Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Effect of NK-cell depletion on post-therapy intra-tumoral CD8+ T-cell activity

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Activation Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: CD8+ T- and NK-cell numbers and activity in post-therapy tumors, TDLN and lungs. Mice bearing advanced (500–600 mm3) primary tumors were treated with a single injection of IL-12+GM-CSF microspheres (day 0). Primary tumors, TDLN and lungs were analyzed for CD8+ T- and NK-cell quantity (cells/gram tumor) and activity (% IFNγ and perforin-positive cells) on indicated days. Day 0 data were obtained prior to treatment. a Primary tumors. Relative fold-change in tumor-infiltrating CD8+ T- and CD3- CD49b + NKG2D + NK-cell numbers is shown on the left ordinate. Percent cells positive for IFNγ or perforin are shown on the right ordinate. The differences between day 0 versus day 1 or 3 IFNγ + CD8+ T-cells were significant (P < 0.005). The differences between day 0 versus day 1 or 3 perforin + CD8+ T-cells were also significant (P < 0.05). The difference between day 0 versus day 3 perforin + NK-cells was significant (P = 0.007). Error bars = S.E., n = 6–13 per group. b TDLN. Cell quantity and activity were determined as in a. Two TDLN were analyzed per mouse. Error bars = S.E., n = 6–13 mice per group. c Lungs. CD8, CD94 exon 1A, IFNγ and perforin mRNA levels in whole lung extracts were quantified by real-time PCR. Relative fold-change compared to pre-therapy (day 0) levels is shown. The increase in CD94 exon 1A mRNA on day 3 was highly significant compared to other days (P < 0.0014). The increases in IFNγ and perforin mRNA (days 1–3) were also significant compared to other time points (P < 0.006). Error bars = S.E., n = 4 mice per group

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Activity Assay, Injection, Real-time Polymerase Chain Reaction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Effect of T-cell depletion on post-therapy NK-cell infiltration. Mice bearing advanced (500–600 mm3) primary tumors were treated with IL-12 + GM-CSF microspheres. The depletion groups received the relevant antibodies on days -4 and -1 prior to treatment. Relative fold-changes in CD3- CD49b + NKG2D + NK-cell numbers were quantified in pre-therapy (day 0) and post-therapy (day 3) primary tumors (cells/gram tumor) and TDLN (cells/lymph node) by flow cytometry. Relative fold-changes in lung CD94 exon 1A mRNA was quantified by real-time PCR analysis. The difference between days 0 and 3 NK-cell numbers in primary tumors was significant in the no depletion group (P = 0.021). The differences between days 0 and 3 in CD8 or CD8+ CD4 depletion groups were not significant (P > 0.7). The differences between days 0 and 3 in no depletion and CD8+ depletion groups were significant in the TDLN and lungs (P < 0.05). Error bars = S.E., n = 5–6 mice per group

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Time-dependent depletion of CD8+ T- and NK-cells in IL-12+GM-CSF microsphere-treated mice. Mice bearing advanced (500–600 mm3) subcutaneous primary tumors were treated with IL-12+GM-CSF microspheres (day 0) and primary tumors were surgically excised 7–8 days after treatment. Mice were monitored for 3 weeks after surgery and lung tumor burden was determined as described in “Materials and methods”. In some groups CD8+ T- or NK-cells were depleted starting on days -1, 4 or 10 via antibody administration. Tumors of control mice (untreated) were excised upon reaching 500–600 mm3 without treatment and lung tumor burden was analyzed 3 weeks after surgery. a CD8+ T-cell panel. Effect of CD8+ T-cell depletion on lung tumor burden. The differences between untreated control and all other groups were significant (P < 0.005). The difference between no depletion and day −1 groups was significant (P = 0.00034). The differences between no depletion and day 4 or day 10 depletion groups were not significant (P > 0.13). Error bars = S.E., n = 21 for no depletion group and eight for all other groups. b NK-cell panel. Effect of NK-cell depletion on lung tumor burden. The differences between control untreated and all other groups were significant (P < 0.03). The difference between no depletion and days -1, 4 or 10 depletion groups was significant (P < 0.004). Error bars = S.E., n = 20 for no depletion group and 8 for all others

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques:

Journal: Infection and Immunity

Article Title: Antibodies to Conserved Pneumococcal Antigens Correlate with, but Are Not Required for, Protection against Pneumococcal Colonization Induced by Prior Exposure in a Mouse Model

doi: 10.1128/iai.73.10.7043-7046.2005

Figure Lengend Snippet: FIG. 1. Intranasal colonization with S. pneumoniae TIGR4:7F4 in normal (C57BL6/J [BL6]) and antibody-deficient (MT) mice previously exposed to saline or S. pneumoniae TIGR4:144 (T144), treated with rifampin, and directly challenged (groups 1 and 2 and groups 5 and 6) or challenged after depletion of CD4 cells (groups 3 and 7) or CD8 cells (group 4). A dashed line indicates the limit of detection; solid lines indicate group medians. P values refer to Mann-Whitney tests for differences in the distributions of CFU/nasal wash between groups.

Article Snippet: As a control for the effect of such a monoclonal antibody, group 4 received the same dose of rat anti-mouse CD8 monoclonal IgG2a (purified from hybridoma 53-6.72; American Type Culture Collection) on the same schedule.

Techniques: Saline, MANN-WHITNEY

Journal: Infection and Immunity

Article Title: Antibodies to Conserved Pneumococcal Antigens Correlate with, but Are Not Required for, Protection against Pneumococcal Colonization Induced by Prior Exposure in a Mouse Model

doi: 10.1128/iai.73.10.7043-7046.2005

Figure Lengend Snippet: FIG. 2. Relationship between colonization intensities (expressed as log10 CFU/nasal wash) and antibodies to pneumococcal surface pro- tein A (PspA IgG), pneumococcal surface adhesin A (PsaA IgG), and cell wall polysaccharide (CWPS Ig) (all expressed as log2 ELISA units/ ml). Correlation coefficients shown in the upper corners of each plot are all statistically significant at a P value of 0.01 or less.

Article Snippet: As a control for the effect of such a monoclonal antibody, group 4 received the same dose of rat anti-mouse CD8 monoclonal IgG2a (purified from hybridoma 53-6.72; American Type Culture Collection) on the same schedule.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Live-cell imaging reveals the relative contributions of antigen-presenting cell subsets to thymic central tolerance

doi: 10.1038/s41467-019-09727-4

Figure Lengend Snippet: Both conventional DC subsets induce negative selection of OT-I and OT-II SPs responding to RIP-mOVA and RIP-OVA hi TRAs. a Sequential gating of live CD45 + cells from a digested CD11c-EYFP thymus to detect the indicated APC subsets. Histograms show EYFP and MHC-II levels for the indicated APC subsets. b EYFP mean fluorescence intensity (MFI) of Sirpα + cDC2 and Sirpα − cDC1. Flow-cytometry data averaged from three CD11c-EYFP mice, stained independently. Data points represent mice, and bars show mean ± SEM. c 2PM volume from three perspectives, showing two activated OT-I CD8SPs (red mask, SP1 and SP2) interacting with cDC2 (yellow) and cDC1 (gray), respectively. Scale bar is 10 µm. d Frequency of activated OT-I CD8SP and OT-II CD4SP thymocytes contacting DCs, that interacted with cDC2 or cDC1 on RIP-mOVA or RIP-OVA hi thymic slices. Graph shows mean + SEM. Data are compiled from experiments analyzed in Fig. . Analyzed by t tests with multiple comparison correction, p -values: * < 0.05, *** < 0.001. ns not significant. e mTEC hi (green), cDC2 (red), and cDC1 (blue) were sorted from RIP-mOVA and RIP-OVA hi thymi and cultured with CFSE-labeled splenic OT-I CD8 + T cells. WT splenocytes ± OVAp served as positive and negative control APCs. Histograms show CFSE dilution in Vα2 + Vβ5 + CD8 + cells after incubation with APCs for 72 h, the gate shows the percent of cells that proliferated. Dashed line shows the CFSE profile for T cells cultured with WT splenocytes in the absence of OVAp, and gray shading shows that of T cells cultured with OVAp-pulsed WT splenocytes. Data are representative of two independent experiments per condition, and graphs depict mean ± SEM of the percent proliferation after incubation with indicated APCs for triplicate wells. Source data are provided as a Source Data file. See also Supplementary Figs. ,

Article Snippet: Antibodies directed against the following mouse markers were used: CD3 (145–2C11, Tonbo Biosciences 60–0031), CD4 (RM4–5, BioLegend 100559; GK1.5, BioXCell BE0003–1), CD8 (53–6.7, Tonbo Biosciences 75–0081; 53.6.72, BioXCell BE0004), CD11b (M1/70, BioLegend 101204; M1/70, BioXCell BE0007), CD11c (N418, BioLegend 117322), CD19 (6D5, BioLegend 115534), CD25 (PC61, BioLegend 102024; PC61.5.3, BioXCell BE0012), CD45 (30-F11, BioLegend 103138), CD45.1 (A20, BioLegend 110714), CD69 (H1.2F3, BioLegend 104504), CD80 (16–10A1, BioLegend 104724), B220 (RA3–6B2, BioLegend 103232; RA3.3A1/6.1, BioXCell BE0067), AIRE (5H12, eBioscience 53–5934–82), EpCAM (G8.8, BioLegend 118206), F4/80 (BM8, BioLegend 123116), Gr-1 (RB6–8C5, BioLegend 108410; RB6–8C5, BioXCell BE0075), I-A/I-E (M5/114.15.2, BioLegend 107628), NK1.1 (PK136, BioLegend 108716), PDCA1 (eBio927, eBioScience 12–3172–82), Sirpα (P84, BioLegend 144008), TER-119 (TER-119, BioLegend 116210; TER-119, BioXCell BE0183), Vα2 (B20.1, BioLegend 127818), Vβ5 (MR9–4, BioLegend 139504), and XCR1 (ZET, BioLegend 148212).

Techniques: Selection, Fluorescence, Flow Cytometry, Staining, Comparison, Cell Culture, Labeling, Negative Control, Incubation