Rapid and Highly Efficient Genetic Transformation and Application of Interleukin-17B Expressed in Duckweed as Mucosal Vaccine Adjuvant

• Published in: Biomolecules (Basel, Switzerland) Vol. 12; no. 12; p. 1881
• Main Authors: Tan, Xiao, Chen, Shuang, Fang, Yang, Liu, Penghui, Hu, Zhubin, Jin, Yanling, Yi, Zhuolin, He, Kaize, Li, Xing, Zhao, Leyi, Wang, Hongning, Zhao, Hai
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01-12-2022; MDPI
• Subjects: adjuvant; Adjuvants, Immunologic - pharmacology; Adjuvants, Vaccine; Animals; Aquatic plants; Araceae - genetics; Araceae - metabolism; Biology; Biopharmaceuticals; Biotechnology industry; Bronchitis; Chickens; Cytokines; Cytokines - metabolism; duckweed; Efficiency; Farm buildings; Farming; Gene expression; Genetic aspects; Genetic transformation; IBV; Identification and classification; Immune response (humoral); Immunoglobulin A; Interleukin-17 - genetics; Interleukin-17 - metabolism; interleukin-17B; Janus Kinases - genetics; Lemna minor; Lemnaceae; Medical research; Mucosal immunity; NF-κB protein; Oral administration; Proteins; Signal Transduction; STAT Transcription Factors - genetics; Transformation, Genetic; Transgenic plants; Vaccination; Vaccines; China; Lemna minor; interleukin-17B; transgenic plants; IBV; adjuvant; duckweed
• ISSN: 2218-273X; 2218-273X
• DOI: 10.3390/biom12121881

Molecular farming utilizes plants as a platform for producing recombinant biopharmaceuticals. Duckweed, the smallest and fastest growing aquatic plant, is a promising candidate for molecular farming. However, the efficiency of current transformation methods is generally not high in duckweed. Here, we developed a fast and efficient transformation procedure in
ZH0403, requiring 7-8 weeks from screening calluses to transgenic plants with a stable transformation efficiency of 88% at the DNA level and 86% at the protein level. We then used this transformation system to produce chicken interleukin-17B (chIL-17B). The plant-produced chIL-17B activated the NF-κB pathway, JAK-STAT pathway, and their downstream cytokines in DF-1 cells. Furthermore, we administrated chIL-17B transgenic duckweed orally as an immunoadjuvant with mucosal vaccine against infectious bronchitis virus (IBV) in chickens. Both IBV-specific antibody titer and the concentration of secretory immunoglobulin A (sIgA) were significantly higher in the group fed with chIL-17B transgenic plant. This indicates that the duckweed-produced chIL-17B enhanced the humoral and mucosal immune responses. Moreover, chickens fed with chIL-17B transgenic plant demonstrated the lowest viral loads in different tissues among all groups. Our work suggests that cytokines are a promising adjuvant for mucosal vaccination through the oral route. Our work also demonstrates the potential of duckweed in molecular farming.