Acta Parasitologica

, Volume 64, Issue 2, pp 236–245 | Cite as

A Survey on the Adjuvant Role of Naloxone Alone or Combined with Alum in Vaccination Against Fasciolosis in BALB/c Mice

  • Hakim Azizi
  • Hadi Mirzaeei
  • Amin Bagheri
  • Ali BaziEmail author
  • Ali Khamesipour
  • Hajar Yaghoobi
  • Aliyar Mirzapour
  • Mehrdad Khatami
  • Samira Elikaee
Original Paper



Fasciolosis is a zoonotic parasitic disease imposing a heavy load of livestock losses worldwide.


We aimed to evaluate immune-stimulatory effects of naloxone (NLX), an opioid receptor antagonist, in combination with alum in mice vaccinated with excretory–secretory antigens (E/S) of Fasciola hepatica.


8-week-old female BALB/c mice were subcutaneously vaccinated using E/S antigens of F. hepatica. Experimental groups (14 mice per group) included: vaccine (E/S antigen), alum vaccine (E/S antigen plus alum), NLX vaccine (E/S antigen plus NLX), and alum–NLX vaccine (E/S antigen plus a mixture of alum–NLX). The control group was infused with PBS. Lymphocyte proliferation and the levels of IFN-γ, IL-4, IgG2a, IgG1, and total IgG were measured.


Mice vaccinated with NLX or alum–NLX adjuvants showed significantly higher rates of lymphocyte proliferation, IFN-γ, total IgG, and IgG2a levels. The mice that were injected with alum showed a significantly higher concentration of IL-4. Ratios of IFN-γ/Il-4 and IgG2a/IgG1 were significantly higher in the NLX and alum–NLX groups in comparison with the groups vaccinated either with alum or without any adjuvant. A significantly higher protection rate (62.5%) was seen in mice vaccinated with the alum–NLX adjuvant compared to the other groups.


NLX can be effective in conferring cellular immunity and protection against F. hepatica. It is recommended to consider this agent as a potential adjuvant in vaccines against fasciolosis.


Fasciolosis Adjuvant Aluminum sulfate Naloxone Vaccine 



This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. This research was supported by Deputy of research of Zabol University of medical sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All experiments were performed according to the Animal Care and Use Protocol of Zabol University of Medical Sciences. The study was approved by Ethical Committee of Zabol University of Medical Sciences.


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Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2019

Authors and Affiliations

  • Hakim Azizi
    • 1
  • Hadi Mirzaeei
    • 2
  • Amin Bagheri
    • 3
  • Ali Bazi
    • 4
    Email author
  • Ali Khamesipour
    • 5
  • Hajar Yaghoobi
    • 6
  • Aliyar Mirzapour
    • 7
  • Mehrdad Khatami
    • 8
  • Samira Elikaee
    • 9
  1. 1.Department of Medical Parasitology, School of MedicineZabol University of Medical SciencesZabolIran
  2. 2.Department of Medical Genetics, School of MedicineZabol University of Medical SciencesZabolIran
  3. 3.Faculty of Veterinary Medicine, Sciences and Research BranchIslamic Azad UniversityTehranIran
  4. 4.Clinical Research Development UnitZabol University of Medical SciencesZabolIran
  5. 5.Skin and Leprosy Research CenterTehran University of Medical SciencesTehranIran
  6. 6.Cellular and Molecular Research CenterShahrekord University of Medical SciencesShahrekordIran
  7. 7.Department of Medical Parasitology, School of MedicineShahid Beheshti University of Medical SciencesTehranIran
  8. 8.School of MedicineBam University of Medical SciencesBamIran
  9. 9.Department of Medical Parasitology and Mycology, School of Public HealthTehran University of Medical SciencesTehranIran

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