Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat)

다국어 초록 (Multilingual Abstract)

Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model.
Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat.
Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR.
Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of −2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected.
Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.

참고문헌 (Reference)

1 Williams JA, "Vector design for improved DNA vaccine efficacy, safety and production" 1 (1): 225-249, 2013

2 Kusumawati A, "Vaccine against Jembrana disease virus infection : a summary findings" 11 (11): 68-73, 2015

3 Zhu Y, "Transcription elongation factor P-TEFb is required for HIV-1 tat transactivation in vitro" 11 (11): 2622-2632, 1997

4 Fröhlich E, "The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles" 7 : 5577-5591, 2012

5 Suwiti NK, "The phenomenon Jembrana disease and bovine immunodeficiency viruses in Bali cattle" 1 (1): 21-25, 2009

6 Porebski BT, "Structural and dynamic properties that govern the stability of an engineered fibronectin type III domain" 28 (28): 67-78, 2015

7 Zhao K, "Preparation and immunological effectiveness of a Swine influenza DNA vaccine encapsulated in PLGA microspheres" 27 (27): 178-186, 2010

8 Zhao K, "Preparation and efficacy of Newcastle disease virus DNA vaccine encapsulated in PLGA nanoparticles" 8 (8): e82648-, 2013

9 Xu Q, "Preparation and characterization of negatively charged poly(lactic-coglycolic acid)microspheres" 98 (98): 2377-2389, 2009

10 Avadi MR, "Preparation and characterization of insulin nanoparticles using chitosan and Arabic gum with ionic gelation method" 6 (6): 58-63, 2010

11 Ravi Kumar MN, "Preparation and characterization of cationic PLGA nanospheres as DNA carriers" 25 (25): 1771-1777, 2004

12 Amir Kalvanagh P, "Preparation and characterization of PLGA nanoparticles containing plasmid DNA encoding human IFN-lambda-1/IL-29" 18 (18): 156-167, 2019

13 Lúcio M, "Polymeric versus lipid nanoparticles : comparative study of nanoparticulate systems as indomethacin carriers" 4 (4): 95-109, 2015

14 Danhier F, "PLGA-based nanoparticles : an overview of biomedical applications" 161 (161): 505-522, 2012

15 Mancebo HS, "P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro" 11 (11): 2633-2644, 1997

16 Mohanraj VJ, "Nanoparticles-a review" 5 (5): 561-573, 2006

17 Chen H, "Jembrana disease virus Tat can regulate human immunodeficiency virus(HIV)long terminal repeat-directed gene expression and can substitute for HIV Tat in viral replication" 74 (74): 2703-2713, 2000

18 Mura S, "Influence of surface charge on the potential toxicity of PLGA nanoparticles towards Calu-3 cells" 6 : 2591-2605, 2011

19 Ishak J, "In vitro evaluation of chitosan-DNA plasmid complex encoding Jembrana disease virus env-tm protein as a vaccine candidate" 63 (63): 7-16, 2019

20 Obeng-Adjei N, "Immunogenicity of novel consensus-based DNA vaccines against hepatitis B core antigen" 186 (186): 106-101, 2011

21 Laddy DJ, "Immunogenicity of novel consensus-based DNA vaccines against avian influenza" 25 (25): 2984-2989, 2007

22 Cheng-Mayer C, "Host range, replicative, and cytopathic properties of human immunodeficiency virus type 1 are determined by very few amino acid changes in tat and gp120" 65 (65): 6931-6941, 1991

23 Unsunnidhal L, "Expression of gag-CA gene of Jembrana disease virus with cationic liposomes and chitosan nanoparticle delivery systems as DNA vaccine candidates" 30 (30): 15-36, 2019

24 Boyoglu S, "Enhanced delivery and expression of a nanoencapsulated DNA vaccine vector for respiratory syncytial virus" 5 (5): 463-472, 2009

25 Palocci C, "Endocytic pathways involved in PLGA nanoparticle uptake by grapevine cells and role of cell wall and membrane in size selection" 36 (36): 1917-1928, 2017

26 Agustini NLP, "Effication test of Jembrana disease vaccine" 27 (27): 1-16, 2009

27 Gao L, "Drug nanocrystals for the formulation of poorly soluble drugs and its application as a potential drug delivery system" 10 (10): 845-862, 2008

28 Starodubova S, "Creation of DNA vaccine vector based on codon-optimized gene of rabies virus glycoprotein(G protein)with consensus amino acid sequence" 50 (50): 328-331, 2016

29 Marshall NF, "Control of RNA polymerase II elongation potential by a novel carboxyl-terminal domain kinase" 271 (271): 27176-27183, 1996

30 Porebski BT, "Consensus protein design" 29 (29): 245-251, 2016

31 Haas J, "Codon usage limitation in the expression of HIV-1 envelope glycoprotein" 6 (6): 315-324, 1996

32 Kusumawati A, "Clinical and pathological perspectives of Jembrana disease virus infection : a review" 11 (11): 1221-1225, 2014

33 Zhao K, "Chitosan-coated poly(lactic-co-glycolic)acid nanoparticles as an efficient delivery system for Newcastle disease virus DNA vaccine" 9 : 4609-4619, 2014

34 Huang T, "Chitosan-DNA nanoparticles enhanced the immunogenicity of multivalent DNA vaccination on mice against Trueperella pyogenes infection" 16 (16): 8-, 2018

35 Chen H, "Characterization of the Jembrana disease virus tat gene and the cis-and trans-regulatory elements in its long terminal repeats" 73 (73): 658-666, 1999

36 Xu K, "Broad humoral and cellular immunity elicited by a bivalent DNA vaccine encoding HA and NP genes from an H5N1 virus" 24 (24): 45-56, 2011

37 Tanaya IWM, "Bio-molecular study of Jembrana virus : as basic development of tissue culture vaccine" 8 (8): 187-202, 2016

38 Smith CA, "An RNA-binding chameleon" 6 (6): 1067-1076, 2000