Original Article

Synergistic Anti-Leishmanial Activities of Morphine and Imiquimod on Leishmania infantum (MCAN/ES/98/LIM-877)

Abstract

Background: This study was performed to evaluate in vitro and in vivo Leishmanicidal potential of morphine (Mph), imiquimod (IQ), and their combination.Methods: Leishmania infantum promastigote and amastigote assays were performed at the presence of 0.015–150µM Mph, 0.04–416µM IQ, and their combination. The inhibition effects of these drugs on promastigotes were evaluated after 24, 48, and 72h. The cytotoxic effects of the drugs were evaluated by MTT as well as flow cytometry after 72h. We explored the therapeutic effects of Mph and IQ in BALB/c mice at the end of the treatment using parasite load de­termination and cytokine assay. One group of mice received Mph for three weeks before infection.Results: The results of promastigote and amastigote assays showed the cytotoxic effects of the drugs at low concentra­tions. The cytotoxic effects were higher on promastigotes than amastigotes (p< 0.05). There was a negative correlation between drug concentration and amastigote/promastigote viability. Imiquimod alone or combined with Mph showed remarkable cytotoxic effects at all concentrations (p< 0.05). Flow cytometry results revealed apoptosis in the parasite following exposure to the drug combinations. Accordingly, the reduction of parasite loads in the spleen and liver was observed (p< 0.05) with simultaneous increases in IFN-γ and IL-4. We believe that the in vivo leishmanicidal effect was mediated by Mph through IL-4 and by IQ through both IL-4 and IFN-γ.Conclusion: Results pointed out the promising effects of Mph and IQ at low concentrations, especially when combined.
1. Goto H, Lindoso JAL (2010) Current diagno¬-sis and treatment of cutaneous and mu-cocutaneous leishmaniasis. Expert Rev Anti Infect Ther. 8(4): 419–433.
2. Dantas-Torres F (2010) Review of" Hu-man-Animal Medicine-Clinical Approach¬es to Zoonoses and Other Shared Health Risks" by Peter M. Rabinowitz and Lisa A. Conti (eds.). Parasite Vectors. 3(1): 20.
3. Mohebali M (2013) Visceral leishmaniasis in Iran: Review of the Epidemiologi¬cal and Clinical Features. Iran J Par¬a¬sitol. 8 (3): 348–358.
4. Mohebali M, Moradi-Asl E, Rassi Y (2018) Geographic distribution and spatial anal¬ysis of Leishmania infantum infection in domestic and wild animal reservoir hosts of zoonotic visceral leishmania¬sis in Iran: A systematic review. J Vector Borne Dis. 55(3): 173–180.
5. Barrett MP, Croft SL (2012) Management of trypanosomiasis and leishmaniasis. Br Med Bull. 104(1): 175–196.
6. Moradi-Asl E, Rassi Y, Adham D, Hanafi-Bojd AA, Saghafipour A, Rafizadeh S (2018) Spatial distribution of sand flies (Diptera: Psychodidae; Larroussius group), the vectors of visceral leish¬man¬iasis in Northwest of Iran. Asian Pac J Trop Bi-omed. 8(9): 425–430.
7. Cianciulli A, Porro C, Calvello R, Trotta T, Panaro MA (2018) Resistance to apop-tosis in Leishmania infantum-infected hu¬man macrophages: a critical role for anti-apoptotic Bcl-2 protein and cellu¬lar IAP1/ 2. Int J Clin Exp Med. 18(2): 251–261.
8. Gupta P, Srivastav S, Saha S, Das PK, Ukil A (2016) Leishmania donovani inhib¬its macrophage apoptosis and pro-in¬flam-matory response through AKT-me-diated regulation of β-catenin and FOXO-1. Cell Death Dis. 23(11): 1815–1826.
9. Donovan MJ, Maciuba BZ, Mahan CE, McDowell MA (2009) Leishmania infection inhibits cyclo¬heximide-in¬duced mac¬rophage apoptosis in a strain-de¬pend¬ent manner. Exp Parasitol. 12 (1): 58–64.
10. Ouellette M, Drummelsmith J, Papado-pou¬lou B (2004) Leishmaniasis: drugs in the clinic, resistance and new devel-op¬ments. Drug Resist Updat. 7(4–5): 257–266.
11. Moreira W, Leprohon P, Ouellette M (2011) Tolerance to drug-induced cell death fa¬vours the acquisition of multidrug re¬sistance in Leishmania. Cell Death Dis. 2(9): e201.
12. Doroodgar M, Delavari M, Doroodgar M, Abbasi A, Taherian AA, Doroodgar A (2016) Tamoxifen induces apoptosis of Leishmania major promastigotes in vitro. Korean J Parasitol. 54(1): 9–14.
13. Erfan MB, Mohebali M, Kazemi-Rad E, Hajjaran H, Edrissian G, Mamishi S, Saffar M, Raoofian R, Heidari M (2013) Downregulation of calcineurin gene is associated with Glucantime® resiatance in Leishmania infantum. Iran J Parasi-tol. 8(3): 359–366.
14. Alavi NR, Fazaeli A, Pejman B, Ansari H, Fouladi B, Khamesipour A (2009) The efficacy of morphine on murine (BALB/ c) cutaneous leishmaniasis. Iran J In¬fect Dis Trop Med. 14(46): 15–22.
15. Tubaro E, Borelli G, Croce C, Cavallo G, Santiangeli C (1983) Effect of mor-phine on resistance to infection. Int J Infect Dis. 148(4): 656–666.
16. Singhal PC, Bhaskaran M, Patel J, Patel K, Kasinath BS, Duraisamy S, Franki N, Reddy K, Kapasi AA (2002) Role of p 38 mitogen-activated protein ki¬nase phos¬phorylation and Fas-Fas lig¬and in-ter¬ac¬tion in morphine-induced macro-phage apopto¬sis. J Immunol. 168(8): 4025–4033.
17. Malik AA, Radhakrishnan N, Reddy K, Smith AD, Singhal PC (2002) Mor-phine-induced macrophage apoptosis modu¬lates migration of macrophages: use of in vitro model of urinary tract infection. J En¬dourol. 16(8): 605–610.
18. Hemmi H, Kaisho T, Takeuchi O, Sato S, Sanjo H, Hoshino K, Horiuchi T, Tomi-zawa H, Takeda K, Akira S (2002) Small anti-viral compounds activate immune cells via the TLR7 MyD88–dependent signaling pathway. Nat Immunol. 3(2): 196–200.
19. Kim SJ, Park GH, Kim D, Lee J, Min H, Wall E, Lee CJ, Simon MI, Lee SJ, Han SK (2011) Analysis of cellular and be-havioral responses to imiquimod re-veals a unique itch pathway in transi¬ent re¬ceptor potential vanilloid 1 (TRPV1)-express¬ing neurons. Proc Natl Acad Sci U S A. 108(8): 3371–3376.
20. Urosevic M, Oberholzer PA, Maier T, Hafner J, Laine E, Slade H, Benning¬hoff B, Burg G, Dummer R (2004) Imiquimod treatment induces expres¬sion of opioid growth factor receptor: a novel tumor an¬tigen induced by interferon-α? Clin Can¬cer Res. 10(15): 4959–4970.
21. Jacob S, Berman B, Nassiri M, Vincek V (2003) Topical application of imiquimod 5% cream to keloids alters expression genes associated with apoptosis. Br J Der-matol. 149(Suppl. 66): 62–65.
22. Honoré S, Garin YJ, Sulahian A, Gangneux JP, Derouin F (1998) Influ¬ence of the host and parasite strain in a mouse mod¬el of visceral Leishmania in¬fantum infec¬tion. FEMS Immunol Med Micro¬biol. 21 (3): 231–239.
23. Rolão N, Cortes S, Gomes-Pereira S, Campino L (2007) Leishmania infan-tum: mixed T-helper-1/T-helper-2 im-mune re¬sponse in experimentally in-fect¬ed BALB/ c mice. Exp Parasitol. 11 (3): 270–276.
24. González-De la Fuente S, Peiró-Pastor R, Rastrojo A, Moreno J, Carrasco-Ramiro F, Requena JM, Aguado B (2017) Rese-quencing of the Leishma¬nia infantum (strain JPCM5) genome and de novo as-sembly into 36 contigs. Sci Rep. (1): 1–10.
25. Dehkordi NM, Ghaffarifar F, Hassan ZM, Heydari FE (2013) In vitro and in vivo studies of anti leishmanial effect of ar-temether on Leishmania infantum. Jun-dishapur J Microbiol. 6(5): e6379.
26. Molaie S, Ghaffarifar F, Dalimi A, Zuhair MH, Sharifi Z (2019) Evaluation of syn¬ergistic therapeutic effect of shark car¬ti¬lage extract with artemisinin and glu¬can¬time on visceral leishmaniasis in BALB/ c mice. Iran J Basic Med Sci. 22(2): 146–153.
27. Ebrahimisadr P, Ghaffarifar F, Horton J, Dalimi A, Sharifi Z (2018) Apoptotic ef¬fect of morphine, imiquimod and nalmefene on promastigote, infected and uninfected macrophages with amastigote of Leishmania major by flow cytome¬try. Iran J Pharm Res. 17(3): 986–994.
28. Jabari J, Ghaffarifar F, Horton J, Dalimi A, Sharifi Z (2019) Evaluation of mor-phine with imiquimod as opioid growth fac¬tor receptor or nalmefene as opioid blocking drug on leishmaniasis caused by Leishmania major in vitro. Iran J Parasi¬tol. 14(3): 394–403.
29. Ebrahimisadr P, Ghaffarifar F, Hassan ZM (2013) In-vitro evaluation of an-tileish¬manial activity and toxicity of ar-teme¬ther with focus on its apoptotic ef-fect. Iran J Pharm Res. 12(4): 903–909.
30. Molaie S, Ghaffarifar F, Hasan ZM, Da-limi A (2019) Enhancement effect of shark cartilage extract on treatment of Leishmania infantum with artemisinin and glucantime and evaluation of kill-ing factors and apoptosis in-vitro con-dition. Iran J Pharm Res. 18(2): 887–902.
31. Schriefer A, Wilson ME, Carvalho EM (2008) Recent developments leading to-ward a paradigm switch in the diag¬nos-tic and therapeutic approach to human leish¬maniasis. Curr Opin Infect Dis. 21(5): 483–488.
32. Corral MJ, González-Sánchez E, Cuquerella M, Alunda JM (2014) In vitro syn¬er¬gistic effect of amphotericin B and al¬licin on Leishmania donovani and L. in¬fantum. Antimicrob Agents Chemoth¬er. 58 (3): 1596–602.
33. Sundar S, Goyal N (2007) Molecular mech¬anisms of antimony resistance in Leish¬mania. J Med Microbiol. 56(2): 143–153.
34. Ghaffarpasand F, Akbarzadeh A, Heiran HR, Karimi AA, Akbarzadeh A, Ghoba¬difar MA (2016) Effect of topical Mor¬phine on cutaneous leishmaniasis in an animal model: a preliminary report. Iran Red Cres-cent Med J. 18(5): e24402.
35. Singal P, Singh PP (2005) Leishmania do-novani amastigote component-induced col-ony-stimulating factor production by mac¬rophages: modulation by mor¬phine. Mi¬crobes Infect. 7(2): 148–156.
36. Lysle DT, Fecho K, Maslonek KA, Dykstra LA (1995) Evidence for the involve¬ment of macrophage-derived nitric ox¬ide in the immunomodulatory effect of morphine and aversive Pavlovian con¬ditioning. The Brain Immune Axis and Substance Abuse. Springer Publishing. North Carolina, US, pp. 141–147.
37. Heussler VT, Küenzi P, Rottenberg S (2001) Inhibition of apoptosis by in¬tra¬cellular pro-tozoan parasites. Int J Par¬asitol. 31(11): 1166–1176.
38. DaMata JP, Mendes BP, Maciel-Lima K, Menezes CAS, Dutra WO, Sousa LP (2015) Distinct macrophage fates after in vitro infection with different species of Leishmania: induction of apoptosis by Leishmania (Leishmania) ama¬zonen-sis, but not by Leishmania (Viannia) guy¬a-nen¬sis. PLoS One. 10(10): e0141196.
39. Ruhland A, Kima PE (2009) Activation of PI3K/Akt signaling has a dominant neg-ative effect on IL-12 production by mac-rophages infected with Leishmania ama-zonensis promastigotes. Exp Parasitol. 122 (1): 28–36.
40. Kardeh S, Ashkani-Esfahani S, Alizadeh AM (2014) Paradoxical action of reac-tive oxygen species in creation and ther¬apy of cancer. Eur J Pharmacol. 735: 150–168.
41. Singh PP, Singal P (2007) Morphine-in¬duced neuroimmunomodulation in mu¬rine vis-ceral leishmaniasis: the role (s) of cyto-kines and nitric oxide. J Neu¬roimmune Pharmacol. 2(4): 338–351.
42. Poonawala T, Levay-Young BK, Hebbel RP, Gupta K (2005) Opioids heal is-chem¬ic wounds in the rat. Wound Repair Re¬gen. 13(2): 165–174.
43. Sacks D, Noben-Trauth N (2002) The im-munology of susceptibility and re-sistance to Leishmania major in mice. Nat Rev Immunol. 2(11): 845–858.
44. Oldfield V, Keating GM, Perry CM (2005) Imiquimod. Am J Clin Dermatol. 6(3): 195–200.
45. Geisse J, Caro I, Lindholm J, Golitz L, Stampone P, Owens M (2004) Imiquimod 5% cream for the treatment of super¬fi¬cial basal cell carcinoma: results from two phase III, randomized, vehicle-con¬trolled studies. J Am Acad Dermatol. 50(5): 722–733.
46. Firooz A, Khamesipour A, Ghoorchi MH, Nassiri-Kashani M, Eskandari SE, Khata¬mi A, Hooshmand B, Gorouhi F, Rashighi-Firoozabadi M, Dowlati Y (2006) Imiquimod in combination with meglumine antimoniate for cutaneous leishmaniasis: a randomized assessor-blind controlled trial. Arch Dermatol. 142(12): 1575–1579.
47. Barati M, Mohebali M, Alimohammadian MH, Khmesipour A, Keshavarz H, Ak-houndi B, Zarei Z (2015) Double-blind randomized efficacy field trial of alum precipitated autoclaved Leishmania ma-jor (Alum-ALM) vaccine mixed with bcg plus imiquimod vs. placebo control group.
Iran J Parasitol. 10(3): 351–359.
48. Sauder DN (2000) Immunomodulatory and pharmacologic properties of imiquimod. J Am Acad Dermatol. 43(1): S6–11.
49. Spellberg B, Edwards Jr JE (2001) Type 1/Type 2 immunity in infectious dis¬eas-es. Clin Infect Dis. 32(1): 76–102.
50. Huang SJ, Hijnen D, Murphy GF, Kupper TS, Calarese AW, Mollet IG (2009) Imiquimod enhances IFN-γ production and effector function of T cells in¬fil¬trat-ing human squamous cell carcinomas of the skin. J Invest Dermatol. 129(11): 2676–2685.
51. Robinson CM, O’Dee D, Hamilton T, Nau GJ (2010) Cytokines involved in inter-feron-γ production by human macro-phag¬es. J Innate Immun. 2(1): 56–65.
52. Panaro MA, Acquafredda A, Lisi S, Lofru¬mento D, Mitolo V, Sisto M, Fasanella A, Trotta T, Bertani F, Consenti B, Bran¬donisio O (2001) Nitric oxide production by macrophages of dogs vaccinated with killed Leishmania infantum pro¬mastigotes. Comp Immunol Microbiol Infect Dis. 24 (3): 187–195.
53. Holzmuller P, Sereno D, Cavaleyra M, Mangot I, Daulouede S, Vincendeau P, Lemesre JL (2002) Nitric oxide-medi¬at-ed proteasome-dependent oligonucle-oso¬mal DNA fragmentation in Leishmania amazonensis amastigotes. Infect Immun. 70(7): 3727–3735.
54. Ghaffarifar F, Heydari FE, Dalimi A, Hassan ZM, Delavari M, Mikaeiloo H (2015) Evaluation of apoptotic and an-tileishmanial activities of Artemisinin on promastigotes and BALB/C mice in-fected with Leishmania major. Iran J Parasitol. 10(2): 258–267.
55. Rassi Y, Jalali M, Vatandoost H (2000) Susceptibility status of Ph. pa¬patasi to DDT in Arsanjan county in Fras Prov-ince, Iran. Iran J Public Health. 29(1–4): 21–23.
56. Rassi Y, Javadian E, Jalali M, Motazedian MH, Vatandoost H (2004) Investiga¬tion on zoonotic cutaneous leishmani¬a¬sis, south¬ern Iran. Iran J Public Health. 33(1): 31–35.
57. Rassi Y, Javadian E, Amin M, Rafiza¬deh S, Vatandoost H, Motazedian H (2006) Meriones libycus, the principal reservoir of zoonotic cutaneous leish-maniasis in southern Iran. Eastern Medi¬terr Health J. 12(3/4): 474–477.
58. Yaghoobi-Ershadi MR, Akhavan AA, Ja-hanifard E, Vatandoost H, Amin Gh, Moosa¬vi L, Zahraei Ramazani AR, Abdoli H, Arandian MH (2006) Re¬pel¬lency ef-fect of Myrtle essential oil and DEET against Phlebotomus pap¬tasi Sco¬poli, the main vector of zoono¬tic cu¬ta¬neous leish¬maniasis under la¬boratory con-ditions. Iran J Public Health. 35(3): 7–13.
59. Moosa-Kazemi SH, Shayeghi M, Vatan-doost H, Sadeghi MT, Javadian E, Mot-abar M, Hosseini MR, Abtahi M (2009) High performance thin layer chromatog¬raphy analysis of deltame-thrin residue on the impregnated bed nets during a Leish¬maniasis control pro¬gram in Iran. Iran J Arthropod Borne Dis. 3(1): 1–7.
60. Oshaghi MA, Ravasan NM, Javadian E, Rassi Y, Sadraei J, Enayati AA, Vatan-doost H, Zare Z, Emami SN (2009) Ap-plication of predictive de¬gree day model for field development of sandfly vectors of visceral leish¬maniasis in northwest of Iran. J Vector Borne Dis. 46(4): 247–255.
61. Aghaei Afshar A, Rassi Y, Sharifi I, Abai MR, Oshaghi MA, Yaghoobi-Ershadi MR, Vatandoost H (2011) Sus¬ceptibility sta-tus of Phlebotomus pa¬pa¬tasi and P. ser-genti (Diptera: Psy¬chodi¬dae) to DDT and Deltame¬thrin in a fo¬cus of Cutane-ous Leish¬maniasis after earthquake strike in Bam, Iran. Iran J Arthropod Borne Dis. 5(2): 32–41.
62. Saeidi Z, Vatandoost H, Akhavan AA, Yaghoobi-Ershadi MR, Rassi Y, Sheikh Z, Arandian MH, Jafari R, Sanei-Dehkor¬di AR (2012) Baseline suscepti¬bility of a wild strain of Phlebotomus papatasi (Diptera: Psy¬chodidae) to DDT and pyrethroids in an endemic focus of zoonotic cutane¬ous leishmaniasis in Iran. Pest Manag SCi. 68(5): 669–675.
63. Veysi A, Vatandoost H, Yaghoobi-Er-shadi MR, Arandian MH, Jafari R, Hosseini M, Abdoli H, Rassi Y, Hei¬dari K, Sadjadi A, Fadaei R, Rama¬zanpour J, Aminian K, Shirzadi MR, Akhavan AA (2012) Com¬parative study on the effec-tiveness of Couma¬vec and zinc phos-phide in con¬trolling zoonotic cutaneous leishmaniasis in a hyperendemic focus in central Iran. J Arthropod Borne Dis. 6(1): 18–27.
64. Veysi A, Vatandoost H, Arandian MH, Jafa¬ri R, Yaghoobi-Ershadi MR, Rassi Y, Akha¬van AA (2013) Labora¬tory eval-uation of a rodenticide-insecticide, Coumavec®, against Rhombomys opi-mus, the main res¬er¬voir host of zoono¬tic cutaneouse leish¬maniasis in Iran. J Ar-thropod Borne Dis. 7(2): 188–193.
65. Saeidi Z, Vatandoost H, Akhavan AA, Yaghoobi-Ershadi MR, Rassi Y, Aran-dian MH, Jafari R (2013) Baseline in-secticide susceptibility data of Phleboto¬mus papatasi in Iran. J Vec¬tor-Borne Dis. 50: 57–61.
66. Aghai Afshar A, Vatandoost H, Shari¬fi I, Rassi Y, Abai MR, Oshaghi MA, Yaghoobi-Ershadi MR, Rafizadeh S (2013) First determination of impact and outcome indicators following in-door re¬sidual spraying (IRS) with del-tamethrin in a new focus of anthro-ponotic cutane¬ous leishmaniasis (ACL) in Iran. Asian Pac J Trop Dis. 3(1): 5–9.
67. Aghai Afshar A, Rassi Y, Sharifi I, Vatan-doost H, Mollaie HR, Oshaghi MA, Abai MR, Rafizadeh S (2014) First report on natural Leishmania in¬fec¬tion of Phleboto¬mus sergenti due Leishma¬nia tropica by high resolution melting curve method in South-eastern Iran. Asian Pac J Trop Med. 7(2): 93–96.
68. Akhavan AA, Veisi A, Arandian MH, Vatandoost H, Yaghoobi-Ershadi MR, Hosseini M, Abdoli H, Heidari K, Sad-jadi A, Fa¬daei R, Ramazanpour J, Amini¬an K, Shir¬zadi MR, Jafari R (2014) Field evaluation of phostoxin and zinc phos¬phide for the control of zoono¬tic cutane¬ous leishman¬iasis in a hy-perendemic ar¬ea, central Iran. J Vector- Borne Dis. 51(4): 307–312.
69. Jalilnavaz MR, Abai MR, Vatandoost H, Mohebali M, Akhavan AA, Zarei Z, Ra-fizadeh S, Bakhshi H, Rassi Y (2016) Application of flumethrin pour-on on res¬ervoir dogs and its efficacy against sand flies in endemic focus of visceral leishmaniasis, Meshkinshahr, Iran. J Ar¬thropod Borne Dis. 10(1): 78–86.
70. Hazratian T, Vatandoost H, Oshaghi MA, Yaghoobi-Ershadi MR, Fallah E, Ra-fiza¬deh S, Shirzadi MR, Shayeghi M, Akbarzadeh K, Rassi Y (2016) Di-versity of sand flies (Diptera: Psycho-didae) in endemic focus of visceral leishmaniasis in Azar shahr district, east Azarbaijan Province, North West of Iran. J Arthro¬pod Borne Dis. 10(3): 328–334.
71. Sofizadeh A, Vatandoost H, Rassi Y, Hanafi-Bojd AA, Rafizade S (2016) Spatial Anal¬yses of the relation be¬tween rodent’s ac¬tive burrows and in¬cidence of zoonotic cutaneous leish¬maniasis in Go¬lestan Prov¬ince, North¬eastern of Iran. J Arthropod Borne Dis. 10(4): 569–576.
72. Saghafipour A, Vatandoost H, Zahraei-Ram-azani AR, Yaghoobi-Ershadi MR, Rassi Y, Shirzadi MR, Akhavan AA (2017) Spa¬tial distribu¬tion of phlebotomine sand flies species (Dip¬tera: Psychodidae) in Qom Prov¬ince, central Iran. J Med Entoml. 54 (1): 35–43.
73. Saghafipour A, Vatandoost H, Zahraei-Ram-azani AR, Yaghoobi-Ershadi MR, Kara-mi Jooshin M, Rassi Y, Shirzadi MR, Akhavan AA, Hanafi-Bojd AA (2016b) Epidemio¬logical Study on Cu¬taneous Leish¬man¬iasis in an Endemic Area of Qom Province, Central Iran. J Arthro-pod Borne Dis. 11(3): 403–413.
74. Sofizadeh A, Rassi Y, Vatandoost H, Hanafi-Bojd AA, Mollalo A, Rafiza¬deh S, Akha¬van AA (2016b) Predict¬ing the distribu¬tion of Phlebotomus papatasi (Diptera: Psychodidae), the primary vec¬tor of zo¬onotic cutaneous leishmaniasis, in Go¬lestan Province of Iran using Eco¬logical Niche Modeling: Comparison of MaxEnt and GARP Models. J Med En¬tomol. 54 (2): 312–320.
75. Veysi A, Vatandoost H, Yaghoobi-Ershadi MR, Jafari R, Arandian MH, Hosseini M, Fadaei R, Ramazanpour J, Heidari K, Sad¬jadi A, Shirzadi MR, Akhavan AA (2016) Rodenticide com¬parative effect of klerat ® and zinc phos¬phide for controlling zo¬onotic cu¬taneous leishmaniasis in cen¬tral Iran. Iran J Par-asitol. 11(4): 471–479.
76. Saghafipour A, Vatandoost H, Zahraei-Ram-azani AR, Yaghoobi-Ershadi MR, Rassi Y, Jooshin M, Shirzadi MR, Akhavan AA (2017) Control of zo¬onotic cutaneous leish¬maniasis vector, Phlebotomus papa-tasi, using attractive toxic sugar baits (ATSB). PLoS One. 12(4): e0173558.
77. Shirani-Bidabadi L, Zahraei-Ramazani AR, Yaghoobi-Ershadi MR, Rassi Y, Akha¬van AA, Oshaghi MA, Enayati AA, Saeidi Z, Jafari R, Vatandoost H (2017) Assessing the insecticide sus-cep¬tibility status of field population of Phlebotomus papa¬tasi (Diptera: Psy-chodidae) in a hyperendem¬ic area of zo-onotic cutaneous leishmaniasis in Esfa-han Province, central Iran. Acta Trop. 176: 316–322.
78. Arzamani K, Vatandoost H, Rassi Y, Abai MR, Akhavan AA, Alavinia M, Ak¬bar-zadeh K, Mohebali M, Rfizadeh S (2017) Susceptibility status of wild pop¬ulation of Phlebotomus sergenti (Dip¬tera: Psy-cho¬didae) to different imagi¬cides in an endemic focus of cu¬taneous leishmania-sis in northeast of Iran. J Vec¬tor Borne Dis. 54(3): 282–286.
79. Moradiasl E, Rassi Y, Hanafi-Bojd AA, Vatandoost H, Saghafipour A, Adham D, Aabasgolizadeh N, Omidi Oskouei A, Sadeghi H (2018) The re¬lationship be¬tween climatic factors and the prev¬a-lence of visceral leish¬maniasis in Northwest of Iran. Intern J Pediatrics. 2(50): 7169–7178.
80. Vatandoost H, Nejati J, Saghafipour A, Zahraei-Ramazani A (2018) Geo¬graphic and ecological features of phlebotomine sand flies (Diptera: Psy¬chodidae) as leish¬maniasis in Central Iran. J Parasitic Dis. 42(1): 43–49.
81. Karimian F, Vatandoost H, Rassi Y, Ma-leki-Ravasan N, Choubdar N, Koosha M, Arzamani K, Moradi-Asl E, Veysi A, Ali¬pour H, Shirani M, Oshaghi MA (2018) Wsp-based analy¬sis of Wolbach-ia strains associated with Phlebotomus papatasi and P. sergenti (Diptera: Psy-chodidae) main cutane¬ous leishmaniasis vectors. Pathog Glob Health. 112(3): 152–160.
82. Arzamani K, Vatandoost H, Rassi Y, Akha¬van AA, Abai MR, Alavinia M, Akbar¬zadeh K, Mohebali M, Rafiza¬deh S (2018) Richness and diversity of phlebotomine sand flies (Diptera: Psy-chodidae) in North Khorasan Prov¬ince, northeast of Iran. J Arthropod Borne Dis. 12(3): 232–239.
83. Karimian F, Vatandoost H, Rassi Y, Ma-leki-Ravasan N, Mohebali M, Shi¬razi MH, Koosha M, Choubdar N, Oshaghi MA (2019) Aerobic midgut microbio¬ta of sand fly vectors of zo¬onotic vis¬ceral leishman¬iasis from northern Iran, a step toward finding potential para¬transgenic candi¬dates. Parasite Vectors. 12(1): 1–2.
84. Yaghoobi-Ershadi MR, Akhavan AA, Shir¬zadi MR, Rassi Y, Khamesipour A, Hanafi AA, Vatandoost H (2019) Conducting in¬ternational di¬ploma course on leishmani¬asis and its control in the Islamic Repub¬lic of Iran. J Arthropod Borne Dis. 13 (3): 234–242.
85. Rassi Y, Moradi-Asl E, Vatandoost H, Ab¬azari M, Saghafipour A (2020) In-secti¬cide susceptibility status of wild population of Phlebotomus kandelakii and Phleboto¬mus perfiliewi transcau¬ca-sicus collected from visceral leish¬mani-asis endemic foci in northwestern Iran. J Arthropod Borne Dis. 14(3): 277–285.
86. Yousefi S, Zahraei-Ramazani AR, Rassi Y, Vatandoost H, Yaghoobi-Ershadi MR, Af¬latoonian MR, Akha¬van AA, Aghaei-Af¬shar A, Amin M, Paksa A (2020) Evalu¬ation of different attrac¬tive traps for cap¬turing sand flies (Dip¬tera: Psychodidae) in an endemic area of Leishmaniasis, South¬east of Iran. J Ar-thropod Borne Dis. 14 (2): 202–213.
87. Moradi-Asl E, Mohebali M, Rassi Y, Vatan-doost H, Saghafipour A (2020) Environ-mental variables associated with distri-bution of canine visceral leishman¬iasis in dogs in Ardabil Prov¬ince, Northwestern Iran. Iran J Public Health. 49(6): 1033–1044.
88. Shirani-Bidabadi L, Zahraei-Ramazani AR, Yaghoobi-Ershadi MR, Akhavan AA, Oshaghi MA, Enayati AA, Rassi Y, Gholampour F, Shareghi N, Madreseh E, Vatandoost H (2020) Mon¬itoring of Laboratory Reared of Phlebotomus pa-patasi (Diptera: Psy¬chodidae), main vec¬tor of zoonotic cu¬ta¬neous leishmaniasis to different imagi¬cides in hyper endemic areas, Esfahan Province, Iran. J Arthro¬pod Borne Dis. 14(1): 116–125.
89. Mozaffari E, Vatandoost H, Rassi Y, Mohe-bali M, Akhavan AA, Moradi-Asl E, Za¬rei Z, Zahrai-Ramazani A, Ghor¬bani E (2020) Epidemiology of visceral leishman-iasis with emphasis on the dy¬namic activ¬ity of sand flies in an im¬portant endemic focus of disease in Northwestern Iran. J Arthropod Borne Dis. 14(1): 97–105.
90. Rassi Y, Asadollahi H, Abai MR, Kayedi MH, Vatandoost H (2020) Ef¬ficiency of two capture methods provid¬ing live sand flies and assess¬ment the suscep-tibility sta¬tus of Phlebotomus pa¬patasi (Dip-tera: Psy¬chodidae) in the foci of Cuta-neous Leish¬maniasis, Lorestan Province, Western Iran. J Arthropod Borne Dis. 14(4): 408–415.
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IssueVol 15 No 2 (2021) QRcode
SectionOriginal Article
DOI https://doi.org/10.18502/jad.v15i2.7492
Keywords
Morphine; Imiquimod; Synergism effect; Leishmania infantum; Iran

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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Ghaffarifar F, Foroutan M, Molaei S, Moradi-Asl E. Synergistic Anti-Leishmanial Activities of Morphine and Imiquimod on Leishmania infantum (MCAN/ES/98/LIM-877). J Arthropod Borne Dis. 2021;15(2):236-254.