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Differential Immune Response against Recombinant Leishmania donovani Peroxidoxin 1 and Peroxidoxin 2 Proteins in BALB/c Mice.

Daifalla NS, Bayih AG, Gedamu L - J Immunol Res (2015)

Bottom Line: We also evaluated the effect of coadministration of TLR agonists (CpG ODN and GLA-SE) on the antigen-specific immune response.Immunization with recombinant LdPxn1 alone induced a predominantly Th2 type immune response that is associated with the production of high level of IgG1 and no IgG2a isotype while rLdPxn2 resulted in a mixed Th1/Th2 response characterized by the production of antigen-specific IgG2a in addition to IgG1 isotype.TLR agonists also enhanced a more polarized Th 1 type immune response against rLdPxn2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of Calgary, Room 374, 2500 University Drive NW, Calgary, AB, Canada T2N 1N4 ; The Forsyth Institute, Cambridge, MA 02142, USA.

ABSTRACT
We assessed the immune response against recombinant proteins of two related, albeit functionally different, peroxidoxins from Leishmania donovani: peroxidoxin 1 (LdPxn1) and peroxidoxin 2 (LdPxn2) in BALB/c mice. We also evaluated the effect of coadministration of TLR agonists (CpG ODN and GLA-SE) on the antigen-specific immune response. Immunization with recombinant LdPxn1 alone induced a predominantly Th2 type immune response that is associated with the production of high level of IgG1 and no IgG2a isotype while rLdPxn2 resulted in a mixed Th1/Th2 response characterized by the production of antigen-specific IgG2a in addition to IgG1 isotype. Antigen-stimulated spleen cells from mice that were immunized with rLdPxn1 produced low level of IL-10 and IL-4 and no IFN-γ, whereas cells from mice immunized with rLdPxn2 secreted high level of IFN-γ, low IL-4, and no IL-10. Coadministration of CpG ODN or GLA-SE with rLdPxn1 skewed the immune response towards a Th 1 type as indicated by robust production of IgG2a isotype. Furthermore, the presence of TLR agonists together with rLdPxn1 antigen enhanced the production of IFN-γ and to a lesser extent of IL-10. TLR agonists also enhanced a more polarized Th 1 type immune response against rLdPxn2.

No MeSH data available.


Cytokine responses in lymph node cells of rLdPxn1 and rLdPxn2 immunized mice. Mice were immunized subcutaneously three times at three-week intervals with rLdPxn1 or rLdPxn2 with or without CpG ODN or GLA-SE. Four weeks after the last immunization, cells from pooled lymph nodes were stimulated in vitro with the respective antigen (10 μg/mL) or ConA (5 μg/mL). The release of IFN-γ and IL-10 in mice immunized with rLdPxn1 or rLdPxn2 was measured in supernatants after 72 hr of in vitro stimulation at 37°C. Results are presented as the amount of IFN-γ (ng/mL) and IL-10 (ng/mL) for rLdPxn1 (a) and rLdPxn2 (b).
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fig4: Cytokine responses in lymph node cells of rLdPxn1 and rLdPxn2 immunized mice. Mice were immunized subcutaneously three times at three-week intervals with rLdPxn1 or rLdPxn2 with or without CpG ODN or GLA-SE. Four weeks after the last immunization, cells from pooled lymph nodes were stimulated in vitro with the respective antigen (10 μg/mL) or ConA (5 μg/mL). The release of IFN-γ and IL-10 in mice immunized with rLdPxn1 or rLdPxn2 was measured in supernatants after 72 hr of in vitro stimulation at 37°C. Results are presented as the amount of IFN-γ (ng/mL) and IL-10 (ng/mL) for rLdPxn1 (a) and rLdPxn2 (b).

Mentions: There was no spontaneous release of cytokines by unstimulated lymph node cells in any of the groups (Figures 4(a) and 4(b)). No detectable cytokine was released by lymph node cells from mice immunized with the recombinant proteins alone (Figures 4(a) and 4(b)). However, immunization of mice with the recombinant proteins in the presence of TLR agonists resulted in the production of a high level of IFN-γ and a low level of IL-10 (Figures 4(a) and 4(b)). While coadministration of CpG ODN triggered the production of similar amounts of IFN-γ in mice immunized with rLdPxn1 or rLdPxn2, GLA-SE stimulated the release of more IFN-γ in the group receiving rLdPxn2. The amount of IFN-γ produced by lymph node cells of mice from this group (Figure 4(b)) was more than 3-fold higher than the level produced by lymph node cells from mice immunized with rLdPxn1 alone (Figure 4(a)). Interestingly, CpG ODN and GLA-SE triggered the production of lower amount of IL-10 in mice receiving rLdPxn1 as compared to rLdPxn2 (Figures 4(a) and 4(b)). Lymph node cells stimulated with ConA mitogen produced comparable levels of cytokines (Figures 4(a) and 4(b) in sets) with the exception of the group immunized with rLdPxn2 alone which produces lower level of cytokines (Figure 4(b) in set). The results of cytokine analyses in lymph node cells demonstrate that rLdPxn1 and rLdPxn2 can stimulate lymph node cells of immunized mice to produce cytokines only in the presence of adjuvants. The results also show that CpG ODN and GLA-SE adjuvants favor a Th1 type response against the two antigens as indicated by the high IFN-γ/IL-10 ratios (Table 1). Moreover, the results show that while coadministration of GLA-SE exerts comparable effect on both antigens as indicated by comparable ratios of IFN-γ/IL-10 (Table 1), CpG ODN induces stronger Th1 in mice receiving rLdPxn1 as compared to rLdPxn2 (IFN-γ/IL-10 ratio of 24.74 and 10.83 for rLdPxn1 and rLdPxn2, resp.) (Table 1).


Differential Immune Response against Recombinant Leishmania donovani Peroxidoxin 1 and Peroxidoxin 2 Proteins in BALB/c Mice.

Daifalla NS, Bayih AG, Gedamu L - J Immunol Res (2015)

Cytokine responses in lymph node cells of rLdPxn1 and rLdPxn2 immunized mice. Mice were immunized subcutaneously three times at three-week intervals with rLdPxn1 or rLdPxn2 with or without CpG ODN or GLA-SE. Four weeks after the last immunization, cells from pooled lymph nodes were stimulated in vitro with the respective antigen (10 μg/mL) or ConA (5 μg/mL). The release of IFN-γ and IL-10 in mice immunized with rLdPxn1 or rLdPxn2 was measured in supernatants after 72 hr of in vitro stimulation at 37°C. Results are presented as the amount of IFN-γ (ng/mL) and IL-10 (ng/mL) for rLdPxn1 (a) and rLdPxn2 (b).
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Related In: Results  -  Collection

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fig4: Cytokine responses in lymph node cells of rLdPxn1 and rLdPxn2 immunized mice. Mice were immunized subcutaneously three times at three-week intervals with rLdPxn1 or rLdPxn2 with or without CpG ODN or GLA-SE. Four weeks after the last immunization, cells from pooled lymph nodes were stimulated in vitro with the respective antigen (10 μg/mL) or ConA (5 μg/mL). The release of IFN-γ and IL-10 in mice immunized with rLdPxn1 or rLdPxn2 was measured in supernatants after 72 hr of in vitro stimulation at 37°C. Results are presented as the amount of IFN-γ (ng/mL) and IL-10 (ng/mL) for rLdPxn1 (a) and rLdPxn2 (b).
Mentions: There was no spontaneous release of cytokines by unstimulated lymph node cells in any of the groups (Figures 4(a) and 4(b)). No detectable cytokine was released by lymph node cells from mice immunized with the recombinant proteins alone (Figures 4(a) and 4(b)). However, immunization of mice with the recombinant proteins in the presence of TLR agonists resulted in the production of a high level of IFN-γ and a low level of IL-10 (Figures 4(a) and 4(b)). While coadministration of CpG ODN triggered the production of similar amounts of IFN-γ in mice immunized with rLdPxn1 or rLdPxn2, GLA-SE stimulated the release of more IFN-γ in the group receiving rLdPxn2. The amount of IFN-γ produced by lymph node cells of mice from this group (Figure 4(b)) was more than 3-fold higher than the level produced by lymph node cells from mice immunized with rLdPxn1 alone (Figure 4(a)). Interestingly, CpG ODN and GLA-SE triggered the production of lower amount of IL-10 in mice receiving rLdPxn1 as compared to rLdPxn2 (Figures 4(a) and 4(b)). Lymph node cells stimulated with ConA mitogen produced comparable levels of cytokines (Figures 4(a) and 4(b) in sets) with the exception of the group immunized with rLdPxn2 alone which produces lower level of cytokines (Figure 4(b) in set). The results of cytokine analyses in lymph node cells demonstrate that rLdPxn1 and rLdPxn2 can stimulate lymph node cells of immunized mice to produce cytokines only in the presence of adjuvants. The results also show that CpG ODN and GLA-SE adjuvants favor a Th1 type response against the two antigens as indicated by the high IFN-γ/IL-10 ratios (Table 1). Moreover, the results show that while coadministration of GLA-SE exerts comparable effect on both antigens as indicated by comparable ratios of IFN-γ/IL-10 (Table 1), CpG ODN induces stronger Th1 in mice receiving rLdPxn1 as compared to rLdPxn2 (IFN-γ/IL-10 ratio of 24.74 and 10.83 for rLdPxn1 and rLdPxn2, resp.) (Table 1).

Bottom Line: We also evaluated the effect of coadministration of TLR agonists (CpG ODN and GLA-SE) on the antigen-specific immune response.Immunization with recombinant LdPxn1 alone induced a predominantly Th2 type immune response that is associated with the production of high level of IgG1 and no IgG2a isotype while rLdPxn2 resulted in a mixed Th1/Th2 response characterized by the production of antigen-specific IgG2a in addition to IgG1 isotype.TLR agonists also enhanced a more polarized Th 1 type immune response against rLdPxn2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of Calgary, Room 374, 2500 University Drive NW, Calgary, AB, Canada T2N 1N4 ; The Forsyth Institute, Cambridge, MA 02142, USA.

ABSTRACT
We assessed the immune response against recombinant proteins of two related, albeit functionally different, peroxidoxins from Leishmania donovani: peroxidoxin 1 (LdPxn1) and peroxidoxin 2 (LdPxn2) in BALB/c mice. We also evaluated the effect of coadministration of TLR agonists (CpG ODN and GLA-SE) on the antigen-specific immune response. Immunization with recombinant LdPxn1 alone induced a predominantly Th2 type immune response that is associated with the production of high level of IgG1 and no IgG2a isotype while rLdPxn2 resulted in a mixed Th1/Th2 response characterized by the production of antigen-specific IgG2a in addition to IgG1 isotype. Antigen-stimulated spleen cells from mice that were immunized with rLdPxn1 produced low level of IL-10 and IL-4 and no IFN-γ, whereas cells from mice immunized with rLdPxn2 secreted high level of IFN-γ, low IL-4, and no IL-10. Coadministration of CpG ODN or GLA-SE with rLdPxn1 skewed the immune response towards a Th 1 type as indicated by robust production of IgG2a isotype. Furthermore, the presence of TLR agonists together with rLdPxn1 antigen enhanced the production of IFN-γ and to a lesser extent of IL-10. TLR agonists also enhanced a more polarized Th 1 type immune response against rLdPxn2.

No MeSH data available.