Journal: The Journal of Experimental Medicine

Article Title: Host conditioning with IL-1β improves the antitumor function of adoptively transferred T cells

doi: 10.1084/jem.20181218

Figure Lengend Snippet: IL-1β endows CD8 + T cells with an effector-like gene signature. (A) RNA-seq analysis of vehicle- and IL-1β–exposed OT-I cells isolated from the draining LNs on day 4. Heatmap of selected genes associated with effector and memory differentiation is shown ( n = 3). (B and C) GSEA plots showing the enrichment of effector-associated genes in IL-1β– versus vehicle-exposed OT-I cells as described in A. NES, normalized enrichment score; FDR, false discovery rate. (D) Representative contour plots showing T-bet and Eomes expression in vehicle- and IL-1β–exposed OT-I cells isolated from the draining LNs at indicated time points post OVA/LPS immunization. FTY720 was injected i.p. daily starting from day 2. Plots were gated on the live CD8 + Vα2 + CD45.1 + CD45.2 – population. (E) Kinetics of T-bet/Eomes ratio as described in D ( n = 3). (F) Representative contour plots showing Gzm B and CD62L expression as described in D. (G) Kinetics of Gzm B expression as described in D. (D–G) Representative of two independent experiments (error bars, SD).

Article Snippet: Mice of the following strains were obtained from Taconic Farms: C57BL/6 OT-I Rag2 −/− CD45.1, C57BL/6 OT-I Rag2 −/− CD45.2, C57BL/6 OT-I Rag2 −/− Il1r1 −/− CD45.2, C57BL/6, C57BL/6 CD45.1/2, and C57BL/6 Il1r1 −/− .

Techniques: RNA Sequencing Assay, Isolation, Expressing, Injection

Journal: The Journal of Experimental Medicine

Article Title: Host conditioning with IL-1β improves the antitumor function of adoptively transferred T cells

doi: 10.1084/jem.20181218

Figure Lengend Snippet: Differential enhancements of T cell tissue accumulation and function by IL-1β. (A) Absolute cell number and frequency of apoptotic cells (viability dye + population) of vehicle (white bars)– and IL-1β (black bars)–exposed OT-I cells in the draining LNs on day 2 ( n = 5). (B) Representative contour plots and histograms showing the expression of indicated molecules and proliferation dye in vehicle- and IL-1β–exposed OT-I cells as described in A. Plots were gated on the live, CD8 + Vα2 + CD45.1 + CD45.2 – population. (C and D) Absolute cell number and frequency of OT-I cells expressing Gzm B isolated from indicated tissues on days 3–7 from vehicle (white circles)– and IL-1β (black circles)–treated mice ( n = 3). (E) A schematic illustrating the induction of a primary OT-I response by OVA/LPS. Naive OT-I cells were transferred to a congenic host on day −1, followed by OVA/LPS immunization on day 0 and four daily injections of FTY720 along with vehicle or IL-1β on days 1–4. Tissues were harvested on day 6. (F and G) Absolute cell number and frequency of OT-I cells expressing Gzm B isolated on day 6 from indicated tissues of vehicle (white bars)– and IL-1β (black bars)–treated mice as described in E ( n = 5). Data are representative of three (A and B) or two (C–G) independent experiments (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant; error bars, SD).

Article Snippet: Mice of the following strains were obtained from Taconic Farms: C57BL/6 OT-I Rag2 −/− CD45.1, C57BL/6 OT-I Rag2 −/− CD45.2, C57BL/6 OT-I Rag2 −/− Il1r1 −/− CD45.2, C57BL/6, C57BL/6 CD45.1/2, and C57BL/6 Il1r1 −/− .

Techniques: Expressing, Isolation

Journal: The Journal of Experimental Medicine

Article Title: Host conditioning with IL-1β improves the antitumor function of adoptively transferred T cells

doi: 10.1084/jem.20181218

Figure Lengend Snippet: IL-1β–exposed CD8 + T cells show superior trafficking and survival in peripheral tissues. (A) A schematic illustrating the priming of adoptively transferred naive CD45.1 + OT-I cells and CD45.2 + OT-I cells by OVA/LPS under the influence of FTY720 in primary hosts (CD45.1 + CD45.2 + ) treated with vehicle or IL-1β, respectively. Vehicle (CD45.1 + )– and IL-1β (CD45.2 + )–exposed OT-I cells were isolated on day 4 from the draining LNs, mixed at equal numbers (shown in the contour plot; CD8 + Vα2 + gated), labeled with the eFluor 450 proliferation dye, and transferred to a secondary host that had been immunized with OVA/LPS 4 d earlier. Tissues were harvested 14 and 62 h after the second cell transfer ( n = 5). (B) Representative contour plots showing the relative abundance of vehicle- and IL-1β–exposed OT-I cells in the draining LNs and liver 14 and 62 h after the transfer as described in A. Plots were gated on the live CD8 + Vα2 + population. (C) Absolute cell number of the two OT-I populations in the draining LNs and liver as described in A. (D) Dilution of the eFluor 450 proliferation dye for vehicle (gray)– and IL-1β (red)–exposed OT-I cells in the draining LNs and liver as described in A. (E) Frequency of apoptotic cells (viability dye–positive and/or annexin V–positive populations) of the two OT-I populations in the draining LNs and liver as described in A. (F) Ratio of IL-1β– to vehicle-exposed OT-I cells in the draining LNs and liver 14 and 62 h after the transfer as described in A. Data are representative of two independent experiments (**, P < 0.01; ****, P < 0.0001; ns, not significant; error bars, SD).

Article Snippet: Mice of the following strains were obtained from Taconic Farms: C57BL/6 OT-I Rag2 −/− CD45.1, C57BL/6 OT-I Rag2 −/− CD45.2, C57BL/6 OT-I Rag2 −/− Il1r1 −/− CD45.2, C57BL/6, C57BL/6 CD45.1/2, and C57BL/6 Il1r1 −/− .

Techniques: Isolation, Labeling

Journal: The Journal of Experimental Medicine

Article Title: Host conditioning with IL-1β improves the antitumor function of adoptively transferred T cells

doi: 10.1084/jem.20181218

Figure Lengend Snippet: Host-mediated enhancements of T cell tissue accumulation by IL-1β. (A) A schematic illustrating the priming of adoptively transferred naive CD45.1 + OT-I cells by OVA/LPS under the influence of FTY720 in a primary host. OT-I cells were isolated on day 4 from the draining LNs, labeled with the eFluor 450 proliferation dye, and transferred to secondary hosts that had been treated with OVA/LPS and vehicle or IL-1β. Tissues were harvested 14 h after the second cell transfer ( n = 5). (B and C) Absolute cell number and frequency of apoptotic OT-I cells in the draining LNs and liver from vehicle (open symbols)– and IL-1β (solid symbols)–treated hosts as described in A. (D) Dilution of the eFluor 450 proliferation dye for OT-I cells in the draining LNs and liver from vehicle (gray)– and IL-1β (red)–treated hosts as described in A. (E) Absolute cell number of WT (white circles) and Il1r1 −/− (KO; black circles) OT-I cells in the draining LNs and liver isolated on day 6 from WT hosts treated with OVA/LPS on day 0 and IL-1β on days 1–4 ( n = 5). (F) Absolute cell number of WT OT-I cells in the draining LNs and liver isolated on day 6 from WT (white squares) and KO (black squares) hosts treated with OVA/LPS on day 0 and IL-1β on days 1–4 ( n = 5). Data are representative of two (A–D) or four (E and F) independent experiments (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant).

Article Snippet: Mice of the following strains were obtained from Taconic Farms: C57BL/6 OT-I Rag2 −/− CD45.1, C57BL/6 OT-I Rag2 −/− CD45.2, C57BL/6 OT-I Rag2 −/− Il1r1 −/− CD45.2, C57BL/6, C57BL/6 CD45.1/2, and C57BL/6 Il1r1 −/− .

Techniques: Isolation, Labeling

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Escape latency during Morris water maze testing. Escape latency was measured in p50 mice during the acquisition phase, where both groups significantly decreased (P < 0.001) their escape latency times over 7 days of testing (p50 +/+ : from mean 36 s to 20 s; p50 −/− mean 46 s to 14 s), but there was no statistical difference (P = 0.39) between p50 −/− and p50 +/+ mice over the 7 day period. A trend is seen on day 3 where p50 −/− mice show shorter escape times.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Swim speed assessment during Morris water maze testing. Swim speed was measured during the acquisition phase in p50 mice. There was no significant change in swim speed over the 7 days for both groups (P = 0.05). However, post hoc testing showed a significant difference (*P = 0.025) between strains on day 4 of testing where p50 −/− mice swam slower than control mice.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Control

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Search strategy assignments during Morris water maze testing. Search strategies were assigned for each mouse during the acquisition phase. Search strategies were categorized into three main groups: Spatial, Non-spatial Systematic, and Peripheral Looping. The data (right panel) show a disturbance in the formation of spatial memory in the p50 −/− mice as represented by fluctuations in their usage of spatial search strategies. p50 +/+ mice, on the other hand (left panel) show an expected gradual and steady increase in the proficient use of this hippocampal-dependent spatial strategy over time, which is consistent with several other studies. Chi-square tests showed several significant associations (P < 0.05) between strain and the adopted search strategies (days 2, 3, 4, and 7).

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Summary of in vivo search strategy data: Percentage of use over time during acquisition training.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: In Vivo

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Path length determination during Morris water maze testing. Path lengths (i.e., distance travelled) were measured for p50 mice during the acquisition phase. Over this period, a significant effect was seen for both groups over time (P < 0.001). Post hoc analyses also showed a significant difference (*P = 0.014) between strains on day 3 where the p50 −/− mice showed a shorter distance travelled.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Time in target quadrant during Morris water maze testing. The time in target quadrant was measured during the three day retention phase. The p50 −/− mice showed a highly significant (P = 0.009) decrease relative to the p50 +/+ mice for this measurement on the 8 th day of testing (i.e., first day of the retention phase). The experiment was conducted for 2 minutes (120 s) and so the dashed line drawn at 30 seconds indicates chance levels.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Passes over the missing platform during Morris water maze testing. Passes over the missing platform were measured during the retention phase. p50 −/− mice were found to pass over the missing platform significantly less frequently (P = 0.029) than p50 +/+ mice on day 8.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Annulus crossing index during Morris water maze testing. The annulus crossing index was measured during the retention phase. The p50 −/− mice showed a significant (P = 0.01) decrease relative to the p50 +/+ mice.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: Basal synaptic transmission in CA1 hippocampal slices. A: Input–output relations were measured as a function of increased voltage. We found no significant difference between p50 −/− and p50 +/+ strains (P = 0.109) when assessing input–output curves. However, we did find a significant main effect of voltage on EPSP amplitude response (P < 0.001). In addition, we found a significant interaction between strain and stimulation voltage (P = 0.004). B: We also compared paired-pulse responses, an indicator of presynaptic function, and found no significant difference (P = 0.34) between strains using an interstimulus interval of 50 ms

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Transmission Assay

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: LTP evaluation in hippocampal slices. LTP was reliably induced in both groups in CA1 hippocampus immediately after high frequency stimulation (HFS - black arrow). Early LTP time points (1, 2, and 3 hours - 5 min sampling intervals) show no significant difference in LTP responses between the two groups (P > 0.05). However, a significant decrease (*P < 0.05) was found in EPSP slope in the p50 −/− hippocampal slices (triangles) vs. p50 +/+ (circles) slices at 4, 5, and 6 hour time points (5 m sampling intervals) indicating deficits in late LTP. Scale bar: 2 mV, 10 ms.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Sampling

Journal: BMC Neuroscience

Article Title: NF-κB p50 subunit knockout impairs late LTP and alters long term memory in the mouse hippocampus

doi: 10.1186/1471-2202-13-45

Figure Lengend Snippet: NF-κB subunit expression in hippocampal and cerebellum NF-κB p50 mouse samples. Representative Western blots are shown indicating that NF-κB p50 subunit expression was relatively absent in hippocampal and cerebellum samples from p50 −/− mice following LTP experiments.

Article Snippet: Two month-old NF-κB p50 male knockout mice (B6;129P2-Nfkb1 tm1Bal /J) (p50 −/− ) and the recommended male controls (B6129PF2/J) (p50 +/+ ) were purchased from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Western Blot