Journal: Nature Communications

Article Title: Cationic crosslinked carbon dots-adjuvanted intranasal vaccine induces protective immunity against Omicron-included SARS-CoV-2 variants

doi: 10.1038/s41467-023-38066-8

Figure Lengend Snippet: a The percentages of Tfh (CD4 + CXCR5 + PD-1 + ) and RBD-specific GCB (B220 + GL7 + CD95 + RBD + ) cells in the CLN were analyzed with FCM at 28 days after the last immunization. b , c Lung CD8 + and CD4 + T cells were evaluated for IFN-γ generation by FCM upon restimulation with RBD peptide pools or irrelevant antigens. b Representative FCM plots (left) and quantification (right) of IFN-γ-expressing lung CD8 + T cells. d The proportion of antigen-experienced (CD44 + ) CD4 + and CD8 + T cells expressing CD69 and CD103 in lung tissues. e The percentage of CD103 + DCs coexpressing CD86 and MHC II in the lungs. f Upper left: t-SNE maps were created by concentrating CD3 + T cells from the BAL fluid of the vaccinated mice. Analysis was carried out with default FlowJo V.10 software. Upper right and bottom: heatmap projections of CD4, CD8, CD44, CD69, and CD103 expression on t-SNE maps from three independent mice ( n = 3). Red and black hashed circles are indicators of antigen-experienced CD4 + and CD8 + T RM cells in the BAL fluid, respectively. g Absolute numbers of CD8 + T cells producing IFN-γ (left) and TNF-α (right) in the BAL after ex vivo restimulation with RBD peptide pools or irrelevant antigen at 4 weeks after the last immunization. h Quantification of the percentages (left) and the numbers (right) of MHC II + AM in the BAL fluid. Data were displayed with floating bars in ( a – e , g , and h ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–6 mice in each group in ( a – e , g , and h ). P values in a , d , e , h were calculated with One-way ANOVA followed by Dunnett’s multiple comparisons tests. P values in b , c , and g were calculated with two-way ANOVA followed by Tukey’s multiple comparisons tests. Source data are provided as a Source Data file.

Article Snippet: T cells in untreated, female, 6–8 weeks BALB/c mouse spleen were obtained by a mouse CD3 T-cell enrichment kit (Stemcell).

Techniques: Expressing, Software, Ex Vivo

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Immunodeficient mouse model for human hematopoietic stem cell engraftment and immune system development

doi: 10.1007/978-1-4939-1133-2_18

Figure Lengend Snippet: At 12 weeks post-implant human immune cell chimerism levels can be evaluated in the peripheral blood. Our standard panel includes antibodies specific for mouse CD45, human CD45, human CD3, human CD20, human CD4 and human CD8. (A) The gating strategy for this panel includes first evaluating percentages of human CD45+ and mouse CD45+ cells. (B) Human CD45+ cells are evaluated for expression of human CD3 to enumerate T cells and human CD20 to enumerate B cells and (C) then the T cells populations are further defined by CD4 and CD8 expression. (D) In addition the human thymocytes recovered from the thymic organoid can be evaluated by expression of human CD4 and CD8.

Article Snippet: The buffer can be stored at 4°C Quenching Buffer: Wash Buffer supplemented with 3% fetal bovine serum (FBS, Atlanta Biologicals, Lawrenceville, GA, USA) Human fetal liver and fetal thymus tissue (Advanced Bioscience Resources, Alameda, CA). (See NOTE 1 ) 100 mm plastic petri dishes (BD Falcon, Franklin Lakes, NJ, USA), sterile No. 21 disposable scalpel (Feather Safety Razor Co., LTD, Kita-Ku, Osaka, Japan) 50mL centrifuge tubes (BD Falcon, Franklin Lakes, NJ, USA) Liver digest buffer (Gibco, Life technologies, Grand Island, NY USA) Water bath Parafilm Nutating mixer (VWR international, Randor, PA, USA) Laboratory tape or alternatively rubber bands 85mm tissue grinder homogenizer cup with size 50 metal sieve (Sigma-Aldrich, St Louis, MO, USA) 10mm Syringe Histopaque-1077 (Sigma-Aldrich, St Louis, MO, USA) 2.2 CD3 T cell Depletion Human CD3 microbead kit (Miltenyi Biotech, Auburn, CA, USA) (See NOTE 2 ) MidiMACS Separator (Miltenyi Biotech, Auburn, CA, USA) MACS multistand (Miltenyi Biotech, Auburn, CA, USA) LD columns (Miltenyi Biotech, Auburn, CA, USA) MACS Buffer: PBS supplemented with 2% FBS, 1mM EDTA.

Techniques: Expressing