cd8-depleting ab be0223-a025  (BioXell Inc)

Journal: Nature Communications

Article Title: Progranulin mediates immune evasion of pancreatic ductal adenocarcinoma through regulation of MHCI expression

doi: 10.1038/s41467-021-27088-9

Figure Lengend Snippet: a Representative mIF image demonstrates the intratumoral heterogeneity of PGRN expression in human PDAC, where high and low PGRN-expressing tumor regions were observed in the same specimen. ( n = 8). Representative images are shown. b mIF staining of PGRN (yellow), CD8 (green), and PanCK (red) in human PDAC shows increased CD8 infiltration in low PGRN-expressing tumor areas. ( n = 8). Representative images are shown. c Differential MHCI (HLA-A) expression in PGRN + and PGRN − tumors, and the infiltration of GzmB + CD8 + cells in their corresponding neighborhoods. ( n = 8). Representative images are shown. d Automated computational analysis showing the percentage of MHCI + cells in PGRN + PanCK + and PGRN − PanCK + tumor populations, and the number of CD8 − GzmB − cells in proximity (<50 μM radical distance) of PGRN + MHCI − or PGRN − MHCI + PanCK + tumor cells in human PDAC ( n = 8). Two-tailed Mann–Whitney test. Scale bar unit: μm.

Article Snippet: In the CD8 + T-cell depletion experiments, 25 mg/ml of anti-CD8 depleting Ab (clone 2.43, Bio X Cell) was injected i.p at days −3 and day 1 relative to the starting date of PGRN Ab treatment.

Techniques: Expressing, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Progranulin mediates immune evasion of pancreatic ductal adenocarcinoma through regulation of MHCI expression

doi: 10.1038/s41467-021-27088-9

Figure Lengend Snippet: IHC staining of a T-cell markers CD3 and CD8; cytotoxic markers granzyme B (GzmB), and T-bet in CKP tumors treated with or without PGRN Ab (PAb) or mIg (50 mg/kg). The lower panels show the percentages of positive cells in the whole tumor. ctrl: n = 5; mIg: n = 10; PAb: n = 8. One-way ANOVA, Kruskal–Wallis test. b mIF showing the proportion of CD8 + cells co-expressing cytotoxic markers GzmB or T-bet. n = 4. c IHC staining of apoptotic marker cleaved caspase 3 (Cl casp3) in CKP tumors treated with or without PGRN Ab (PAb) or mIg. The lower panels show the percentages of positive cells in the whole tumor. ctrl: n = 5; mIg: n = 10; PAb: n = 8. One-way ANOVA, Kruskal–Wallis test. d Timeline for treatment of CKP mice with CD8 depleting Ab (aCD8, 25 mg/kg), mouse isotype (mIg) or anti-PGRN antibody (PAb, 50 mg/kg). e Representative pictures of tumors and spleens from CKP mice. f Weight of pancreas of CKP mice upon dissection. mIg: n = 4; PAb: n = 7; aCD8: n = 7; aCD8+ PAb: n = 6. g Quantification of tumor-infiltrating CD8 in the CKP tumors by flow cytometry. One-way ANOVA, Kruskal–Wallis test. IHC staining of h PanCK, CD8, GzmB, and i cleaved casp3 in CKP tumors treated with or without PGRN Ab, mIg and/or CD8 depleting Ab (aCD8). mIg: n = 4; PAb: n = 4; aCD8: n = 7; aCD8+ PAb: n = 6. One-way ANOVA, Kruskal–-Wallis test. Mean ± SD is shown. Scale bar unit: μm.

Article Snippet: In the CD8 + T-cell depletion experiments, 25 mg/ml of anti-CD8 depleting Ab (clone 2.43, Bio X Cell) was injected i.p at days −3 and day 1 relative to the starting date of PGRN Ab treatment.

Techniques: Immunohistochemistry, Expressing, Marker, Dissection, Flow Cytometry

Journal: Nature Communications

Article Title: Progranulin mediates immune evasion of pancreatic ductal adenocarcinoma through regulation of MHCI expression

doi: 10.1038/s41467-021-27088-9

Figure Lengend Snippet: a IHC of MHCI marker H-2Db and MHCII in CKP tumors treated with or without PGRN Ab or mIg. ctrl: n = 5; mIg: n = 10; PGRN Ab (PAb): n = 8. The right panels show the percentages of positive cells in the whole tumor. One-way ANOVA, Kruskal–Wallis test. Mean ± SD is shown. b mIF of PGRN (yellow), MHCI (green), CD8 (purple) and PanCK (red) in PGRN Ab-treated CKP tumors ( n = 8). Intratumoral heterogeneity of PGRN expression was observed, in which PGRN + and PGRN − regions were depicted in the representative tumor. PGRN and MHCI show opposite staining patterns in the PGRN Ab-treated tumor. c mIF showing the differential MHCI and CD8 expression in PGRN + and PGRN − tumor regions. ( n = 8 PAb-treated CKP tumors). Representative images are shown. Scale bar unit: μm.

Article Snippet: In the CD8 + T-cell depletion experiments, 25 mg/ml of anti-CD8 depleting Ab (clone 2.43, Bio X Cell) was injected i.p at days −3 and day 1 relative to the starting date of PGRN Ab treatment.

Techniques: Marker, Expressing, Staining

Journal: Nature Communications

Article Title: Progranulin mediates immune evasion of pancreatic ductal adenocarcinoma through regulation of MHCI expression

doi: 10.1038/s41467-021-27088-9

Figure Lengend Snippet: a Genetic strategy to induce spatially and temporally controlled GP33 expression by tamoxifen-mediated activation of CreERT2 in pancreatic cells harboring mutant Kras G12D and loss of Tp53 . Ptf1a wt/flp ; Kras wt/FSF-G12D ; p53 frt/frt (FKP) mice crossed to Gt(ROSA)26Sor tm3(CAG-Cre/ERT2)Das ( R26 FSF-CAG−CreERT2 ) and Gt(ROSA)26Sor tmloxP-STOP-loxP-GP-IRES-YFP ( R26 LSL-GP ) strains to generate FKPC2GP mice. b Experimental setup for co-culture of GP82, LCMV-gp33-expressing cell line derived from FKPC2GP tumor, and the gp33-reactive T cells isolated from the spleen of P14-TCR-Tg mice. c LCMV-gp33 (GP) expression in GP82 cells treated with tamoxifen (25 μM) or vehicle control DMSO for 2 days was assessed by flow cytometry ( n = 4 independent experiments). Two-tailed Mann–Whitney test. Tam: Tamoxifen. d Cellular PGRN level in GP82 cells treated with or without PGRN Ab or mIg (100 μg/ml) was assessed by flow cytometry ( n = 4 independent experiments). One-way ANOVA, Kruskal–Wallis test. e Surface expression of MHCI marker H-2Db on GP82 cells treated with or without PGRN Ab or mIg (100 μg/ml) was assessed by flow cytometry ( n = 6 independent experiments). One-way ANOVA, Kruskal–Wallis test. f IF staining of MHCI marker H-2Db of GP82 upon treatment with or without PGRN Ab or mIg (100 μg/ml). White arrowheads indicate the membraneous staining of MHCI. ( n = 3 independent experiments). Representative images are shown. g Microscopic images of GP82 cells and LCMV-gp33-reactive T cells (CFSE-labeled, green) after 2 days of co-culture. When anti-MHCI (H-2Db) neutralizing antibody (MHCI Ab, MAb) was included in the treatment, MHCI Ab was added 1 h after PGRN Ab (PAb) treatment. T cells were then added 1 h after MHCI Ab treatment. White arrowheads indicate T-cell clusters accumulated at GP82 cells. ( n = 6 independent experiments). Representative images are shown. h Cytotoxicity level (PI + %) of GP82 cells upon co-culture with LCMV-gp33-reactive T cells ( n = 6 independent experiments performed with LCMV-gp33-reactive T cells isolated from six different mice). T: T cells; Tu: GP82 tumor cells; Tu*GP: LCMV-gp33-induced GP82 tumor cells; PAb: PGRN Ab (100 μg/ml); MAb: MHCI Ab (100 μg/ml). One-way ANOVA, Kruskal–Wallis test. For PAb vs Mab: Two-tailed Mann–Whitney test. i Percentage of CD8 + cells that are positive for cytotoxic markers granzyme B (GzmB), TNF, and IFNg (% in total T cells) assessed by flow cytometer ( n = 6 independent experiments performed with LCMV-gp33-reactive T cells isolated from six different mice). One-way ANOVA, Kruskal–Wallis test. For PAb vs Mab: Two-tailed Mann–Whitney test. Mean + SD are shown. MFI mean fluorescence intensity. Scale bar unit: μm.

Article Snippet: In the CD8 + T-cell depletion experiments, 25 mg/ml of anti-CD8 depleting Ab (clone 2.43, Bio X Cell) was injected i.p at days −3 and day 1 relative to the starting date of PGRN Ab treatment.

Techniques: Expressing, Activation Assay, Mutagenesis, Co-Culture Assay, Derivative Assay, Isolation, Control, Flow Cytometry, Two Tailed Test, MANN-WHITNEY, Marker, Staining, Labeling, Fluorescence

Journal: Nature Communications

Article Title: Progranulin mediates immune evasion of pancreatic ductal adenocarcinoma through regulation of MHCI expression

doi: 10.1038/s41467-021-27088-9

Figure Lengend Snippet: a Timeline for treatment of anti-PGRN antibody (PAb) or mIg (50 mg/kg) in an orthotopic model of GP82 cells in C57BL/6 J mice. b Tumors and spleens of mIg-treated ( n = 4) and PGRN Ab-treated ( n = 4) mice with orthotopic GP82 transplantation and intravenous injection of LCMV-gp33-reactive T cells freshly isolated from P14-TCR-Tg mice. c Tumor growth was assessed by ultrasound imaging and presented as a fold change in tumor volume before and after PGRN Ab (PAb) or mIg treatment started. d Tumors were digested into disaggregated cells and stained for T-cell infiltration. Flow cytometric analysis showing the percentage of tumor-infiltrating T (CD3 + ) cells and the exogenously injected LCMV-gp33-reactive (CD45.1 + ) T cells, in tumors treated with PGRN Ab ( n = 4) or mIg ( n = 4). Two-tailed Mann–Whitney test. e Flow cytometric analysis showing the expression of cytotoxic markers granzyme B (GzmB), TNF, and IFNg on CD3 + T cells in tumors with PGRN Ab ( n = 4) or mIg ( n = 4). Two-tailed Mann–Whitney test. f IHC staining of PGRN, MHCI marker H-2Db, CD8, GzmB, and cleaved caspase 3 (Cl Casp3) in orthotopic GP82 tumors with PGRN Ab ( n = 4) or mIg ( n = 4). The lower panels how the percentages of positive cells in the whole tumors quantified by Definiens. Two-tailed Mann–Whitney test. Mean ± SD is shown. Scale bar unit: μm.

Article Snippet: In the CD8 + T-cell depletion experiments, 25 mg/ml of anti-CD8 depleting Ab (clone 2.43, Bio X Cell) was injected i.p at days −3 and day 1 relative to the starting date of PGRN Ab treatment.

Techniques: Transplantation Assay, Injection, Isolation, Imaging, Staining, Two Tailed Test, MANN-WHITNEY, Expressing, Immunohistochemistry, Marker

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Immune Modulation to Improve Survival of Viral Pneumonia in Mice

doi: 10.1165/rcmb.2020-0241OC

Figure Lengend Snippet: Antibodies

Article Snippet: CD8-depleting Ab , Bioxell , BE0223-A025.

Techniques:

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Immune Modulation to Improve Survival of Viral Pneumonia in Mice

doi: 10.1165/rcmb.2020-0241OC

Figure Lengend Snippet: Pam2-ODN pretreatment reduces SeV-induced lung CD8 + T cells. ( A ) Differential Giemsa staining of BAL cells from mice challenged with SeV with or without Pam2-ODN pretreatment. ( B ) Flow cytometry for CD8 + T cells from disaggregated mouse lungs 11 days after SeV infection with or without Pam2-ODN. ( C ) Lung CD8 + T cells 11 days after SeV challenge in mice pretreated with PBS or Pam2-ODN. Data shown are from one representative experiment of three independent experiments for A and of five independent experiments for B and C . * P < 0.05 compared with PBS+SeV, Student’s t test.

Article Snippet: CD8-depleting Ab , Bioxell , BE0223-A025.

Techniques: Staining, Flow Cytometry, Infection

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Immune Modulation to Improve Survival of Viral Pneumonia in Mice

doi: 10.1165/rcmb.2020-0241OC

Figure Lengend Snippet: Pam2-ODN treatment reduces CD8 + T-cell–associated SeV-induced immunopathology. ( A ) Experimental outline, ( B ) survival, and ( C ) percentage of CD8 + T cells from disaggregated mouse lungs 10 days after SeV challenge following pretreatment with PBS or Pam2-ODN and with or without CD8 + T cells depleted on day 8 of SeV challenge. ( D ) Lung histology 10 days after SeV challenged with or without Pam2-ODN treatment and/or CD8 + T cells. Data shown are one representative experiment of two independent experiments. Scale bars, 100 μm. n = 16 mice/group for survival in experiment A and n = 4 mice/group in experiment B . **** P < 0.0001 compared with PBS in C , *** P < 0.0005 compared with PBS in B and C , † P < 0.05 compared with CD8 Ab-PBS and * P < 0.05 compared with PBS. Mouse survival analysis in B were analyzed using log-rank, Mantel-Cox test. One-way ANOVA with multiple comparisons was used in C . Ab = antibody.

Article Snippet: CD8-depleting Ab , Bioxell , BE0223-A025.

Techniques:

Journal: Molecular Therapy

Article Title: Targeting a Pre-existing Anti-transgene T Cell Response for Effective Gene Therapy of MPS-I in the Mouse Model of the Disease

doi: 10.1016/j.ymthe.2019.04.014

Figure Lengend Snippet: IDUA-IMM Mice Develop an Anti-IDUA Immune Response (A) MPS-I mice were either left untreated (naive, n = 10) or immunized with rhIDUA in Incomplete Freund’s Adjuvant (IDUA-IMM, n = 21). At 35 days after immunization, mice were sacrificed and the concentration of anti-IDUA IgGs in serum was measured by ELISA. Each dot represents one mouse; error bars indicate mean ± SEM. ****p ≤ 0.00005, Mann-Whitney test. (B) Numbers of B and plasma cells actively secreting anti-IDUA IgGs were quantified by ELISPOT assay as spot-forming units (SFU)/3.5 × 10 5 total cells isolated from the spleen and BM of mice. Values for mean ± SEM of naive (n = 4) and IDUA-IMM (n = 6) MPS-I mice are shown. *p ≤ 0.05, Mann-Whitney test. (C) Number of IDUA-specific CD8 + T cells was measured by ELISPOT assay as SFU/10 5 CD8 + T cells isolated from the spleen of treated mice. Each dot represents one mouse; error bars indicate mean ± SEM of naive (n = 4) and IDUA-IMM (n = 6) MPS-I mice. **p ≤ 0.005, Mann-Whitney test. (D) Total splenocytes were stimulated with rhIDUA (Aldurazyme, 10 μg/mL), and cytokine concentration was measured in culture supernatants at day 4 by Bioplex assay. Results are expressed as fold increase of cytokine secretion in stimulated compared to not-stimulated conditions. Values for mean ± SEM of naive (n = 4) and IDUA-IMM (n = 6) MPS-I mice are shown. **p ≤ 0.005, Mann-Whitney test.

Article Snippet: In some experiments, IDUA-IMM MPS-I mice were i.v. injected with depleting anti-CD8 Ab (clone YTS169.4, BioXcell, 250 μg/mouse) 1 day before transplantation of transduced cells.

Techniques: Adjuvant, Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Clinical Proteomics, Enzyme-linked Immunospot, Isolation, BioPlex Assay

Journal: Molecular Therapy

Article Title: Targeting a Pre-existing Anti-transgene T Cell Response for Effective Gene Therapy of MPS-I in the Mouse Model of the Disease

doi: 10.1016/j.ymthe.2019.04.014

Figure Lengend Snippet: Pre-existing Anti-IDUA Immunity Prevents Engraftment of IDUA-Transduced HSCs in an Antigen-Specific Manner (A) Naive and IDUA-IMM MPS-I mice were lethally irradiated and i.v. injected with LV.IDUA (IDUA-GT)- or LV-OVA (OVA-GT)-transduced autologous BM-derived HSCs (10 6 cells/mouse). (B) Percentages of survival for naive MPS-I mice transplanted with LV.IDUA-transduced HSCs (naive IDUA-GT, n = 5), naive MPS-I mice transplanted with LV.OVA-transduced HSCs (naive OVA-GT, n = 2), IDUA-IMM MPS-I mice transplanted with LV.IDUA-transduced HSCs (IDUA-IMM IDUA-GT, n = 11), IDUA-IMM MPS-I mice transplanted with LV.OVA-transduced HSCs (IDUA-IMM OVA-GT, n = 4), and irradiated and not transplanted MPS-I mice (Irrad Ctrl, n = 8). (C) At 12 weeks after transplantation, vector copy number was quantified by digital droplet PCR (ddPCR) in BM cells (VCN BM) of treated mice. Each dot represents one mouse; error bars indicate mean ± SEM. **p ≤ 0.005, Mann-Whitney test. (D) The concentration of anti-IDUA IgGs in serum was measured by ELISA before and after transplantation. Results are shown as mean ± SEM. (E) At 12 weeks after transplantation, the number of anti-IDUA IgG-secreting cells was quantified by ELISPOT assay as SFU/3.5 × 10 5 total splenocytes. Values for mean ± SEM of naive (n = 5) and IDUA-IMM OVA-GT (n = 4) MPS-I mice are shown. *p ≤ 0.05, Mann-Whitney test. (F) At 12 weeks after transplantation, the number of IDUA-specific CD8 + cells was measured by ELISPOT assay as SFU/10 5 CD8 + T cells isolated from the spleen of treated mice. Each dot represents one mouse; error bars indicate mean ± SEM of naive (n = 5) and IDUA-IMM (n = 4) MPS-I mice. *p ≤ 0.05, Mann-Whitney test.

Article Snippet: In some experiments, IDUA-IMM MPS-I mice were i.v. injected with depleting anti-CD8 Ab (clone YTS169.4, BioXcell, 250 μg/mouse) 1 day before transplantation of transduced cells.

Techniques: Irradiation, Injection, Derivative Assay, Transplantation Assay, Plasmid Preparation, MANN-WHITNEY, Concentration Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Isolation

Journal: Molecular Therapy

Article Title: Targeting a Pre-existing Anti-transgene T Cell Response for Effective Gene Therapy of MPS-I in the Mouse Model of the Disease

doi: 10.1016/j.ymthe.2019.04.014

Figure Lengend Snippet: CD8 + T Cell Depletion Rescues Engraftment of IDUA-Transduced HSCs in Immunized MPS-I Mice (A) Naive and IDUA-IMM MPS-I mice left untreated or treated with anti-CD8 depleting mAb were lethally irradiated and transplanted with LV.IDUA-transduced (10 6 cells/mouse) autologous BM-derived HSCs. (B) At the time of transplantation, the absolute number of CD8 + T cells was quantified in the peripheral blood of treated mice by fluorescence-activated cell sorting (FACS) staining. Each dot represents one mouse; error bars indicate mean ± SEM. *p ≤ 0.05, Mann-Whitney test. (C) Percentages of survival for naive mice transplanted with LV.IDUA-transduced cells (naive, n = 5), IDUA-IMM MPS-I mice transplanted with LV.IDUA-transduced cells untreated (IDUA-IMM n = 3) or treated with anti-CD8 mAb (IDUA-IMM anti-CD8, n = 8), and irradiated and not-transplanted MPS-I mice (Irrad Ctrl, n = 3). (D) At 12 weeks after transplantation, vector copy number was quantified by ddPCR in BM cells (VCN BM) of treated mice. Each dot represents absolute VCN of a single mouse normalized on the mean of VCN of the naive group; error bars indicate mean ± SEM. *p ≤ 0.05, Mann-Whitney test. (E) The concentration of anti-IDUA IgGs was measured by ELISA in the sera of treated MPS-I mice before and after transplantation. Results are shown as mean ± SEM.

Article Snippet: In some experiments, IDUA-IMM MPS-I mice were i.v. injected with depleting anti-CD8 Ab (clone YTS169.4, BioXcell, 250 μg/mouse) 1 day before transplantation of transduced cells.

Techniques: Irradiation, Derivative Assay, Transplantation Assay, Fluorescence, FACS, Staining, MANN-WHITNEY, Plasmid Preparation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy

Article Title: Targeting a Pre-existing Anti-transgene T Cell Response for Effective Gene Therapy of MPS-I in the Mouse Model of the Disease

doi: 10.1016/j.ymthe.2019.04.014

Figure Lengend Snippet: Pre-treatment with T Cell Depleting Agents Partially Rescues Engraftment of IDUA-Transduced HSCs in Immunized MPS-I Mice (A) Naive and IDUA-IMM MPS-I mice left untreated or treated with Fludarabine or anti-CD3 depleting mAb were lethally irradiated and transplanted with LV-IDUA-transduced BM-derived HSCs isolated from autologous mice (IDUA-GT, 10 6 cells/mouse). (B) At the time of transplantation, the absolute number of CD8 + T cells was quantified in peripheral blood of treated mice by FACS staining. Each dot represents one mouse; error bars indicate mean ± SEM. *p ≤ 0.05, Mann-Whitney test. (C) At 12 weeks after transplantation, vector copy number was quantified by ddPCR in BM cells (VCN BM) of treated and control MPS-I mice. Each dot represents absolute VCN of a single mouse normalized on the mean of VCN of the naive group; error bars indicate mean ± SEM. *p ≤ 0.05, Mann-Whitney test. (D) At 12 weeks after transplantation, the number of IDUA-specific CD8 + T cells was quantified by ELISPOT assay as SFU/10 5 CD8 + T cells isolated from the spleen of treated mice. The number of IDUA-specific CD8 + T cells was correlated to the VCN of a single mouse normalized on the mean of VCN of the naive untreated group VCN data. Each dot represents one mouse.

Article Snippet: In some experiments, IDUA-IMM MPS-I mice were i.v. injected with depleting anti-CD8 Ab (clone YTS169.4, BioXcell, 250 μg/mouse) 1 day before transplantation of transduced cells.

Techniques: Irradiation, Derivative Assay, Isolation, Transplantation Assay, Staining, MANN-WHITNEY, Plasmid Preparation, Control, Enzyme-linked Immunospot

Journal: Molecular Therapy

Article Title: Targeting a Pre-existing Anti-transgene T Cell Response for Effective Gene Therapy of MPS-I in the Mouse Model of the Disease

doi: 10.1016/j.ymthe.2019.04.014

Figure Lengend Snippet: Viral Infection Simulation during ERT Induces Anti-IDUA CD8 + T Cells Able to Eliminate LV.IDUA-Transduced HSCs MPS-I mice received 5 weekly i.v. injections of rhIDUA alone (ERT, n = 5) or in combination with poly(I:C) on weeks 2 and 3 (ERT-poly(I:C), n = 6). Mice were then lethally irradiated and transplanted with LV.IDUA-transduced autologous BM-derived HSCs (10 6 cells/mouse). (A) At 12 weeks after transplantation, vector copy number was quantified by ddPCR in BM cells (VCN BM) of treated mice. Each dot represents the absolute VCN of a single mouse normalized on the mean of VCN of the ERT group; error bars indicate mean ± SEM. *p ≤ 0.05, Mann-Whitney test. (B) At 12 weeks after transplantation, the number of IDUA-specific CD8 + T cells was quantified by ELISPOT assay as SFU/10 5 CD8 + T cells isolated from the spleen of treated mice. Each dot represents one mouse; error bars indicate mean ± SEM. (C) Sera from ERT-poly(I:C)-treated mice were collected before (PRE) and 12 weeks after gene therapy (POST), and IDUA enzymatic activity was quantified fluorometrically. Dashed line indicates the threshold of positivity obtained as mean + 2SD of IDUA enzymatic activity in the sera of n = 17 untreated MPS-I mice (2.41 pmol/1 h/1 μL). IDUA enzymatic activity of each single mouse is shown.

Article Snippet: In some experiments, IDUA-IMM MPS-I mice were i.v. injected with depleting anti-CD8 Ab (clone YTS169.4, BioXcell, 250 μg/mouse) 1 day before transplantation of transduced cells.

Techniques: Infection, Irradiation, Derivative Assay, Transplantation Assay, Plasmid Preparation, MANN-WHITNEY, Enzyme-linked Immunospot, Isolation, Activity Assay