Expression of Phlebotomus papatasi Salivary Protein 15 (PpSP15) in COS-7 Cells

Mahboubeh  Fatemi; Fatemeh  Ghaffarifar; Elham  Gholami; Mehdi  Mohebali; Ali  Khamesipour; Mohammad Ali  Oshaghi; Yavar  Rassi; Alireza  Zahraei-Ramazani; Amir Ahmad  Akhavan

doi:10.18502/jad.v18i4.19340

Mahboubeh Fatemi Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Fatemeh Ghaffarifar Department of Medical Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Elham Gholami Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran
Mehdi Mohebali Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Ali Khamesipour Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Ali Oshaghi Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Yavar Rassi Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Alireza Zahraei-Ramazani Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Amir Ahmad Akhavan Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

DOI: https://doi.org/10.18502/jad.v18i4.19340

Keywords: Leishmania major; Phlebotomus papatasi; PpSP15; pcDNA 3.1; PEI

Abstract

Background: Cutaneous leishmaniasis (CL) is a neglected tropical infection and the most prevalent vector-borne dis-ease in Iran. There is no approved human vaccine and current treatments are restricted; some drugs are expensive and have notable side effects. Therefore, the need for the development of a safe and effective vaccine that can be produced at a low cost remains urgent. It has been shown that vaccinating animals with salivary gland homogenate or saliva com¬ponents of sand flies protected against Leishmania infection. In this study, we aimed to prepare a mammalian expres¬sion vector encoding Phlebotomus papatasi salivary protein 15 (PpSP15) intended to be used as a DNA vaccine in our forthcoming studies.

Methods: In this study, we designed and constructed pcDNA3. 1, a constitutive mammalian expression vector, to en-code the immunogenic protein PpSP15. The presence of the target gene was confirmed by enzymatic digestion and se-quencing. The mammalian COS-7 cells were transfected with the pcDNA3.1 vector and the expression of PpSP15 pro-tein was then examined in the cell line using Western Blotting analysis.

Results: Restriction enzyme digestion and sequencing revealed the correctly constructed pcDNA3.1-PpSP15. After the transfection of the COS-7 cell line with pcDNA3.1-PpSP15 using Linear Polyethylenimine, the PpSP15 protein expres-sion was confirmed by western blot analysis using anti-His antibody.

Conclusion: A high expression level of PpSP15 protein in COS-7 cells was achieved after the transfection of COS-7 cells, using cationic Linear Polyethylenimine. In subsequent research, this recombinant plasmid is supposed to be uti-lized as a candidate DNA vaccine to find its immunity induction in susceptible animal models.