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Differential innate immune activation by four different CpG-ODNs in porcine and murine cells: Implications for
Giuliana Vedelago1, Luz M Palacios1, Sofía D Castell1
1Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Bioquímica Clínica. Haya de la Torre y Medina Allende, X5000HUA Córdoba, Argentina; CONICET, CIBICI. Haya de la Torre y Medina Allende, X5000HUA, Córdoba, Argentina.
Abstract:
Synthetic oligodeoxynucleotides containing unmethylated cytosine-guanine motifs (CpG-ODNs) are immunostimulants widely used as vaccine adjuvants. This study evaluated the innate immune responses induced by four CpG-ODNs: [2395 (class C), 1826, 1018, and 2007 (class B)] in porcine and murine cells, representing species of veterinary and preclinical interest, respectively. Porcine peripheral blood mononuclear cells and murine splenocytes were stimulated in vitro with each CpG-ODN, and the mRNA expression of IL-6, IL-12p40, IFN-β, IFN-γ, TNF, and TLR9 was assessed by RT-qPCR. In porcine cells, only CpG-ODN 2395 significantly upregulated IL-6, IL-12p40, and TLR9. In murine cells, all CpG-ODNs induced IL-6; CpG-ODNs 2395, 1826, and 1018 upregulated IL-12p40; and only CpG-ODN 2395 induced TLR9. Stimulation with CpG-ODNs 2395 and 1018 enhanced IFN-β and IFN-γ expression in porcine cells. In contrast, murine cells showed increased IFN-β levels in response to CpG-ODNs 2395 and 1826, and upregulated IFN-γ following stimulation with CpG-ODNs 2395, 1826, and 1018. TNF expression was exclusively induced in porcine cells by CpG-ODN 1018, 2395, and 2007. Notably, CpG-ODN 1826 exhibited species-specific activity in mice, inducing IL-6, IL-12p40, IFN-β, and IFN-γ. These findings reveal significant interspecies differences in CpG-ODN-induced immune responses, emphasizing the importance of species-specific considerations when selecting CpG-ODNs as vaccine adjuvants for preclinical and veterinary applications.
Insights
Synthetic oligodeoxynucleotides (CpG-ODNs) are vaccine adjuvants. This study found significant species-specific differences in immune responses between porcine and murine cells stimulated with various CpG-ODNs, impacting adjuvant selection.
Area of Science:
- Immunology
- Vaccinology
- Molecular Biology
Background:
- Synthetic oligodeoxynucleotides containing unmethylated cytosine-guanine motifs (CpG-ODNs) are potent immunostimulants.
- CpG-ODNs are widely utilized as vaccine adjuvants to enhance immune responses.
- Understanding species-specific immune activation is crucial for effective vaccine development.
Purpose of the Study:
- To evaluate and compare the innate immune responses induced by different classes of CpG-ODNs in porcine and murine cells.
- To assess the expression of key immune-related genes (IL-6, IL-12p40, IFN-β, IFN-γ, TNF, TLR9) following CpG-ODN stimulation.
- To identify species-specific patterns of immune activation relevant for veterinary and preclinical applications.
Main Methods:
- Porcine peripheral blood mononuclear cells and murine splenocytes were stimulated in vitro with four distinct CpG-ODNs (2395, 1826, 1018, 2007).
- Messenger RNA (mRNA) expression levels of target genes were quantified using quantitative reverse transcription polymerase chain reaction (RT-qPCR).
- Immune responses were analyzed comparatively between porcine and murine cell models.
Main Results:
- CpG-ODN 2395 significantly upregulated IL-6, IL-12p40, and TLR9 in porcine cells, while all tested CpG-ODNs induced IL-6 in murine cells.
- CpG-ODN 1826 demonstrated species-specific activity in mice, inducing multiple immune mediators.
- Significant interspecies variations were observed in the induction of IFN-β, IFN-γ, and TNF across different CpG-ODNs and cell types.
Conclusions:
- CpG-ODN-induced immune responses exhibit significant interspecies differences between pigs and mice.
- The selection of appropriate CpG-ODNs as vaccine adjuvants requires careful consideration of the target species.
- These findings highlight the importance of species-specific evaluation for optimizing adjuvant efficacy in veterinary and preclinical settings.
