Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

Silveira, Marcelle Moura; Conceição, Fabricio Rochedo; Mendonça, Marcelo; Moreira, Gustavo Marçal Schmidt Garcia; Cunha, Carlos Eduardo Pouey da; Rizzi, Caroline; Hartwig, Daiane Drawanz; Moreira, Angelita da Silveira; Vendrusculo, Claire Tondo; Moreira, Ângela Nunes

doi:10.1590/1678-4324-2020190090

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Article - Human and Animal Health • Braz. arch. biol. technol. 63 • 2020 • https://doi.org/10.1590/1678-4324-2020190090 linkcopy

Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

Authorship SCIMAGO INSTITUTIONS RANKINGS

DNA vaccines have been evaluated as an option to prevent several diseases. In this study, the capacity of the xanthan biopolymer to improve the DNA vaccines immune response, administered intramuscularly, was evaluated. The experimental vaccines consisted of genes encoding fragments of the proteins LigA and LigB of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Copenhageni strain Fiocruz L1-130. The humoral immune response was evaluated by indirect ELISA. Cytokine expression levels were determined by RT-qPCR. Compared to the control group, the IgG antibody levels of animals immunized with pTARGET/ligAni and pTARGET/ligBrep plasmids associated with xanthan biopolymer were significantly higher than the control group. Additionally, there was a significant increase in IL-17 expression in animals vaccinated with pTARGET/ligBrep and xanthan.

Keywords:
Recombinant DNA vaccine; xanthan gum; immunologic adjuvant

Authorship

person Marcelle Moura Silveira

0000-0001-5104-5039

person Fabricio Rochedo Conceição

0000-0002-1072-4374

person Marcelo Mendonça

schoolFederal Rural University of Pernambuco, Academic Unit of Garanhuns, Garanhuns, Pernambuco, Brazil. Federal Rural University of PernambucoBrazilGaranhuns, Pernambuco, BrazilFederal Rural University of Pernambuco, Academic Unit of Garanhuns, Garanhuns, Pernambuco, Brazil.

0000-0001-7415-457X

person Gustavo Marçal Schmidt Garcia Moreira

0000-0002-3790-6557

person Carlos Eduardo Pouey da Cunha

0000-0001-9199-0764

person Caroline Rizzi

0000-0001-5810-9592

person Daiane Drawanz Hartwig

schoolFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. schoolFederal University of Pelotas, Department of Microbiology and Parasitology, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Department of Microbiology and Parasitology, Pelotas, Rio Grande do Sul, Brazil.

0000-0003-3604-0832

person Angelita da Silveira Moreira

schoolFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. schoolFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil.

0000-0001-8711-6521

person Claire Tondo Vendrusculo

schoolFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. schoolFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil.

0000-0003-0357-5988

person Ângela Nunes Moreira *

schoolFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. schoolFederal University of Pelotas, Department of Nutrition, Nutrition School, Pelotas, Rio Grande do Sul, Brazil.Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Department of Nutrition, Nutrition School, Pelotas, Rio Grande do Sul, Brazil.

0000-0001-5347-8559

person*Correspondence:
emailangelanmoreira@yahoo.com.br; Tel.: +55 (53)-32757583.

Conflicts of Interest:

The authors declare no conflict of interest.

SCIMAGO INSTITUTIONS RANKINGS

Federal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Technological Development Centre, Biotechnology Unit, Pelotas, Rio Grande do Sul, Brazil.

Federal Rural University of Pernambuco, Academic Unit of Garanhuns, Garanhuns, Pernambuco, Brazil. Federal Rural University of PernambucoBrazilGaranhuns, Pernambuco, BrazilFederal Rural University of Pernambuco, Academic Unit of Garanhuns, Garanhuns, Pernambuco, Brazil.

Federal University of Pelotas, Department of Microbiology and Parasitology, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Department of Microbiology and Parasitology, Pelotas, Rio Grande do Sul, Brazil.

Federal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil. Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Science, Pelotas, Rio Grande do Sul, Brazil.

Federal University of Pelotas, Department of Nutrition, Nutrition School, Pelotas, Rio Grande do Sul, Brazil.Federal University of PelotasBrazilPelotas, Rio Grande do Sul, BrazilFederal University of Pelotas, Department of Nutrition, Nutrition School, Pelotas, Rio Grande do Sul, Brazil.

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Figure 1 Evaluation of the humoral immune response induced by the DNA vaccines using xanthan biopolymer as adjuvant. Panels A, B, and C represent IgG antibody levels after the first, second, and third dose, respectively, for groups 1 and 2, treated with plasmid pTARGET/ligAni. Panels D, E, and F represent IgG antibody levels after the first, second, and third doses, respectively, for groups 3 and 4, which received the pTARGET/ligBrep plasmid. Control groups were inoculated with plasmid pTARGET/ligAni and pTARGET/ligBrep without adjuvant. Data were analysed using t Sudent test and asterisks (*) represent statistical difference to the control group (P < 0.05).

Thumbnail

Figure 2 Relative cytokine gene expression. Gene expression of IL-4, IL-12, IL-17, IFN-γ, and TGF-β was measured in PBMCs of the mice groups 1 and 2 treated with plasmid pTARGET/ligAni, (A), and PBMCs of the mice groups 3 and 4, which received the pTARGET/ligBrep plasmid, (B). The mice groups 2 and 4 were treated with the recombinant plasmids plus biopolymer xanthan as adjuvant. Relative gene expression was determined by qPCR and calculated using the 2-ΔΔCt method with E correction, using gadph mRNA expression as reference gene. Data were analysed using Tukey test and asterisks (**) represent statistical difference to the control group (P < 0.01).

imageFigure 1 Evaluation of the humoral immune response induced by the DNA vaccines using xanthan biopolymer as adjuvant. Panels A, B, and C represent IgG antibody levels after the first, second, and third dose, respectively, for groups 1 and 2, treated with plasmid pTARGET/ligAni. Panels D, E, and F represent IgG antibody levels after the first, second, and third doses, respectively, for groups 3 and 4, which received the pTARGET/ligBrep plasmid. Control groups were inoculated with plasmid pTARGET/ligAni and pTARGET/ligBrep without adjuvant. Data were analysed using t Sudent test and asterisks (*) represent statistical difference to the control group (P < 0.05). open_in_new

imageFigure 2 Relative cytokine gene expression. Gene expression of IL-4, IL-12, IL-17, IFN-γ, and TGF-β was measured in PBMCs of the mice groups 1 and 2 treated with plasmid pTARGET/ligAni, (A), and PBMCs of the mice groups 3 and 4, which received the pTARGET/ligBrep plasmid, (B). The mice groups 2 and 4 were treated with the recombinant plasmids plus biopolymer xanthan as adjuvant. Relative gene expression was determined by qPCR and calculated using the 2-ΔΔCt method with E correction, using gadph mRNA expression as reference gene. Data were analysed using Tukey test and asterisks (**) represent statistical difference to the control group (P < 0.01). open_in_new

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