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mouse anti guinea pig cd8  (Bio-Rad)


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    Structured Review

    Bio-Rad mouse anti guinea pig cd8
    Increased frequencies of CD4 + CD44 + T cells and <t>CD8</t> + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.
    Mouse Anti Guinea Pig Cd8, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+guinea+pig+cd8/Mouse+anti+Guinea+Pig+CD8/pmc11979635-244-14-20
    Average 93 stars, based on 24 article reviews
    mouse anti guinea pig cd8 - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes"

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2025.1568258

    Increased frequencies of CD4 + CD44 + T cells and CD8 + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.
    Figure Legend Snippet: Increased frequencies of CD4 + CD44 + T cells and CD8 + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.

    Techniques Used: Infection

    Increased frequencies of CD8 + CD103 + T cells and CD8 + CRTAM T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD103 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CRTAM T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.
    Figure Legend Snippet: Increased frequencies of CD8 + CD103 + T cells and CD8 + CRTAM T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD103 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CRTAM T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Techniques Used: Infection

    Increased frequencies of CD4 + Ki-67 T cells and CD8 + Ki-67 T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.
    Figure Legend Snippet: Increased frequencies of CD4 + Ki-67 T cells and CD8 + Ki-67 T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Techniques Used: Infection

    Increased frequencies of CD4 + IFN-γ T cells and CD8 + IFN-γ T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals.
    Figure Legend Snippet: Increased frequencies of CD4 + IFN-γ T cells and CD8 + IFN-γ T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals.

    Techniques Used: Infection

    Related Articles

    Flow Cytometry:

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes
    Article Snippet: .. Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), and anti-mouse CD103 (clone 2E7, Biolegend) ( ). ..

    Article Title: Therapeutic prime/pull vaccination of HSV-2-infected guinea pigs with the ribonucleotide reductase 2 (RR2) protein and CXCL11 chemokine boosts antiviral local tissue-resident and effector memory CD4 + and CD8 + T cells and protects against recurrent genital herpes.
    Article Snippet: Following acute herpes simplex virus type 2 (HSV-2) infection, the virus undergoes an asymptomatic latent infection of sensory neurons of dorsal root ganglia (DRG).. Chemical and physical stress cause intermittent virus reactivation from latently infected DRG and recurrent virus shedding in the genital mucosal epithelium causing genital herpes in symptomatic patients.. While T cells appear to play a role in controlling virus reactivation from DRG and reducing the severity of recurrent genital herpes, the mechanisms for recruiting these T cells into DRG and the vaginal mucosa (VM) remain to be fully elucidated.

    Article Title: Therapeutic Prime/Pull Vaccination of HSV-2 Infected Guinea Pigs with the Ribonucleotide Reductase 2 (RR2) Protein and CXCL11 Chemokine Boosts Antiviral Local Tissue-Resident and Effector Memory CD4 + and CD8 + T Cells and Protects Against Recurrent Genital Herpes
    Article Snippet: .. Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), hamster anti-mouse PD-1 clone J43, BD Biosciences, San Jose, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), anti-mouse CXCR3 (clone CXCR3-173, Biolegend) and anti-mouse CD103 (clone 2E7, Biolegend). ..



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    Increased frequencies of CD4 + CD44 + T cells and <t>CD8</t> + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.
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    Figure 1. BCG-vaccination elicits CD1b-restricted, PIM6-specific T cell-responses. Four weeks after BCG-vaccination PBMCs were isolated. Autologous, CD1b-expressing APCs and non-adherent lymphocytes were stained with CFSE and stimulated with mycobacterial antigens. CFSE dilution was analyzed by flow-cytometry after five days. (A) Seven guinea pigs were control-treated. Eight guinea pigs were BCG-vaccinated. After four weeks, T cell-proliferation in response to the indicated mycobacterial antigen was assessed. The graphs on the left depict the gating strategy. In the diagrams, black bars represent the group mean as determined by nested-data-analysis. Error bars represent the standard error of the mean. Asterisks indicate the level of significance as determined by nested-t-test in comparison to the medium control. (B) CFSE-low, PIM6-reactive lymphocytes were stained for a general T cell-marker, CD4 and <t>CD8.</t> The small panel at the left shows the gating strategy. The larger pseudo-color graph representatively shows the CD4- and CD8-distribution. The dot blot on the right shows the relative distribution for T cell-markers in PIM6-reactive cells four weeks after BCG-vaccination. Red symbols represent the normal T cell- and CD4-CD8-distribution in PBMCs from naïve control guinea pigs. Asterisks indicate the level of significance as determined by unpaired t-test. (C) Lymphocytes were stimulated with PIM6 as described above. After 24 h RNA was harvested. Cytokine-transcript levels were determined by qRT-PCR in relation to β-Actin. Open bars represent non-stimulated controls, black bars represent transcript levels after PIM6-stimulation. The left panel shows the results for BCG-vaccinated animals, the right panel shows the upregulation of IFNγ, GM-CSF and IL17 for naïve, non-immunized animals. Black bars represent the group mean. Error bars indicate the standard error of the mean. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test in comparison to non-stimulated medium controls. (D) IFNγ-transcript levels were assessed in the presence of CD1b-blocking or isotype-matched control antibodies. Individually colored circles represent transcript levels of eight BCG-vaccinated guinea pigs four weeks after vaccination. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test as indicated.
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    Image Search Results


    Increased frequencies of CD4 + CD44 + T cells and CD8 + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.

    Journal: Frontiers in Immunology

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes

    doi: 10.3389/fimmu.2025.1568258

    Figure Lengend Snippet: Increased frequencies of CD4 + CD44 + T cells and CD8 + CD44 + T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus- based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + CD44 + T cells detected in the DRG, VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD44 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, **** = P≤0.0001, ns, non significant.

    Article Snippet: Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), and anti-mouse CD103 (clone 2E7, Biolegend) ( ).

    Techniques: Infection

    Increased frequencies of CD8 + CD103 + T cells and CD8 + CRTAM T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD103 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CRTAM T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Journal: Frontiers in Immunology

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes

    doi: 10.3389/fimmu.2025.1568258

    Figure Lengend Snippet: Increased frequencies of CD8 + CD103 + T cells and CD8 + CRTAM T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CD103 + T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + CRTAM T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Article Snippet: Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), and anti-mouse CD103 (clone 2E7, Biolegend) ( ).

    Techniques: Infection

    Increased frequencies of CD4 + Ki-67 T cells and CD8 + Ki-67 T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Journal: Frontiers in Immunology

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes

    doi: 10.3389/fimmu.2025.1568258

    Figure Lengend Snippet: Increased frequencies of CD4 + Ki-67 T cells and CD8 + Ki-67 T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + Ki-67 T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. * = P<0.05, ** = P≤0.01, *** = P≤0.001, ns, non significant.

    Article Snippet: Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), and anti-mouse CD103 (clone 2E7, Biolegend) ( ).

    Techniques: Infection

    Increased frequencies of CD4 + IFN-γ T cells and CD8 + IFN-γ T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals.

    Journal: Frontiers in Immunology

    Article Title: Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protection against recurrent genital herpes

    doi: 10.3389/fimmu.2025.1568258

    Figure Lengend Snippet: Increased frequencies of CD4 + IFN-γ T cells and CD8 + IFN-γ T cells in the DRG and VM of HSV-2 infected guinea pigs following therapeutic immunization with five adenovirus-based vaccine candidates: (A) Representative FACS data (left panel) and average frequencies (right panel) of CD4 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals. (B) Representative FACS data (left panel) and average frequencies (right panel) of CD8 + IFN-γ T cells detected in the DRG, and VM of vaccinated and mock-vaccinated animals.

    Article Snippet: Vaginal mucosa cells and splenocytes were analyzed by flow cytometry using the following antibodies: mouse anti-guinea pig CD8 (clone MCA752F, Bio-Rad Laboratories, Hercules, CA), mouse anti-guinea pig CD4 (clone MCA749PE, Bio-Rad Laboratories), anti-mouse CRTAM (clone 11-5, Biolegend, San Diego, CA), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD69 (clone H1.2F3, BD Biosciences, San Jose, CA), and anti-mouse CD103 (clone 2E7, Biolegend) ( ).

    Techniques: Infection

    Figure 1. BCG-vaccination elicits CD1b-restricted, PIM6-specific T cell-responses. Four weeks after BCG-vaccination PBMCs were isolated. Autologous, CD1b-expressing APCs and non-adherent lymphocytes were stained with CFSE and stimulated with mycobacterial antigens. CFSE dilution was analyzed by flow-cytometry after five days. (A) Seven guinea pigs were control-treated. Eight guinea pigs were BCG-vaccinated. After four weeks, T cell-proliferation in response to the indicated mycobacterial antigen was assessed. The graphs on the left depict the gating strategy. In the diagrams, black bars represent the group mean as determined by nested-data-analysis. Error bars represent the standard error of the mean. Asterisks indicate the level of significance as determined by nested-t-test in comparison to the medium control. (B) CFSE-low, PIM6-reactive lymphocytes were stained for a general T cell-marker, CD4 and CD8. The small panel at the left shows the gating strategy. The larger pseudo-color graph representatively shows the CD4- and CD8-distribution. The dot blot on the right shows the relative distribution for T cell-markers in PIM6-reactive cells four weeks after BCG-vaccination. Red symbols represent the normal T cell- and CD4-CD8-distribution in PBMCs from naïve control guinea pigs. Asterisks indicate the level of significance as determined by unpaired t-test. (C) Lymphocytes were stimulated with PIM6 as described above. After 24 h RNA was harvested. Cytokine-transcript levels were determined by qRT-PCR in relation to β-Actin. Open bars represent non-stimulated controls, black bars represent transcript levels after PIM6-stimulation. The left panel shows the results for BCG-vaccinated animals, the right panel shows the upregulation of IFNγ, GM-CSF and IL17 for naïve, non-immunized animals. Black bars represent the group mean. Error bars indicate the standard error of the mean. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test in comparison to non-stimulated medium controls. (D) IFNγ-transcript levels were assessed in the presence of CD1b-blocking or isotype-matched control antibodies. Individually colored circles represent transcript levels of eight BCG-vaccinated guinea pigs four weeks after vaccination. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test as indicated.

    Journal: Scientific reports

    Article Title: Phosphatidylinositolmannoside vaccination induces lipid-specific Th1-responses and partially protects guinea pigs from Mycobacterium tuberculosis challenge.

    doi: 10.1038/s41598-023-45898-3

    Figure Lengend Snippet: Figure 1. BCG-vaccination elicits CD1b-restricted, PIM6-specific T cell-responses. Four weeks after BCG-vaccination PBMCs were isolated. Autologous, CD1b-expressing APCs and non-adherent lymphocytes were stained with CFSE and stimulated with mycobacterial antigens. CFSE dilution was analyzed by flow-cytometry after five days. (A) Seven guinea pigs were control-treated. Eight guinea pigs were BCG-vaccinated. After four weeks, T cell-proliferation in response to the indicated mycobacterial antigen was assessed. The graphs on the left depict the gating strategy. In the diagrams, black bars represent the group mean as determined by nested-data-analysis. Error bars represent the standard error of the mean. Asterisks indicate the level of significance as determined by nested-t-test in comparison to the medium control. (B) CFSE-low, PIM6-reactive lymphocytes were stained for a general T cell-marker, CD4 and CD8. The small panel at the left shows the gating strategy. The larger pseudo-color graph representatively shows the CD4- and CD8-distribution. The dot blot on the right shows the relative distribution for T cell-markers in PIM6-reactive cells four weeks after BCG-vaccination. Red symbols represent the normal T cell- and CD4-CD8-distribution in PBMCs from naïve control guinea pigs. Asterisks indicate the level of significance as determined by unpaired t-test. (C) Lymphocytes were stimulated with PIM6 as described above. After 24 h RNA was harvested. Cytokine-transcript levels were determined by qRT-PCR in relation to β-Actin. Open bars represent non-stimulated controls, black bars represent transcript levels after PIM6-stimulation. The left panel shows the results for BCG-vaccinated animals, the right panel shows the upregulation of IFNγ, GM-CSF and IL17 for naïve, non-immunized animals. Black bars represent the group mean. Error bars indicate the standard error of the mean. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test in comparison to non-stimulated medium controls. (D) IFNγ-transcript levels were assessed in the presence of CD1b-blocking or isotype-matched control antibodies. Individually colored circles represent transcript levels of eight BCG-vaccinated guinea pigs four weeks after vaccination. Asterisks depict the level of significance as determined by Wilcoxon-matched-pair-signed-ranked-test as indicated.

    Article Snippet: CFSE-negative lymphocytes were further characterized by a triple-staining using allophycocyanin-conjugated mouse-anti-guinea-pig-T cell-antibody (AbD Serotec, Germany); phycoerythrin-conjugated mouse-anti-guineapig-CD4 antibody (AbD Serotec) and biotinylated mouse-anti-guinea-pig-CD8 antibody (kindly provided by Dr. Hubert Schäfer, Robert Koch-Institut, Germany).

    Techniques: Isolation, Expressing, Staining, Flow Cytometry, Control, Comparison, Marker, Dot Blot, Quantitative RT-PCR, Blocking Assay