سامانه مدیریت نشریات علمی دانشگاه علوم پزشکی مشهد

Aghaei, Maryam, Khanahmad, Hossein, Jalali, Akram, Aghaei, Shahrzad, Narimani, Manizheh, Hosseini, Sayed Mohsen, Namdar, Fatemeh, Hejazi, Hossein. (1400). Effect of transgenic Leishmania major expressing mLLO-Bax-Smac fusion gene in the apoptosis of the infected macrophages. , 24(12), 1666-1675. doi: 10.22038/ijbms.2021.56960.12701
Maryam Aghaei; Hossein Khanahmad; Akram Jalali; Shahrzad Aghaei; Manizheh Narimani; Sayed Mohsen Hosseini; Fatemeh Namdar; Hossein Hejazi. "Effect of transgenic Leishmania major expressing mLLO-Bax-Smac fusion gene in the apoptosis of the infected macrophages". , 24, 12, 1400, 1666-1675. doi: 10.22038/ijbms.2021.56960.12701
Aghaei, Maryam, Khanahmad, Hossein, Jalali, Akram, Aghaei, Shahrzad, Narimani, Manizheh, Hosseini, Sayed Mohsen, Namdar, Fatemeh, Hejazi, Hossein. (1400). 'Effect of transgenic Leishmania major expressing mLLO-Bax-Smac fusion gene in the apoptosis of the infected macrophages', , 24(12), pp. 1666-1675. doi: 10.22038/ijbms.2021.56960.12701
Aghaei, Maryam, Khanahmad, Hossein, Jalali, Akram, Aghaei, Shahrzad, Narimani, Manizheh, Hosseini, Sayed Mohsen, Namdar, Fatemeh, Hejazi, Hossein. Effect of transgenic Leishmania major expressing mLLO-Bax-Smac fusion gene in the apoptosis of the infected macrophages. , 1400; 24(12): 1666-1675. doi: 10.22038/ijbms.2021.56960.12701

Effect of transgenic Leishmania major expressing mLLO-Bax-Smac fusion gene in the apoptosis of the infected macrophages

Iranian Journal of Basic Medical Sciences
مقاله 7، دوره 24، شماره 12، اسفند 2021، صفحه 1666-1675    اصل مقاله (1.03 M)
نوع مقاله: Original Article
شناسه دیجیتال (DOI): 10.22038/ijbms.2021.56960.12701
نویسندگان
Maryam Aghaei1؛ Hossein Khanahmad2؛ Akram Jalali3؛ Shahrzad Aghaei4؛ Manizheh Narimani5؛ Sayed Mohsen Hosseini6؛ Fatemeh Namdar5؛ Hossein Hejazi* 1، 7
1Skin Diseases and Leishmaniasis Research Centre, Isfahan University of Medical Sciences, Isfahan, Iran
2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
4Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
5Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
6Department of Biostatistics & Epidemiology, School of Public Health, Isfahan University of Medical Sciences, Isfahan, Iran
7Skin Diseases and Leishmaniasis Research Center, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده
Objective(s): Leishmaniasis is a complex infection against which no confirmed vaccine has been reported so far. Transgenic expression of proteins involved in macrophage apoptosis-like BAX through the parasite itself accelerates infected macrophage apoptosis and prevents Leishmania differentiation. So, in the present research, the impact of the transgenic Leishmania major including mLLO-BAX-SMAC proapoptotic proteins was assayed in macrophage apoptosis acceleration. 
Materials and Methods: The coding sequence mLLO-Bax-Smac was designed and integrated into the pLexyNeo2 plasmid. The designed sequence was inserted under the 18srRNA locus into the L. major genome using homologous recombination. Then, mLLO-BAX-SMAC expression was studied using the Western blot, and the transgenic parasite pathogenesis was investigated compared with wild-type L. major in vitro and also in vivo. 
Results: Western blot and PCR results approved mLLO-BAX-SMAC expression and proper integration of the mLLO-Bax-Smac fragment under the 18srRNA locus of L. major, respectively. The flow cytometry results revealed faster apoptosis of transgenic Leishmania-infected macrophages compared with wild-type parasite-infected macrophages. Also, the mild lesion with the less parasitic burden of the spleen was observed only in transgenic Leishmania-infected mice. The delayed progression of leishmaniasis was obtained in transgenic strain-injected mice after challenging with wild-type Leishmania. 
Conclusion: This study recommended transgenic L. major including mLLO-BAX-SMAC construct as a pilot model for providing a protective vaccine against leishmaniasis. 
کلیدواژه‌ها
Homologous recombination؛ Integration؛ Leishmaniasis؛ Transfection؛ Vaccine
مراجع
1. Doudi M, Hejazi SH, Razavi MR, Narimani M, Khanjani S, Eslami G. Comparative molecular epidemiology of Leishmania major and Leishmania tropica by PCR-RFLP technique in hyper endemic cities of Isfahan and Bam, Iran. Med Sci Monit 2010;1:530-535.
2. Georgiadou SP, Makaritsis KP, Dalekos GN. Leishmaniasis revisited: current aspects on epidemiology, diagnosis and treatment. J Transl Int Med 2015;3:43-50.
3. Dubie T, Mohammed Y. Review on the role of host immune response in protection and immunopathogenesis during cutaneous leishmaniasis infection. J Immunol Res 2020;2020: 2496713.
4. Samarasinghe SR, Samaranayake N, Kariyawasam UL, Siriwardana YD, Imamura H, Karunaweera ND. Genomic insights into virulence mechanisms of Leishmania donovani: evidence from an atypical strain. BMC Genomics 2018;19:1-18.
5. Ivens AC, Peacock CS, Worthey EA, Murphy L, Aggarwal G, Berriman M, et al. The genome of the kinetoplastid parasite, Leishmania major. Science. 2005;309:436-442.
6. Sacks D, Kamhawi S. Molecular aspects of parasite-vector and vector-host interactions in leishmaniasis. Annu Rev  Microbiol 2001;55:453-483.
7. Assreuy J, Cunha FQ, Epperlein M, Noronha‐Dutra A, O’Donnell CA, Liew FY, et al. Production of nitric oxide and superoxide by activated macrophages and killing of Leishmania major. Eur J Immunol 1994;24:672-676.
8. Alexander J, Russell DG. The interaction of Leishmania species with macrophages.  Adv Parasitol 1992;31:175-254.
9. Chow SH, Deo P, Naderer T. Macrophage cell death in microbial infections. Cell Microbiol 2016;18:466-474.
10. Filardy AA, Costa-da-Silva AC, Koeller CM, Guimarães-Pinto K, Ribeiro-Gomes FL, Lopes MF, et al. Infection with Leishmania major induces a cellular stress response in macrophages. PLoS One. 2014;9:e85715.
11. Conceição‐Silva F, Hahne M, Schröter M, Louis J, Tschopp J. The resolution of lesions induced by Leishmania major in mice requires a functional Fas (APO‐1, CD 95) pathway of cytotoxicity. Eur J Immunol 1998;28:237-245.
12. Chakour R, Guler R, Bugnon M, Allenbach C, Garcia I, Mauël J, et al. Both the Fas ligand and inducible nitric oxide synthase are needed for control of parasite replication within lesions in mice infected with Leishmania major whereas the contribution of tumor necrosis factor is minimal. Infect Immun 2003;71:5287-5295.
13. Kamir D, Zierow S, Leng L, Cho Y, Diaz Y, Griffith J, et al. A Leishmania ortholog of macrophage migration inhibitory factor modulates host macrophage responses. J Immunol 2008;180:8250-8261.
14. Akarid K, Arnoult D, Micic‐Polianski J, Sif J, Estaquier J, Ameisen JC. Leishmania major‐mediated prevention of programmed cell death induction in infected macrophages is associated with the repression of mitochondrial release of cytochrome c. J Leukoc Biol 2004;76:95-103.
15. Oltval ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. cell 1993;74:609-619.
16. Apte SS, Mattei M-G, Olsen BR. Mapping of the human BAX gene to chromosome 19q13. 3–q13. 4 and isolation of a novel alternatively spliced transcript, BAXδ. Genomics 1995;26:592-594.
17. Huret J-L, Ahmad M, Arsaban M, Bernheim A, Cigna J, Desangles F, et al. Atlas of genetics and cytogenetics in oncology and haematology in 2013. Nucleic Acids Res 2012;41:920-924.
18. Weng C, Li Y, Xu D, Shi Y, Tang H. Specific cleavage of Mcl-1 by caspase-3 in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in Jurkat leukemia T cells. J Biol Chem2005;280:10491-10500.
19. Cartron P-F, Oliver L, Martin S, Moreau C, LeCabellec M-T, Jezequel P, et al. The expression of a new variant of the pro-apoptotic molecule Bax, Baxψ, is correlated with an increased survival of glioblastoma multiforme patients. Hum Mol Genet 2002;11:675-687.
20. McNeish I, Bell S, McKay T, Tenev T, Marani M, Lemoine N. Expression of Smac/DIABLO in ovarian carcinoma cells induces apoptosis via a caspase-9-mediated pathway. Exp Cell Res 2003;286:186-198.
21. Moshrefi M, Spotin A, Kafil HS, Mahami-Oskouei M, Baradaran B, Ahmadpour E, et al. Tumor suppressor p53 induces apoptosis of host lymphocytes experimentally infected by Leishmania major, by activation of Bax and caspase-3: A possible survival mechanism for the parasite. Parasitol Res 2017;116:2159-2166.
22. Cianciulli A, Porro C, Calvello R, Trotta T, Panaro MA. Resistance to apoptosis in Leishmania infantum-infected human macrophages: a critical role for anti-apoptotic Bcl-2 protein and cellular IAP1/2. Clin Exp Med 2018;18:251-261.
23. Lasjerdi Z, Ghanbarian H, Yeganeh SM, Tabaei SJS, Mohebali M, Taghipour N, et al. Comparative Expression Profile Analysis of Apoptosis-Related miRNA and Its Target Gene in Leishmania major Infected Macrophages. Iran J Parasitol 2020;15:332.
24. Dorlo TP, Balasegaram M, Beijnen JH, de Vries PJ. Miltefosine: a review of its pharmacology and therapeutic efficacy in the treatment of leishmaniasis. J Antimicrob Chemother 2012;67:2576-2597.
25. Gholamrezaei M, Rouhani S, Mohebali M, Mohammadi-Yeganeh S, Hoseini MHM, Haghighi A, et al. MicroRNAs expression induces apoptosis of macrophages in response to Leishmania major (MRHO/IR/75/ER): An in-vitro and in-vivo study. Iran J Parasitol 2020;15:475.
26. Beattie L, Evans K, Kaye P, Smith D. Transgenic Leishmania and the immune response to infection. Parasite Immunol 2008;30:255-266.
27. Aghaei M, Khanahmad H, Aghaei S, Hosseini SM, Farahmand M, Hejazi SH. Evaluation of transgenic Leishmania infantum expressing mLLO‐BAX‐SMAC in the apoptosis of the infected macrophages in vitro and in vivo. Parasite Immunol 2020;42:e12726.
28. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: Cold spring harbor laboratory pres. 1989.
29. Maniatis T, Fritsch Et. andJ. Sambrook. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY; 1982.
30. Field AE, Wagage S, Conrad SM, Mosser DM. Reduced pathology following infection with transgenic Leishmania major expressing murine CD40 ligand. Infect Immun 2007;75:3140-3149.
31. Cuervo P, De Jesus JB, Saboia-Vahia L, Mendonça-Lima L, Domont GB, Cupolillo E. Proteomic characterization of the released/secreted proteins of Leishmania (Viannia) braziliensis promastigotes. J Proteomics 2009;73:79-92.
32. Sambrook J, Fritsch E, Maniatis T. Molecular cloning a laboratory manual second edition vols. 1 2 and 3. molecular cloning: a laboratory manual, second edition, vols. 1989;1(2).
33. Datta AR, Wentz BA, Russell J. Cloning of the listeriolysin O gene and development of specific gene probes for Listeria monocytogenes. Appl Environ Microbiol 1990;56:3874-3877.
34. Singh N, Gupta R, Jaiswal AK, Sundar S, Dube A. Transgenic Leishmania donovani clinical isolates expressing green fluorescent protein constitutively for rapid and reliable ex vivo drug screening. J Antimicrob Chemother 2009;64:370-374.
35. Rieger AM, Nelson KL, Konowalchuk JD, Barreda DR. Modified annexin V/propidium iodide apoptosis assay for accurate assessment of cell death. J Vis Exp 2011:50;2597.
36. Rabhi I, Rabhi S, Ben-Othman R, Rasche A, Daskalaki A, Trentin B, et al. Transcriptomic signature of Leishmania infected mice macrophages: a metabolic point of view. PLoS Negl Trop Dis 2012;6:1763.
37. Melby PC, Yang Y-Z, Cheng J, Zhao W. Regional differences in the cellular immune response to experimental cutaneous or visceral infection with Leishmania donovani. Infect Immun 1998;66:18-27.
38. Saljoughian N, Taheri T, Zahedifard F, Taslimi Y, Doustdari F, Bolhassani A, et al. Development of novel prime-boost strategies based on a tri-gene fusion recombinant L. tarentolae vaccine against experimental murine visceral leishmaniasis. PLoS Negl Trop Dis 2013;7:2174.
39. Hara H, Kawamura I, Nomura T, Tominaga T, Tsuchiya K, Mitsuyama M. Cytolysin-dependent escape of the bacterium from the phagosome is required but not sufficient for induction of the Th1 immune response against Listeria monocytogenes infection: distinct role of listeriolysin O determined by cytolysin gene replacement. Infect Immun 2007;75:3791-3801.
40. Grode L, Seiler P, Baumann S, Hess J, Brinkmann V, Eddine AN, et al. Increased vaccine efficacy against tuberculosis of recombinant Mycobacterium bovis bacille Calmette-Guerin mutants that secrete listeriolysin. J Clin Invest 2005;115:2472-2479.
41. Yadava A, Ockenhouse CF. Effect of codon optimization on expression levels of a functionally folded malaria vaccine candidate in prokaryotic and eukaryotic expression systems. Infect Immun 2003;71:4961-4969.
42. Kianmehr A, Golavar R, Rouintan M, Mahrooz A, Fard-Esfahani P, Oladnabi M, et al. Cloning and expression of codon-optimized recombinant darbepoetin alfa in Leishmania tarentolae T7-TR. Protein exp Purif 2016;118:120-125.
43. Glomski IJ, Gedde MM, Tsang AW, Swanson JA, Portnoy DA. The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells. J Cell Biol 2002;156:1029-1038.
44. Esseiva AC, Chanez AL, Bochud‐Allemann N, Martinou JC, Hemphill A, Schneider A. Temporal dissection of Bax‐induced events leading to fission of the single mitochondrion in Trypanosoma brucei. EMBO Rep 2004;5:268-273.
45. Oliveira SC, Splitter GA. CD8+ type 1 CD44hi CD45 RBlo T lymphocytes control intracellular Brucella abortus infection as demonstrated in major histocompatibility complex class I‐and class II‐deficient mice. Eur J Immunol 1995;25:2551-2557.
46. Conrad SM, Strauss-Ayali D, Field AE, Mack M, Mosser DM. Leishmania-derived murine monocyte chemoattractant protein 1 enhances the recruitment of a restrictive population of CC chemokine receptor 2-positive macrophages. Infect Immun 2007;75:653-665.
47. Dousti A, Davoudi N. Vaccination of C57BL/6 mice with transgenic Leishmania Major. 2008.

آمار
تعداد مشاهده مقاله: 746
تعداد دریافت فایل اصل مقاله: 486