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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.


Effects of non-CpG ODN on late activation events. Spleen cells were incubated with the different combinations of ODNs and OVA as indicated. For MHCII expression measurements cells were harvested and stained for flow cytometry after 4 days (a). Medians and interquartile ranges of 9 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. For proliferation studies EdU was added on day 1 and left for incorporation for another day. The percentage of divided cells was then measured by flow cytometric analysis of EdU incorporation ((b), (c)). Results shown are from a single experiment; identical results were observed when EdU was added after 2 days.
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fig2: Effects of non-CpG ODN on late activation events. Spleen cells were incubated with the different combinations of ODNs and OVA as indicated. For MHCII expression measurements cells were harvested and stained for flow cytometry after 4 days (a). Medians and interquartile ranges of 9 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. For proliferation studies EdU was added on day 1 and left for incorporation for another day. The percentage of divided cells was then measured by flow cytometric analysis of EdU incorporation ((b), (c)). Results shown are from a single experiment; identical results were observed when EdU was added after 2 days.

Mentions: Interestingly, the expression of MHCII did not reflect our observations with CD69. Non-CpG ODN triggered the increase of MHCII in the absence of OVA (both low and high concentrations of Control and high concentrations of Inhibitor) but had no effects in the presence of OVA (Figure 2(a)). As expected, proliferation of T cells, expressed as the percentage of cells that incorporated the nucleotide analogue EdU, was only observed in the presence of OVA (Figure 2(b)). Proliferation of both T and B cells reflected the pattern of early activation marker expression (Figure 2(c)). Thus, non-CpG ODN alone induced moderate increase of B cell MHCII expression and synergized with OVA to increase the number of dividing B cells.


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)

Effects of non-CpG ODN on late activation events. Spleen cells were incubated with the different combinations of ODNs and OVA as indicated. For MHCII expression measurements cells were harvested and stained for flow cytometry after 4 days (a). Medians and interquartile ranges of 9 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. For proliferation studies EdU was added on day 1 and left for incorporation for another day. The percentage of divided cells was then measured by flow cytometric analysis of EdU incorporation ((b), (c)). Results shown are from a single experiment; identical results were observed when EdU was added after 2 days.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Effects of non-CpG ODN on late activation events. Spleen cells were incubated with the different combinations of ODNs and OVA as indicated. For MHCII expression measurements cells were harvested and stained for flow cytometry after 4 days (a). Medians and interquartile ranges of 9 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. For proliferation studies EdU was added on day 1 and left for incorporation for another day. The percentage of divided cells was then measured by flow cytometric analysis of EdU incorporation ((b), (c)). Results shown are from a single experiment; identical results were observed when EdU was added after 2 days.
Mentions: Interestingly, the expression of MHCII did not reflect our observations with CD69. Non-CpG ODN triggered the increase of MHCII in the absence of OVA (both low and high concentrations of Control and high concentrations of Inhibitor) but had no effects in the presence of OVA (Figure 2(a)). As expected, proliferation of T cells, expressed as the percentage of cells that incorporated the nucleotide analogue EdU, was only observed in the presence of OVA (Figure 2(b)). Proliferation of both T and B cells reflected the pattern of early activation marker expression (Figure 2(c)). Thus, non-CpG ODN alone induced moderate increase of B cell MHCII expression and synergized with OVA to increase the number of dividing B cells.

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.