anti cd122  (Bio X Cell)

Journal: bioRxiv

Article Title: Microbiota promote enhanced CD39 expression in γδ intraepithelial lymphocytes through the activation of TCR and IL-15 signaling

doi: 10.1101/2025.03.22.644616

Figure Lengend Snippet: (A) ELISA of IL-15RC in WT and γδ HYP jejunum. (B) Frequency and (C) MFI of CD122 + γδ IELs among CD39 populations. WT CD39 neg or CD39 pos γδ IELs were sorted and stimulated in vitro with 2 or 100 ng/ml IL-15RC for 72 h. (D) The frequency of CD39 neg γδ IELs that became CD39 pos after stimulation. (E) CD39 MFI of sorted CD39 pos γδ IELs following IL-15RC treatment. WT and γδ HYP mice were injected i.p. daily with isotype control or anti-CD122 for 72 h. (F) Representative flow plots showing CD122 and CD39 expression on γδ IELs and (G) frequency of CD39 neg , CD39 int , and CD39 hi γδ IELs. All data shown as mean ± SEM from at least 2 independent experiments. (A-C,F,G) Each data point represents an individual mouse. n=5-6. (D,E) γδ IELs were pooled from multiple mice; each data point represents an individual replicate. n=3. Statistical analysis: (A) Unpaired student’s t-test, (B,C,G) two-way ANOVA with Tukey’s post hoc test, (D,E) one-way ANOVA with Tukey’s post hoc test. **P<0.01, ***P<0.001, #P<0.0001.

Article Snippet: Mice were injected with 0.45 mg anti-CD122 (TM-β1, BioXCell) or IgG2b isotype control (LTF-2, BioXCell) i.p daily for 3 days prior to the experiment.

Techniques: Enzyme-linked Immunosorbent Assay, In Vitro, Injection, Control, Expressing

Journal: bioRxiv

Article Title: Microbiota promote enhanced CD39 expression in γδ intraepithelial lymphocytes through the activation of TCR and IL-15 signaling

doi: 10.1101/2025.03.22.644616

Figure Lengend Snippet: (A) Representative flow plots and (B) MFI of TCRγδ showing TCR internalization among CD39 γδ IEL populations from WT and γδ HYP mice. WT CD39 neg γδ IELs were sorted and stimulated in vitro with 100 ng/ml IL-15RC in the presence or absence of anti-CD3 for 72 h. (C) Representative flow plots and frequency of (D) CD39 neg γδ IELs that became CD39 pos and (E) CD122 + γδ IELs following stimulation. WT and γδ HYP mice were injected i.p. daily with either isotype control or anti-TCRγδ antibody for 48 h. (F) Representative flow plots showing intracellular (IntC) versus extracellular (ExtC) TCRγδ after antibody treatment. (G) Frequency of CD122 + γδ IELs among CD39 populations and (H) frequency of CD39 neg , CD39 int , and CD39 hi γδ IELs in γδ IELs with IntC vs ExtC TCR. All data shown as mean ± SEM from at least 2 independent experiments. (B,G,H) Each data point represents an individual mouse. n=5-6. (C-E) γδ IELs were pooled from multiple mice, each data point represents an individual replicate. n=3. Statistical analysis: (B,G,H) two-way ANOVA with Tukey’s post hoc test, (D,E) one-way ANOVA with Tukey’s post hoc test. *P<0.05, **P<0.01, ***P<0.001, #P<0.0001.

Article Snippet: Mice were injected with 0.45 mg anti-CD122 (TM-β1, BioXCell) or IgG2b isotype control (LTF-2, BioXCell) i.p daily for 3 days prior to the experiment.

Techniques: In Vitro, Injection, Control

Journal: bioRxiv

Article Title: Microbiota promote enhanced CD39 expression in γδ intraepithelial lymphocytes through the activation of TCR and IL-15 signaling

doi: 10.1101/2025.03.22.644616

Figure Lengend Snippet: Mice were administered broad-spectrum antibiotics (Abx) in drinking water for 6 weeks. (A) Representative flow plots and frequency of (B) CD122 + γδ IELs among CD39 populations and (C) CD39 neg , CD39 int , and CD39 hi γδ IELs. (D) ELISA of IL-15RC in jejunal lysates. (E) Morphometric analysis of jejunal γδ IELs in untreated (untx) and Abx-treated mice. (F) WT and γδ HYP mice were administered either vancomycin (vanco) or neomycin (neo) in drinking water for 6 weeks and the frequency of CD39 γδ IEL populations is shown. All data shown as mean ± SEM from at least 2 independent experiments. (B-F) Each data point represents an individual mouse. n=4-6. Statistical analysis: (B,C,F) Two-way ANOVA with Tukey’s post hoc test, (D,E) one-way ANOVA with Tukey’s post hoc test. *P<0.05, **P<0.01, ***P<0.001, #P<0.0001.

Article Snippet: Mice were injected with 0.45 mg anti-CD122 (TM-β1, BioXCell) or IgG2b isotype control (LTF-2, BioXCell) i.p daily for 3 days prior to the experiment.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Microbiota promote enhanced CD39 expression in γδ intraepithelial lymphocytes through the activation of TCR and IL-15 signaling

doi: 10.1101/2025.03.22.644616

Figure Lengend Snippet: (A) WT and γδ HYP mice were injected i.p. daily with isotype control or anti-CD122 for 72 h. Morphometric analysis of γδ IELs in WT and γδ HYP jejunum. (B) Representative flow plot and (C) MFI of Bcl-2 among CD39 γδ IEL populations. WT and γδ HYP IELs were isolated and cultured in vitro with 100 ng/ml IL-15RC in the presence or absence of anti-CD122 for 24 h. (D) Frequency of CD39 γδ IEL populations and (E) Bcl-2 MFI among CD39 neg , CD39 int , and CD39 hi γδ IELs. All data shown as mean ± SEM from at least 2 independent experiments. Each data point represents an individual mouse. n=4-6. Statistical analysis: (A) One-way ANOVA with Tukey’s post hoc test, (C-E)) two-way ANOVA with Tukey’s post hoc test. *P<0.05, **P<0.01, ***P<0.001, #P<0.0001.

Article Snippet: Mice were injected with 0.45 mg anti-CD122 (TM-β1, BioXCell) or IgG2b isotype control (LTF-2, BioXCell) i.p daily for 3 days prior to the experiment.

Techniques: Injection, Control, Isolation, Cell Culture, In Vitro

Journal: bioRxiv

Article Title: Microbiota promote enhanced CD39 expression in γδ intraepithelial lymphocytes through the activation of TCR and IL-15 signaling

doi: 10.1101/2025.03.22.644616

Figure Lengend Snippet: WT and γδ hvp mice were injected i.p with either isotype control or anti-CD3 for 16 h. (A) Percent capacity or dependency of different metabolic pathways among CD39 populations. (B) Frequency and (C) CD39 MFI of CD39 ne 0 , CD39 int , and CD39 hi yS IELs from WT and γδ HYP mice. (D) Representative flow plots showing CD122 and CD39 expression following treatment. (E) Frequency and (F) puromycin MFI among CD122 and/or CD39-expressing y8 IELs. All data shown as mean ± SEM from at least 2 independent experiments. Each data point represents an individual mouse. n=6. Statistical analysis: (A-C,E,F) Two-way ANOVA with Tukey’s post hoc test. **P<0.01, ***P<0.001, #P<0.0001.

Article Snippet: Mice were injected with 0.45 mg anti-CD122 (TM-β1, BioXCell) or IgG2b isotype control (LTF-2, BioXCell) i.p daily for 3 days prior to the experiment.

Techniques: Injection, Control, Expressing

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