of species, isolate source, and Leishmania RNA virus-1. Trop Med Health. 2019; 11; 47:25. [DOI:10.1186/s41182-019-0153-x]
28. Ronet C, Beverley SM, Fasel N. Muco-cutaneous leishmaniasis in the New World. Landes Biosci. 2011; 2 (6): 547-52. [DOI:10.4161/viru.2.6.17839]
29. de Araújo Soares RM, dos Santos AL, Bonaldo MC, de Andrade AF, Alviano CS, Angluster J, et al. Leishmania (Leishmania) amazonensis: differential expression of proteinases and cell-surface polypeptides in avirulent and virulent promastigotes. Exp Parasitol. 2003; 104 (3-4): 104-112. [DOI:10.1016/S0014-4894(03)00135-8]
30. Pereira BAS, Britto C, Alves CR. Expression of infection-related genes in parasites and host during murine experimental infection with Leishmania amazonensis. Microb Pathog. 2012; 52 (2): 101-8. [DOI:10.1016/j.micpath.2011.11.004]
31. Aronson N, Herwaldt BL, Libman M, Pearson R, Lopez-Velez R, Weina P, et al. Diagnosis and treatment of Leishmaniasis: clinical practice guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH). Clin Infect Dis. 2016; 63 (12): e202-64. [DOI:10.1093/cid/ciw670]
32. Convit J, Ulrich M, Pérez M, Hung J, Castillo J, Rojas H, et al. Atypical cutaneous leishmaniasis in Central America: possible interaction between infectious and environmental elements. Trans R Soc Trop Med Hyg. 2005; 99 (1): 13-17. [DOI:10.1016/j.trstmh.2004.02.005]
33. Schönian G, Akuffo H, Lewin S, Maasho K, Nylén S, Pratlong F, et al. Genetic variability within the species Leishmania aethiopica does not correlate with clinical variations of cutaneous leishmaniasis. Mol Biochem Parasitol. 2000; 106 (2): 239-248. [DOI:10.1016/S0166-6851(99)00216-9]
34. Oryan A, Shirian S, Tabandeh M-R, Hatam G-R, Randau G, Daneshbod Y. Genetic diversity of Leishmania major strains isolated from different clinical forms of cutaneous leishmaniasis in southern Iran based on minicircle kDNA. Infect Genet Evol. 2013; 19: 226-231. [DOI:10.1016/j.meegid.2013.07.021]
35. Hide M, Bañuls AL. Polymorphisms of cpb multicopy genes in the Leishmania donovani complex. Trans R Soc Trop Med Hyg. 2008, 102 (2): 105-106. [DOI:10.1016/j.trstmh.2007.09.013]
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.