Journal: Cell reports

Article Title: Temporal dynamics of immune cell transcriptomics in brain metastasis progression influenced by gut microbiome dysbiosis

doi: 10.1016/j.celrep.2025.115356

Figure Lengend Snippet:

Article Snippet: Following viability stain, cells were blocked in the same manner as described for CITE-seq preparation, using FcR Blocking Reagent (Miltenyi Biotec, 130-092-575) and then stained with one or a combination of the following antibodies: rat anti-mouse CD3e-647 (BioLegend, 155609, KT3.1.1), hamster anti-mouse TCR γ/δ-PE (BioLegend, 118107, GL3), rat anti-mouse CD117(c-kit)-PE-Cy7 (BioLegend, 105813, 2B8), mouse anti-mouse CX3CR1–421 (BioLegend, 149023, SA011F11), rat anti-mouse CD192(CCR2)-510 (BioLegend, 150617, SA203G11), rat anti-mouse CD3-SparkBlue550 (BioLegend, 100259, 17A2), rat anti-mouse CD8-FITC (BioLegend, 100706, 53–6.7), rat anti-mouse CD4–711 (BioLegend, 100549, RM4–5), rat anti-mouse Ly-6C-785 (BioLegend, 128041, HK1.4), rat anti-mouse Ly-6G-650 (BioLegend, 127641, 1A8), rat anti-mouse CD186(CXCR6)-PerCP/Cyanine5.5 (BioLegend, 151120, SA051D1), rat anti-mouse CD11b-605 (BioLegend, 101237, M1/70), rat anti-mouse CD45-PacificBlue (BioLegend, 103125, 30-F11), rat anti-mouse CD45-PerCP/Cyanine5.5 (BioLegend, 103131, 30-F11), rat anti-CD3-FITC (BioLegend, 100203, 17A2), hamster anti-mouse TCR γ/δ-PE-Cy7 (BioLegend, 118123, GL3), rat anti-mouse CD279(PD-1)-421 (BioLegend, 135217, 29F.1A12), rat anti-CD25-PE (BioLegend, 102007, PC61), rat anti-CD4-APC-Cy7 (Tonbo, 25-0042-U025, RM4–5), rat anti-mouse CD8a-710 (Tonbo, 80-0081-U025, 53–6.7), rat anti-mouse CD3-PacificBlue (BioLegend, 100213, 17A2), rat anti-mouse CD4–570 (BioLegend, 100541, RM4–5), rat anti-mouse CD8a-APC-Cy7 (BioLegend, 100713, 53–6.7), hamster anti-mouse TCR γ/δ−711 (BioLegend, 118149, GL3), mouse anti-mouse NK-1.1-SparkRed718 (BioLegend, 156533, S17016D), rat anti-mouse CD25–750 (BioLegend, 102077, PC61), mouse anti-mouse CD159a(NKG2A B6 )-PE (BioLegend, 142803, 16A11), rat anti-mouse CD107a(LAMP-1)-PE-Cy7 (BioLegend, 121619, 1D4B), rat anti-mouse/human CD11b-650 (BioLegend, 101259, M1/70), rat anti-mouse CD8a-647 (BioLegend, 100727, 53–6.7), rat anti-mouse CD279(PD-1)-421 (BioLegend, 135221, 29F.1A12), rat anti-mouse FOXP3–647 (BioLegend, 126407, MF-14).

Techniques: In Vivo, Recombinant, Blocking Assay, Red Blood Cell Lysis, Multiplex Assay, Sequencing, Software, Amplification, Fluorescence

Journal: Cell reports

Article Title: Temporal dynamics of immune cell transcriptomics in brain metastasis progression influenced by gut microbiome dysbiosis

doi: 10.1016/j.celrep.2025.115356

Figure Lengend Snippet:

Article Snippet: Flow: rat anti-CD4-APC-Cy7 (RM4–5) , Tonbo , Cat#25–0042-U025; RRID: N/A.

Techniques: In Vivo, Recombinant, Blocking Assay, Red Blood Cell Lysis, Multiplex Assay, Sequencing, Software, Amplification, Fluorescence

Journal: eBioMedicine

Article Title: Intranasal recombinant protein subunit vaccine targeting TLR3 induces respiratory tract IgA and CD8 T cell responses and protects against respiratory virus infection

doi: 10.1016/j.ebiom.2025.105615

Figure Lengend Snippet: Intranasal administration of CAF09b adjuvanted SARS-CoV-2 spike protein elicits higher respiratory tract immune responses than the licenced vaccine mRNA-1273 . Mice were immunised with two doses of SARS-CoV-2 spike HexaPro trimer (5 μg of protein) formulated in CAF09b adjuvant given intranasally (i.n./i.n.) or the licenced vaccine mRNA-1273 (Spikevax) vaccine (1 μg of mRNA given intramuscularly (i.m/i.m.)). a) Serum IgG antibody responses against spike protein. b) IgA antibody responses against spike protein measured in serum, lungs, nasal-associated lymphoid tissue (NALT), and nasal washes. Mice were injected i.v. with anti-CD45.2 to distinguish between circulating CD45+ (IV+) and tissue resident CD45− (IV−) cells c) CD8 T cell responses, as identified by gating on live+CD8+CD44+ cells binding a spike-specific tetramer (VNFNFNGL), were measured by flow cytometry. CD8 T cell responses were assessed systemically (IV+) in spleen (left panel), and locally (IV−) in lungs (middle panel) and NALT (right panel). Data represent two independent experiments with n = 3 (naïve) or 8–9 (vaccinated) mice per group. To obtain enough cells for analysis of CD8 T cell responses in NALT, each data point displays a pool of three mice. d) CD4 T cell responses, measured by gating for spike tetramer S62-76 (VTWFHAIHVSGTNGT) on live+CD8−CD19−CD4+CD62L−CD44+ cells, was assessed systemically (IV+) in spleen (left panel), and locally (IV−) in lungs (middle panel) and NALT (right panel). The experiment was performed once and data represent n = 3 (naïve) or n = 8–9 (vaccinated) mice per group. Mean ± SEM is displayed. Statistically significant differences are indicated by ∗, ∗∗, ∗∗∗ or ∗∗∗∗ (Student t-test, p < 0.05, 0.01, 0.001 or 0.0001, respectively). There were no statistically significant differences among groups unless indicated.

Article Snippet: One million cells were treated with Fc-block (BD Biosciences) and stained in PBS +1% FBS with cocktails of antibodies against the following surface proteins: CD45.2 FITC (104, BD 553772), CD4 APC-Cy7 (RM4-5, 1:600, eBioscience 47-0042-82), CD4 BV786 (GK1.5, 1:200, BD 563331), CD8a PerCP-Cy5.5 (53-6.7, 1:600, eBioscience 45-0081-82), CD8a BV421 (53-6.7, 1:600, Biolegend 100738), CD44 APC (IM7, 1:600, BD 559250), CD44 APC-Cy7 (IM7, 1:200, eBioscience 47-0441-82), CD62L PerCP-Cy5.5 (MEL-14, 1:200, BD 560513), CD19 PE-Cy7 (1D3, 1:600, BD 552854), CD11c PE-Cy7 (HL3, 1:200, BD 558079), and I-A/I-E BV605 (M5/114.15.2, 1:200, BD 563413).

Techniques: Adjuvant, Injection, Binding Assay, Flow Cytometry