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Oct4 promoted cytoplasmic translocation of c-Myc. A Heatmap of c-Myc-differentially expressed genes (DEGs, change relative to the mean) in vector-transfected cMSCs and Oct4-overexpressed cMSCs. The top 10 highly expressed proangiogenic factors are listed. Heatmap colors indicate directionality (red: increased; blue: decreased). B RNAs extracted from vector-transfected cMSCs and Oct4-overexpressed cMSCs were subjected to real-time RT-PCR. Significantly higher levels of c-Myc target factors were detected in the Oct4-overexpressed cMSCs than in the vector-transfected cMSCs. C The c-Myc null cMSCs were transfected with Oct4 or a control vector followed by real-time RT-PCR to confirm no significant difference in the expression of c-Myc. D Tube formation ability of cMSCs transfected with Oct4 or control vector. All data are the means ± SEM; statistical significance was evaluated using the unpaired two-tailed Student’s t test with Welch’s correction. E Typical images of capillary-like net-work formation of cMSCs from these two groups. F The siRNA-transfected cells expressed significantly lower levels of VEGF signaling signals than the control. All data are the means ± SEM, and independent samples t test was used. G The mRNA expression of Oct4 and c-Myc in the cytoplasm and the nuclei of c-Myc null cMSCs were detected by real-time RT-PCR. GAPDH was served as positive control. GAPDH and c-Myc were mainly expressed in the cytoplasm, while significantly higher levels of Oct4 were detected in the nuclei than in the cytoplasm. P value for the difference between groups was calculated by the nonparametric Kruskal–Wallis test followed by a Dunn multiple comparisons test. H , RNAs extracted from cytoplasm and nuclei of cMSCs transfected with Oct4, Oct4 siRNA, or the vector were subjected to real-time RT-PCR. Significantly higher levels of c-Myc were detected in the cytoplasm than in the nuclei. All data are the means ± SEM, and independent samples t test was used ( n = 10, each group). I Western blotting revealed similar protein levels of c-Myc expression in the cell lysates. oe Oct4-transfected cMSCs showed more expression of cytoplasmic c-Myc than vector-transfected cMSCs, but Oct4 siRNA reversed this trend of expression, disclosing the dense expression of this factor in the nuclei. K Chromatin immunoprecipitation (ChIP) assay for the binding of Oct4 to c-Myc promoter. <t>Anti-IgG</t> was used as a negative control, anti-RNA-polymerase II was used as a positive control
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1) Product Images from "Oct4 cooperates with c-Myc to improve mesenchymal-to-endothelial transition and myocardial repair of cardiac-resident mesenchymal stem cells"

Article Title: Oct4 cooperates with c-Myc to improve mesenchymal-to-endothelial transition and myocardial repair of cardiac-resident mesenchymal stem cells

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-022-03120-7

Oct4 promoted cytoplasmic translocation of c-Myc. A Heatmap of c-Myc-differentially expressed genes (DEGs, change relative to the mean) in vector-transfected cMSCs and Oct4-overexpressed cMSCs. The top 10 highly expressed proangiogenic factors are listed. Heatmap colors indicate directionality (red: increased; blue: decreased). B RNAs extracted from vector-transfected cMSCs and Oct4-overexpressed cMSCs were subjected to real-time RT-PCR. Significantly higher levels of c-Myc target factors were detected in the Oct4-overexpressed cMSCs than in the vector-transfected cMSCs. C The c-Myc null cMSCs were transfected with Oct4 or a control vector followed by real-time RT-PCR to confirm no significant difference in the expression of c-Myc. D Tube formation ability of cMSCs transfected with Oct4 or control vector. All data are the means ± SEM; statistical significance was evaluated using the unpaired two-tailed Student’s t test with Welch’s correction. E Typical images of capillary-like net-work formation of cMSCs from these two groups. F The siRNA-transfected cells expressed significantly lower levels of VEGF signaling signals than the control. All data are the means ± SEM, and independent samples t test was used. G The mRNA expression of Oct4 and c-Myc in the cytoplasm and the nuclei of c-Myc null cMSCs were detected by real-time RT-PCR. GAPDH was served as positive control. GAPDH and c-Myc were mainly expressed in the cytoplasm, while significantly higher levels of Oct4 were detected in the nuclei than in the cytoplasm. P value for the difference between groups was calculated by the nonparametric Kruskal–Wallis test followed by a Dunn multiple comparisons test. H , RNAs extracted from cytoplasm and nuclei of cMSCs transfected with Oct4, Oct4 siRNA, or the vector were subjected to real-time RT-PCR. Significantly higher levels of c-Myc were detected in the cytoplasm than in the nuclei. All data are the means ± SEM, and independent samples t test was used ( n = 10, each group). I Western blotting revealed similar protein levels of c-Myc expression in the cell lysates. oe Oct4-transfected cMSCs showed more expression of cytoplasmic c-Myc than vector-transfected cMSCs, but Oct4 siRNA reversed this trend of expression, disclosing the dense expression of this factor in the nuclei. K Chromatin immunoprecipitation (ChIP) assay for the binding of Oct4 to c-Myc promoter. Anti-IgG was used as a negative control, anti-RNA-polymerase II was used as a positive control
Figure Legend Snippet: Oct4 promoted cytoplasmic translocation of c-Myc. A Heatmap of c-Myc-differentially expressed genes (DEGs, change relative to the mean) in vector-transfected cMSCs and Oct4-overexpressed cMSCs. The top 10 highly expressed proangiogenic factors are listed. Heatmap colors indicate directionality (red: increased; blue: decreased). B RNAs extracted from vector-transfected cMSCs and Oct4-overexpressed cMSCs were subjected to real-time RT-PCR. Significantly higher levels of c-Myc target factors were detected in the Oct4-overexpressed cMSCs than in the vector-transfected cMSCs. C The c-Myc null cMSCs were transfected with Oct4 or a control vector followed by real-time RT-PCR to confirm no significant difference in the expression of c-Myc. D Tube formation ability of cMSCs transfected with Oct4 or control vector. All data are the means ± SEM; statistical significance was evaluated using the unpaired two-tailed Student’s t test with Welch’s correction. E Typical images of capillary-like net-work formation of cMSCs from these two groups. F The siRNA-transfected cells expressed significantly lower levels of VEGF signaling signals than the control. All data are the means ± SEM, and independent samples t test was used. G The mRNA expression of Oct4 and c-Myc in the cytoplasm and the nuclei of c-Myc null cMSCs were detected by real-time RT-PCR. GAPDH was served as positive control. GAPDH and c-Myc were mainly expressed in the cytoplasm, while significantly higher levels of Oct4 were detected in the nuclei than in the cytoplasm. P value for the difference between groups was calculated by the nonparametric Kruskal–Wallis test followed by a Dunn multiple comparisons test. H , RNAs extracted from cytoplasm and nuclei of cMSCs transfected with Oct4, Oct4 siRNA, or the vector were subjected to real-time RT-PCR. Significantly higher levels of c-Myc were detected in the cytoplasm than in the nuclei. All data are the means ± SEM, and independent samples t test was used ( n = 10, each group). I Western blotting revealed similar protein levels of c-Myc expression in the cell lysates. oe Oct4-transfected cMSCs showed more expression of cytoplasmic c-Myc than vector-transfected cMSCs, but Oct4 siRNA reversed this trend of expression, disclosing the dense expression of this factor in the nuclei. K Chromatin immunoprecipitation (ChIP) assay for the binding of Oct4 to c-Myc promoter. Anti-IgG was used as a negative control, anti-RNA-polymerase II was used as a positive control

Techniques Used: Translocation Assay, Plasmid Preparation, Transfection, Quantitative RT-PCR, Expressing, Two Tailed Test, Positive Control, Western Blot, Chromatin Immunoprecipitation, Binding Assay, Negative Control

2) Product Images from "An Antibody from Single Human VH-rearranging Mouse Neutralizes All SARS-CoV-2 Variants Through BA.5 by Inhibiting Membrane Fusion"

Article Title: An Antibody from Single Human VH-rearranging Mouse Neutralizes All SARS-CoV-2 Variants Through BA.5 by Inhibiting Membrane Fusion

Journal: Science Immunology

doi: 10.1126/sciimmunol.add5446

Immunizing the V H 1-2/Vκ1-33-rearranging mouse model with SARS-CoV-2 spike or RBD elicited multiple V H 1-2/Vκ1-33 antibodies. ( A ) Immunization scheme. Prime plus boost immunizations were performed at an interval of four weeks. ( B ) Binding curves showing reactivity of sera to SARS-CoV-2 spike, RBD, NTD and SARS-CoV-1 spike protein. The upper panel showed the sera from the SARS-CoV-2 Spike immunized mice at week 0, 2 and 6. The bottom panel showed the sera from VHH7-RBD immunized mice. Data were mean ± SD of three mice. ( C ) Table showed the V H 1-2/Vκ1-33 antibodies isolated from SARS-CoV-2 spike-specific or RBD-specific IgG + B cells. The antibody sequences and sequence features were shown in Table S3.
Figure Legend Snippet: Immunizing the V H 1-2/Vκ1-33-rearranging mouse model with SARS-CoV-2 spike or RBD elicited multiple V H 1-2/Vκ1-33 antibodies. ( A ) Immunization scheme. Prime plus boost immunizations were performed at an interval of four weeks. ( B ) Binding curves showing reactivity of sera to SARS-CoV-2 spike, RBD, NTD and SARS-CoV-1 spike protein. The upper panel showed the sera from the SARS-CoV-2 Spike immunized mice at week 0, 2 and 6. The bottom panel showed the sera from VHH7-RBD immunized mice. Data were mean ± SD of three mice. ( C ) Table showed the V H 1-2/Vκ1-33 antibodies isolated from SARS-CoV-2 spike-specific or RBD-specific IgG + B cells. The antibody sequences and sequence features were shown in Table S3.

Techniques Used: Binding Assay, Mouse Assay, Isolation, Sequencing

SP1-77 inhibited ACE2-mediated S1 dissociation on trypsin-treated VSV-SARS-CoV-2 Atto 565 viruses. ( A ) Histogram of the number of Atto 565 molecules on VSV-SARS-CoV-2-Atto 565 determined by single molecule counting. ( B ) Histogram of averages of peak distribution determined by a Gaussian fit of 3 independent experiments. p values were accessed by unpaired two-tail t-test. Data were shown in Table S7. Treatments prior to adsorption included none (control), incubation with 1 μg/mL trypsin for 30 min at 37°C, treatment without or with trypsin followed by an incubation with 0.5 μM of recombinant ACE2 for 10 min at 37°C, or treatment with trypsin then incubated with 100 ng/mL SP1-77 IgG or SP1-77 Fab for 1 hour at 37°C followed by incubation with ACE2. ( C ) Schematic representation of proposed mechanism of SP1-77 inhibition of SARS-CoV-2 infection. Left panel: Without antibody treatments, the spike protein on the viral surface bound to the ACE2 receptor on the infected cell surface. Membrane fusion was activated either by TMPRSS2 protease on the cell membrane or by cathepsin L protease following endocytosis. Cleavage at the S2’ site by these proteases led to dissociation of the S1 subunit, which exposed the fusion peptide on the S2, facilitating viral-host membrane fusion and viral entry into the infected cells. Middle panel: Pre-treatment of the virus with SP1-77, a non-ACE2-blocking antibody, did not appreciably impact binding of viruses to the cell surface and their endocytosis. However, SP1-77 greatly inhibited the dissociation of S1 subunit, thereby, blocking activation of the fusion peptide and membrane fusion. Right panel: Pre-treatment of the virus with VHH7-5-82, an ACE2-blocking antibody, prevented binding of the virus to the cell surface.
Figure Legend Snippet: SP1-77 inhibited ACE2-mediated S1 dissociation on trypsin-treated VSV-SARS-CoV-2 Atto 565 viruses. ( A ) Histogram of the number of Atto 565 molecules on VSV-SARS-CoV-2-Atto 565 determined by single molecule counting. ( B ) Histogram of averages of peak distribution determined by a Gaussian fit of 3 independent experiments. p values were accessed by unpaired two-tail t-test. Data were shown in Table S7. Treatments prior to adsorption included none (control), incubation with 1 μg/mL trypsin for 30 min at 37°C, treatment without or with trypsin followed by an incubation with 0.5 μM of recombinant ACE2 for 10 min at 37°C, or treatment with trypsin then incubated with 100 ng/mL SP1-77 IgG or SP1-77 Fab for 1 hour at 37°C followed by incubation with ACE2. ( C ) Schematic representation of proposed mechanism of SP1-77 inhibition of SARS-CoV-2 infection. Left panel: Without antibody treatments, the spike protein on the viral surface bound to the ACE2 receptor on the infected cell surface. Membrane fusion was activated either by TMPRSS2 protease on the cell membrane or by cathepsin L protease following endocytosis. Cleavage at the S2’ site by these proteases led to dissociation of the S1 subunit, which exposed the fusion peptide on the S2, facilitating viral-host membrane fusion and viral entry into the infected cells. Middle panel: Pre-treatment of the virus with SP1-77, a non-ACE2-blocking antibody, did not appreciably impact binding of viruses to the cell surface and their endocytosis. However, SP1-77 greatly inhibited the dissociation of S1 subunit, thereby, blocking activation of the fusion peptide and membrane fusion. Right panel: Pre-treatment of the virus with VHH7-5-82, an ACE2-blocking antibody, prevented binding of the virus to the cell surface.

Techniques Used: Single Molecule Counting, Adsorption, Incubation, Recombinant, Inhibition, Infection, Blocking Assay, Binding Assay, Activation Assay

3) Product Images from "Humanized antibody potently neutralizes all SARS-CoV-2 variants by a novel mechanism"

Article Title: Humanized antibody potently neutralizes all SARS-CoV-2 variants by a novel mechanism

Journal: bioRxiv

doi: 10.1101/2022.06.26.497634

Immunizing the mouse model with SARS-CoV-2 spike or RBD elicited potent and broad V H 1-2/Vκ1-33 antibodies. ( A ) Immunization scheme. Prime plus boost immunizations were performed at an interval of four weeks. ( B ) Table shows the V H 1-2/Vκ1-33 antibodies isolated from SARS-CoV-2 spike-specific or RBD-specific IgG + B cells. The antibody sequences and sequence features were shown in table S2 . ( C ) Table shows the neutralization activities of three mAbs against VOCs and VOIs in pseudovirus neutralization assays. Experiments were done in 293T/ACE2 cells. The data are also shown in fig. S4, A and B . Data are representative of 2 biologically independent experiments for most VOCs and VOIs, but one experiment for BA.2 and BA.3. Each independent experiment contains 2 technical replicates. IC 50 values were color-coded based on the key shown at the right. ( D ) Table shows the neutralization activities of three mAbs against VOCs in PRNT live virus neutralization assays. Data are representative of one independent experiment with 2 technical replicates. IC 50 and IC 90 values were color-coded based on the key shown at the right.
Figure Legend Snippet: Immunizing the mouse model with SARS-CoV-2 spike or RBD elicited potent and broad V H 1-2/Vκ1-33 antibodies. ( A ) Immunization scheme. Prime plus boost immunizations were performed at an interval of four weeks. ( B ) Table shows the V H 1-2/Vκ1-33 antibodies isolated from SARS-CoV-2 spike-specific or RBD-specific IgG + B cells. The antibody sequences and sequence features were shown in table S2 . ( C ) Table shows the neutralization activities of three mAbs against VOCs and VOIs in pseudovirus neutralization assays. Experiments were done in 293T/ACE2 cells. The data are also shown in fig. S4, A and B . Data are representative of 2 biologically independent experiments for most VOCs and VOIs, but one experiment for BA.2 and BA.3. Each independent experiment contains 2 technical replicates. IC 50 values were color-coded based on the key shown at the right. ( D ) Table shows the neutralization activities of three mAbs against VOCs in PRNT live virus neutralization assays. Data are representative of one independent experiment with 2 technical replicates. IC 50 and IC 90 values were color-coded based on the key shown at the right.

Techniques Used: Isolation, Sequencing, Neutralization, Plaque Reduction Neutralization Test

Immunizing the mouse model with SARS-CoV-2 spike or RBD elicited V H 1-2/Vκ1-33 antibodies. ( A ) Binding curves showing reactivity of sera to SARS-CoV-2 related antigens. The upper panel shows the sera from the SARS-CoV-2 Spike immunized mice at week 0, 2 and 6. The bottom panel shows the sera from VHH7-RBD immunized mice. Data were mean ± SD of three mice. ( B ) Gating strategy for single cell sorting of SARS-CoV2 spike-specific or RBD-specific IgG + B cells after immunization. The upper panel showed the mice immunized with SARS-CoV2 spike plus poly I:C adjuvant. The bottom panels showed the mice immunized with poly I:C (control) or VHH7-RBD plus poly I:C adjuvant. ( C ) Table shows the binding affinities of nine mAbs to the SARS-CoV-2 spike protein, RBD, NTD. Data are representative of two independent experiments. EC 50 values were color-coded based on the key shown at the right.
Figure Legend Snippet: Immunizing the mouse model with SARS-CoV-2 spike or RBD elicited V H 1-2/Vκ1-33 antibodies. ( A ) Binding curves showing reactivity of sera to SARS-CoV-2 related antigens. The upper panel shows the sera from the SARS-CoV-2 Spike immunized mice at week 0, 2 and 6. The bottom panel shows the sera from VHH7-RBD immunized mice. Data were mean ± SD of three mice. ( B ) Gating strategy for single cell sorting of SARS-CoV2 spike-specific or RBD-specific IgG + B cells after immunization. The upper panel showed the mice immunized with SARS-CoV2 spike plus poly I:C adjuvant. The bottom panels showed the mice immunized with poly I:C (control) or VHH7-RBD plus poly I:C adjuvant. ( C ) Table shows the binding affinities of nine mAbs to the SARS-CoV-2 spike protein, RBD, NTD. Data are representative of two independent experiments. EC 50 values were color-coded based on the key shown at the right.

Techniques Used: Binding Assay, Mouse Assay, FACS

Single molecule calibration of VSV-SARS-CoV-2-Atto565 virus. ( A ) Example image of Atto 565 fluorescence of virus adsorbed for 10 minutes on a glass coverslip coated with poly-D-lysine coated slides and imaged and imaged with spinning disc confocal microscopy. Photobleaching (B), used to determine the intensity of single Atto 565 dye intensities allowed to establish that ~20-40 dyes were attached to each virus. ( C ) Histogram and ( D ) data summary of the number of Atto 565 molecules on VSV-SARS-CoV-2-Atto 565 determined by single molecule counting. Treatments prior to adsorption included none (control), incubation with 1 µg/mL trypsin for 30 min at 37°C, treatment without or with trypsin followed by an incubation with 0.5 µM of recombinant ACE2 for 10 min at 37°C, or treatment with trypsin then incubated with 100 ng/mL SP1-77 IgG or SP1-77 Fab for 1 hour at 37°C followed by incubation with ACE2. ( D ) Statistics are averages of peak distribution determined by a Gaussian fit of 3 independent experiments. ( E ) Infection assays showing neutralization of VSV-SARS-CoV-2-Atto 565 infection with SP1-77 IgG or SP1-77 Fab. A soluble eGFP reporter genetically encoded into this VSV chimera allowed for infection to be determined by fluorescence imaging on a spinning disc confocal microscope as shown in the representative examples obtained 7 hr post infection. Cell outlines were obtained with a WGA-Alexa 647 membrane stain applied to cells immediately prior to fixation. Each condition was measured with 3 independent experiments.
Figure Legend Snippet: Single molecule calibration of VSV-SARS-CoV-2-Atto565 virus. ( A ) Example image of Atto 565 fluorescence of virus adsorbed for 10 minutes on a glass coverslip coated with poly-D-lysine coated slides and imaged and imaged with spinning disc confocal microscopy. Photobleaching (B), used to determine the intensity of single Atto 565 dye intensities allowed to establish that ~20-40 dyes were attached to each virus. ( C ) Histogram and ( D ) data summary of the number of Atto 565 molecules on VSV-SARS-CoV-2-Atto 565 determined by single molecule counting. Treatments prior to adsorption included none (control), incubation with 1 µg/mL trypsin for 30 min at 37°C, treatment without or with trypsin followed by an incubation with 0.5 µM of recombinant ACE2 for 10 min at 37°C, or treatment with trypsin then incubated with 100 ng/mL SP1-77 IgG or SP1-77 Fab for 1 hour at 37°C followed by incubation with ACE2. ( D ) Statistics are averages of peak distribution determined by a Gaussian fit of 3 independent experiments. ( E ) Infection assays showing neutralization of VSV-SARS-CoV-2-Atto 565 infection with SP1-77 IgG or SP1-77 Fab. A soluble eGFP reporter genetically encoded into this VSV chimera allowed for infection to be determined by fluorescence imaging on a spinning disc confocal microscope as shown in the representative examples obtained 7 hr post infection. Cell outlines were obtained with a WGA-Alexa 647 membrane stain applied to cells immediately prior to fixation. Each condition was measured with 3 independent experiments.

Techniques Used: Fluorescence, Confocal Microscopy, Single Molecule Counting, Adsorption, Incubation, Recombinant, Infection, Neutralization, Imaging, Microscopy, Whole Genome Amplification, Staining

4) Product Images from "Identification of Immunodominant Antigens From a First-Generation Vaccine Against Cutaneous Leishmaniasis"

Article Title: Identification of Immunodominant Antigens From a First-Generation Vaccine Against Cutaneous Leishmaniasis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2022.825007

Humoral and cellular immune response in immunized mice. Vaccination scheme (A) . Anti- Leishmania IgG antibodies (B) using serum samples dilution (1:500) obtained 3 weeks after each immunization. Anti- Leishmania IgG1 and IgG2a (C) antibodies using serum samples dilution (1:1,000) obtained 3 weeks after boost. Results are presented as the mean ± standard error of mean (SEM) of optical density (O.D.) values from biological replicates obtained from 5 animals per groups. Cytokine levels of IFN-γ (D) , IL-4 (E) , and IL-10 (F) measured by enzyme-linked immunosorbent assay (ELISA) using supernatant of splenocytes stimulated with TLA for 72 h 3 weeks after boost; results are presented as the mean ± SEM of cytokine concentration (pg) from biological replicates obtained from 5 animals per group. Cell proliferation assay (G) determined by MTT on stimulated splenocytes with TLA for 72 h 3 weeks after boost; results are presented as the mean ± SEM of the cell proliferation index (ratio of unstimulated control) from biological replicates obtained from 5 animals per group. Asterisks indicate significant differences among groups: * p
Figure Legend Snippet: Humoral and cellular immune response in immunized mice. Vaccination scheme (A) . Anti- Leishmania IgG antibodies (B) using serum samples dilution (1:500) obtained 3 weeks after each immunization. Anti- Leishmania IgG1 and IgG2a (C) antibodies using serum samples dilution (1:1,000) obtained 3 weeks after boost. Results are presented as the mean ± standard error of mean (SEM) of optical density (O.D.) values from biological replicates obtained from 5 animals per groups. Cytokine levels of IFN-γ (D) , IL-4 (E) , and IL-10 (F) measured by enzyme-linked immunosorbent assay (ELISA) using supernatant of splenocytes stimulated with TLA for 72 h 3 weeks after boost; results are presented as the mean ± SEM of cytokine concentration (pg) from biological replicates obtained from 5 animals per group. Cell proliferation assay (G) determined by MTT on stimulated splenocytes with TLA for 72 h 3 weeks after boost; results are presented as the mean ± SEM of the cell proliferation index (ratio of unstimulated control) from biological replicates obtained from 5 animals per group. Asterisks indicate significant differences among groups: * p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Proliferation Assay, MTT Assay

5) Product Images from "Exploring the Potential Effects and Mechanisms of Asarum sieboldii Radix Essential Oil for Treatment of Asthma"

Article Title: Exploring the Potential Effects and Mechanisms of Asarum sieboldii Radix Essential Oil for Treatment of Asthma

Journal: Pharmaceutics

doi: 10.3390/pharmaceutics14030558

Number of inflammatory cells, including macrophages, neutrophils, eosinophils, lymphocytes and total cells, in BALF ( A ) and serum IgE and IgG2a levels ( B ). Results are presented as mean ± standard error of the mean. # p
Figure Legend Snippet: Number of inflammatory cells, including macrophages, neutrophils, eosinophils, lymphocytes and total cells, in BALF ( A ) and serum IgE and IgG2a levels ( B ). Results are presented as mean ± standard error of the mean. # p

Techniques Used:

6) Product Images from "Centrin-deficient Leishmania mexicana confers protection against New World cutaneous leishmaniasis"

Article Title: Centrin-deficient Leishmania mexicana confers protection against New World cutaneous leishmaniasis

Journal: NPJ Vaccines

doi: 10.1038/s41541-022-00449-1

LmexCen −/− immunization mediates protection against challenge with Lmex WT in BALB/c mice. a Schematic of the vaccination study design for BALB/c mice. b Ear lesion thickness of vaccinated and control mice measured for 10 wpc. c – f Representative vaccinated ear ( c ), challenged ear of vaccinated mouse ( d ), PBS-inoculated ear of control mouse ( e ), and challenged ear of control mouse ( f ) at 10 wpc. g – j H E staining of cross-section of representative vaccinated ear ( g ), challenged ear of vaccinated mouse ( h ), PBS-inoculated ear of control mouse ( i ), and challenged ear of control mouse ( j ) at 10 wpc. Scale bar for g – j : 100,000 μm. k , l Parasitic burdens in the ear ( k ) and draining lymph nodes ( l ) at 10 wpc. m – p Cytokine levels of IL-12 ( m ), IFN-γ ( n ), IL-10 ( o ), and IL-4 ( p ) in the lymph nodes of vaccinated and control mice stimulated with L. mexicana antigen at 10 wpc. q , r Ratio between IFN-γ/IL-10 ( q ) and IFN-γ/IL-4 ( r ) levels in the lymph nodes of vaccinated and control mice at 10 wpc. s Ratio of log titers of IgG2a/IgG1 antibodies in the serum of vaccinated and control mice measured for 10 wpc with Lmex WT. Data show one representative experiment out of four independent experiments and show mean ± SEM, N = 10 for each group at each time point. Unpaired two-tailed Student’s t -test was performed to compare statistical significance at each time point. A P -value
Figure Legend Snippet: LmexCen −/− immunization mediates protection against challenge with Lmex WT in BALB/c mice. a Schematic of the vaccination study design for BALB/c mice. b Ear lesion thickness of vaccinated and control mice measured for 10 wpc. c – f Representative vaccinated ear ( c ), challenged ear of vaccinated mouse ( d ), PBS-inoculated ear of control mouse ( e ), and challenged ear of control mouse ( f ) at 10 wpc. g – j H E staining of cross-section of representative vaccinated ear ( g ), challenged ear of vaccinated mouse ( h ), PBS-inoculated ear of control mouse ( i ), and challenged ear of control mouse ( j ) at 10 wpc. Scale bar for g – j : 100,000 μm. k , l Parasitic burdens in the ear ( k ) and draining lymph nodes ( l ) at 10 wpc. m – p Cytokine levels of IL-12 ( m ), IFN-γ ( n ), IL-10 ( o ), and IL-4 ( p ) in the lymph nodes of vaccinated and control mice stimulated with L. mexicana antigen at 10 wpc. q , r Ratio between IFN-γ/IL-10 ( q ) and IFN-γ/IL-4 ( r ) levels in the lymph nodes of vaccinated and control mice at 10 wpc. s Ratio of log titers of IgG2a/IgG1 antibodies in the serum of vaccinated and control mice measured for 10 wpc with Lmex WT. Data show one representative experiment out of four independent experiments and show mean ± SEM, N = 10 for each group at each time point. Unpaired two-tailed Student’s t -test was performed to compare statistical significance at each time point. A P -value

Techniques Used: Mouse Assay, Staining, Two Tailed Test

7) Product Images from "Cruzipain Sulfotopes-Specific Antibodies Generate Cardiac Tissue Abnormalities and Favor Trypanosoma cruzi Infection in the BALB/c Mice Model of Experimental Chagas Disease"

Article Title: Cruzipain Sulfotopes-Specific Antibodies Generate Cardiac Tissue Abnormalities and Favor Trypanosoma cruzi Infection in the BALB/c Mice Model of Experimental Chagas Disease

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2021.814276

IgG1 and IgG2a humoral immune responses in sera from BALB/c mice exposure of GlcNAc6S by immunization with Cz, C-T, and its desulfated counterparts. Determination of IgG2a/IgG1 ratios. IgG1 and IgG2a isotype levels were measured in sera from Cz IM and dCz IM (A) and C-T IM and dC-T IM (B) by ELISA. In each case, the antigen adsorbed in the plate is Cz. The bars represent the averages of duplicate determinations, and the error bars indicate the SD. (A) ▪ p
Figure Legend Snippet: IgG1 and IgG2a humoral immune responses in sera from BALB/c mice exposure of GlcNAc6S by immunization with Cz, C-T, and its desulfated counterparts. Determination of IgG2a/IgG1 ratios. IgG1 and IgG2a isotype levels were measured in sera from Cz IM and dCz IM (A) and C-T IM and dC-T IM (B) by ELISA. In each case, the antigen adsorbed in the plate is Cz. The bars represent the averages of duplicate determinations, and the error bars indicate the SD. (A) ▪ p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

Immunization of BALB/c mice with GlcNAc6S-BSA in the absence of infection. (A) Ultrastructural analysis of cardiac tissue of BSA-GlcNAc6S IM , BSA-GlcNAc IM , and BSA-Phy-Sol IM . Cardiac muscle BSA-GlcNAc6S IM showing severe abnormalities (C) in comparison with normal controls BSA-PhySol (A) and BSA-GlcNAc (B) by transmission electron-microscopy. Magnification of muscle tissue longitudinal sections (7000 ×). At a major magnification, ultrastructural alterations were evidenced in a section of BSA-GlcNAc6S IM [20000 × (D, H)12,000 × (G)], in comparison with the micrograph corresponding to BSA-GlcNAc IM (F) with slight changes and to the normal structure in the BSA-PhySol control mice (E). Determination of immune response to synthetic GlcNAc6S. (B) Humoral immune response in BSA-GlcNAc6S IM . Isotype profile levels in BSA-GlcNAc6S IM and BSA-GlcNAc IM by indirect ELISA, using C-T and dC-T as plate antigens. Control mice were immunized with BSA-IFA. The bars represent the average of the determinations by duplicates and their SD. Black bars, dark grey bars, and light grey bars represent BSA-GlcNAc6S IM , BSA-GlcNAc IM , and BSA, respectively, on C-T antigen. White bars represent BSA-GlcNAc6S on dC-T antigen. • p = 0.0079 vs. IgG2a in BSA-NAcGlc6S IM ; p = 0.0079 vs. IgG1 in BSA-GlcNAc IM ; ♦ p = 0.0159 vs. IgG2b from BSA-GlcNAc IM . * C-T vs. dC-T in plate: p = 0.0079 IgG1 (C-T) vs. IgG1 (dC-T); p = 0.0119 IgG2a (C-T) vs. IgG2a (dC-T); p = 0.0079 IgG2b (C-T) vs. IgG2b (dC-T). (C) T-dependent cellular response obtained with GlcNAc6S. Delayed type hypersensitivity (DTH) was measured in dorsal metatarsal hind-limb of control (BSA IM ) ( n = 5), BSA-GlcNAc6S IM ( n = 5), and BSA-GlcNAc IM ( n = 5), 24 and 48 h after intradermal inoculation of BSA-GlcNAc6S( ID ), BSA-GlcNAc ( ID ), and BSA ( ID ). The bars represent the average of the dorsal metatarsal hind-limb indurations thick with their SD. Dark grey bars, light grey bars, and white bars represent BSA-GlcNAc6S (ID) , BSA-GlcNAc (ID) , and BSA (ID) intradermic antigen, respectively. ▪ p = 0.0079 vs. BSA-GlcNAc ( ID ) in BSA-GlcNAc6S IM ; ○ p = 0.0357 vs. BSA-GlcNAc6S ( ID ) in control mice. The oval scheme in the left part of the figure represents strategy 2.
Figure Legend Snippet: Immunization of BALB/c mice with GlcNAc6S-BSA in the absence of infection. (A) Ultrastructural analysis of cardiac tissue of BSA-GlcNAc6S IM , BSA-GlcNAc IM , and BSA-Phy-Sol IM . Cardiac muscle BSA-GlcNAc6S IM showing severe abnormalities (C) in comparison with normal controls BSA-PhySol (A) and BSA-GlcNAc (B) by transmission electron-microscopy. Magnification of muscle tissue longitudinal sections (7000 ×). At a major magnification, ultrastructural alterations were evidenced in a section of BSA-GlcNAc6S IM [20000 × (D, H)12,000 × (G)], in comparison with the micrograph corresponding to BSA-GlcNAc IM (F) with slight changes and to the normal structure in the BSA-PhySol control mice (E). Determination of immune response to synthetic GlcNAc6S. (B) Humoral immune response in BSA-GlcNAc6S IM . Isotype profile levels in BSA-GlcNAc6S IM and BSA-GlcNAc IM by indirect ELISA, using C-T and dC-T as plate antigens. Control mice were immunized with BSA-IFA. The bars represent the average of the determinations by duplicates and their SD. Black bars, dark grey bars, and light grey bars represent BSA-GlcNAc6S IM , BSA-GlcNAc IM , and BSA, respectively, on C-T antigen. White bars represent BSA-GlcNAc6S on dC-T antigen. • p = 0.0079 vs. IgG2a in BSA-NAcGlc6S IM ; p = 0.0079 vs. IgG1 in BSA-GlcNAc IM ; ♦ p = 0.0159 vs. IgG2b from BSA-GlcNAc IM . * C-T vs. dC-T in plate: p = 0.0079 IgG1 (C-T) vs. IgG1 (dC-T); p = 0.0119 IgG2a (C-T) vs. IgG2a (dC-T); p = 0.0079 IgG2b (C-T) vs. IgG2b (dC-T). (C) T-dependent cellular response obtained with GlcNAc6S. Delayed type hypersensitivity (DTH) was measured in dorsal metatarsal hind-limb of control (BSA IM ) ( n = 5), BSA-GlcNAc6S IM ( n = 5), and BSA-GlcNAc IM ( n = 5), 24 and 48 h after intradermal inoculation of BSA-GlcNAc6S( ID ), BSA-GlcNAc ( ID ), and BSA ( ID ). The bars represent the average of the dorsal metatarsal hind-limb indurations thick with their SD. Dark grey bars, light grey bars, and white bars represent BSA-GlcNAc6S (ID) , BSA-GlcNAc (ID) , and BSA (ID) intradermic antigen, respectively. ▪ p = 0.0079 vs. BSA-GlcNAc ( ID ) in BSA-GlcNAc6S IM ; ○ p = 0.0357 vs. BSA-GlcNAc6S ( ID ) in control mice. The oval scheme in the left part of the figure represents strategy 2.

Techniques Used: Mouse Assay, Infection, Transmission Assay, Electron Microscopy, Indirect ELISA, Immunofluorescence

Treatment of BALB/c mice by passive transference of purified IgG-GlcNAc6S in the presence or absence of T. cruzi sublethal challenge. (A) Ultrastructural alterations of cardiac muscle tissue in mice treated with purified mouse IgG-GlcNAc6S, in the absence of infection. The ultrastructural morphologic analysis was performed by electron microscopy of transmission. Ultramicrograph of cardiac tissue from control mice (A), IgG-pre-imm- TM , mouse treated with purified IgG-pre-imm from mouse sera (B); cardiac tissue of mice treated with purified IgG-GlcNAc6S, IgG-GlcNAc6S- TM (C–E). (A–C, E) (20,000×); in (D), (12000 ×). Mitochondria of cardiac tissue from IgG-pre-imm- TM (F) or IgG-GlcNAc6S-(G). In (F) and (G), (50,000×). Acute phase of infection from BALB/c mice, treated with IgG-GlcNAc6S after a sublethal dose of T. cruzi . (B) Parasitemia (parasite number/ml of peripheral blood) and (C) survival percentage (%) from control of infection and treated with IgG-pre-imm and IgG-GlcNAc6S ( n = 5 mice per group). The challenge was performed on day 1 from the experiment and the exogenous antibody transference via intraperitoneal administration was performed on days 2, 4, 6, 9, 12, and 17. Error bars indicate SD. Blue line, black line, and dotted black line represent IgG-BSA-GlcNAc6S- TM , IgG-pre-imm- TM , and control groups, respectively. The oval schemes in the upper and medium parts of the figure represent strategy 3 part of passive immunization and passive immunization followed by challenge with trypomastigotes, respectively.
Figure Legend Snippet: Treatment of BALB/c mice by passive transference of purified IgG-GlcNAc6S in the presence or absence of T. cruzi sublethal challenge. (A) Ultrastructural alterations of cardiac muscle tissue in mice treated with purified mouse IgG-GlcNAc6S, in the absence of infection. The ultrastructural morphologic analysis was performed by electron microscopy of transmission. Ultramicrograph of cardiac tissue from control mice (A), IgG-pre-imm- TM , mouse treated with purified IgG-pre-imm from mouse sera (B); cardiac tissue of mice treated with purified IgG-GlcNAc6S, IgG-GlcNAc6S- TM (C–E). (A–C, E) (20,000×); in (D), (12000 ×). Mitochondria of cardiac tissue from IgG-pre-imm- TM (F) or IgG-GlcNAc6S-(G). In (F) and (G), (50,000×). Acute phase of infection from BALB/c mice, treated with IgG-GlcNAc6S after a sublethal dose of T. cruzi . (B) Parasitemia (parasite number/ml of peripheral blood) and (C) survival percentage (%) from control of infection and treated with IgG-pre-imm and IgG-GlcNAc6S ( n = 5 mice per group). The challenge was performed on day 1 from the experiment and the exogenous antibody transference via intraperitoneal administration was performed on days 2, 4, 6, 9, 12, and 17. Error bars indicate SD. Blue line, black line, and dotted black line represent IgG-BSA-GlcNAc6S- TM , IgG-pre-imm- TM , and control groups, respectively. The oval schemes in the upper and medium parts of the figure represent strategy 3 part of passive immunization and passive immunization followed by challenge with trypomastigotes, respectively.

Techniques Used: Mouse Assay, Purification, Infection, Electron Microscopy, Transmission Assay

General scheme of immunization from BALB/c mice to study: the input of sulfotopes (GlcNAc6S) in cardiac muscle tissue immunopathology and immune responses, in absence of T. cruzi infection, and the participation of the GlcNAc6S during the infection by T. cruzi. The scheme details the immunization plans performed with the different immunogens and the passive transference of IgG-GlcNAc6S. The immunization scheme with C-T and dC-T was performed in BALB/c female mice (6–8 weeks old; n = 10 animals per group). Five weekly doses of C-T or dC-T (10 μg each), emulsified with IFA (v/v, 1/1) were subcutaneously administered. Control groups received injections with either PBS or with PBS-containing reaction media for solvolysis and were all subjected to the same procedure conditions than the desulfated immunogens plus IFA ( Acosta et al., 2008 ). The immunization scheme with BSA-GlcNAc and BSA-GlcNAc6S was also performed in BALB/c female mice (6–8 weeks old; n = 12 animals per group). Five weekly doses of BSA-GlcNAc or BSA-GlcNAc6S (10 μg each), emulsified with IFA (v/v, 1/1) were subcutaneously administered. Control group mice were immunized with BSA+IFA following the same scheme. In both schemes of the immunization, half of the immunized animals with C-T/dC-T; BSA-GlcNAc6S/BSA-GlcNAc and the respective controls were euthanized to evaluate the immune response and perform a tissue analysis, while the other half of immunized mice were submitted to sublethal challenge with 2 × 10 3 trypomastigotes from Tulahuen 2 (Tul 2) strain, 14 days after the last immunization dose, respectively. Parasitemia was determined each 2–3 days up to negative counting in all groups, and deaths were daily recorded. All the survived mice were subjected to euthanasia in each scheme. For passive transference of antibody assay, BALB/c female mice (6–8 weeks old) were separated in three groups ( n = 5 mice per group) and received 6 doses of 10 μg from purified IgGs; each were administered by intraperitoneally in days 2, 4, 6, 9, 12, and 17. The groups were treated with IgG-GlcNAc6S, purified from mice sera of BSA-GlcNAc6S IM , and purified IgG-pre-imm. Control groups received Phy-Sol administered intraperitoneally. On the other hand, BALB/c female mice (6–8 weeks old) were separated in three groups ( n = 5 mice per group) and were intraperitoneally infected with 500 T. cruzi blood trypomastigotes, Tul 2 strain, the day before to the start of the passive administration for the antibody scheme. A total of 6 doses of 10 μg from purified IgGs each were administered intraperitoneally in days 2, 4, 6, 9, 12, and 17. The mice groups were treated with IgG-GlcNAc6S (IgG-GlcNAc6S- TM ), IgG-pre-imm (IgG-pre-imm- TM ), and Phy-Sol. Control groups received Phy-Sol administered intraperitoneally. Parasitemia was monitored each 2–3 days until the fourth day after the last dose of IgGs was administered. Deaths were daily recorded. Euthanasia was performed on days 21 and 50 for tissue analysis and survival recording, respectively. The sublethal infective doses have been selected both by previous experience in the laboratory group and based on bibliography ( Lu et al., 2008 ).
Figure Legend Snippet: General scheme of immunization from BALB/c mice to study: the input of sulfotopes (GlcNAc6S) in cardiac muscle tissue immunopathology and immune responses, in absence of T. cruzi infection, and the participation of the GlcNAc6S during the infection by T. cruzi. The scheme details the immunization plans performed with the different immunogens and the passive transference of IgG-GlcNAc6S. The immunization scheme with C-T and dC-T was performed in BALB/c female mice (6–8 weeks old; n = 10 animals per group). Five weekly doses of C-T or dC-T (10 μg each), emulsified with IFA (v/v, 1/1) were subcutaneously administered. Control groups received injections with either PBS or with PBS-containing reaction media for solvolysis and were all subjected to the same procedure conditions than the desulfated immunogens plus IFA ( Acosta et al., 2008 ). The immunization scheme with BSA-GlcNAc and BSA-GlcNAc6S was also performed in BALB/c female mice (6–8 weeks old; n = 12 animals per group). Five weekly doses of BSA-GlcNAc or BSA-GlcNAc6S (10 μg each), emulsified with IFA (v/v, 1/1) were subcutaneously administered. Control group mice were immunized with BSA+IFA following the same scheme. In both schemes of the immunization, half of the immunized animals with C-T/dC-T; BSA-GlcNAc6S/BSA-GlcNAc and the respective controls were euthanized to evaluate the immune response and perform a tissue analysis, while the other half of immunized mice were submitted to sublethal challenge with 2 × 10 3 trypomastigotes from Tulahuen 2 (Tul 2) strain, 14 days after the last immunization dose, respectively. Parasitemia was determined each 2–3 days up to negative counting in all groups, and deaths were daily recorded. All the survived mice were subjected to euthanasia in each scheme. For passive transference of antibody assay, BALB/c female mice (6–8 weeks old) were separated in three groups ( n = 5 mice per group) and received 6 doses of 10 μg from purified IgGs; each were administered by intraperitoneally in days 2, 4, 6, 9, 12, and 17. The groups were treated with IgG-GlcNAc6S, purified from mice sera of BSA-GlcNAc6S IM , and purified IgG-pre-imm. Control groups received Phy-Sol administered intraperitoneally. On the other hand, BALB/c female mice (6–8 weeks old) were separated in three groups ( n = 5 mice per group) and were intraperitoneally infected with 500 T. cruzi blood trypomastigotes, Tul 2 strain, the day before to the start of the passive administration for the antibody scheme. A total of 6 doses of 10 μg from purified IgGs each were administered intraperitoneally in days 2, 4, 6, 9, 12, and 17. The mice groups were treated with IgG-GlcNAc6S (IgG-GlcNAc6S- TM ), IgG-pre-imm (IgG-pre-imm- TM ), and Phy-Sol. Control groups received Phy-Sol administered intraperitoneally. Parasitemia was monitored each 2–3 days until the fourth day after the last dose of IgGs was administered. Deaths were daily recorded. Euthanasia was performed on days 21 and 50 for tissue analysis and survival recording, respectively. The sublethal infective doses have been selected both by previous experience in the laboratory group and based on bibliography ( Lu et al., 2008 ).

Techniques Used: Mouse Assay, Infection, Immunofluorescence, Purification

Native Cz was obtained from T. cruzi ; C-T was purified from Cz. The immunogenic pairs Cz/dCz and C-T/dC-T were obtained by chemical desulfation process. The latter were used in strategy 1. In Strategy 2, the synthetic immunogens GlcNAc6S/GlcNAc coupled to BSA, as carrier proteins were used. Both strategies 1 and 2 evidenced both the humoral and cellular-specific immune responses and the ultrastructural pathogenesis in the cardiac muscle tissue, after in vivo exposition to the sulfotope. In mice, pre-exposed to the sulfotope (strategies 1 and 2) and sublethally challenged, the infection showed to be favored, with elevated parasitemias. Finally, in strategy 3, passive treatment with antibodies obtained in strategy 2 was performed. This treatment has demonstrated that IgG-GlcNAc6S, by in vivo exposition, have generated ultrastructural alterations in heart tissue and the administration of IgG-GlcNAc6S postsublethal challenge favored parasitic infection. , mannose; , GlcNAc; , SO3; , BSA; , IgG-GlcNAc6S.
Figure Legend Snippet: Native Cz was obtained from T. cruzi ; C-T was purified from Cz. The immunogenic pairs Cz/dCz and C-T/dC-T were obtained by chemical desulfation process. The latter were used in strategy 1. In Strategy 2, the synthetic immunogens GlcNAc6S/GlcNAc coupled to BSA, as carrier proteins were used. Both strategies 1 and 2 evidenced both the humoral and cellular-specific immune responses and the ultrastructural pathogenesis in the cardiac muscle tissue, after in vivo exposition to the sulfotope. In mice, pre-exposed to the sulfotope (strategies 1 and 2) and sublethally challenged, the infection showed to be favored, with elevated parasitemias. Finally, in strategy 3, passive treatment with antibodies obtained in strategy 2 was performed. This treatment has demonstrated that IgG-GlcNAc6S, by in vivo exposition, have generated ultrastructural alterations in heart tissue and the administration of IgG-GlcNAc6S postsublethal challenge favored parasitic infection. , mannose; , GlcNAc; , SO3; , BSA; , IgG-GlcNAc6S.

Techniques Used: Purification, In Vivo, Mouse Assay, Infection, Generated

8) Product Images from "Immunogenicity and Protective Potential of Mucosal Vaccine Formulations Based on Conserved Epitopes of Influenza A Viruses Fused to an Innovative Ring Nanoplatform in Mice and Chickens"

Article Title: Immunogenicity and Protective Potential of Mucosal Vaccine Formulations Based on Conserved Epitopes of Influenza A Viruses Fused to an Innovative Ring Nanoplatform in Mice and Chickens

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2021.772550

Evaluation of the immunogenicity and protective potential of LAH- and 3M2e-bearing nanorings against heterosubtypic H5N8 HPAIV infection in chickens. Chickens received at 2-week intervals three intranasal administrations of H5N9 LAH and 3M2e peptides fused to nanorings either mucosally (‘muc.’) with Montanide™ gel (MG) or intramuscularly (‘i.m.’) with Montanide™ ISA (ISA). The non-vaccinated control group received only PBS. Eighteen days after the third immunization, serum (A) anti-LAH or (B) anti-M2e IgG titers or mucosal (C) anti-LAH IgG and IgA or (D) anti-M2e IgG or IgA titers in the lachrymal secretions (LS) were dosed. The titer of each chicken sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P
Figure Legend Snippet: Evaluation of the immunogenicity and protective potential of LAH- and 3M2e-bearing nanorings against heterosubtypic H5N8 HPAIV infection in chickens. Chickens received at 2-week intervals three intranasal administrations of H5N9 LAH and 3M2e peptides fused to nanorings either mucosally (‘muc.’) with Montanide™ gel (MG) or intramuscularly (‘i.m.’) with Montanide™ ISA (ISA). The non-vaccinated control group received only PBS. Eighteen days after the third immunization, serum (A) anti-LAH or (B) anti-M2e IgG titers or mucosal (C) anti-LAH IgG and IgA or (D) anti-M2e IgG or IgA titers in the lachrymal secretions (LS) were dosed. The titer of each chicken sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P

Techniques Used: Infection

Evaluation of the immunopotentiator effect of nanorings and P97c on LAH- and M2e-specific systemic and mucosal humoral responses in mice. Mice received at 2-week intervals three intranasal administrations of LAH or 3M2e peptides fused to nanorings and/or P97c or, as controls, free LAH or 3M2e peptides. Preparations without P97c were admixed with Montanide™ gel (MG). Mice receiving P97c nanorings (N-P97c), naked nanorings (N + MG), P97c or MG were included as negative control groups. Two weeks after the third immunization, (A) anti-LAH or (B) anti-M2e IgG titers in the serum, and anti-LAH (C) IgG and (E) IgA or anti-M2e (D) IgG and (F) IgA titers in the bronchoalveolar lavage fluids (BAL) were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P
Figure Legend Snippet: Evaluation of the immunopotentiator effect of nanorings and P97c on LAH- and M2e-specific systemic and mucosal humoral responses in mice. Mice received at 2-week intervals three intranasal administrations of LAH or 3M2e peptides fused to nanorings and/or P97c or, as controls, free LAH or 3M2e peptides. Preparations without P97c were admixed with Montanide™ gel (MG). Mice receiving P97c nanorings (N-P97c), naked nanorings (N + MG), P97c or MG were included as negative control groups. Two weeks after the third immunization, (A) anti-LAH or (B) anti-M2e IgG titers in the serum, and anti-LAH (C) IgG and (E) IgA or anti-M2e (D) IgG and (F) IgA titers in the bronchoalveolar lavage fluids (BAL) were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P

Techniques Used: Mouse Assay, Negative Control, Enzyme-linked Immunosorbent Assay

Comparison of specific serum and mucosal humoral responses in mice immunized with LAH- and M2e-bearing nanorings instilled separately or in combination. Mice received at 2-week intervals three intranasal administrations of N-LAH and N-3M2e nanorings admixed with Montanide™ gel (MG) instilled separately or in combination. Control groups received intranasal administrations of free LAH or 3M2e peptides admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, (A) anti-LAH or (B) anti-M2e IgG titers in the serum, and anti-LAH (C) IgG and (E) IgA or anti-M2e (D) IgG and (F) IgA titers in the bronchoalveolar lavage fluids (BAL) were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P
Figure Legend Snippet: Comparison of specific serum and mucosal humoral responses in mice immunized with LAH- and M2e-bearing nanorings instilled separately or in combination. Mice received at 2-week intervals three intranasal administrations of N-LAH and N-3M2e nanorings admixed with Montanide™ gel (MG) instilled separately or in combination. Control groups received intranasal administrations of free LAH or 3M2e peptides admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, (A) anti-LAH or (B) anti-M2e IgG titers in the serum, and anti-LAH (C) IgG and (E) IgA or anti-M2e (D) IgG and (F) IgA titers in the bronchoalveolar lavage fluids (BAL) were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

IgG subclass composition of LAH-specific serum humoral response in mice immunized with N-LAH nanorings instilled alone or in combination with N-3M2e nanorings. Mice received at 2-week intervals three intranasal administrations of N-LAH nanorings admixed with Montanide™ gel (MG) instilled alone or in combination with N-3M2e nanorings. Control groups received intranasal administrations of free LAH peptide admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, serum anti-LAH (A) IgG1, (B) IgG2a, (C) IgG2b, and (D) IgG3 titers were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P
Figure Legend Snippet: IgG subclass composition of LAH-specific serum humoral response in mice immunized with N-LAH nanorings instilled alone or in combination with N-3M2e nanorings. Mice received at 2-week intervals three intranasal administrations of N-LAH nanorings admixed with Montanide™ gel (MG) instilled alone or in combination with N-3M2e nanorings. Control groups received intranasal administrations of free LAH peptide admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, serum anti-LAH (A) IgG1, (B) IgG2a, (C) IgG2b, and (D) IgG3 titers were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

IgG subclass composition of M2e-specific serum humoral response in mice immunized with N-3M2e nanorings instilled alone or in combination with N-LAH nanorings. Mice received at 2-week intervals three intranasal administrations of N-3M2e nanorings admixed with Montanide™ gel (MG) instilled alone or in combination with N-LAH nanorings. Control groups received intranasal administrations of free 3M2e peptide admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, serum anti-M2e (A) IgG1, (B) IgG2a, (C) IgG2b, and (D) IgG3 titers were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P
Figure Legend Snippet: IgG subclass composition of M2e-specific serum humoral response in mice immunized with N-3M2e nanorings instilled alone or in combination with N-LAH nanorings. Mice received at 2-week intervals three intranasal administrations of N-3M2e nanorings admixed with Montanide™ gel (MG) instilled alone or in combination with N-LAH nanorings. Control groups received intranasal administrations of free 3M2e peptide admixed with MG or vaccine vehicle (MG), or subcutaneous administrations of UV-inactivated PR8 strain (iPR8) admixed with MG. Two weeks after the third immunization, serum anti-M2e (A) IgG1, (B) IgG2a, (C) IgG2b, and (D) IgG3 titers were determined by ELISA. The titer of each mouse sample is presented, including the geometric mean with 95% confidence interval. *, statistically significant difference ( P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

9) Product Images from "Cudraxanthone D Ameliorates Psoriasis-like Skin Inflammation in an Imiquimod-Induced Mouse Model via Inhibiting the Inflammatory Signaling Pathways"

Article Title: Cudraxanthone D Ameliorates Psoriasis-like Skin Inflammation in an Imiquimod-Induced Mouse Model via Inhibiting the Inflammatory Signaling Pathways

Journal: Molecules

doi: 10.3390/molecules26196086

Effects of CD on serum immunoglobulin levels and infiltration of MPO-associated cells. After the in vivo experiments, the mice were sacrificed, after which their blood and skin were harvested. ( a , b ) The serum levels of tumor necrosis factor (TNF)-α, IgG2a, and MPO were measured via sandwich enzyme-linked immunosorbent assay (ELISA). Each data point represents the mean ± SEM of the two independent samples. * p
Figure Legend Snippet: Effects of CD on serum immunoglobulin levels and infiltration of MPO-associated cells. After the in vivo experiments, the mice were sacrificed, after which their blood and skin were harvested. ( a , b ) The serum levels of tumor necrosis factor (TNF)-α, IgG2a, and MPO were measured via sandwich enzyme-linked immunosorbent assay (ELISA). Each data point represents the mean ± SEM of the two independent samples. * p

Techniques Used: In Vivo, Mouse Assay, Sandwich ELISA, Enzyme-linked Immunosorbent Assay

10) Product Images from "Poor CD4+ T Cell Immunogenicity Limits Humoral Immunity to P. falciparum Transmission-Blocking Candidate Pfs25 in Humans"

Article Title: Poor CD4+ T Cell Immunogenicity Limits Humoral Immunity to P. falciparum Transmission-Blocking Candidate Pfs25 in Humans

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2021.732667

Assessment of murine and human antibodies’ effector functions, avidity and cross-reactivity. (A) Isotype profiles of murine Pfs25-specific antibody responses on d84 were assessed by ELISA. Responses are shown for IgG1, IgG2a and IgG2b. (B) Isotype profiles of human serum antibody responses were assessed by ELISA. Responses are shown for immunization groups 2B and 2C, assessed at d84 for IgG1, IgG2, IgG3, IgG4 and IgM. (C) Avidities of murine and human serum IgG responses at day 84 were assessed by NaSCN-displacement Pfs25 ELISA and are reported as the molar (M) concentration of NaSCN required to reduce the starting OD in the ELISA by 50% (IC50) Differences in avidity were assessed by Mann-Whitney test. Each symbol represents an individual response (ns- not significant). (D) Endpoint anti-Pfs28 IgG titers were assessed by ELISA on human serum samples isolated on d0 (n = 16) and d74 (n = 16) or murine samples (n = 6) obtained on d74 post ChAd63/MVA Pfs25-IMX313 vaccination. Murine serum sample acquired 2 weeks post boost immunization with recombinant Pfs28 protein in Alum, was used as positive control. Error bars show SEM.
Figure Legend Snippet: Assessment of murine and human antibodies’ effector functions, avidity and cross-reactivity. (A) Isotype profiles of murine Pfs25-specific antibody responses on d84 were assessed by ELISA. Responses are shown for IgG1, IgG2a and IgG2b. (B) Isotype profiles of human serum antibody responses were assessed by ELISA. Responses are shown for immunization groups 2B and 2C, assessed at d84 for IgG1, IgG2, IgG3, IgG4 and IgM. (C) Avidities of murine and human serum IgG responses at day 84 were assessed by NaSCN-displacement Pfs25 ELISA and are reported as the molar (M) concentration of NaSCN required to reduce the starting OD in the ELISA by 50% (IC50) Differences in avidity were assessed by Mann-Whitney test. Each symbol represents an individual response (ns- not significant). (D) Endpoint anti-Pfs28 IgG titers were assessed by ELISA on human serum samples isolated on d0 (n = 16) and d74 (n = 16) or murine samples (n = 6) obtained on d74 post ChAd63/MVA Pfs25-IMX313 vaccination. Murine serum sample acquired 2 weeks post boost immunization with recombinant Pfs28 protein in Alum, was used as positive control. Error bars show SEM.

Techniques Used: Enzyme-linked Immunosorbent Assay, Concentration Assay, MANN-WHITNEY, Isolation, Recombinant, Positive Control

11) Product Images from "Diesel exhaust particles increase nasal symptoms and IL-17A in house dust mite-induced allergic mice"

Article Title: Diesel exhaust particles increase nasal symptoms and IL-17A in house dust mite-induced allergic mice

Journal: Scientific Reports

doi: 10.1038/s41598-021-94673-9

Serum immunoglobulin levels in each group. ( A ) Total IgE, ( B ) HDM-specific IgE, ( C ) HDM-specific IgG1, and ( D ) HDM-specific IgG2a. HDM and DEP co-exposed group showing suppressed levels of HDM-specific IgE and IgG 1 . Data are expressed as mean ± standard error mean. * p
Figure Legend Snippet: Serum immunoglobulin levels in each group. ( A ) Total IgE, ( B ) HDM-specific IgE, ( C ) HDM-specific IgG1, and ( D ) HDM-specific IgG2a. HDM and DEP co-exposed group showing suppressed levels of HDM-specific IgE and IgG 1 . Data are expressed as mean ± standard error mean. * p

Techniques Used:

12) Product Images from "Diesel exhaust particles increase nasal symptoms and IL-17A in house dust mite-induced allergic mice"

Article Title: Diesel exhaust particles increase nasal symptoms and IL-17A in house dust mite-induced allergic mice

Journal: Scientific Reports

doi: 10.1038/s41598-021-94673-9

Serum immunoglobulin levels in each group. ( A ) Total IgE, ( B ) HDM-specific IgE, ( C ) HDM-specific IgG1, and ( D ) HDM-specific IgG2a. HDM and DEP co-exposed group showing suppressed levels of HDM-specific IgE and IgG 1 . Data are expressed as mean ± standard error mean. * p
Figure Legend Snippet: Serum immunoglobulin levels in each group. ( A ) Total IgE, ( B ) HDM-specific IgE, ( C ) HDM-specific IgG1, and ( D ) HDM-specific IgG2a. HDM and DEP co-exposed group showing suppressed levels of HDM-specific IgE and IgG 1 . Data are expressed as mean ± standard error mean. * p

Techniques Used:

13) Product Images from "Viability Status-Dependent Effect of Bifidobacterium longum ssp. longum CCM 7952 on Prevention of Allergic Inflammation in Mouse Model"

Article Title: Viability Status-Dependent Effect of Bifidobacterium longum ssp. longum CCM 7952 on Prevention of Allergic Inflammation in Mouse Model

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2021.707728

Intranasal administration of untreated and heat-treated Bl 7952 differently attenuates the OVA-induced cellular and humoral immune responses. Effect of PBS (PBS/OVA, black box), untreated Bl 7952 (UN Bl 7952/OVA, blue box) and heat-treated Bl 7952 (HT Bl 7952/OVA, orange box) strain on cytokine production was determined in supernatants of OVA-stimulated splenocytes isolated from bacteria or PBS pre-treated and OVA-sensitized and challenged mice. (A) IL-4, (B) IL-5, (C) IL-10 and (D) IFN- γ cytokines are expressed as pg/ml. Levels of OVA-specific (E) IgE as % of β -hexosaminidase release by RBL assay, (F) IgG1, (G) IgG2a, (H) IgA and (I) total IgA antibodies were measured in sera of experimental mice. Values are expressed as boxplot with mean and min to max value bars; each dot represents a single mouse. Significant difference between PBS/OVA and bifidobacteria-treated/OVA experimental groups was calculated by one-way ANOVA with Dunnett’s multiple comparison test *p
Figure Legend Snippet: Intranasal administration of untreated and heat-treated Bl 7952 differently attenuates the OVA-induced cellular and humoral immune responses. Effect of PBS (PBS/OVA, black box), untreated Bl 7952 (UN Bl 7952/OVA, blue box) and heat-treated Bl 7952 (HT Bl 7952/OVA, orange box) strain on cytokine production was determined in supernatants of OVA-stimulated splenocytes isolated from bacteria or PBS pre-treated and OVA-sensitized and challenged mice. (A) IL-4, (B) IL-5, (C) IL-10 and (D) IFN- γ cytokines are expressed as pg/ml. Levels of OVA-specific (E) IgE as % of β -hexosaminidase release by RBL assay, (F) IgG1, (G) IgG2a, (H) IgA and (I) total IgA antibodies were measured in sera of experimental mice. Values are expressed as boxplot with mean and min to max value bars; each dot represents a single mouse. Significant difference between PBS/OVA and bifidobacteria-treated/OVA experimental groups was calculated by one-way ANOVA with Dunnett’s multiple comparison test *p

Techniques Used: Isolation, Mouse Assay

14) Product Images from "Fentanyl conjugate vaccine by injected or mucosal delivery with dmLT or LTA1 adjuvants implicates IgA in protection from drug challenge"

Article Title: Fentanyl conjugate vaccine by injected or mucosal delivery with dmLT or LTA1 adjuvants implicates IgA in protection from drug challenge

Journal: NPJ Vaccines

doi: 10.1038/s41541-021-00329-0

Significant correlations between anti-FEN IgG antibody analyses by serum ELISAs and ASC ELISPOT. a Correlations between indicated IgG ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants IM prime/boost or prime/boost/boost experiments. b Correlations between indicated IgG ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants prime IM and mucosal (SL or IN) booster experiments. c Correlations between indicated IgA ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants prime IM and mucosal (SL or IN) booster experiments. ELISPOT data graphed as bone marrow ASC per 1e6 cells (log10). ELISA data graphed as serum IgG EU/ml (log2). Spearman correlations P values and correlation coefficient ( r ) indicated on each graph.
Figure Legend Snippet: Significant correlations between anti-FEN IgG antibody analyses by serum ELISAs and ASC ELISPOT. a Correlations between indicated IgG ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants IM prime/boost or prime/boost/boost experiments. b Correlations between indicated IgG ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants prime IM and mucosal (SL or IN) booster experiments. c Correlations between indicated IgA ELISAs or ELISPOT results with coating antigen indicated in parenthesis using compiled data from FEN-CRM ± adjuvants prime IM and mucosal (SL or IN) booster experiments. ELISPOT data graphed as bone marrow ASC per 1e6 cells (log10). ELISA data graphed as serum IgG EU/ml (log2). Spearman correlations P values and correlation coefficient ( r ) indicated on each graph.

Techniques Used: Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay

Reduction in fentanyl-induced antinociception and brain tissue distribution in dmLT and LTA1 vaccinated animals compared to alum or antigen alone groups. Groups of Balb/c mice ( n = 5–20) were left naïve or immunized on weeks 0, 3, and 6 with FEN-CRM IM or with adjuvant and route combinations as performed previously. These included 1500 μg alum IM, 0.1–1 μg dmLT IM, 0.1 μg dmLT IM + 5 μg dmLT SL boosters (dmLT SL), or 5 μg LTA1 IM + 5 μg LTA1 IN boosters (LTA1 IN). FEN-CRM was administered at 5 μg for IM delivery and 9–10 μg for mucosal delivery. Blood was collected on week 6, 8, and 10 for serum analyses and mice challenged with fentanyl doses during weeks 9–12. Nociception tests were conducted using tail flick and hotplate assays with tissue taken for drug quantification. a Schematic of immunization schedule. b Compiled serum anti-FEN (FEN-BSA) serum IgG ELISA from week 6, 8, and 10 tailbleeds. c % maximum possible effect (MPE) from tail flick antinociception evaluation. d % MPE from hotplate antinociception evaluation. e Fentanyl concentrations in brain and serum after a final 0.1 mg/kg fentanyl challenge. F Correlation between brain fentanyl levels or tail flick antinociception %MPE vs anti-FEN serum IgG for immunized groups. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P
Figure Legend Snippet: Reduction in fentanyl-induced antinociception and brain tissue distribution in dmLT and LTA1 vaccinated animals compared to alum or antigen alone groups. Groups of Balb/c mice ( n = 5–20) were left naïve or immunized on weeks 0, 3, and 6 with FEN-CRM IM or with adjuvant and route combinations as performed previously. These included 1500 μg alum IM, 0.1–1 μg dmLT IM, 0.1 μg dmLT IM + 5 μg dmLT SL boosters (dmLT SL), or 5 μg LTA1 IM + 5 μg LTA1 IN boosters (LTA1 IN). FEN-CRM was administered at 5 μg for IM delivery and 9–10 μg for mucosal delivery. Blood was collected on week 6, 8, and 10 for serum analyses and mice challenged with fentanyl doses during weeks 9–12. Nociception tests were conducted using tail flick and hotplate assays with tissue taken for drug quantification. a Schematic of immunization schedule. b Compiled serum anti-FEN (FEN-BSA) serum IgG ELISA from week 6, 8, and 10 tailbleeds. c % maximum possible effect (MPE) from tail flick antinociception evaluation. d % MPE from hotplate antinociception evaluation. e Fentanyl concentrations in brain and serum after a final 0.1 mg/kg fentanyl challenge. F Correlation between brain fentanyl levels or tail flick antinociception %MPE vs anti-FEN serum IgG for immunized groups. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P

Techniques Used: Mouse Assay, Tail Flick Test, Enzyme-linked Immunosorbent Assay

Anti-FEN antibody affinity, IgG2a, and IgA isotypes contributed to protection from fentanyl challenge in dmLT and LTA1 immunized animals. Serum samples from fentanyl challenge experiments were further characterized for anti-FEN immunity. a IgG antibody binding affinity calculated as % IgG binding to FEN-TT in the presence of a chaotropic agent (week 6). b Compiled serum anti-FEN (FEN-TT) serum IgG1 (week 8), IgG2a (week 8), IgA (week 10) ELISA from tailbleeds. c Ratio of serum anti-FEN (FEN-TT) IgG1/IgG2a. (Zero values for IgG2a were replaced with a 1.) d Correlation between hotplate antinociception vs IgG2a or IgG binding affinity for 0.1 and 1 μg dmLT IM immunization groups only. e Correlation between brain fentanyl levels or tail flick antinociception %MPE vs anti-FEN serum IgA (week 10) for immunized groups. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P
Figure Legend Snippet: Anti-FEN antibody affinity, IgG2a, and IgA isotypes contributed to protection from fentanyl challenge in dmLT and LTA1 immunized animals. Serum samples from fentanyl challenge experiments were further characterized for anti-FEN immunity. a IgG antibody binding affinity calculated as % IgG binding to FEN-TT in the presence of a chaotropic agent (week 6). b Compiled serum anti-FEN (FEN-TT) serum IgG1 (week 8), IgG2a (week 8), IgA (week 10) ELISA from tailbleeds. c Ratio of serum anti-FEN (FEN-TT) IgG1/IgG2a. (Zero values for IgG2a were replaced with a 1.) d Correlation between hotplate antinociception vs IgG2a or IgG binding affinity for 0.1 and 1 μg dmLT IM immunization groups only. e Correlation between brain fentanyl levels or tail flick antinociception %MPE vs anti-FEN serum IgA (week 10) for immunized groups. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P

Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Tail Flick Test

Immune responses to intramuscular immunization with FEN-BSA are enhanced with dmLT adjuvant. Groups of Balb/c mice ( n = 4) were left naïve or immunized intramuscularly with 8 μg FEN-BSA ± 1 μg dmLT on weeks 0 and 4. Two-weeks later (week 6) serum, spleen, or bone-marrow were collected for antibody evaluations. a Schematic of immunization schedule identifying week of immunizations (IM) and sacrifice for blood, spleen, and bone-marrow sample collections (SAC). b Raw ELISA data for serum anti-BSA or anti-FEN IgG expressed as 405 nm optical density (OD) absorbance. For anti-FEN IgG, plates were coated with FEN-TT. c Compiled ELISA units (EU)/ml of indicated serum anti-BSA and anti-FEN IgG titers assessed by ELISAs graphed on a log2 scale. d Compiled EU/ml of indicated serum anti-FEN IgG1 or IgG2a. e Raw ELISA data for serum anti-FEN IgA and IgM. f Representative images for ASCs IgG/IgA ELISPOT results using splenocytes (Sp) or bone marrow (BM) cells and plates coated with FEN-TT. Blue spots indicate IgG ASCs and red spots IgA ASCs. g Compiled ASC per 10 6 cells from spleens or bone marrow. Bars at mean + s.e.m. with significance indicated as (***) for P ≤ 0.001 (*) for P ≤ 0.05 by one-way ANOVA with Bonferroni’s multiple comparisons test.
Figure Legend Snippet: Immune responses to intramuscular immunization with FEN-BSA are enhanced with dmLT adjuvant. Groups of Balb/c mice ( n = 4) were left naïve or immunized intramuscularly with 8 μg FEN-BSA ± 1 μg dmLT on weeks 0 and 4. Two-weeks later (week 6) serum, spleen, or bone-marrow were collected for antibody evaluations. a Schematic of immunization schedule identifying week of immunizations (IM) and sacrifice for blood, spleen, and bone-marrow sample collections (SAC). b Raw ELISA data for serum anti-BSA or anti-FEN IgG expressed as 405 nm optical density (OD) absorbance. For anti-FEN IgG, plates were coated with FEN-TT. c Compiled ELISA units (EU)/ml of indicated serum anti-BSA and anti-FEN IgG titers assessed by ELISAs graphed on a log2 scale. d Compiled EU/ml of indicated serum anti-FEN IgG1 or IgG2a. e Raw ELISA data for serum anti-FEN IgA and IgM. f Representative images for ASCs IgG/IgA ELISPOT results using splenocytes (Sp) or bone marrow (BM) cells and plates coated with FEN-TT. Blue spots indicate IgG ASCs and red spots IgA ASCs. g Compiled ASC per 10 6 cells from spleens or bone marrow. Bars at mean + s.e.m. with significance indicated as (***) for P ≤ 0.001 (*) for P ≤ 0.05 by one-way ANOVA with Bonferroni’s multiple comparisons test.

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Immune responses to prime/boost intramuscular immunization with FEN-CRM 197 are enhanced in magnitude and longevity with dmLT and LTA1 adjuvants. Groups of Balb/c mice ( n = 3–5) were left naïve or immunized IM with 5 μg FEN-CRM ± 150 μg alum, 0.1 μg dmLT, or 5 μg LTA1 on weeks 0 and 3. Two-weeks later (week 5) or six-weeks later (week 9) serum, spleen, and bone-marrow was collected for antibody evaluations. a Schematic of the immunization schedule. b Raw ELISA data for week 5 serum anti-CRM or anti-FEN serum IgG expressed as absorbance. Two anti-FEN ELISA coating antigens were used as indicated by FEN-BSA or FEN-TT. c Compiled week 5 and 9 serum anti-CRM or anti-FEN (coating antigen indicated) serum IgG. d Representative anti-FEN (FEN-TT coating antigen) IgG/IgA ELISPOT images from bone-marrow of week 5 immunized mice. Blue spots indicate IgG and red spots IgA ASCs. e Compiled anti-FEN IgG ASC per 1e6 cells from bone marrow. f Anti-FEN IgG memory B cells per 10 6 cells differentiated in culture from immunized splenocytes from week 9 post immunization. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P
Figure Legend Snippet: Immune responses to prime/boost intramuscular immunization with FEN-CRM 197 are enhanced in magnitude and longevity with dmLT and LTA1 adjuvants. Groups of Balb/c mice ( n = 3–5) were left naïve or immunized IM with 5 μg FEN-CRM ± 150 μg alum, 0.1 μg dmLT, or 5 μg LTA1 on weeks 0 and 3. Two-weeks later (week 5) or six-weeks later (week 9) serum, spleen, and bone-marrow was collected for antibody evaluations. a Schematic of the immunization schedule. b Raw ELISA data for week 5 serum anti-CRM or anti-FEN serum IgG expressed as absorbance. Two anti-FEN ELISA coating antigens were used as indicated by FEN-BSA or FEN-TT. c Compiled week 5 and 9 serum anti-CRM or anti-FEN (coating antigen indicated) serum IgG. d Representative anti-FEN (FEN-TT coating antigen) IgG/IgA ELISPOT images from bone-marrow of week 5 immunized mice. Blue spots indicate IgG and red spots IgA ASCs. e Compiled anti-FEN IgG ASC per 1e6 cells from bone marrow. f Anti-FEN IgG memory B cells per 10 6 cells differentiated in culture from immunized splenocytes from week 9 post immunization. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Immune responses to FEN-CRM 197 can be generated by sublingual or intranasal booster immunizations containing either dmLT or LTA1 adjuvants. Groups of Balb/c mice ( n = 6) were left naïve or primed by 5 μg FEN-CRM ± 0.1 μg of dmLT or 5 μg of LTA1 by IM delivery. Mice were boosted mucosally on weeks 3 and 6 with 9–10 μg FEN-CRM ± 5 μg dmLT SL or 9 μg FEN-CRM ± 5 μg LTA1 IN, both in 30 μl. On week 8, serum and bone-marrow were collected for immune response evaluation. a Schematic of immunization schedule. b Compiled serum anti-CRM and anti-FEN (FEN-BSA, FEN-TT) serum IgG ELISAs. c Compiled serum anti-FEN (FEN-TT) serum IgG1 or IgG2a ELISAs. d Representative anti-FEN (TT-BSA coated) IgG/IgA ELISPOT images from bone-marrow. Blue spots indicate IgG and red spots IgA ASCs. d Compiled serum anti-FEN (FEN-BSA) IgA ELISA. e Compiled anti-FEN IgG ASC per 10 6 cells from bone-marrow. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P
Figure Legend Snippet: Immune responses to FEN-CRM 197 can be generated by sublingual or intranasal booster immunizations containing either dmLT or LTA1 adjuvants. Groups of Balb/c mice ( n = 6) were left naïve or primed by 5 μg FEN-CRM ± 0.1 μg of dmLT or 5 μg of LTA1 by IM delivery. Mice were boosted mucosally on weeks 3 and 6 with 9–10 μg FEN-CRM ± 5 μg dmLT SL or 9 μg FEN-CRM ± 5 μg LTA1 IN, both in 30 μl. On week 8, serum and bone-marrow were collected for immune response evaluation. a Schematic of immunization schedule. b Compiled serum anti-CRM and anti-FEN (FEN-BSA, FEN-TT) serum IgG ELISAs. c Compiled serum anti-FEN (FEN-TT) serum IgG1 or IgG2a ELISAs. d Representative anti-FEN (TT-BSA coated) IgG/IgA ELISPOT images from bone-marrow. Blue spots indicate IgG and red spots IgA ASCs. d Compiled serum anti-FEN (FEN-BSA) IgA ELISA. e Compiled anti-FEN IgG ASC per 10 6 cells from bone-marrow. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P

Techniques Used: Generated, Mouse Assay, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay

Immune responses to prime/boost/boost intramuscular FEN-CRM197 immunization are enhanced in magnitude with dmLT and LTA1 adjuvants. Groups of Balb/c mice ( n = 3–5) were left naïve or immunized IM with 5 μg FEN-CRM ± 150 μg alum, 0.1 μg dmLT, or 5 μg LTA1 on weeks 0, 3, and 6. Two-weeks later (week 8) serum and bone-marrow was collected for antibody evaluations. a Schematic of immunization schedule. b Compiled Week 5 and Week 9 serum anti-CRM or anti-FEN (coating antigen indicated) serum IgG. c Compiled EU/ml of indicated serum anti-FEN serum IgG1, or IgG2a ELISAs. d Representative anti-FEN (TT-BSA coated) IgG/IgA ELISPOT images from bone-marrow of week 8 immunized mice. Blue spots indicate IgG and red spots IgA ASCs. e Compiled anti-FEN IgG ASC per 10 6 cells from bone-marrow. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P
Figure Legend Snippet: Immune responses to prime/boost/boost intramuscular FEN-CRM197 immunization are enhanced in magnitude with dmLT and LTA1 adjuvants. Groups of Balb/c mice ( n = 3–5) were left naïve or immunized IM with 5 μg FEN-CRM ± 150 μg alum, 0.1 μg dmLT, or 5 μg LTA1 on weeks 0, 3, and 6. Two-weeks later (week 8) serum and bone-marrow was collected for antibody evaluations. a Schematic of immunization schedule. b Compiled Week 5 and Week 9 serum anti-CRM or anti-FEN (coating antigen indicated) serum IgG. c Compiled EU/ml of indicated serum anti-FEN serum IgG1, or IgG2a ELISAs. d Representative anti-FEN (TT-BSA coated) IgG/IgA ELISPOT images from bone-marrow of week 8 immunized mice. Blue spots indicate IgG and red spots IgA ASCs. e Compiled anti-FEN IgG ASC per 10 6 cells from bone-marrow. Bars at mean + s.e.m. with significance determined by ANOVA paired with Bonferroni post-hoc test as shown (* P

Techniques Used: Mouse Assay, Enzyme-linked Immunospot

15) Product Images from "Probiotic Bacteria and Their Cell Walls Induce Th1-Type Immunity Against Salmonella Typhimurium Challenge"

Article Title: Probiotic Bacteria and Their Cell Walls Induce Th1-Type Immunity Against Salmonella Typhimurium Challenge

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2021.660854

Probiotic and their CW modulate mucosal antibody response against S. Typhimurium. (A, B) Total and Salmonella -specific sIgA levels in intestinal fluid of uninfected and S. Typhimurium infected mice. (C) Salmonella -specific IgG titer in serum of uninfected and S. Typhimurium infected mice. (D, E) Salmonella -specific IgG subclasses determined in serum of uninfected (D) and S. Typhimurium infected (E) mice. Data are shown as mean ± SEM. Data are pooled from two independent experiments (A, B) or representative of three independent experiments (C–E) with 3-6 mice per group. P values were calculated with one-way ANOVA with Šidak correction for multiple comparisons (A–C) and non-parametric Mann-Whitney test (D, E) . * P
Figure Legend Snippet: Probiotic and their CW modulate mucosal antibody response against S. Typhimurium. (A, B) Total and Salmonella -specific sIgA levels in intestinal fluid of uninfected and S. Typhimurium infected mice. (C) Salmonella -specific IgG titer in serum of uninfected and S. Typhimurium infected mice. (D, E) Salmonella -specific IgG subclasses determined in serum of uninfected (D) and S. Typhimurium infected (E) mice. Data are shown as mean ± SEM. Data are pooled from two independent experiments (A, B) or representative of three independent experiments (C–E) with 3-6 mice per group. P values were calculated with one-way ANOVA with Šidak correction for multiple comparisons (A–C) and non-parametric Mann-Whitney test (D, E) . * P

Techniques Used: Infection, Mouse Assay, MANN-WHITNEY

Probiotic and their CW reveal similar modulatory effect on the immune system during S. Typhimurium infection. Principal component analysis (PCA) plots of uninfected (A) and S. Typhimurium infected (B) mice. Variables analyzed: phagocytosis, sIgA, Salmonella-specific sIgA, IgG, IgG1, IgG2a, footpad swelling, IFN-γ, TNF-α, IL-12p70, IL-6, and IL-10.
Figure Legend Snippet: Probiotic and their CW reveal similar modulatory effect on the immune system during S. Typhimurium infection. Principal component analysis (PCA) plots of uninfected (A) and S. Typhimurium infected (B) mice. Variables analyzed: phagocytosis, sIgA, Salmonella-specific sIgA, IgG, IgG1, IgG2a, footpad swelling, IFN-γ, TNF-α, IL-12p70, IL-6, and IL-10.

Techniques Used: Infection, Mouse Assay

16) Product Images from "A2 adenosine receptor contributes to the development of cow’s milk protein allergy via regulating regulatory T cells"

Article Title: A2 adenosine receptor contributes to the development of cow’s milk protein allergy via regulating regulatory T cells

Journal: Iranian Journal of Basic Medical Sciences

doi: 10.22038/IJBMS.2021.57614.12812

Specific antibodies in ovalbumin (OVA)-treated mice. The blood was collected from mice. Serum was separated, and the levels of total immunoglobulin E (IgE, A), OVA-specific IgE (B), specific IgA (C), specific IgG1 (D) and specific IgG2a (E) were quantified by ELISA kits. Data represent the means ± SD of individual mice (n=8) in the group. ** P
Figure Legend Snippet: Specific antibodies in ovalbumin (OVA)-treated mice. The blood was collected from mice. Serum was separated, and the levels of total immunoglobulin E (IgE, A), OVA-specific IgE (B), specific IgA (C), specific IgG1 (D) and specific IgG2a (E) were quantified by ELISA kits. Data represent the means ± SD of individual mice (n=8) in the group. ** P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

17) Product Images from "A2 adenosine receptor contributes to the development of cow’s milk protein allergy via regulating regulatory T cells"

Article Title: A2 adenosine receptor contributes to the development of cow’s milk protein allergy via regulating regulatory T cells

Journal: Iranian Journal of Basic Medical Sciences

doi: 10.22038/IJBMS.2021.57614.12812

Specific antibodies in ovalbumin (OVA)-treated mice. The blood was collected from mice. Serum was separated, and the levels of total immunoglobulin E (IgE, A), OVA-specific IgE (B), specific IgA (C), specific IgG1 (D) and specific IgG2a (E) were quantified by ELISA kits. Data represent the means ± SD of individual mice (n=8) in the group. ** P
Figure Legend Snippet: Specific antibodies in ovalbumin (OVA)-treated mice. The blood was collected from mice. Serum was separated, and the levels of total immunoglobulin E (IgE, A), OVA-specific IgE (B), specific IgA (C), specific IgG1 (D) and specific IgG2a (E) were quantified by ELISA kits. Data represent the means ± SD of individual mice (n=8) in the group. ** P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

18) Product Images from "Daily full spectrum light exposure prevents food allergy-like allergic diarrhea by modulating vitamin D3 and microbiota composition"

Article Title: Daily full spectrum light exposure prevents food allergy-like allergic diarrhea by modulating vitamin D3 and microbiota composition

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-021-00213-8

Impact of phototherapy on immunoglobulin production in food allergy (FA). Serum levels of total or OVA-specific IgE ( a ), IgG1 ( b ), and IgG2a ( c ) in the FA (black bar) and phototherapy (gray bar) group were measured by ELISA. Values are presented as the mean ± SD of at least eight individuals in each group. *, **, P
Figure Legend Snippet: Impact of phototherapy on immunoglobulin production in food allergy (FA). Serum levels of total or OVA-specific IgE ( a ), IgG1 ( b ), and IgG2a ( c ) in the FA (black bar) and phototherapy (gray bar) group were measured by ELISA. Values are presented as the mean ± SD of at least eight individuals in each group. *, **, P

Techniques Used: Enzyme-linked Immunosorbent Assay

19) Product Images from "Daily full spectrum light exposure prevents food allergy-like allergic diarrhea by modulating vitamin D3 and microbiota composition"

Article Title: Daily full spectrum light exposure prevents food allergy-like allergic diarrhea by modulating vitamin D3 and microbiota composition

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-021-00213-8

Impact of phototherapy on immunoglobulin production in food allergy (FA). Serum levels of total or OVA-specific IgE ( a ), IgG1 ( b ), and IgG2a ( c ) in the FA (black bar) and phototherapy (gray bar) group were measured by ELISA. Values are presented as the mean ± SD of at least eight individuals in each group. *, **, P
Figure Legend Snippet: Impact of phototherapy on immunoglobulin production in food allergy (FA). Serum levels of total or OVA-specific IgE ( a ), IgG1 ( b ), and IgG2a ( c ) in the FA (black bar) and phototherapy (gray bar) group were measured by ELISA. Values are presented as the mean ± SD of at least eight individuals in each group. *, **, P

Techniques Used: Enzyme-linked Immunosorbent Assay

20) Product Images from "Reduction of Allergic Lung Disease by Mucosal Application of Toxoplasma gondii-Derived Molecules: Possible Role of Carbohydrates"

Article Title: Reduction of Allergic Lung Disease by Mucosal Application of Toxoplasma gondii-Derived Molecules: Possible Role of Carbohydrates

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.612766

TLA reduces systemic IL-4 and serum IgE-levels in a co-application model. (A) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and ovalbumin (OVA) restimulation of spleen cells from mice treated as in Figure 2A . (B) Levels of OVA-specific antibodies IgG1 and IgG2a in serum collected at the beginning and at the end of the experiment. (C) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 experiment with 5 mice per group (PBS/PBS) or 1 representative experiment from 2 independent experiments with 5 mice per group (PBS/OVA and TLA/OVA). Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; OD , optical density; nd , not detectable; ns , not significant; *P
Figure Legend Snippet: TLA reduces systemic IL-4 and serum IgE-levels in a co-application model. (A) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and ovalbumin (OVA) restimulation of spleen cells from mice treated as in Figure 2A . (B) Levels of OVA-specific antibodies IgG1 and IgG2a in serum collected at the beginning and at the end of the experiment. (C) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 experiment with 5 mice per group (PBS/PBS) or 1 representative experiment from 2 independent experiments with 5 mice per group (PBS/OVA and TLA/OVA). Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; OD , optical density; nd , not detectable; ns , not significant; *P

Techniques Used: Mouse Assay

TLA reduces allergic airway inflammation in a co-application model. (A) , Experimental design. (B) Airway hyperresponsiveness in response to methacholine. (C) Differential cell count in bronchoalveolar lavage (BALF). (D) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (E) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (F) PAS or hematoxylin and eosin (H E)-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (G) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and OVA restimulation of lung cells. Graphs show results from 1 experiment with 5 mice per group ( Figure 2B , PBS/PBS group in Figures 2C–G ) or from 1 representative experiment from 2 independent experiments with 5 mice per group (groups PBS/OVA and TLA/OVA, Figures 2C–G ). Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; OVA , ovalbumin; i.n. , intranasal; i.p. , intraperitoneal; macro , macrophages; eos , eosinophils; neutro , neutrophils; lympho , lymphocytes; nd , not detectable; ns , not significant; OD , optical density; *P
Figure Legend Snippet: TLA reduces allergic airway inflammation in a co-application model. (A) , Experimental design. (B) Airway hyperresponsiveness in response to methacholine. (C) Differential cell count in bronchoalveolar lavage (BALF). (D) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (E) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (F) PAS or hematoxylin and eosin (H E)-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (G) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and OVA restimulation of lung cells. Graphs show results from 1 experiment with 5 mice per group ( Figure 2B , PBS/PBS group in Figures 2C–G ) or from 1 representative experiment from 2 independent experiments with 5 mice per group (groups PBS/OVA and TLA/OVA, Figures 2C–G ). Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; OVA , ovalbumin; i.n. , intranasal; i.p. , intraperitoneal; macro , macrophages; eos , eosinophils; neutro , neutrophils; lympho , lymphocytes; nd , not detectable; ns , not significant; OD , optical density; *P

Techniques Used: Cell Counting, Histopathology, Staining, Mouse Assay

Therapeutic treatment with native or heat-inactivated TLA reduces systemic type 2 responses. (A) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and ovalbumin (OVA) restimulation of spleen cells from mice treated as in Figure 4A . (B) Levels of OVA-specific antibodies IgG1 and IgG2a in serum collected at the beginning and at the end of the experiment. (C) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group. Error bars show mean ± SEM. nd , not detectable; TLA , tachyzoites lysate antigen; hiTLA , heat-inactivated TLA; ns , not significant; OD , optical density; *P
Figure Legend Snippet: Therapeutic treatment with native or heat-inactivated TLA reduces systemic type 2 responses. (A) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and ovalbumin (OVA) restimulation of spleen cells from mice treated as in Figure 4A . (B) Levels of OVA-specific antibodies IgG1 and IgG2a in serum collected at the beginning and at the end of the experiment. (C) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group. Error bars show mean ± SEM. nd , not detectable; TLA , tachyzoites lysate antigen; hiTLA , heat-inactivated TLA; ns , not significant; OD , optical density; *P

Techniques Used: Mouse Assay

Therapeutic treatment with native or heat-inactivated TLA reduces allergic airway inflammation. (A) Experimental design. (B) Differential cell count in bronchoalveolar lavage (BALF). (C) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (D) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (E) PAS or H E-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (F) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and OVA restimulation of lung cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group. Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; hiTLA , heat-inactivated; OVA, ovalbumin; i.n., intranasal; i.p., intraperitoneal; macro, macrophages; eos, eosinophils; neutro, neutrophils; lympho, lymphocytes; n.s., not significant; OD, optical density; *P
Figure Legend Snippet: Therapeutic treatment with native or heat-inactivated TLA reduces allergic airway inflammation. (A) Experimental design. (B) Differential cell count in bronchoalveolar lavage (BALF). (C) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (D) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (E) PAS or H E-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (F) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and OVA restimulation of lung cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group. Error bars show mean ± SEM. TLA , tachyzoites lysate antigen; hiTLA , heat-inactivated; OVA, ovalbumin; i.n., intranasal; i.p., intraperitoneal; macro, macrophages; eos, eosinophils; neutro, neutrophils; lympho, lymphocytes; n.s., not significant; OD, optical density; *P

Techniques Used: Cell Counting, Histopathology, Staining, Mouse Assay

Prophylactic treatment with TLA fails to prevent allergic airway inflammation. (A) Experimental design. (B) Differential cell counts in bronchoalveolar lavage (BALF). (C) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (D) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (E) PAS or hematoxylin and eosin (H E)-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (F) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and ovalbumin (OVA) restimulation of lung cells. (G) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and OVA restimulation of spleen cells. (H) Levels of OVA-specific antibody IgG2a in serum collected at the beginning and at the end of the experiment. (I) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group ( Figures 1B–I ). Error bars show mean ± SEM. TLA, tachyzoites lysate antigen; OVA, ovalbumin; i.n., intranasal; i.p., intraperitoneal; macro, macrophages; eos, eosinophils; neutro, neutrophils; lympho , lymphocytes; OD, optical density; nd , not detectable; ns , not significant; ***P
Figure Legend Snippet: Prophylactic treatment with TLA fails to prevent allergic airway inflammation. (A) Experimental design. (B) Differential cell counts in bronchoalveolar lavage (BALF). (C) Levels of OVA-specific antibody IgG2a in BALF collected at the end of the experiment. (D) Average histopathology score and number of Periodic acid-Schiff (PAS)-positive goblet cells of lung sections. (E) PAS or hematoxylin and eosin (H E)-stained lung sections from 1 representative example from each group (n = 5); scale bars, 100 µm. (F) Levels of IL-4, IL-5, IL-10, IL-13, and IFN-γ after medium and ovalbumin (OVA) restimulation of lung cells. (G) Levels of IL-4, IL-5, IL-10, and IFN-γ after medium and OVA restimulation of spleen cells. (H) Levels of OVA-specific antibody IgG2a in serum collected at the beginning and at the end of the experiment. (I) Release of β-hexosaminidase by rat basophil leukemia (RBL) cells. Graphs show results from 1 representative experiment from 2 independent experiments with 5 mice per group ( Figures 1B–I ). Error bars show mean ± SEM. TLA, tachyzoites lysate antigen; OVA, ovalbumin; i.n., intranasal; i.p., intraperitoneal; macro, macrophages; eos, eosinophils; neutro, neutrophils; lympho , lymphocytes; OD, optical density; nd , not detectable; ns , not significant; ***P

Techniques Used: Histopathology, Staining, Mouse Assay

21) Product Images from "SLy2‐overexpression impairs B‐cell development in the bone marrow and the IgG response towards pneumococcal conjugate‐vaccine"

Article Title: SLy2‐overexpression impairs B‐cell development in the bone marrow and the IgG response towards pneumococcal conjugate‐vaccine

Journal: Immunity, Inflammation and Disease

doi: 10.1002/iid3.413

Prevenar13 (PCV13)‐specific immune globulin levels in the serum of mice. IgM, IgG 1 , IgG 2 , and IgG 3 titers were determined by ELISA at Day 0 before immunization and 7, 14 and 21 days post‐immunization with PCV13. Graphs depict the optical density (OD) at 450 nm measurement wavelength over time (570 nm reference wavelength). All sera were incubated on one plate to allow for direct comparison of the time points. Curves represent n = 8‐11 mice per genotypes from two independently performed experiments. Error bars display the mean ± SEM . Significance was determined with two‐way ANOVA with a p value of less than 0.05 was considered statistically significant (* p
Figure Legend Snippet: Prevenar13 (PCV13)‐specific immune globulin levels in the serum of mice. IgM, IgG 1 , IgG 2 , and IgG 3 titers were determined by ELISA at Day 0 before immunization and 7, 14 and 21 days post‐immunization with PCV13. Graphs depict the optical density (OD) at 450 nm measurement wavelength over time (570 nm reference wavelength). All sera were incubated on one plate to allow for direct comparison of the time points. Curves represent n = 8‐11 mice per genotypes from two independently performed experiments. Error bars display the mean ± SEM . Significance was determined with two‐way ANOVA with a p value of less than 0.05 was considered statistically significant (* p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Incubation

Surface immune globulin (Ig) class‐switch and antibody secretion of B cells upon in vitro stimulation. (A) The proliferation rate of isolated splenic B cells after 48 h of stimulation with lipopolysaccharide (LPS) only or LPS + interleukin‐4 (IL‐4) is shown. In addition, percentages of IgA + , IgG 2ab + , IgG 3 + (LPS, upper panel) and IgM + , IgD + , IgG 1 + (LPS + IL‐4, lower panel) B cells are depicted. (B) After 48 h of stimulation, immune globulins in the cell culture supernatants were quantified by LEGENDplex analysis. Graphs depict the mean fluorescent index (MFI) of PE‐fluorescence, which corresponds to the concentration of either IgM, IgA, IgG 2a , IgG 2b , IgG 1 , or IgG 3 in the supernatants. In every assay, an unstimulated (US) control was included. Data was assessed within three independent experiments using n = 5–9 mice per genotype. Significances were determined by one‐way analysis of variance (ANOVA) with a p value of less than 0.05 considered as statistically significant (* p
Figure Legend Snippet: Surface immune globulin (Ig) class‐switch and antibody secretion of B cells upon in vitro stimulation. (A) The proliferation rate of isolated splenic B cells after 48 h of stimulation with lipopolysaccharide (LPS) only or LPS + interleukin‐4 (IL‐4) is shown. In addition, percentages of IgA + , IgG 2ab + , IgG 3 + (LPS, upper panel) and IgM + , IgD + , IgG 1 + (LPS + IL‐4, lower panel) B cells are depicted. (B) After 48 h of stimulation, immune globulins in the cell culture supernatants were quantified by LEGENDplex analysis. Graphs depict the mean fluorescent index (MFI) of PE‐fluorescence, which corresponds to the concentration of either IgM, IgA, IgG 2a , IgG 2b , IgG 1 , or IgG 3 in the supernatants. In every assay, an unstimulated (US) control was included. Data was assessed within three independent experiments using n = 5–9 mice per genotype. Significances were determined by one‐way analysis of variance (ANOVA) with a p value of less than 0.05 considered as statistically significant (* p

Techniques Used: In Vitro, Isolation, Cell Culture, Fluorescence, Concentration Assay, Mouse Assay

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    Becton Dickinson mouse anti human igg1
    Elicitation and functionality of S1/S2- and RBD-specific antibodies in COVID-19 patients according to SARS-CoV-2 viremia. ( A ) <t>Anti-S1/S2</t> IgG (AU/ml) were assessed by means of a commercially available ELISA (LIAISON SARS-CoV-2 S1/S2 IgG, DiaSorin) in aviremic and viremic COVID-19 patients (n = 18 and n = 22). ( B ) Anti-RBD Ig (AUC) were assessed in aviremic and viremic COVID-19 patients by means of an in-house ELISA (n = 18 and n = 22 for total Ig; n = 17 and n = 21 for IgM, IgA, <t>IgG1</t> and IgG3). ( C ) Neutralization activity was measured by incubating pseudoviruses bearing the D614G SARS-CoV-2 Spike with serial dilutions of plasma for 1 h at 37 °C before infecting 293 T-ACE2 target cells. Neutralization half maximal inhibitory serum dilution (ID 50 ) values were determined using a normalized non-linear regression. ( D ) %ADCC in the presence of plasma at a 1/500 dilution. CEM.NKr parental cells were mixed at a 1:1 ratio with CEM.NKr-Spike cells and were used as target cells. PBMCs from uninfected donors were used as effector cells in a FACS-based ADCC assay. ( E ) Heatmap of correlations between SARS-CoV-2 viremia and humoral response. Median and interquartile range (IQR) are shown for each group of patients. S1/S2-specific total IgG, RBD-specific total Abs, IgM, IgA, IgG1 and IgG3 were negatively correlated with SARS-CoV-2 viremia (Spearman r = − 0.67, − 0.58, − 0.54, − 0.35, − 0.51 and − 0.34; p-value = 0.000003, 0.0001, 0.0005, 0.032, 0.001, 0.04, respectively). Mann–Whitney U test and Spearman’s correlation test, *statistical significance at p-value
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    Elicitation and functionality of S1/S2- and RBD-specific antibodies in COVID-19 patients according to SARS-CoV-2 viremia. ( A ) Anti-S1/S2 IgG (AU/ml) were assessed by means of a commercially available ELISA (LIAISON SARS-CoV-2 S1/S2 IgG, DiaSorin) in aviremic and viremic COVID-19 patients (n = 18 and n = 22). ( B ) Anti-RBD Ig (AUC) were assessed in aviremic and viremic COVID-19 patients by means of an in-house ELISA (n = 18 and n = 22 for total Ig; n = 17 and n = 21 for IgM, IgA, IgG1 and IgG3). ( C ) Neutralization activity was measured by incubating pseudoviruses bearing the D614G SARS-CoV-2 Spike with serial dilutions of plasma for 1 h at 37 °C before infecting 293 T-ACE2 target cells. Neutralization half maximal inhibitory serum dilution (ID 50 ) values were determined using a normalized non-linear regression. ( D ) %ADCC in the presence of plasma at a 1/500 dilution. CEM.NKr parental cells were mixed at a 1:1 ratio with CEM.NKr-Spike cells and were used as target cells. PBMCs from uninfected donors were used as effector cells in a FACS-based ADCC assay. ( E ) Heatmap of correlations between SARS-CoV-2 viremia and humoral response. Median and interquartile range (IQR) are shown for each group of patients. S1/S2-specific total IgG, RBD-specific total Abs, IgM, IgA, IgG1 and IgG3 were negatively correlated with SARS-CoV-2 viremia (Spearman r = − 0.67, − 0.58, − 0.54, − 0.35, − 0.51 and − 0.34; p-value = 0.000003, 0.0001, 0.0005, 0.032, 0.001, 0.04, respectively). Mann–Whitney U test and Spearman’s correlation test, *statistical significance at p-value

    Journal: Scientific Reports

    Article Title: Association between SARS-CoV-2 RNAemia and dysregulated immune response in acutely ill hospitalized COVID-19 patients

    doi: 10.1038/s41598-022-23923-1

    Figure Lengend Snippet: Elicitation and functionality of S1/S2- and RBD-specific antibodies in COVID-19 patients according to SARS-CoV-2 viremia. ( A ) Anti-S1/S2 IgG (AU/ml) were assessed by means of a commercially available ELISA (LIAISON SARS-CoV-2 S1/S2 IgG, DiaSorin) in aviremic and viremic COVID-19 patients (n = 18 and n = 22). ( B ) Anti-RBD Ig (AUC) were assessed in aviremic and viremic COVID-19 patients by means of an in-house ELISA (n = 18 and n = 22 for total Ig; n = 17 and n = 21 for IgM, IgA, IgG1 and IgG3). ( C ) Neutralization activity was measured by incubating pseudoviruses bearing the D614G SARS-CoV-2 Spike with serial dilutions of plasma for 1 h at 37 °C before infecting 293 T-ACE2 target cells. Neutralization half maximal inhibitory serum dilution (ID 50 ) values were determined using a normalized non-linear regression. ( D ) %ADCC in the presence of plasma at a 1/500 dilution. CEM.NKr parental cells were mixed at a 1:1 ratio with CEM.NKr-Spike cells and were used as target cells. PBMCs from uninfected donors were used as effector cells in a FACS-based ADCC assay. ( E ) Heatmap of correlations between SARS-CoV-2 viremia and humoral response. Median and interquartile range (IQR) are shown for each group of patients. S1/S2-specific total IgG, RBD-specific total Abs, IgM, IgA, IgG1 and IgG3 were negatively correlated with SARS-CoV-2 viremia (Spearman r = − 0.67, − 0.58, − 0.54, − 0.35, − 0.51 and − 0.34; p-value = 0.000003, 0.0001, 0.0005, 0.032, 0.001, 0.04, respectively). Mann–Whitney U test and Spearman’s correlation test, *statistical significance at p-value

    Article Snippet: The following biotinylated antibodies were used: goat anti-human kappa and lambda light chain for total antibodies (Bethyl Laboratories, Inc.), rabbit monoclonal anti-human IgM and IgA (Abnova), mouse anti-human IgG1 (BD Biosciences) and IgG3 (Southern Biotech); followed by avidin-HRP (ThermoFischer Scientific) for 30 min at RT.

    Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, FACS, ADCC Assay, MANN-WHITNEY