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Non-CpG Oligonucleotides Exert Adjuvant Effects by Enhancing Cognate B Cell-T Cell Interactions, Leading to B Cell Activation, Differentiation, and Isotype Switching.

Herbáth M, Papp K, Erdei A, Prechl J - J Immunol Res (2015)

Bottom Line: We found that both of the tested non-CpG ODN exerted significant immunomodulatory effects on early T cell and on late B cell activation events.Importantly, a synergism between non-CpG effects and T cell help acting on B cells was observed, resulting in enhanced IgG production following cognate T cell-B cell interactions.We propose that non-CpG ODN may perform as better adjuvants when a strong antigen-independent immune activation, elicited by CpG ODNs, is undesirable.

View Article: PubMed Central - PubMed

Affiliation: MTA-ELTE Immunology Research Group, 1/C Pázmány Péter Sétány, Budapest 1117, Hungary.

ABSTRACT
Natural and synthetic nucleic acids are known to exert immunomodulatory properties. Notably, nucleic acids are known to modulate immune function via several different pathways and various cell types, necessitating a complex interpretation of their effects. In this study we set out to compare the effects of a CpG motif containing oligodeoxynucleotide (ODN) with those of a control and an inhibitory non-CpG ODN during cognate B cell-T cell interactions. We employed an antigen presentation system using splenocytes from TCR transgenic DO11.10 mice and the ovalbumin peptide recognized by the TCR as model antigen. We followed early activation events by measuring CD69 expression, late activation by MHC class II expression, cell division and antibody production of switched, and nonswitched isotypes. We found that both of the tested non-CpG ODN exerted significant immunomodulatory effects on early T cell and on late B cell activation events. Importantly, a synergism between non-CpG effects and T cell help acting on B cells was observed, resulting in enhanced IgG production following cognate T cell-B cell interactions. We propose that non-CpG ODN may perform as better adjuvants when a strong antigen-independent immune activation, elicited by CpG ODNs, is undesirable.

No MeSH data available.


Non-CpG ODN enhances CD69 expression in T and B cells engaging in cognate interaction. DO11.10 splenocytes were incubated with different combinations of ODNs and OVA; after 24 hours CD69 expression was measured on T and B cell populations by flow cytometry ((a)-(b)). Medians and interquartile ranges of 13 independent experiments are shown. Asterisks indicate significant difference from the group receiving no ODN at all, within the respective OVA treatment group. MFI: mean fluorescence intensity; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.
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fig1: Non-CpG ODN enhances CD69 expression in T and B cells engaging in cognate interaction. DO11.10 splenocytes were incubated with different combinations of ODNs and OVA; after 24 hours CD69 expression was measured on T and B cell populations by flow cytometry ((a)-(b)). Medians and interquartile ranges of 13 independent experiments are shown. Asterisks indicate significant difference from the group receiving no ODN at all, within the respective OVA treatment group. MFI: mean fluorescence intensity; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.

Mentions: In order to extend earlier observations showing costimulation of T cells by non-CpG ODN [13], we utilized an antigen presentation system based on the transgenic expression of ovalbumin specific T cell receptor. Helper CD4 positive T cells from the DO11.10 mouse strain recognize an ovalbumin peptide sequence (referred to as OVA from here on). This peptide, displayed on MHCII of APCs served as TCR stimulus in our experiments. We used suspensions of splenocytes, which due to their abundance B cells (around 50% of all splenic white blood cells) strongly contribute as APC, especially when peptides that are taken up by pinocytosis and require no further processing are used as antigen. The concentration of OVA was set to 25 nM, which confers suboptimal T cell activation (Figure 1(a)) and thus enabled us to observe modulation of the outcome of antigen presentation. Along the same lines of thought, modulating ODNs were used at 0.25 µM and 2.5 µM concentrations, at which non-CpG ODNs (Inhibitor and Control) were found to only modestly activate B cells, in contrast to CpG ODN (Figure 1(b)).


Non-CpG Oligonucleotides Exert Adjuvant Effects by Enhancing Cognate B Cell-T Cell Interactions, Leading to B Cell Activation, Differentiation, and Isotype Switching.

Herbáth M, Papp K, Erdei A, Prechl J - J Immunol Res (2015)

Non-CpG ODN enhances CD69 expression in T and B cells engaging in cognate interaction. DO11.10 splenocytes were incubated with different combinations of ODNs and OVA; after 24 hours CD69 expression was measured on T and B cell populations by flow cytometry ((a)-(b)). Medians and interquartile ranges of 13 independent experiments are shown. Asterisks indicate significant difference from the group receiving no ODN at all, within the respective OVA treatment group. MFI: mean fluorescence intensity; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4562091&req=5

fig1: Non-CpG ODN enhances CD69 expression in T and B cells engaging in cognate interaction. DO11.10 splenocytes were incubated with different combinations of ODNs and OVA; after 24 hours CD69 expression was measured on T and B cell populations by flow cytometry ((a)-(b)). Medians and interquartile ranges of 13 independent experiments are shown. Asterisks indicate significant difference from the group receiving no ODN at all, within the respective OVA treatment group. MFI: mean fluorescence intensity; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.
Mentions: In order to extend earlier observations showing costimulation of T cells by non-CpG ODN [13], we utilized an antigen presentation system based on the transgenic expression of ovalbumin specific T cell receptor. Helper CD4 positive T cells from the DO11.10 mouse strain recognize an ovalbumin peptide sequence (referred to as OVA from here on). This peptide, displayed on MHCII of APCs served as TCR stimulus in our experiments. We used suspensions of splenocytes, which due to their abundance B cells (around 50% of all splenic white blood cells) strongly contribute as APC, especially when peptides that are taken up by pinocytosis and require no further processing are used as antigen. The concentration of OVA was set to 25 nM, which confers suboptimal T cell activation (Figure 1(a)) and thus enabled us to observe modulation of the outcome of antigen presentation. Along the same lines of thought, modulating ODNs were used at 0.25 µM and 2.5 µM concentrations, at which non-CpG ODNs (Inhibitor and Control) were found to only modestly activate B cells, in contrast to CpG ODN (Figure 1(b)).

Bottom Line: We found that both of the tested non-CpG ODN exerted significant immunomodulatory effects on early T cell and on late B cell activation events.Importantly, a synergism between non-CpG effects and T cell help acting on B cells was observed, resulting in enhanced IgG production following cognate T cell-B cell interactions.We propose that non-CpG ODN may perform as better adjuvants when a strong antigen-independent immune activation, elicited by CpG ODNs, is undesirable.

View Article: PubMed Central - PubMed

Affiliation: MTA-ELTE Immunology Research Group, 1/C Pázmány Péter Sétány, Budapest 1117, Hungary.

ABSTRACT
Natural and synthetic nucleic acids are known to exert immunomodulatory properties. Notably, nucleic acids are known to modulate immune function via several different pathways and various cell types, necessitating a complex interpretation of their effects. In this study we set out to compare the effects of a CpG motif containing oligodeoxynucleotide (ODN) with those of a control and an inhibitory non-CpG ODN during cognate B cell-T cell interactions. We employed an antigen presentation system using splenocytes from TCR transgenic DO11.10 mice and the ovalbumin peptide recognized by the TCR as model antigen. We followed early activation events by measuring CD69 expression, late activation by MHC class II expression, cell division and antibody production of switched, and nonswitched isotypes. We found that both of the tested non-CpG ODN exerted significant immunomodulatory effects on early T cell and on late B cell activation events. Importantly, a synergism between non-CpG effects and T cell help acting on B cells was observed, resulting in enhanced IgG production following cognate T cell-B cell interactions. We propose that non-CpG ODN may perform as better adjuvants when a strong antigen-independent immune activation, elicited by CpG ODNs, is undesirable.

No MeSH data available.