Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
Autor: | Mina Ebrahimi-Rad, Sara Mohammadzadeh, Parastoo Ehsani, Hamideh Ofoghi |
---|---|
Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
Macrophage colony-stimulating factor
HBsAg medicine.medical_treatment Pharmaceutical Science Context (language use) Bone Marrow Cells RM1-950 Biology Hepatitis b surface antigen Protein Engineering Cell Line Mice Immune system Antigen Adjuvants Immunologic Mouse Granulocyte vaccine Drug Discovery Tobacco medicine Animals plant-produced Pharmacology hbsag Mice Inbred BALB C Hepatitis B Surface Antigens Macrophage Colony-Stimulating Factor Granulocyte-Macrophage Colony-Stimulating Factor gm-csf General Medicine Plants Genetically Modified Recombinant Proteins co-expression Complementary and alternative medicine Immunology Molecular Medicine Therapeutics. Pharmacology Adjuvant |
Zdroj: | Pharmaceutical Biology, Vol 57, Iss 1, Pp 669-675 (2019) |
ISSN: | 1744-5116 1388-0209 |
Popis: | Context: The co-delivery of adjuvant and antigen has shown to be more effective for targeting the immune response than antigen alone. Therefore, designing an efficient bicistronic system is more assuring for production of both elements in the same tobacco cells as a plant model system. Objective: Comparing the efficient transient co-expression of hepatitis B surface antigen (HBsAg) and mouse granulocyte macrophage colony stimulating factor (mGM-CSF) in tobacco leaves by designing either mono or bicistronic cassettes. Materials and methods: Four expression cassettes containing tobacco etch virus (TEV) leader sequence were constructed with and without above genes in different orders. The cassettes were transferred into tobacco, Nicotiana tabacum L. (Solanaceae), leaves by agroinfiltration technique. The expression levels were compared using ELISA and western blotting and bioactivity of cytokine was assessed by in vitro proliferation of mouse GM-CSF-responsive progenitor cells. Results: Agroinfiltrated leaves contained recombinant HBsAg protein at 20–50 ng/mg and mGM-CSF at 0.2–4 ng/mg in both nonglycosylated and glycosylated forms. The highest expression obtained in HBsAg and mGM-CSF monocistronic co-agroinfiltrated leaves. The expression of mGM-CSF was 1.1 and 0.2 ng/mg in two different orders of bicistronic cassettes. The growth frequency of GM progenitors was approximately 1/187 cells for standard rGM-CSF and 3.2 times less activity for the plant produced. Discussion and conclusions: The recombinant mGM-CSF was produced less in bicistronic cassette than other forms; however, co-presenting of both vaccine candidate and adjuvant is confirmed and could be promising for amelioration of plant expression system as a means for vaccine production. |
Databáze: | OpenAIRE |
Externí odkaz: | |
Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |