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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.


Schematic representation of the immunization protocol. Six- to 8-week-old female BALB/c mice were immunized s.c. in the hind foot pad with recombinant LdPxn1 or LdPxn2 protein (25 ug/mouse) with or without CpG ODN (50 ug/mouse) or GLA-SE (20 ug/mouse) adjuvants. Mice were boosted twice in 3 weeks interval. Sera were collected at each time of injection. Four weeks after the last boost mice were euthanized and sera, lymph nodes, and spleens were collected. Samples were used for antibody and cytokine analysis using standard protocols.
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fig2: Schematic representation of the immunization protocol. Six- to 8-week-old female BALB/c mice were immunized s.c. in the hind foot pad with recombinant LdPxn1 or LdPxn2 protein (25 ug/mouse) with or without CpG ODN (50 ug/mouse) or GLA-SE (20 ug/mouse) adjuvants. Mice were boosted twice in 3 weeks interval. Sera were collected at each time of injection. Four weeks after the last boost mice were euthanized and sera, lymph nodes, and spleens were collected. Samples were used for antibody and cytokine analysis using standard protocols.

Mentions: Immunization protocol is schematically represented in Figure 2. Mice were randomly divided into groups of three and were immunized subcutaneously (s.c.) with recombinant LdPxn1, rLdPxn1 plus CpG ODN, rLdPxn1 plus GLA-SE, rLdPxn2, rLdPxn2 plus CpG ODN, and rLdPxn2 plus GLA-SE. Recombinant proteins, CpG ODN 1826 (Coley Pharmaceutical Group, Canada) and GLA-SE (Infectious Disease Research Institute, Seattle, USA), were given at 25, 50, and 20 μg/mouse, respectively. Two booster injections were given in three-week interval. Sera were isolated from blood (collected every three weeks starting from the time of first immunization (week 0) until the time of euthanization) and stored at –20°C. Mice were euthanized four weeks after the last boost, and lymph node and spleens were aseptically harvested and processed for the isolation of single cell suspensions. The isolated lymph node and spleen cells were used for in vitro antigen stimulation experiments.


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)

Schematic representation of the immunization protocol. Six- to 8-week-old female BALB/c mice were immunized s.c. in the hind foot pad with recombinant LdPxn1 or LdPxn2 protein (25 ug/mouse) with or without CpG ODN (50 ug/mouse) or GLA-SE (20 ug/mouse) adjuvants. Mice were boosted twice in 3 weeks interval. Sera were collected at each time of injection. Four weeks after the last boost mice were euthanized and sera, lymph nodes, and spleens were collected. Samples were used for antibody and cytokine analysis using standard protocols.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4562178&req=5

fig2: Schematic representation of the immunization protocol. Six- to 8-week-old female BALB/c mice were immunized s.c. in the hind foot pad with recombinant LdPxn1 or LdPxn2 protein (25 ug/mouse) with or without CpG ODN (50 ug/mouse) or GLA-SE (20 ug/mouse) adjuvants. Mice were boosted twice in 3 weeks interval. Sera were collected at each time of injection. Four weeks after the last boost mice were euthanized and sera, lymph nodes, and spleens were collected. Samples were used for antibody and cytokine analysis using standard protocols.
Mentions: Immunization protocol is schematically represented in Figure 2. Mice were randomly divided into groups of three and were immunized subcutaneously (s.c.) with recombinant LdPxn1, rLdPxn1 plus CpG ODN, rLdPxn1 plus GLA-SE, rLdPxn2, rLdPxn2 plus CpG ODN, and rLdPxn2 plus GLA-SE. Recombinant proteins, CpG ODN 1826 (Coley Pharmaceutical Group, Canada) and GLA-SE (Infectious Disease Research Institute, Seattle, USA), were given at 25, 50, and 20 μg/mouse, respectively. Two booster injections were given in three-week interval. Sera were isolated from blood (collected every three weeks starting from the time of first immunization (week 0) until the time of euthanization) and stored at –20°C. Mice were euthanized four weeks after the last boost, and lymph node and spleens were aseptically harvested and processed for the isolation of single cell suspensions. The isolated lymph node and spleen cells were used for in vitro antigen stimulation experiments.

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.