southern biotech 1070 05 Search Results


human ads hrp southern biotech  (SouthernBiotech)
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SouthernBiotech human ads hrp southern biotech
Human Ads Hrp Southern Biotech, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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goat anti mouse igg1 hrp conjugate  (SouthernBiotech)
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igg2b horseradish peroxidase southern biotech  (SouthernBiotech)
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SouthernBiotech igg2b horseradish peroxidase southern biotech
Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
Igg2b Horseradish Peroxidase Southern Biotech, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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horseradish peroxidase conjugated antibody  (SouthernBiotech)
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Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
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goat anti rabbit  (SouthernBiotech)
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SouthernBiotech goat anti rabbit
Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
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room temperature  (SouthernBiotech)
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SouthernBiotech room temperature
Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
Room Temperature, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rat anti mouse ige  (SouthernBiotech)
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SouthernBiotech rat anti mouse ige
Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
Rat Anti Mouse Ige, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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5 w ytterbium-doped fibre laser ylg-1070-5lp  (IPG Photonics)
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IPG Photonics 5 w ytterbium-doped fibre laser ylg-1070-5lp
Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum <t>IgG1</t> (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.
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igg2a  (SouthernBiotech)
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NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific <t>IgG</t> antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01
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ig hrp conjugated secondary antibodies  (SouthernBiotech)
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NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific <t>IgG</t> antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01
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horseradish peroxidase hrp  (SouthernBiotech)
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NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific <t>IgG</t> antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01
Horseradish Peroxidase Hrp, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hpr-conjugated goat anti-mouse secondary anti-igg, #1030-05, anti-igg1 #1070-05, anti-igg2a #1080-05, anti-igm #1020-05  (SouthernBiotech)
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SouthernBiotech hpr-conjugated goat anti-mouse secondary anti-igg, #1030-05, anti-igg1 #1070-05, anti-igg2a #1080-05, anti-igm #1020-05
NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific <t>IgG</t> antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01
Hpr Conjugated Goat Anti Mouse Secondary Anti Igg, #1030 05, Anti Igg1 #1070 05, Anti Igg2a #1080 05, Anti Igm #1020 05, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum IgG1 (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Cell metabolism

Article Title: T-bet + B cells accumulate in adipose tissue and exacerbate metabolic disorder during obesity.

doi: 10.1016/j.cmet.2022.07.002

Figure Lengend Snippet: Figure 3. Expansion of T-bet+ CD11c+ B cells in diet-induced obesity depends on iNKT cells (A) Flow cytometry of lymphocytes from WT or Cd1d1/ mice fed the NCD or HFD, presented with the t-SNE algorithm. See also Figure S4. Color in top row cell maps indicates expression of CD19: blue (low) to red (high). Second row cell maps indicate subgating of B cell cluster 1 (blue) and 2 (red). Histograms show expression of T-bet (third row) and CD11c (bottom row) by clusters identified in t-SNE maps: B cell cluster 1 (blue) and 2 (red). (B–D) Representative flow cytometry of splenic (B) and quantification of WAT and splenic (C and D) T-bet+ CD11c+ cell frequency of total B cells. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). (E) Flow cytometric quantification of numbers of NK1.1+ iNKT cells in WAT from WT mice fed an NCD (gray) or HFD (red). (F and G) ELISA measure of serum IgG1 (F), IgG2c (G), and IgM (H) antibodies from WT or Cd1d1/ mice fed the NCD or HFD. There is a correlation between frequency of splenic T-bet+ CD11c+ B cells and IgG1 (F), IgG2c (G), and IgM (H), assessed by flow cytometry and ELISA, respectively. WT NCD (white) and HFD (black) or Cd1d1/ NCD (light blue) and HFD (dark blue). Dots represent individual mice. Data pooled from 7 (A–D), 2 (E), or 3 (F and G) experiments with at least 3 mice/group. Mean ± SEM; two-way ANOVA (B and C) or Student’s t test and Student’s paired t test (D–F). Lines indicate linear regression trend with coefficient of determination (r2); *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER CD3 BV510 Biolegend 317332; RRID: AB_2561943 Va24-Ja18 Brilliant Violet 421 Biolegend 342915; RRID: AB_2564004 CD15 FITC Biolegend 301904; RRID: AB_314196 T-bet BV785 Biolegend 644835; RRID: AB_2721566 Tbet PECy7 Biolegend 644824; RRID: AB_2561761 Isotype MOPC-21; IgG1 k BV785 Biolegend 400169; RRID: AB_11219601 CD11c BV650 Biolegend 301637; RRID: AB_2562231 CD69 BV650 Biolegend 310933; RRID: AB_2561783 CD27 APC Clone O323 Tonbo 20-0279-T025; RRID: AB_2621569 CD27 Clone LG.3A10 Biolegend 124211; RRID: AB_1236460 IgD BV421 Biolegend 348225; RRID: AB_2561618 Total capture Invitrogen PA1-85986 IgG Horseradish Peroxidase Southern Biotech 1030-05 IgM Horseradish Peroxidase Southern Biotech 1020-05 IgG1 Horseradish Peroxidase Southern Biotech 1070-05 IgG2b Horseradish Peroxidase Southern Biotech 1090-05 IgG2c Horseradish Peroxidase Southern Biotech 1078-05 IgG3 Horseradish Peroxidase Southern Biotech 1100-05 IgA Horseradish Peroxidase Southern Biotech 1040-05 IgE Horseradish Peroxidase Southern Biotech 1110-05 Total capture Southern Biotech 2010-01 IgG Horseradish Peroxidase Jackson ImmunoResearch N/A IgM Horseradish Peroxidase Southern Biotech 2020-05 IgG1 Horseradish Peroxidase Southern Biotech 9052-05 IgG2 Horseradish Peroxidase Southern Biotech 9060-05 IgG3 Horseradish Peroxidase Southern Biotech 9210-05 IgG4 Horseradish Peroxidase Southern Biotech 9200-05 IgA Horseradish Peroxidase Jackson ImmunoResearch N/A IgE Horseradish Peroxidase Southern Biotech 9160-05 Rabbit Serum Fisher; Sigma ICN1935780; R4505-500 Goat Serum Sigma G6767 Biological samples Human samples San Antonio Plastic Surgery Institute/Christus Santa Rosa Hospital N/A Chemicals, peptides, and recombinant proteins Alpha Galactosyl-Ceramide KRN7000 Enzo Life Sciences; Avanti Polar lipids Cat# BML-SL232; 867000P-1mg Resiquimod Sigma SML0196-10MG CD1d-tetramers NIH Tetramer Core Facility N/A Brefeldin A (GolgiPlug) Biolegend 420601 FBS Cytiva SH30088.03 L-Glutamine Fisher 25-005-CI RPMI 1640 medium Fisher MT10040CV Collagenase Sigma C5138-5G Streptavidin-Horseradish Peroxidase Fisher; BD Pharmingen SNN2004 (nowS911); 554066 Glucose Sigma G8769-100ML Formalin Fisher 22-050-104 OCT Fisher 23-730-571 (Continued on next page) e2 Cell Metabolism 34, 1121–1136.e1–e6, August 2, 2022

Techniques: Flow Cytometry, Expressing, Cytometry, Enzyme-linked Immunosorbent Assay

NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific IgG antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01

Journal: Brazilian Journal of Microbiology

Article Title: Nano-multilamellar lipid vesicles (NMVs) enhance protective antibody responses against Shiga toxin (Stx2a) produced by enterohemorrhagic Escherichia coli strains (EHEC)

doi: 10.1007/s42770-018-0035-0

Figure Lengend Snippet: NMV-OVA induce high antigen-specific titers and long-lasting serum antibody response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of the vaccine formulations: OVA protein, NMV-OVA, and NMVs. Serum was collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose. On day 91 after the last dose, mice were inoculated with a boost of the OVA antigen (25 μg), except for the control group (NMV). a Schematic representation of the vaccination protocol. b, c Anti-OVA-specific IgG antibody titers from sera of 14 days after each dose analyzed by ELISA (b) or 60, 90, and 105 days after the third dose (c). The antibody titers were determined by ELISA and the data correspond to mean values ± SD of individual experiments carried out in duplicate. Titers values (MEAN + 2 × SD) of the values obtained in the control group (NMV) were subtracted from the titer values determined for the other immunization groups. Statistical differences were determined by One-way ANOVA with Bonferroni post-test ***p < 0.001. Intergroup analysis carried out with Student’s t test was applied with •••p < 0.001 and ••p < 0.01

Article Snippet: In parallel, plaques were incubated with HRP-conjugated anti-mouse monoclonal IgG1 or IgG2a (SOUTHERN BIOTECH—catalog number 1070-05 and 1080-05) under the same conditions.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Protective immunity to Stx2 conferred by immunization with NMV-rStx2B. a Schematic representation of the tested vaccination regimen. BALB/c mice were immunized s.c. with three doses of each vaccine formulation: rStx2B or NMV-rStx2B. Blood samples were collected 14 days after the administration of each dose, and the challenge was carried out with Stx2a toxin 21 days after the last dose. b–c) Analysis of the serum Stx2B-specific total IgG and IgG subclass responses 14 days following each dose (b) or 14, 60, 90, and 105 days after the third dose (c). The values represent the mean ± SD (n = 10 mice/group) for each group. Statistical differences were determined by one-way ANOVA with Bonferroni’s post-hoc test in the analysis considering the doses and the subclasses of each group (***p < 0.001, *p < 0.05) and by t test (•••p < 0.001, ••p < 0.01) in the intergroup analysis. d In vitro neutralization of the Stx2 toxicity. Vero cells were exposed to the recombinant Stx2 holotoxin (1 × CD50–0.75 ng) previously incubated with diluted tested serum samples. Values correspond to the average of two independent experiments performed in duplicates. e Protective immunity conferred by vaccination with NMV-rStx2B. Mice (n = 10/group) were challenged with the toxin Stx2 (1 × LD100 = 3 ng) on the 21st day after the last immunization dose. The non-immunized group was not submitted to any treatment. Statistical analyses were carried out with the Mantel-Cox test (***p < 0.001)

Journal: Brazilian Journal of Microbiology

Article Title: Nano-multilamellar lipid vesicles (NMVs) enhance protective antibody responses against Shiga toxin (Stx2a) produced by enterohemorrhagic Escherichia coli strains (EHEC)

doi: 10.1007/s42770-018-0035-0

Figure Lengend Snippet: Protective immunity to Stx2 conferred by immunization with NMV-rStx2B. a Schematic representation of the tested vaccination regimen. BALB/c mice were immunized s.c. with three doses of each vaccine formulation: rStx2B or NMV-rStx2B. Blood samples were collected 14 days after the administration of each dose, and the challenge was carried out with Stx2a toxin 21 days after the last dose. b–c) Analysis of the serum Stx2B-specific total IgG and IgG subclass responses 14 days following each dose (b) or 14, 60, 90, and 105 days after the third dose (c). The values represent the mean ± SD (n = 10 mice/group) for each group. Statistical differences were determined by one-way ANOVA with Bonferroni’s post-hoc test in the analysis considering the doses and the subclasses of each group (***p < 0.001, *p < 0.05) and by t test (•••p < 0.001, ••p < 0.01) in the intergroup analysis. d In vitro neutralization of the Stx2 toxicity. Vero cells were exposed to the recombinant Stx2 holotoxin (1 × CD50–0.75 ng) previously incubated with diluted tested serum samples. Values correspond to the average of two independent experiments performed in duplicates. e Protective immunity conferred by vaccination with NMV-rStx2B. Mice (n = 10/group) were challenged with the toxin Stx2 (1 × LD100 = 3 ng) on the 21st day after the last immunization dose. The non-immunized group was not submitted to any treatment. Statistical analyses were carried out with the Mantel-Cox test (***p < 0.001)

Article Snippet: In parallel, plaques were incubated with HRP-conjugated anti-mouse monoclonal IgG1 or IgG2a (SOUTHERN BIOTECH—catalog number 1070-05 and 1080-05) under the same conditions.

Techniques: Formulation, In Vitro, Neutralization, Recombinant, Incubation

NMV-rStx2B vaccine formulation improves magnitude and quality of the Stx2-specifi humoral immune response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of rStx2B or NMV-rStx2B. Ninety-one days after the last dose, the animals were inoculated with a 25-μg dose of the rStx2B protein, except for the control group (NMV). Sera were collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose and analyzed by ELISA assay (as described in the Methodology section), and the anti-rstx2 titers were measured. a Schematic representation of the vaccination protocol. b, c Evaluation of anti-rStx2B IgG response from sera harvested 14 days after each vaccine dose (b) or 60, 90, and 105 days after the last dose (c). The antibody titers were determined by ELISA and the values obtained for the control group (NMV) were subtracted from the titers determined for the experimental groups (MEAN + 2 × SD). d Antibody affinity of the sera collected at different days after the third dose. Bars represent the molar concentrations of ammonium thiocyanate required to dissociate 50% of the antibodies bound to Stx2B. e Stx2 Neutralizing activity of the anti-Stx2B antibodies against the Stx2-induced cytotoxic effects in cell culture. The maximum dilution required to preserve 50% of the viable cells were determined for each serum sample. The data correspond to the mean values ± SE of the individual values for each group in duplicate. Statistical differences are indicated by brackets: ***p < 0.001, **p < 0.01, *p < 0.05, for one-way ANOVA with Bonferroni’s post-test for the analysis as a function of the day each dose inoculation •••p < 0.001, ••p < 0.01, •p < 0.05 (t test) for the intergroup analysis

Journal: Brazilian Journal of Microbiology

Article Title: Nano-multilamellar lipid vesicles (NMVs) enhance protective antibody responses against Shiga toxin (Stx2a) produced by enterohemorrhagic Escherichia coli strains (EHEC)

doi: 10.1007/s42770-018-0035-0

Figure Lengend Snippet: NMV-rStx2B vaccine formulation improves magnitude and quality of the Stx2-specifi humoral immune response. Female BALB/c mice (n = 5/group) were subcutaneously immunized with three doses of rStx2B or NMV-rStx2B. Ninety-one days after the last dose, the animals were inoculated with a 25-μg dose of the rStx2B protein, except for the control group (NMV). Sera were collected on days 7, 14, 21, and 28 after each dose and at 60, 90, and 105 days after the last dose and analyzed by ELISA assay (as described in the Methodology section), and the anti-rstx2 titers were measured. a Schematic representation of the vaccination protocol. b, c Evaluation of anti-rStx2B IgG response from sera harvested 14 days after each vaccine dose (b) or 60, 90, and 105 days after the last dose (c). The antibody titers were determined by ELISA and the values obtained for the control group (NMV) were subtracted from the titers determined for the experimental groups (MEAN + 2 × SD). d Antibody affinity of the sera collected at different days after the third dose. Bars represent the molar concentrations of ammonium thiocyanate required to dissociate 50% of the antibodies bound to Stx2B. e Stx2 Neutralizing activity of the anti-Stx2B antibodies against the Stx2-induced cytotoxic effects in cell culture. The maximum dilution required to preserve 50% of the viable cells were determined for each serum sample. The data correspond to the mean values ± SE of the individual values for each group in duplicate. Statistical differences are indicated by brackets: ***p < 0.001, **p < 0.01, *p < 0.05, for one-way ANOVA with Bonferroni’s post-test for the analysis as a function of the day each dose inoculation •••p < 0.001, ••p < 0.01, •p < 0.05 (t test) for the intergroup analysis

Article Snippet: In parallel, plaques were incubated with HRP-conjugated anti-mouse monoclonal IgG1 or IgG2a (SOUTHERN BIOTECH—catalog number 1070-05 and 1080-05) under the same conditions.

Techniques: Formulation, Control, Enzyme-linked Immunosorbent Assay, Activity Assay, Cell Culture