Journal: Nature

Article Title: PGE 2 limits effector expansion of tumour-infiltrating stem-like CD8 + T cells

doi: 10.1038/s41586-024-07254-x

Figure Lengend Snippet: a , Experimental design for b – f . b , Flow cytometric plots of CD8 + T cells from tdLNs and tumours from the indicated days. c , d , Numbers of expanded OT-I CD8 + T cells in tdLNs ( c ) and tumours ( d ) at indicated time points ( n = 6). e , f , Analysis of CD44 and CXCR6 expression in Cd4 cre Ptger2 −/− Ptger4 fl/fl OT-I cells. e , Flow cytometry plots. f , Subset frequencies ( n = 6). g – j , Effect of CD122/CD132 blockade on OT-I T cell expansion in tumours. g – j , Effect of anti-CD122 and anti-CD132 (anti-CD122/CD132) treatment on OT-I TIL expansion in WT mice with control or Ptgs1/Ptgs2 −/− BRAF V600E -OVA tumours or with MC38-OVA tumours, analysed 11 days after tumour transplantation. g , h , Flow cytometry plots ( g ) and OT-I TIL numbers ( h ) in BRAF V600E -OVA tumours ( n = 6). i , j , Flow cytometry plots ( i ) and OT-I TIL numbers ( j ) in MC38-OVA tumours ( n = 10). k , Experimental design for l and m with MC38-OVA tumours. l , Flow cytometry plot (left) and quantification (right) of OT-I TILs at day 10 ( n = 6). m , Flow cytometry plots showing the population size of TIM-3 + CXCR6 + cells among control and Cd122 −/− Cd4 cre Ptger2 −/− Ptger4 fl/fl OT-I TILs. n , WT mice received 1 × 10 3 naive OT-I T cells or 1 × 10 3 naive Cd4 c re Ptger2 −/− Ptger4 fl/fl OT-I T cells intravenously (i.v.) and were transplanted s.c. with 2 × 10 6 MC38-OVA cells before analysis of tumour growth over time ( n = 10). Asterisk indicates that termination criteria were reached. Data in c , d , f , h , j , l and n are pooled from two ( c , d , h , l ) or three ( f , j , n ) independent experiments and depicted as box plots extending from the 25th to 75th percentiles with the median as the centre and the whiskers corresponding to minimum and maximum values ( c , d , h , j , l ) or shown as the mean ± s.e.m. ( f , n ). Plots in b , e , g , i , l and m show data for 1 sample representative of n = 6 samples analysed in 2 ( b , g , l , m ) or 3 ( e , i ) independent experiments. P values are from paired t -tests ( l ), one-way ANOVA with Tukey’s multiple-comparison test ( c , d ) or Dunnett’s multiple-comparison test ( h , j ), or two-way ANOVA with Bonferroni’s correction for multiple testing ( n ).

Article Snippet: For blockade of IL-2Rβ and IL-2Rγc, mice received i.p. injections of 150 µl anti-mouse CD122 (300 µg per mouse, TM-Beta 1, BioXCell) and anti-mouse CD132 (300 µg per mouse, 3E12, BioXCell) antibodies on days 6 and 7 after tumour cell transplantation.

Techniques: Expressing, Flow Cytometry, Transplantation Assay, Comparison

Journal: EBioMedicine

Article Title: Intradermal vaccination prevents anti-MOG autoimmune encephalomyelitis in macaques

doi: 10.1016/j.ebiom.2019.08.052

Figure Lengend Snippet: Cytokine levels in plasma of treated and control animals. a) Heatmap showing cytokine levels in the plasma of all animals at several time points after sensitisation with rhMOG/IFA. Cytokine levels are represented by a colour gradient, ranging from yellow (no expression) to deep blue (highest concentration). Hierarchical clustering, represented by dendrograms, was performed at individual and cytokine levels. Four groups of animals were identified and are numbered from 1 to 4 in the text: animals treated with anti-DC-ASGPR-MOG are named T1, T2, and T3, controls treated with anti-DC-ASGPR-PSA, C1, C2, and C3, and untreated non-immunised naïve animals, 1, 2, 3, and 4; timepoints in dpi are numbered from D7 to D37; group I (red), group II (green), group III (purple), group IV (yellow); see also Supplemental table 4). b) Cytokine levels (expressed in log10 of pg/ml) in naïve animals relative to those measured in “EAE incubation” group II, in which there were significantly lower TGFβ1, TGFβ2, and IL-8 levels. c) Cytokine levels measured in “EAE resolution” group III, in which there were significantly higher TGFβ1, TGFβ2 and IL-8 levels in treated animals than naïve macaques. d) Varying cytokine levels between the two groups of treated and control animals and the various timepoints. In green, animals treated with anti-DC-ASGPR-MOG (T); in blue, control animals treated with anti-DC-ASGPR-PSA (C) and timepoints in day (d) post-sensitisation with rhMOG/IFA. The levels of the pro-inflammatory cytokines IL-1β, IFNγ, and TNFα were elevated at 35 dpi in controls but not treated animals. The levels of IL-8, TGFβ1, and TGFβ2 were elevated in treated animals at the last timepoint of 35 dpi, but not in controls. Statistics: exploratory analysis, with no multiple test correction, using the two-tailed unpaired t-test. (ns) p > .05; (*) p ≤ .050; (**) p ≤ .010; (***) p ≤ .0010. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Intracellular staining with anti-IL10-PE (JES3-9D7, BD) and anti-TGFβ1-AF488 (Mouse 9016, R&D system) and extracellular staining with anti-LAP-FITC (TGFβ1) (CH6-17E5.1, Miltenyi Biotec) of macaque PBMCs were also tested but did not permit detection of their targets.

Techniques: Clinical Proteomics, Control, Expressing, Concentration Assay, Incubation, Two Tailed Test