Heterologous Expression of Human IFNγ and Anti-IL17 Antibody in Leishmania tarentolae Promastigote

• Published in: Acta parasitologica Vol. 69; no. 2; pp. 1107 - 1114
• Main Authors: Rouzbahani, Arian Karimi, Hosseini, Seyedeh-Zeinab, Bandehpour, Mojgan, Kazemi, Bahram, Tavasoli, Afsaneh, Mamaghani, Amirreza Javadi, Kheirandish, Farnaz
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.06.2024; Springer Nature B.V
• Subjects: Animal Systematics/Taxonomy/Biogeography; Animals; Antibodies; Antibodies, Protozoan - immunology; Biomedical and Life Sciences; Biomedicine; Deoxyribonucleic acid; DNA; Ecology; Electroporation; Enzyme-linked immunosorbent assay; Genomes; Humans; Immunology; Interferon-gamma - genetics; Interferon-gamma - immunology; Interleukin 17; Interleukin-17 - genetics; Interleukin-17 - immunology; Interleukin-17 - metabolism; Leishmania; Leishmania - genetics; Leishmania - immunology; Leishmaniasis; Lizards - immunology; Lizards - parasitology; Medical Microbiology; Microbiology; mRNA; Original Paper; Parasites; Parasitic diseases; Parasitology; Plasmids - genetics; Recombinant DNA; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Toxicity; Vector-borne diseases; γ-Interferon; IFNγ; Anti-IL17; Leishmanization; Iranian lizard; Iranian lizard Leishmania
• ISSN: 1230-2821; 1896-1851; 1896-1851
• DOI: 10.1007/s11686-024-00822-1

Background Leishmania is an intracellular flagellate protozoan parasite that causes a wide range of clinical diseases in humans. The basis of immunological resistance against leishmaniasis depends on Thl reactions and is within the time period of cytokine function. Methods In this study, human anti-IL17 antibody and IFNγ-producing promastigote were produced to be used in leishmanization. A sequence of light and heavy chains’ gene of anti-IL17 antibody and human IFNγ (hIFNγ) was obtained from the NCBI database and synthesized in the ECORV reaction site in the plasmid pGH, which it’s called pGH–hIFNγ–antiIL17. The synthesized part using the restriction enzyme ECORV was extracted from the plasmid and after purification by electroporation was transferred to Iranian lizard Leishmania (I.L.L). Evaluation of structural presence in the I.L.L genome at the level of DNA and mRNA was assessed. The expressions of hIFNγ and anti-IL17 were evaluated and confirmed using ELISA and western blot analysis. The hIFNγ secreted from the culture medium was collected at high concentrations of 124.36 ± 6.47 pg/mL. Results Targeted gene replacement into the I.L.L genome was successfully performed for the first time using the pGH–hIFNγ–antiIL17 plasmid in an identical replacement process. Stabilized recombinant DNA contains a target gene that has no toxicity to the parasite. Conclusions The effective achievement of producing a recombinant gene was done for the first time by replacing the I.L.L-CPC gene with plasmid pGH-hIFNγ-antiIL17 by targeted gene replacement. This cab can regulate the production of hIFNγ and anti-IL17. This makes it a viable choice for eliminating leishmania. Graphical Abstract