Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

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

Marcelle Moura Silveira

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.

http://orcid.org/0000-0001-5104-5039

Fabricio Rochedo Conceição

http://orcid.org/0000-0002-1072-4374

Marcelo Mendonça

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.

http://orcid.org/0000-0001-7415-457X

Gustavo Marçal Schmidt Garcia Moreira

http://orcid.org/0000-0002-3790-6557

Carlos Eduardo Pouey da Cunha

http://orcid.org/0000-0001-9199-0764

Caroline Rizzi

http://orcid.org/0000-0001-5810-9592

Daiane Drawanz Hartwig

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.

http://orcid.org/0000-0003-3604-0832

Angelita da Silveira Moreira

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.

http://orcid.org/0000-0001-8711-6521

Claire Tondo Vendrusculo

http://orcid.org/0000-0003-0357-5988

Ângela Nunes Moreira ^**Correspondence: angelanmoreira@yahoo.com.br; Tel.: +55 (53)-32757583.

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.

http://orcid.org/0000-0001-5347-8559

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

Conflicts of Interest:

The authors declare no conflict of interest.

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Figures

Figures (2)

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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).

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).

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).

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Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

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[1] *Correspondence: angelanmoreira@yahoo.com.br; Tel.: +55 (53)-32757583.