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The Jackson Laboratory b6 pl
Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in <t>B6.PL/3H9.KI</t> mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.
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1) Product Images from "Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance"

Article Title: Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance

Journal: Journal of Clinical Investigation

doi: 10.1172/JCI200318310

Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in B6.PL/3H9.KI mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.
Figure Legend Snippet: Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in B6.PL/3H9.KI mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.

Techniques Used: Mouse Assay, Staining, Immunohistochemistry, Activation Assay

Loss of B cell tolerance is associated with B7.2 expression on anti-dsDNA B cells. Seven days after cGVHD induction in B6.PL/3H9.KI mice, spleen cells were stained for expression of B7.2, B220, and λ1. Histograms show B7.2 expression on gated B220 + λ1 + cells for mice receiving B6 (thin lines) or I-A b –reactive CD4 + T cells (thick lines). Representative results of four experiments are shown.
Figure Legend Snippet: Loss of B cell tolerance is associated with B7.2 expression on anti-dsDNA B cells. Seven days after cGVHD induction in B6.PL/3H9.KI mice, spleen cells were stained for expression of B7.2, B220, and λ1. Histograms show B7.2 expression on gated B220 + λ1 + cells for mice receiving B6 (thin lines) or I-A b –reactive CD4 + T cells (thick lines). Representative results of four experiments are shown.

Techniques Used: Expressing, Mouse Assay, Staining

2) Product Images from "IL-1?-Mediated Signals Preferentially Drive Conversion of Regulatory T Cells but Not Conventional T Cells into IL-17-Producing Cells"

Article Title: IL-1?-Mediated Signals Preferentially Drive Conversion of Regulatory T Cells but Not Conventional T Cells into IL-17-Producing Cells

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.1001536

Tregs are capable of producing IL-17A in vivo after immunization. IL-1R1 −/− or B6.PL congenic (Thy1.1 + ) Tregs were transferred to IL-1R1 −/− mice that were subsequently immunized with KLH in IFA. A and B , Three days later,
Figure Legend Snippet: Tregs are capable of producing IL-17A in vivo after immunization. IL-1R1 −/− or B6.PL congenic (Thy1.1 + ) Tregs were transferred to IL-1R1 −/− mice that were subsequently immunized with KLH in IFA. A and B , Three days later,

Techniques Used: In Vivo, Mouse Assay, Immunofluorescence

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    The Jackson Laboratory thy1 1
    Kinetics of naive and memory CD4 + T cells in the same mouse. Wildtype mice containing 1.3×10 3 naïve SMARTA <t>(Thy1.1)</t> and 1.3×10 3 memory SMARTA (Ly5a) cells were given LCMV, and the relative abundance of the two SMARTA cell populations was determined by flow cytometry at various times post infection (two mice per time point). A. After gating on CD4 + T cells, the host CD4 + T cells (H), and the naïve and memory SMARTA cells (N M respectively) were distinguished by Thy1.1 and Ly5a staining. The numbers indicate the frequencies of naïve and memory SMARTA cells as a percentage of all CD4 + T cells. B. The average±SE of the percentage of each population among all CD4 + T cells is shown over time. C. The total number of memory or naïve SMARTA CD4 + T cells per spleen is shown (average±SE).
    Thy1 1, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory b6 pl mice
    Induction of antigen specific IL-10 + Tr1 and Treg cells in vivo . ( a ) <t>B6.PL</t> (Thy1.1 + ) mice reconstituted with OT-II TCR transgenic T cells were injected i.v. with class II–restricted OVA 323–339 peptide (OVA) alone, or OVA plus LPS, OVA plus zymosan, or OVA plus curdlan. Four days after challenge, the splenocytes were isolated and expression of Foxp3, IL-17, IFN-γ and IL-10 by CD4 + T Thy1.2 + cell was assessed by intracellular staining and flow cytometry. Data are from one experiment representative of two. ( b ) C57BL/6 or Tlr2 −/− or Il-10 −/− mouse were reconstituted with OT-II TCR transgenic T cells, and on the following day, injected with OVA or OVA plus zymosan. Five days later, splenocytes were isolated and induction of OVA specific Foxp3 + T cells was assessed by intracellular staining and flow cytometry. Means + s. d. of 3 or 4 mice per group. ( c ) Splenocytes from immunized mice described in ( b ) were restimulated with OVA in culture for 48 h and cytokines in the supernatants were analyzed by ELISA. Means + s. d. of 3 or 4 mice per group. * P
    B6 Pl Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory b6 pl
    Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in <t>B6.PL/3H9.KI</t> mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.
    B6 Pl, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory b6 thy1 1 mice
    The number of CD8 + NKT-like cells increased after immunization with GFP-DCs. ( a ) The flow chart shows that C57BL/6 mice were immunized by 2 × 10 6 LPS-pulsed GFP-DCs four times. The splenocytes were then isolated, and panNK cells were enriched using a panNK selection kit. ( b ) The gating strategy used to identify NK and NKT cell subsets is shown. ( c ) The proportion of TCRβ + CD49b + cells in panNK cells and the absolute number of TCRβ + CD49b + cells in naïve mice and GFP-DC-immunized mice were compared. ( d ) The proportion of CD8 + NKT-like cells in TCRβ + CD49b + cells and the absolute number of CD8 + NKT-like cells in naïve mice and GFP-DC-immunized mice were compared. ( e ) We isolated 1 × 10 6 CD49b + CD8 T cells and 1 × 10 7 CD49b − CD8 T cells from B6 CD45.1 mice and <t>B6</t> Thy1.1 mice, respectively, through flow cytometry; the cells were injected into C57BL/6 mice, which were subsequently immunized with 2 × 10 6 GFP-DCs. After 14 days, the recipient mice were again immunized with 2 × 10 6 GFP-DCs. Next, donor splenic cells were analyzed for phenotype, and the CFSE dilution experiment was performed after 3 days. The data represent three independent experiments (n = 8–10).
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    Kinetics of naive and memory CD4 + T cells in the same mouse. Wildtype mice containing 1.3×10 3 naïve SMARTA (Thy1.1) and 1.3×10 3 memory SMARTA (Ly5a) cells were given LCMV, and the relative abundance of the two SMARTA cell populations was determined by flow cytometry at various times post infection (two mice per time point). A. After gating on CD4 + T cells, the host CD4 + T cells (H), and the naïve and memory SMARTA cells (N M respectively) were distinguished by Thy1.1 and Ly5a staining. The numbers indicate the frequencies of naïve and memory SMARTA cells as a percentage of all CD4 + T cells. B. The average±SE of the percentage of each population among all CD4 + T cells is shown over time. C. The total number of memory or naïve SMARTA CD4 + T cells per spleen is shown (average±SE).

    Journal: PLoS Pathogens

    Article Title: Tentative T Cells: Memory Cells Are Quick to Respond, but Slow to Divide

    doi: 10.1371/journal.ppat.1000041

    Figure Lengend Snippet: Kinetics of naive and memory CD4 + T cells in the same mouse. Wildtype mice containing 1.3×10 3 naïve SMARTA (Thy1.1) and 1.3×10 3 memory SMARTA (Ly5a) cells were given LCMV, and the relative abundance of the two SMARTA cell populations was determined by flow cytometry at various times post infection (two mice per time point). A. After gating on CD4 + T cells, the host CD4 + T cells (H), and the naïve and memory SMARTA cells (N M respectively) were distinguished by Thy1.1 and Ly5a staining. The numbers indicate the frequencies of naïve and memory SMARTA cells as a percentage of all CD4 + T cells. B. The average±SE of the percentage of each population among all CD4 + T cells is shown over time. C. The total number of memory or naïve SMARTA CD4 + T cells per spleen is shown (average±SE).

    Article Snippet: C57BL/6 mice congenic for Thy1.1 (B6.PL-Thy1a /CyJ) were purchased from The Jackson Laboratory.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Infection, Staining

    Viral epitopes are presented within hours of infection, and stimulate memory T cell effector functions. Mice that contained approximately 3×10 3 SMARTA/Ly5a CD4 + T cells were infected with LCMV and, 354 days later, were re-challenged intraperitoneally with 2×10 6 PFU LCMV-Armstong. Six hours post-infection, the mice were given 0.25 mg Brefeldin A i.v., and 6 hours later the spleens were harvested and immediately surface stained for CD4, Ly5a, or CD8, then permeabilized and stained for intracellular IFNγ. The cells were not re-stimulated ex vivo with peptide antigen. A. ∼5% of all CD8 + T cells, and ∼1% of all CD4 + T cells, are actively producing IFNγ in response to infection. B. Using the SMARTA cells transferred ∼1 year previously as an indicator of the responsiveness of virus-specific CD4 + memory T cells, ∼14% of LCMV-specific CD4 + memory T cells actively produce IFNγ within 12 hours of virus infection. Data shown are from an individual mouse, and are representative of independent datasets. C. A separate set of naive mice were given CFSE-labeled pooled SMARTA cells (4×10 5 naive SMARTA/Thy1.1 cells and 2×10 4 memory SMARTA/Ly5a T cells). 4 days later, some of the recipient mice were given LCMV. Six hours later, BFA was administered to all mice, and after a further 6 hours splenocytes were harvested. The cells were immediately stained (without peptide re-stimulation) for CD4, Thy1.1, Ly5a and IFNγ, and were analyzed by flow cytometry. Approximately 2% memory SMARTA cells had begun to synthesize IFNγ in response to LCMV infection (top row) but none of those responding memory cells showed any dilution of CFSE signal. The naïve SMARTA cells (bottom row) failed to produce IFNγ at this early time point post-infection, and no sign of cell division was seen. Data are from one of two independent experiments.

    Journal: PLoS Pathogens

    Article Title: Tentative T Cells: Memory Cells Are Quick to Respond, but Slow to Divide

    doi: 10.1371/journal.ppat.1000041

    Figure Lengend Snippet: Viral epitopes are presented within hours of infection, and stimulate memory T cell effector functions. Mice that contained approximately 3×10 3 SMARTA/Ly5a CD4 + T cells were infected with LCMV and, 354 days later, were re-challenged intraperitoneally with 2×10 6 PFU LCMV-Armstong. Six hours post-infection, the mice were given 0.25 mg Brefeldin A i.v., and 6 hours later the spleens were harvested and immediately surface stained for CD4, Ly5a, or CD8, then permeabilized and stained for intracellular IFNγ. The cells were not re-stimulated ex vivo with peptide antigen. A. ∼5% of all CD8 + T cells, and ∼1% of all CD4 + T cells, are actively producing IFNγ in response to infection. B. Using the SMARTA cells transferred ∼1 year previously as an indicator of the responsiveness of virus-specific CD4 + memory T cells, ∼14% of LCMV-specific CD4 + memory T cells actively produce IFNγ within 12 hours of virus infection. Data shown are from an individual mouse, and are representative of independent datasets. C. A separate set of naive mice were given CFSE-labeled pooled SMARTA cells (4×10 5 naive SMARTA/Thy1.1 cells and 2×10 4 memory SMARTA/Ly5a T cells). 4 days later, some of the recipient mice were given LCMV. Six hours later, BFA was administered to all mice, and after a further 6 hours splenocytes were harvested. The cells were immediately stained (without peptide re-stimulation) for CD4, Thy1.1, Ly5a and IFNγ, and were analyzed by flow cytometry. Approximately 2% memory SMARTA cells had begun to synthesize IFNγ in response to LCMV infection (top row) but none of those responding memory cells showed any dilution of CFSE signal. The naïve SMARTA cells (bottom row) failed to produce IFNγ at this early time point post-infection, and no sign of cell division was seen. Data are from one of two independent experiments.

    Article Snippet: C57BL/6 mice congenic for Thy1.1 (B6.PL-Thy1a /CyJ) were purchased from The Jackson Laboratory.

    Techniques: Infection, Mouse Assay, Staining, Ex Vivo, Labeling, Flow Cytometry, Cytometry

    Induction of antigen specific IL-10 + Tr1 and Treg cells in vivo . ( a ) B6.PL (Thy1.1 + ) mice reconstituted with OT-II TCR transgenic T cells were injected i.v. with class II–restricted OVA 323–339 peptide (OVA) alone, or OVA plus LPS, OVA plus zymosan, or OVA plus curdlan. Four days after challenge, the splenocytes were isolated and expression of Foxp3, IL-17, IFN-γ and IL-10 by CD4 + T Thy1.2 + cell was assessed by intracellular staining and flow cytometry. Data are from one experiment representative of two. ( b ) C57BL/6 or Tlr2 −/− or Il-10 −/− mouse were reconstituted with OT-II TCR transgenic T cells, and on the following day, injected with OVA or OVA plus zymosan. Five days later, splenocytes were isolated and induction of OVA specific Foxp3 + T cells was assessed by intracellular staining and flow cytometry. Means + s. d. of 3 or 4 mice per group. ( c ) Splenocytes from immunized mice described in ( b ) were restimulated with OVA in culture for 48 h and cytokines in the supernatants were analyzed by ELISA. Means + s. d. of 3 or 4 mice per group. * P

    Journal: Nature medicine

    Article Title: TLR2 dependent induction of vitamin A metabolizing enzymes in dendritic cells promotes T regulatory responses and inhibits TH-17 mediated autoimmunity

    doi: 10.1038/nm.1925

    Figure Lengend Snippet: Induction of antigen specific IL-10 + Tr1 and Treg cells in vivo . ( a ) B6.PL (Thy1.1 + ) mice reconstituted with OT-II TCR transgenic T cells were injected i.v. with class II–restricted OVA 323–339 peptide (OVA) alone, or OVA plus LPS, OVA plus zymosan, or OVA plus curdlan. Four days after challenge, the splenocytes were isolated and expression of Foxp3, IL-17, IFN-γ and IL-10 by CD4 + T Thy1.2 + cell was assessed by intracellular staining and flow cytometry. Data are from one experiment representative of two. ( b ) C57BL/6 or Tlr2 −/− or Il-10 −/− mouse were reconstituted with OT-II TCR transgenic T cells, and on the following day, injected with OVA or OVA plus zymosan. Five days later, splenocytes were isolated and induction of OVA specific Foxp3 + T cells was assessed by intracellular staining and flow cytometry. Means + s. d. of 3 or 4 mice per group. ( c ) Splenocytes from immunized mice described in ( b ) were restimulated with OVA in culture for 48 h and cytokines in the supernatants were analyzed by ELISA. Means + s. d. of 3 or 4 mice per group. * P

    Article Snippet: Mice C57BL/6, OT-II and B6.PL mice were from Jackson Laboratories.

    Techniques: In Vivo, Mouse Assay, Transgenic Assay, Injection, Isolation, Expressing, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in B6.PL/3H9.KI mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.

    Journal: Journal of Clinical Investigation

    Article Title: Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance

    doi: 10.1172/JCI200318310

    Figure Lengend Snippet: Limited T cell repertoires are activated but fail to enter the B cell follicle. ( a ) Donor Thy1.2 + CD4 cells were identified 7 days after induction of cGVHD in B6.PL/3H9.KI mice by staining with Thy1.2 and CD4. Gated populations of the contour plots represent the expansion in percentage of transferred CD4 + T cells for the indicated cell population. The total number of recovered Thy1.2 + CD4 + splenocytes is indicated above the plot. Representative results from four experiments are shown. ( b ) Donor CD4 + cells were localized by staining spleen sections with Ab’s against Thy1.2 (brown) and B220 (blue). The few B6 CD4 + T cells detected by IHC were scattered throughout the T cell area. Activation of the class II–reactive T cell repertoires resulted in the majority of the cells migrating to T-B interface. The diverse T cell repertoires (K14 and bm12) contained T cells that were also evident in the B cell follicle; however, activation of the limited T cell repertoires (2-2-3/K14 and H2-DM –/– ) did not result in the presence of transferred Thy1.2 + T cells in the follicle. Representative results of four experiments are shown.

    Article Snippet: C57Bl/6J, B6.PL, and H-2bm12 (bm12) mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

    Techniques: Mouse Assay, Staining, Immunohistochemistry, Activation Assay

    Loss of B cell tolerance is associated with B7.2 expression on anti-dsDNA B cells. Seven days after cGVHD induction in B6.PL/3H9.KI mice, spleen cells were stained for expression of B7.2, B220, and λ1. Histograms show B7.2 expression on gated B220 + λ1 + cells for mice receiving B6 (thin lines) or I-A b –reactive CD4 + T cells (thick lines). Representative results of four experiments are shown.

    Journal: Journal of Clinical Investigation

    Article Title: Activation of diverse repertoires of autoreactive T cells enhances the loss of anti-dsDNA B cell tolerance

    doi: 10.1172/JCI200318310

    Figure Lengend Snippet: Loss of B cell tolerance is associated with B7.2 expression on anti-dsDNA B cells. Seven days after cGVHD induction in B6.PL/3H9.KI mice, spleen cells were stained for expression of B7.2, B220, and λ1. Histograms show B7.2 expression on gated B220 + λ1 + cells for mice receiving B6 (thin lines) or I-A b –reactive CD4 + T cells (thick lines). Representative results of four experiments are shown.

    Article Snippet: C57Bl/6J, B6.PL, and H-2bm12 (bm12) mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

    Techniques: Expressing, Mouse Assay, Staining

    The number of CD8 + NKT-like cells increased after immunization with GFP-DCs. ( a ) The flow chart shows that C57BL/6 mice were immunized by 2 × 10 6 LPS-pulsed GFP-DCs four times. The splenocytes were then isolated, and panNK cells were enriched using a panNK selection kit. ( b ) The gating strategy used to identify NK and NKT cell subsets is shown. ( c ) The proportion of TCRβ + CD49b + cells in panNK cells and the absolute number of TCRβ + CD49b + cells in naïve mice and GFP-DC-immunized mice were compared. ( d ) The proportion of CD8 + NKT-like cells in TCRβ + CD49b + cells and the absolute number of CD8 + NKT-like cells in naïve mice and GFP-DC-immunized mice were compared. ( e ) We isolated 1 × 10 6 CD49b + CD8 T cells and 1 × 10 7 CD49b − CD8 T cells from B6 CD45.1 mice and B6 Thy1.1 mice, respectively, through flow cytometry; the cells were injected into C57BL/6 mice, which were subsequently immunized with 2 × 10 6 GFP-DCs. After 14 days, the recipient mice were again immunized with 2 × 10 6 GFP-DCs. Next, donor splenic cells were analyzed for phenotype, and the CFSE dilution experiment was performed after 3 days. The data represent three independent experiments (n = 8–10).

    Journal: Scientific Reports

    Article Title: CD8+NKT-like cells regulate the immune response by killing antigen-bearing DCs

    doi: 10.1038/srep14124

    Figure Lengend Snippet: The number of CD8 + NKT-like cells increased after immunization with GFP-DCs. ( a ) The flow chart shows that C57BL/6 mice were immunized by 2 × 10 6 LPS-pulsed GFP-DCs four times. The splenocytes were then isolated, and panNK cells were enriched using a panNK selection kit. ( b ) The gating strategy used to identify NK and NKT cell subsets is shown. ( c ) The proportion of TCRβ + CD49b + cells in panNK cells and the absolute number of TCRβ + CD49b + cells in naïve mice and GFP-DC-immunized mice were compared. ( d ) The proportion of CD8 + NKT-like cells in TCRβ + CD49b + cells and the absolute number of CD8 + NKT-like cells in naïve mice and GFP-DC-immunized mice were compared. ( e ) We isolated 1 × 10 6 CD49b + CD8 T cells and 1 × 10 7 CD49b − CD8 T cells from B6 CD45.1 mice and B6 Thy1.1 mice, respectively, through flow cytometry; the cells were injected into C57BL/6 mice, which were subsequently immunized with 2 × 10 6 GFP-DCs. After 14 days, the recipient mice were again immunized with 2 × 10 6 GFP-DCs. Next, donor splenic cells were analyzed for phenotype, and the CFSE dilution experiment was performed after 3 days. The data represent three independent experiments (n = 8–10).

    Article Snippet: Mice Both the wild-type and transgenic mouse strains, including B6.GFP mice [C57BL/6-Tg(ACTB-EGFP)1Osb/J mice], B6.OVA mice [C57BL/6-Tg(ACTB-OVA)916Jen/J mice], B6 Thy1.1 mice (B6.PL-Thy1a /CyJ mice), OT-I mice [C57BL/6-Tg(TcraTcrb)1100Mjb/J mice], OT-II mice [B6.Cg-Tg(TcraTcrb)425Cbn/J mice], CD1d−/− mice [B6.129S6-Del(3Cd1d2-Cd1d1)1Sbp/J mice], β2 m−/− mice (B6.129P2-B2mtm1Unc /J mice) and TAP−/− mice (B6.129S2-Tap1tm1Arp /J mice) were obtained from the Jackson Laboratory and bred in specific pathogen-free conditions.

    Techniques: Flow Cytometry, Mouse Assay, Isolation, Selection, Cytometry, Injection