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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.


Related in: MedlinePlus

No synergism between OVA and ODN treatments when cognate T cells are not present. Wild type (wt) or TCR transgenic (tg) splenic cells were incubated with different combinations of ODN and OVA for 1 day ((a), (b)) or 4 days (c). After 1 day CD69 expression was measured on B and T cell populations by flow cytometry ((a), (b)). Cells from the 4-day cultures were washed and then transferred onto nitrocellulose-covered slides that were previously coated with light chain capture antibodies. After 10 h of incubation, slides were washed and spots indicating IgG producing ASCs were detected using fluorescently labeled antibodies and a fluorescent scanner (c). Results represent a single experiment.
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fig4: No synergism between OVA and ODN treatments when cognate T cells are not present. Wild type (wt) or TCR transgenic (tg) splenic cells were incubated with different combinations of ODN and OVA for 1 day ((a), (b)) or 4 days (c). After 1 day CD69 expression was measured on B and T cell populations by flow cytometry ((a), (b)). Cells from the 4-day cultures were washed and then transferred onto nitrocellulose-covered slides that were previously coated with light chain capture antibodies. After 10 h of incubation, slides were washed and spots indicating IgG producing ASCs were detected using fluorescently labeled antibodies and a fluorescent scanner (c). Results represent a single experiment.

Mentions: To further confirm that non-CpG ODN synergize with T cell derived stimuli in enhancing antibody production and isotype switching in B cells, we repeated experiments using wild type (wt) mice. While some OVA specific T cells may be present in naïve wt mice, their numbers are expected to be so low as to be negligible (1 : 10 000 or lower). Indeed, the percentage of CD69 positive T cells was unaffected by the addition of OVA in wt mice (Figure 4(a)). In the absence of OVA, the pattern of CD69 expression in B cells was similar in wt and DO11.10 mice (Figure 4(b)). The higher concentration of non-CpG ODN used here induced modest elevation of CD69 in both strains. The presence of OVA induced further increase only in DO11.10 mice. In a similar manner, the presence of OVA enhanced IgG production only in transgenic mice (Figure 4(c)). Therefore, it is not OVA itself but the interaction of B cells presenting the peptide to T cells in its presence that further increases mutual activation of the cells and enhances antibody production and isotype switching.


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)

No synergism between OVA and ODN treatments when cognate T cells are not present. Wild type (wt) or TCR transgenic (tg) splenic cells were incubated with different combinations of ODN and OVA for 1 day ((a), (b)) or 4 days (c). After 1 day CD69 expression was measured on B and T cell populations by flow cytometry ((a), (b)). Cells from the 4-day cultures were washed and then transferred onto nitrocellulose-covered slides that were previously coated with light chain capture antibodies. After 10 h of incubation, slides were washed and spots indicating IgG producing ASCs were detected using fluorescently labeled antibodies and a fluorescent scanner (c). Results represent a single experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: No synergism between OVA and ODN treatments when cognate T cells are not present. Wild type (wt) or TCR transgenic (tg) splenic cells were incubated with different combinations of ODN and OVA for 1 day ((a), (b)) or 4 days (c). After 1 day CD69 expression was measured on B and T cell populations by flow cytometry ((a), (b)). Cells from the 4-day cultures were washed and then transferred onto nitrocellulose-covered slides that were previously coated with light chain capture antibodies. After 10 h of incubation, slides were washed and spots indicating IgG producing ASCs were detected using fluorescently labeled antibodies and a fluorescent scanner (c). Results represent a single experiment.
Mentions: To further confirm that non-CpG ODN synergize with T cell derived stimuli in enhancing antibody production and isotype switching in B cells, we repeated experiments using wild type (wt) mice. While some OVA specific T cells may be present in naïve wt mice, their numbers are expected to be so low as to be negligible (1 : 10 000 or lower). Indeed, the percentage of CD69 positive T cells was unaffected by the addition of OVA in wt mice (Figure 4(a)). In the absence of OVA, the pattern of CD69 expression in B cells was similar in wt and DO11.10 mice (Figure 4(b)). The higher concentration of non-CpG ODN used here induced modest elevation of CD69 in both strains. The presence of OVA induced further increase only in DO11.10 mice. In a similar manner, the presence of OVA enhanced IgG production only in transgenic mice (Figure 4(c)). Therefore, it is not OVA itself but the interaction of B cells presenting the peptide to T cells in its presence that further increases mutual activation of the cells and enhances antibody production and isotype switching.

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.


Related in: MedlinePlus