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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as <t>CD69</t> + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.
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Image Search Results


(A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as CD69 + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.

Journal: bioRxiv

Article Title: Chlamydia muridarum Causes Persistent Subclinical Infection and Elicits Innate and Adaptive Immune Responses in C57BL/6J, BALB/cJ and J:ARC(S) Mice Following Exposure to Shedding Mice

doi: 10.1101/2024.07.16.603732

Figure Lengend Snippet: (A) Gating strategy defining the immune subpopulations of splenic cells after excluding debris, doublets, and dead cells. Splenic immune cells were first identified as CD45 + cells. B cells were then quantified as CD3 - /B220 + . T cells were identified as CD3 + . CD8 and CD4 T cells were identified by expression of their respective marker, while the subpopulation of T reg was identified as CD4 + /CD25 + /CD127 - . CD4 + and CD8 + T cell activation was identified as CD69 + . Neutrophils and monocytes were quantified as CD11b+, and further distinguished as Ly6G+ and Ly6G–Ly6C+, respectively. Data displayed are from B6 mice at 14 DPC. (B) Infected B6 mice demonstrated increased frequencies of myeloid and lymphoid cell population subtypes when compared to uninfected controls, including monocytes (63 DPC), effector CD4+ T cells (14 and 63 DPC), effector CD8+ T cells (63 DPC) and effector memory CD8+ T cells (14 DPC). Other population increases did not reach statistical significance. Additionally, infected B6 mice had fewer B cells (63 DPC), naïve CD4+ cells (14 DPC), and overall CD8+ T cells (14 FPC). Data presented are frequencies of specific cell populations as compared with the parent population and are displayed as proportions of infected compared to control mice (n=6 mice/group). Dotted line (1.0) indicates no difference in frequency of detection between control and infected mice. Values above this line represent an increased frequency of detection in infected mice and values below this line represent a decreased frequency of detection in infected mice. macro = macrophage; mono = monocyte; neut = neutrophil; EM = effector memory cell; TIC = total immune cells. *=p < 0.05.

Article Snippet: Cells were then stained with the following anti-mouse antibodies: APC/Fire 810 CD3 (17A2, catalog no. 100267, Biolegend), CD11b BV570 (M1/70, catalog no. 101233, Biolegend), Alexa Fluor 700 Ly-6C (HK1.4, catalog no. 128024, Biolegend), PE/Cyanine7 CD127 IL-7Rα (A7R34, catalog no. 135014, Biolegend), PerCP Ly-6G (1A8, catalog no. 127653, Biolegend), Brilliant Violet 421 CD25 (PC61, catalog no. 102034, Biolegend), Brilliant Violet 650 CD45R/B220 (RA3-6B2, catalog no. 103241, Biolegend), BUV661 NK-1.1 (PK136, catalog no. 741477, BD Biosciences, Ashland, OR), BUV496 CD4 (GK1.5, catalog no. 612952, BD Biosciences), BUV805 CD8a (5H10-1, catalog no. 752640, BD Biosciences), BUV737 F4/80 (T45-2342, catalog no. 749283, BD Biosciences), BUV395 CD45 (30-F11, catalog no. 564279, BD Biosciences), FITC CD69 (H1.2F3, catalog no. 35-0691-U100, Tonbo, Freemont, CA, PE CD44 (IM7, catalog no. 50-0441-U100, Tonbo), APC CD62L/L-selectin (MEL-14, catalog no. 20-0621-U100, Tonbo), and Live/Dead Blue (catalog no. L34961, Thermo Fisher Scientific) for discrimination of live and dead cells.

Techniques: Expressing, Marker, Activation Assay, Infection, Control