Journal: Virologica Sinica

Article Title: PD1 + CCR2 + CD8 + T Cells Infiltrate the Central Nervous System during Acute Japanese Encephalitis Virus Infection

doi: 10.1007/s12250-019-00134-z

Figure Lengend Snippet: Marked CD8+ T cells infiltration into the mouse central nervous system during Japanese encephalitis virus (JEV) infection. A, B Mass cytometric analyses of infiltrating immune cells isolated from the brains of PBS-and JEV-infected mice. Each marker used for cell characterization, including CD45, CD19, CD45R, CD3e, CD8a and CD4, was overlaid on a viSNE diagram. Each dot represents a single cell, and the color gradient represents the marker’s intensity. Dot plots demonstrate the marker labeling on living single cells. Representative images of three independent repeats are shown. C Mononuclear cells isolated from the brains of PBS- and JEV- infected mice were stained with indicated antibodies and CD45+TCR-β+ CD8+ T cells were gated. D Statistical analyses of the number and proportion of these CD45+TCR-β+ CD8+ T cells among mononuclear cells isolated from the brains. Data are presented as the mean ± SD of two independent experiments. **P < 0.01 and ***P < 0.001, based on a two-tailed, unpaired Student’s t test.

Article Snippet: The following antibodies were used: rat anti-mouse CD16/32 (mouse BD Fc Block, clone 2.4G2, BD Pharmingen), Fixable Viability Dye eFluor506 (Cat: 65-0866, eBioscience), anti-mouse CD45 Alexa Fluor700 (clone 30-F11, Biolegend), anti-mouse TCR beta APC-eFluor 780 ® (clone H57-597, eBioscience), anti-mouse CCR2-BV421 (clone 203G11, Biolenged), anti-mouse CCR5 (CD195)-APC (clone HM-CCR5, Biolegend), anti-mouse CD8a-PE/Dazzle (clone 53-6.7, Biolegend), anti-mouse PD1-PE (clone RMP1-30, eBioscience), anti-mouse CD45-AF700 (clone 30-F11, Fluidigm), anti-mouse CD19 (clone 6D5, Fluidigm), anti-mouse CD45R (B220, clone RA3-6B2, Fluidigm), anti-mouse CD3e (clone 145-2C11, Fluidigm), anti-mouse CD4 (clone RM4-5, Fluidigm), anti-mouse CD8a (clone 53-6.7, Fluidigm), anti-mouse CD62L (clone MEL-14, Fluidigm), anti-mouse CD44 (clone IM7, Fluidigm), anti-mouse TNF-α (clone MP6-XT22, Fluidigm), anti-mouse IFN-γ (clone XMG1.2, Fluidigm).

Techniques: Virus, Infection, Isolation, Marker, Labeling, Staining, Two Tailed Test

Journal: Virologica Sinica

Article Title: PD1 + CCR2 + CD8 + T Cells Infiltrate the Central Nervous System during Acute Japanese Encephalitis Virus Infection

doi: 10.1007/s12250-019-00134-z

Figure Lengend Snippet: CCL2 plays a role in the migration of CD8+ T cells into the mouse central nervous system after Japanese encephalitis virus (JEV) infection. A Gene expression pattern of chemokines and their co-receptors among severe JE cases in mice, as compared to that in control animals. B, C mRNA levels of chemokines (Ccl2, Ccl3, and Ccl5) and their receptors (Ccr2 and Ccr5) in mouse brain tissues were assessed by RT-qPCR during mild and severe cases of JE after JEV infection. Values were normalized for Gapdh mRNA level. D Mononuclear cells from mouse brain tissues and splenocytes were isolated and CCR2+CD8+ T cells were gated after JEV infection. E Statistical analyses of CCR2+CD8+ T cells in the brains and spleens. Data are presented as the mean ± SD of three or more independent experiments. *P < 0.05 and ***P < 0.001, based on two-tailed, unpaired Student’s t test.

Article Snippet: The following antibodies were used: rat anti-mouse CD16/32 (mouse BD Fc Block, clone 2.4G2, BD Pharmingen), Fixable Viability Dye eFluor506 (Cat: 65-0866, eBioscience), anti-mouse CD45 Alexa Fluor700 (clone 30-F11, Biolegend), anti-mouse TCR beta APC-eFluor 780 ® (clone H57-597, eBioscience), anti-mouse CCR2-BV421 (clone 203G11, Biolenged), anti-mouse CCR5 (CD195)-APC (clone HM-CCR5, Biolegend), anti-mouse CD8a-PE/Dazzle (clone 53-6.7, Biolegend), anti-mouse PD1-PE (clone RMP1-30, eBioscience), anti-mouse CD45-AF700 (clone 30-F11, Fluidigm), anti-mouse CD19 (clone 6D5, Fluidigm), anti-mouse CD45R (B220, clone RA3-6B2, Fluidigm), anti-mouse CD3e (clone 145-2C11, Fluidigm), anti-mouse CD4 (clone RM4-5, Fluidigm), anti-mouse CD8a (clone 53-6.7, Fluidigm), anti-mouse CD62L (clone MEL-14, Fluidigm), anti-mouse CD44 (clone IM7, Fluidigm), anti-mouse TNF-α (clone MP6-XT22, Fluidigm), anti-mouse IFN-γ (clone XMG1.2, Fluidigm).

Techniques: Migration, Virus, Infection, Gene Expression, Control, Quantitative RT-PCR, Isolation, Two Tailed Test

Journal: Virologica Sinica

Article Title: PD1 + CCR2 + CD8 + T Cells Infiltrate the Central Nervous System during Acute Japanese Encephalitis Virus Infection

doi: 10.1007/s12250-019-00134-z

Figure Lengend Snippet: Characteristics of mouse central nervous system-infiltrating CD8+ T cells after Japanese encephalitis virus (JEV) infection. A Detailed analyses of infiltrating CD8+ T cells by mass cytometry. The expression of CD44 and CD62L overlaid on viSNE diagrams is shown (z: color gradient of expression). B Dot plots illustrating the distinction between the expression of TNF-α and IFN-γ on infiltrating CD8+ T cells from JEV-infected brains versus those in control samples. C Flow cytometric analysis of the expression of IFN-γ and granzyme B in CD8+ T cells from PBS- and JEV-infected mouse brains. D PD-1 expression on CD45+TCR-β+CD8+ T cells isolated from mouse brains and spleens. E PD-1 expression on CCR2+CD8+T cells or CCR2−CD8+ T cells in mouse brains after JEV infection. F Differential expression of PD-1 on CCR2+CD8+ T cells and CCR2−CD8+ T cells. Data are presented as the mean ± SD of three or more independent experiments. *P < 0.05, based on two-tailed, unpaired Student’s t test.

Article Snippet: The following antibodies were used: rat anti-mouse CD16/32 (mouse BD Fc Block, clone 2.4G2, BD Pharmingen), Fixable Viability Dye eFluor506 (Cat: 65-0866, eBioscience), anti-mouse CD45 Alexa Fluor700 (clone 30-F11, Biolegend), anti-mouse TCR beta APC-eFluor 780 ® (clone H57-597, eBioscience), anti-mouse CCR2-BV421 (clone 203G11, Biolenged), anti-mouse CCR5 (CD195)-APC (clone HM-CCR5, Biolegend), anti-mouse CD8a-PE/Dazzle (clone 53-6.7, Biolegend), anti-mouse PD1-PE (clone RMP1-30, eBioscience), anti-mouse CD45-AF700 (clone 30-F11, Fluidigm), anti-mouse CD19 (clone 6D5, Fluidigm), anti-mouse CD45R (B220, clone RA3-6B2, Fluidigm), anti-mouse CD3e (clone 145-2C11, Fluidigm), anti-mouse CD4 (clone RM4-5, Fluidigm), anti-mouse CD8a (clone 53-6.7, Fluidigm), anti-mouse CD62L (clone MEL-14, Fluidigm), anti-mouse CD44 (clone IM7, Fluidigm), anti-mouse TNF-α (clone MP6-XT22, Fluidigm), anti-mouse IFN-γ (clone XMG1.2, Fluidigm).

Techniques: Virus, Infection, Mass Cytometry, Expressing, Control, Isolation, Quantitative Proteomics, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: Single Cell Multi-Omics Technology: Methodology and Application

doi: 10.3389/fcell.2018.00028

Figure Lengend Snippet: Current multi-omics methods.

Article Snippet: sc-GEM (genotype single cells genotype, gene expression, DNA methylation) (2016) , Human fibroblast, hIPSC, hESC and NSCLC sample. , Genotype single cells while simultaneously interrogating gene expression and DNA methylation at multiple loci , Cell is captured and lysed on C1 Fluidigm chip. RNA is measured using single cell RT-qPCR and methylation is measured using Single Cell Restriction Analysis of Methylation. , Microfluidic device , Tight coupling between the timing of DNA methylation changes and transcription in individual cells; cells have EGFR mutations show a distinct epigenetic signature. , Cheow et al., .

Techniques: Biomarker Discovery, Single-cell Isolation, Sequencing, Magnetic Beads, Flow Cytometry, Lysis, Methylation Sequencing, Methylation, Expressing, RNA Sequencing, Gene Expression, DNA Methylation Assay, Cell Culture, FACS, Derivative Assay, In Vivo, CpG Methylation Assay, RNA Expression, Isolation, Whole Genome Amplification