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: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Establishment of a humanized Ph + ALL xenotransplant model using anti-CD122-conditioned NOD/SCID mice and intra-bone marrow injection (IBMI) with LPCs from Ph + ALL patients. a Compared with an irradiated non-transplanted control mouse (Ctrl), the mouse transplanted with LPCs and treated with vehicle (Vehicle) exhibited significant splenomegaly at 12 weeks post-transplant. A low-magnification image of human Ph + ALL engraftment in a bone section ( left panel ), May-Giemsa staining ( middle panel ) and fluorescence in situ hybridization (FISH) analysis ( right panel ) of leukemic blasts in the BM of the recipients (Vehicle). b Flow cytometric analysis demonstrated that the BMs of the recipients were efficiently engrafted with human Ph + ALL cells with an aberrant phenotype similar to that in the donor Ph + ALL patients. c Human engraftment of the BM, spleen, liver and kidney of the recipient (Vehicle) was further confirmed by HE staining ( upper panels ) and IHC with anti-hCD19 antibody ( lower panels )

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: Injection, Irradiation, Staining, Fluorescence, In Situ Hybridization

Journal: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Effects of single drug treatment or different combinations of IM, NL and RUX on the engraftment of human Ph + ALL cells in the anti-CD122-conditioned NOD/SCID recipients transplanted with LPCs from Ph + ALL patients. a Experimental design for the in vivo experiments. LPCs sorted from newly diagnosed Ph + ALL patients (N = 6) were transplanted by IBMI into 5-week-old, sub-lethally irradiated (2.1 Gy of total body irradiation from a 60 Co source) and anti-CD122-conditioned NOD/SCID mice. From +14 days post-transplantation, the recipient mice were randomly administered with vehicle (10% NMP-90% PEG 300), IM (100 mg/kg/day), NL (75 mg/kg/day), RUX (30 mg/kg/day), IM (100 mg/kg/day) combined with RUX (30 mg/kg/day), or NL (75 mg/kg/day) combined with RUX (30 mg/kg/day) via oral gavage for 14 days. b The engraftment levels of human Ph + ALL CD45 + cells in the BMs and spleens of mice under different treatment conditions were analyzed by flow cytometry at 8 weeks (N = 6 patients, N = 3 mice per patient per treatment group) and c at 12 weeks post-transplant (N = 6 patients, N = 3 mice per patient per treatment group) or until the mice were moribund

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: In Vivo, Irradiation, Transplantation Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Ruxolitinib/nilotinib cotreatment inhibits leukemia-propagating cells in Philadelphia chromosome-positive ALL

doi: 10.1186/s12967-017-1286-5

Figure Lengend Snippet: Cotreatment with NL and RUX exhibited the most effective anti-LPC effect. LPCs sorted from newly diagnosed Ph + ALL patients (N = 6) were transplanted by IBMI into 5-week-old, sub-lethally irradiated (2.1 Gy of total body irradiation from a 60 Co source) and anti-CD122-conditioned NOD/SCID mice. From +14 days post-transplantation, the recipient mice were randomly administered with vehicle (10% NMP-90% PEG 300), IM (100 mg/kg/day), NL (75 mg/kg/day), RUX (30 mg/kg/day), IM (100 mg/kg/day) combined with RUX (30 mg/kg/day), or NL (75 mg/kg/day) combined with RUX (30 mg/kg/day) via oral gavage for 14 days. a Representative images of splenomegaly in the mice under different treatment conditions and a control NOD/SCID mouse (Ctrl) without receiving the Ph + ALL LPCs transplantation. b Differences in human engraftment were further confirmed by HE staining ( upper panels ) and IHC with anti-hCD19 antibody labeling ( lower panels ) of the spleens in the recipient mice treated with the different drugs and the Ctrl mice. c The engraftment analysis of BCR/ABL -expressing BM cells using a TaqMan-based qRT-PCR assay in the recipient mice at 12 weeks post-transplant. d Representative western blots of phospho-CrkL, phospho-JAK2, and GAPDH in the bone marrow cells of humanized mice transplanted with Ph + ALL LPCs following treatment with vehicle, single agents or a combination of RUX and IM or NL mice, and Ctrl mice. All data from the independent experiments are presented as the mean ± SEM. Significance values: *** P < 0.0001

Article Snippet: Briefly, 5- to 6-week-old NOD/SCID mice (Vital River Laboratories, Beijing, China) were sub-lethally irradiated (2.1 Gy total body irradiation from a 60 Co source) followed by treatment with 200 μg of anti-mouse CD122 monoclonal antibody generated from hybridoma TM-β1 (provided by Dr. T. Tanaka of Hyogo University of Health Sciences, Kobe, Japan) [ ].

Techniques: Irradiation, Transplantation Assay, Staining, Antibody Labeling, Expressing, Quantitative RT-PCR, Western Blot