Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The role of Ly49E receptor expression on murine intraepithelial lymphocytes in intestinal cancer development and progression

doi: 10.1007/s00262-016-1894-6

Figure Lengend Snippet: Tumor-infiltrating T cells and tumor uPA expression in ApcMin/+ Ly49E WT versus ApcMin/+ Ly49E KO mice. a Hematoxylin/eosin- (upper), and CD3-(lower) stained paraffin tumor sections from ApcMin/+ Ly49E WT and ApcMin/+ Ly49E KO mice. Scale bar: 250 µm, ×100 magnification. A graph showing CD3 mean gray value/mm2 according to tumor surface area (mm2) is shown for ApcMin/+ Ly49E WT and ApcMin/+ Ly49E KO mice (n = 6). b Small intestinal tumor-infiltrating IEL subpopulation frequencies in ApcMin/+ Ly49E WT and ApcMin/+ Ly49E KO mice, and small intestinal IEL subpopulation frequencies from control littermates, at 14 weeks of age (mean ± SEM; n = 6). The percentage of TCRαβ and TCRγδ IEL is shown as a fraction of the total numbers of T cells. TCRαβ CD4, TCRαβ CD8αβ and TCRαβ CD8αα IEL subpopulation frequencies are shown as a percentage of the total TCRαβ IELs. TCRγδ DN and TCRγδ CD8αα IEL subpopulation frequencies are shown as a percentage of total TCRγδ IEL. c Tumor uPA expression in tumors from ApcMin/+ Ly49E WT and ApcMin/+ Ly49E KO mice at 14 weeks of age. n.s. not significant

Article Snippet: Antibodies used for immunofluorescent staining were polyclonal rabbit anti-human/mouse CD3 (Dako) and Dylight649 donkey anti-rabbit (BioLegend).

Techniques: Expressing, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The role of Ly49E receptor expression on murine intraepithelial lymphocytes in intestinal cancer development and progression

doi: 10.1007/s00262-016-1894-6

Figure Lengend Snippet: Tumor-infiltrating T cells and tumor uPA expression in AOM-treated Ly49E WT versus Ly49E KO mice. a Hematoxylin/eosin-(H&E)- (upper), and CD3-(lower) stained paraffin tumor sections from AOM-treated Ly49E WT and Ly49E KO mice. Scale bar 250 µm, ×100 magnification. A graph showing CD3 mean gray value/mm2 according to tumor surface area (mm2) is shown for AOM-treated Ly49E WT and Ly49E KO mice (n = 6). b Colon tumor-infiltrating IEL subpopulation frequencies in AOM-treated Ly49E WT and Ly49E KO mice 14–22 weeks following the start of treatment, and colon IEL subpopulation frequencies from untreated Ly49E WT and Ly49E KO mice (mean ± SEM; n = 5 for AOM-treated mice; n = 3 for untreated mice). The percentage of TCRαβ and TCRγδ IEL is shown as a fraction of the total numbers of T cells. TCRαβ CD4, TCRαβ CD8αβ and TCRαβ CD8αα IEL subpopulation frequencies are shown as a percentage of the total TCRαβ IELs. TCRγδ DN and TCRγδ CD8αα IEL subpopulation frequencies are shown as a percentage of the total TCRγδ IEL. c Dot plots are shown for CD4/CD8β versus CD8α expression in colon tumor-infiltrating IEL in AOM-treated Ly49E WT and Ly49E KO mice, and colon IEL from untreated Ly49E WT and Ly49E KO mice. Numbers indicate the percentage of cells in each quadrant. Dot plots are representative for n = 5 AOM-treated mice and n = 3 untreated mice. d Tumor uPA expression in tumors of varying size from AOM-treated Ly49E WT and Ly49E KO mice at 14–22 weeks following the start of treatment. n.s. not significant. Data were analyzed using the nonparametric two-tailed Mann–Whitney U-test or the Kruskall–Wallis test. A p value ≤0.05, (*), a p value ≤0.01 (**) and p value ≤0.001 (***), were considered statistically significant

Article Snippet: Antibodies used for immunofluorescent staining were polyclonal rabbit anti-human/mouse CD3 (Dako) and Dylight649 donkey anti-rabbit (BioLegend).

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

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Preventive effect of tertiary lymphoid structures on lymph node metastasis of lung adenocarcinoma

doi: 10.1007/s00262-022-03353-8

Figure Lengend Snippet: Characterization of tertiary lymphoid structures and tumor-infiltrating lymphocytes. Characterization of tertiary lymphoid structures (TLSs) in lung adenocarcinoma by staining of formalin-fixed, paraffin-embedded tissues (A–D). TLS in lung adenocarcinoma stained by hematoxylin and eosin (A), CD20 (B), CD3 (C) and DC-Lamp (D). Tumor-infiltrating lymphocytes stained by CD3 (E), CD8 (F), CD4 (G) and Foxp3 (H). Histogram of the number of TLSs within low-power field (LPF) (I). Histogram of the number of DC-Lamp+ mature dendritic cells in TLSs (/0.04 mm2) (J)

Article Snippet: The primary antibodies were: mouse monoclonal anti-human CD20cy antibody (clone #L26, dilution 1:200; Dako); rat monoclonal anti-mouse DC-Lamp antibody (clone #1010E1.01, dilution 1:100; eurobio SCIENTIFIC); mouse monoclonal anti-human CD3 antibody (clone #F7.2.38, dilution 1:50; DAKO); mouse monoclonal anti-human CD8 antibody (clone #C8/144B, dilution 1:100; Dako); rabbit monoclonal anti-human CD4 antibody (clone #sp35, dilution 1:100; abcam); mouse monoclonal anti-human Foxp3 antibody (clone #236A/E7, dilution 1:100; eBioscience); rabbit polyclonal anti-granzyme B antibody (dilution 1:200; abcam); mouse monoclonal anti-human NKp46/NCR1 antibody (clone #195,314, dilution 1:200; R&D systems); rabbit polyclonal anti-MIP-3 beta/CCL19 antibody (dilution 1:500; abcam); rabbit polyclonal anti-CCL21 antibody (dilution 1:200; abcam); and rabbit polyclonal anti-CCR7 antibody (dilution 1:200; abcam).

Techniques: Staining, Formalin-fixed Paraffin-Embedded

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Preventive effect of tertiary lymphoid structures on lymph node metastasis of lung adenocarcinoma

doi: 10.1007/s00262-022-03353-8

Figure Lengend Snippet: Clinicopathological characteristics of the patients according to mature/immature TLS

Article Snippet: The primary antibodies were: mouse monoclonal anti-human CD20cy antibody (clone #L26, dilution 1:200; Dako); rat monoclonal anti-mouse DC-Lamp antibody (clone #1010E1.01, dilution 1:100; eurobio SCIENTIFIC); mouse monoclonal anti-human CD3 antibody (clone #F7.2.38, dilution 1:50; DAKO); mouse monoclonal anti-human CD8 antibody (clone #C8/144B, dilution 1:100; Dako); rabbit monoclonal anti-human CD4 antibody (clone #sp35, dilution 1:100; abcam); mouse monoclonal anti-human Foxp3 antibody (clone #236A/E7, dilution 1:100; eBioscience); rabbit polyclonal anti-granzyme B antibody (dilution 1:200; abcam); mouse monoclonal anti-human NKp46/NCR1 antibody (clone #195,314, dilution 1:200; R&D systems); rabbit polyclonal anti-MIP-3 beta/CCL19 antibody (dilution 1:500; abcam); rabbit polyclonal anti-CCL21 antibody (dilution 1:200; abcam); and rabbit polyclonal anti-CCR7 antibody (dilution 1:200; abcam).

Techniques: Mutagenesis, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Maintenance of WT1 expression in tumor cells is associated with a good prognosis in malignant glioma patients treated with WT1 peptide vaccine immunotherapy

doi: 10.1007/s00262-021-02954-z

Figure Lengend Snippet: Summary of IHC staining results of both pre- and post- WT1 vaccination specimens

Article Snippet: The sections were then incubated overnight at 4 °C with anti-human WT1 mouse monoclonal antibody 6F-H2 (DAKO, Carpinteria, CA, USA; diluted 1:50), anti-human Transforming growth factor-β (TGF-β) rabbit polyclonal antibody (Abcam, Cambridge, MA, USA; diluted 1:200), anti–human Ki-67 mouse monoclonal antibody MIB-1 (DAKO; diluted 1:40), anti-human HLA class I-ABC mouse monoclonal antibody EMR8-5 (Hokudo, Sapporo, Japan; diluted 1:100), anti-human programmed death-ligand 1 (PD-L1) rabbit monoclonal antibody ab205921 clone 28–8, (Abcam; diluted 1:100), anti-human CD3 mouse monoclonal antibody F7.2.38 (DAKO; diluted 1:5), anti–human CD4 mouse monoclonal antibody 4B12 (DAKO; diluted 1:80), anti–human CD8 mouse monoclonal antibody C8/14B (Nichirei Biosciences, Tokyo, Japan; undiluted), anti-human CD79α mouse monoclonal antibody JCB117 (DAKO; diluted 1:200), anti-human PD-1 mouse monoclonal antibody clone NAT105 (Abcam; diluted 1:100), anti-human Foxp3 mouse monoclonal antibody 236A/E7 (Abcam; diluted 1:100), or anti IDH1­R132H mouse monoclonal Antibody (FUJIFILM Wako Chemicals, Osaka, Japan; diluted 1:100).

Techniques: Immunohistochemistry

Journal: Nature Communications

Article Title: Topical tacrolimus for the treatment of secondary lymphedema

doi: 10.1038/ncomms14345

Figure Lengend Snippet: ( a ) Representative photographs of mouse tails after surgical excision of superficial/deep collecting lymphatics and treatment with or without topical tacrolimus beginning either 2 weeks (early treatment) or 6 weeks (late treatment) after surgery. Images represent 4 weeks of treatment in the early group and 3 weeks of treatment in the late group. ( b ) Graphical representation of tail volume changes after early (* P =0.021) or late (* P =0.018) treatment with tacrolimus, as compared with controls (red arrows indicate timing of treatment) ( n =8/group). ( c ) Representative cross-sectional histological images (upper panel) of control and early tacrolimus-treated mouse tails harvested 6 weeks after lymphatic ablation. Brackets show soft tissue thickness. Quantification of soft tissue changes (lower panel) after early or late treatment with tacrolimus (* P ≤0.001) ( n =8/group). ( d ) Quantification of whole-blood tacrolimus levels demonstrating immunosuppressive concentrations in systemically treated animals (4 mg kg −1 intraperitoneally daily) and non-immunosuppressive levels in the topically treated group (* P =0.007; n =6/group). ( e ) Representative flow cytometry plots displaying side scatter area (SSA) on the y axis and CD3 + cells (T cells) on the x axis for animals retreated with vehicle control, topical tacrolimus or systemic tacrolimus. ( f ) Quantification of blood T cells for each group. Note significant reduction in T cells only in systemically treated animals (* P =0.012; n =6/group). Experiments were repeated two to three times. All data represent mean±s.d. with P ≤0.05 considered as significant. Two-tailed Student's t -test ( c , d ) and analysis of variance (ANOVA) with post hoc tests ( b , f ).

Article Snippet: Rabbit anti-human/mouse CD3 (A0452; 1:300; Dako, Carpinteria, CA) and Cy3-conjugated mouse anti-αSMA (C6-198; 1:1,000; Sigma-Aldrich).

Techniques: Flow Cytometry, Two Tailed Test

Journal: PLoS ONE

Article Title: 3D investigation shows walls and wall-like structures around human germinal centres, probably regulating T- and B-cell entry and exit

doi: 10.1371/journal.pone.0242177

Figure Lengend Snippet: (A) Alpha-actin + (red) and CD3 + cells (green) relate to each other. The number of T-cells decreases abruptly towards the B-zone where the FRC wall is located. (B) Alpha-actin positive (green) and BCL6 positive (red) cells show proportion of walls towards the germinal centre. BCL6 positive germinal centre cells are situated within the physical boundary. Additionally, the BCL6 and alpha-actin negative surrounding between germinal centre and FRC wall indicates that the follicular mantle is within the compartmentalised area of the walls. (C) Alpha-actin-positive (red) and IgD positive (green) structures reveal orientation of FRC walls. The follicular mantle shows a polar orientation without or with only fragmented FRC walls towards the mantle zone. On the opposite pole, the germinal centre is compartmentalised by smaller FRC walls and is more permeable. The remaining germinal centre is enclosed from both sides by the FRC walls in form of a sufficient physical boundary. This staining combination shows properly that the orientation of the FRC walls shows a coherence with the follicular mantle. The maximum resolution of (A), (B) and (C) was set to 0.13 μm per pixel in the native microscopy dataset.

Article Snippet: We used anti-alpha smooth muscle actin antibody (SP171/Abcam), anti-human BCL6 monoclonal mouse antibody (M7211;Agilent/Dako), anti-human polyclonal rabbit IgD antibody (IR517;Agilent/Dako), anti-human CD3 monoclonal mouse antibody (A0452; Agilent/Dako) and anti-human CD23 monoclonal mouse antibody (M7312; Agilent/Dako).

Techniques: Staining, Microscopy

Journal: Frontiers in Immunology

Article Title: Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy

doi: 10.3389/fimmu.2018.01359

Figure Lengend Snippet: Profile of circulating memory CD8 + T cells and regulatory T cells. Percentages of CD45RO + RA − memory CD8 + T cells (on CD3 + CD8 + T cells) in mesenchymal stromal cell (MSC)-treated patients #1 (blue circles), #2 (orange circles) and in control kidney transplant recipients given basiliximab/low-dose rabbit anti-thymocyte globulin (RATG) induction therapy but not MSC (white histograms) during the follow-up (A) and in MSC-treated patients #3 (green circles) and # 4 (violet circles) and in control kidney transplant recipients given induction therapy with low-dose RATG alone but not MSC (gray histograms) during the follow-up (B) . Panels (C,D) represent profile of percentages of regulatory T cells (Tregs) (Foxp3 + CD127 − on CD4 + CD25 high T cells) during the follow-up in MSC-treated patients #1 and #2 and in control patients given basiliximab/low-dose RATG (white histograms) and in MSC-patients #3 and #4 and in control patients given induction therapy with low-dose RATG alone (gray histograms), respectively. Ratio of Treg/memory CD8 + T cell percentages in MSC-treated patients and in control groups are shown in panels (E,F) . Data from controls are mean ± SEM, * P < 0.05 vs pretransplant values.

Article Snippet: The mix antibodies contained BUV395 mouse anti-human CD45 (clone HI30), BV510 mouse anti-human CD3 (clone HIT3a), APC-R700 mouse anti-human CD27 (clone M-T271), BB700 mouse anti-human CD16 (clone B73.1), BV650 mouse anti-human CD56 (clone B159), BV711 mouse anti-human CD11b (clone D12), BUV737 mouse anti-human CD14 (clone M5E2), PE-CF594 mouse anti-human CD64 (clone 10.1), BV605 mouse anti-human HLA-DR (clone G46-6), and PE-Cy5 mouse anti-human CD33 (clone WM-53), all antibodies were purchased from BD Bioscience.

Techniques:

Journal: Frontiers in Immunology

Article Title: Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy

doi: 10.3389/fimmu.2018.01359

Figure Lengend Snippet: Profile of naïve and transitional B cell counts in the peripheral blood. Number (A) and percentages [on total CD3 − CD19 + B cells (B) ] of naïve IgD + CD27 − B cells in the peripheral blood during the follow-up in mesenchymal stromal cell (MSC)-treated patients #1 (blue circles) and #2 (orange circles) and in control kidney transplant recipients given basiliximab/low-dose rabbit anti-thymocyte globulin (RATG) induction therapy but not MSC (white histograms) during the follow-up (left panels) and in MSC-treated patients #3 (green circles) and #4 (violet circles) and in control kidney transplant recipients given induction therapy with low-dose RATG alone but not MSC (gray histograms) (right panels). Number (C) and percentages [on total CD3 − CD19 + B cells (D) ] of CD24 high CD38 high transitional B cells in MSC-treated patients #1 and #2 and in control patients given basiliximab/low-dose RATG (white histograms) (left panels) and in MSC-patients #3 and #4 and in control patients given induction therapy with low-dose RATG alone (gray histograms) (right panels), during the follow-up. Data from controls are mean ± SEM, P = NS.

Article Snippet: The mix antibodies contained BUV395 mouse anti-human CD45 (clone HI30), BV510 mouse anti-human CD3 (clone HIT3a), APC-R700 mouse anti-human CD27 (clone M-T271), BB700 mouse anti-human CD16 (clone B73.1), BV650 mouse anti-human CD56 (clone B159), BV711 mouse anti-human CD11b (clone D12), BUV737 mouse anti-human CD14 (clone M5E2), PE-CF594 mouse anti-human CD64 (clone 10.1), BV605 mouse anti-human HLA-DR (clone G46-6), and PE-Cy5 mouse anti-human CD33 (clone WM-53), all antibodies were purchased from BD Bioscience.

Techniques:

Journal: Frontiers in Immunology

Article Title: Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy

doi: 10.3389/fimmu.2018.01359

Figure Lengend Snippet: Profile of circulating NK cell subsets. (A) Profile of NK cell counts in mesenchymal stromal cell (MSC)-treated patients ( n = 4), and kidney transplant patients undergoing induction therapy with bas/low dose rabbit anti-thymocyte globulin (RATG) ( n = 6) or with low-dose RATG alone ( n = 5) from 1 to 5 years posttransplant. Data are mean ± SEM. P = NS. (B) Gating strategy for identification of natural killer cell subsets and natural killer T cells. In the CD45 + live singlet cells, the population of CD3 − lymphocytes was gated on the morphology based on SSC and plotted for CD16 and CD56 expression. CD16 + CD56 dim cells positive for CD11b expression and negative for CD27 were identified as cytotoxic NK cells. The CD56 bright CD16 −/+ NK cells were divided into CD27 + and CD27 − subpopulations. NKT cells were defined as CD3 + CD56 + cells. The frequency (% on the gated CD3 − population) of CD56 dim NK cells, CD56 bright NK cells negative or positive for CD27 expression, and of NKT cells (% on CD3 + cells) in patients given MSC infusion and in kidney transplant patients undergoing induction therapy with bas/low dose RATG or with low-dose RATG alone from 1 to 5 years posttransplant are reported in panels (C–F) , respectively. Data are mean ± SEM, * P < 0.05 vs MSC-treated patients and control patients given low-dose RATG at the same time point.

Article Snippet: The mix antibodies contained BUV395 mouse anti-human CD45 (clone HI30), BV510 mouse anti-human CD3 (clone HIT3a), APC-R700 mouse anti-human CD27 (clone M-T271), BB700 mouse anti-human CD16 (clone B73.1), BV650 mouse anti-human CD56 (clone B159), BV711 mouse anti-human CD11b (clone D12), BUV737 mouse anti-human CD14 (clone M5E2), PE-CF594 mouse anti-human CD64 (clone 10.1), BV605 mouse anti-human HLA-DR (clone G46-6), and PE-Cy5 mouse anti-human CD33 (clone WM-53), all antibodies were purchased from BD Bioscience.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy

doi: 10.3389/fimmu.2018.01359

Figure Lengend Snippet: Profile of circulating monocytes. (A) Gating strategy for identification of classical, non-classical, and intermediate monocytes in peripheral blood mononuclear cells. In the CD45 + live singlet cells, the population of CD3 − leukocytes was gated on the morphology based on SSC and plotted for CD14 expression. Classical monocytes were identified as CD64 + cells on gated CD33 + HLADR + cells in the CD14 + CD16 − cell population, intermediate monocytes as CD64 + cells on gated CD33 + HLADR + in the CD14 + CD16 + cell population, and non-classical monocytes as CD64 − cells on gated CD33 + HLADR + cells in the CD14 − CD16 + cell population. Percentages of classical monocytes (% on the gated CD3 − population) and their HLA-DR expression in patients given mesenchymal stromal cell (MSC) infusion ( n = 4) and in kidney transplant patients undergoing induction therapy with bas/low dose rabbit anti-thymocyte globulin (RATG) ( n = 6) or with low-dose RATG alone ( n = 5) from 1 to 5 years posttransplant are reported in panels (B,C) , respectively. Data are mean ± SEM, P = NS. Table in panel (D) reported mean ± SEM of intermediate and non classical monocytes; P = NS.

Article Snippet: The mix antibodies contained BUV395 mouse anti-human CD45 (clone HI30), BV510 mouse anti-human CD3 (clone HIT3a), APC-R700 mouse anti-human CD27 (clone M-T271), BB700 mouse anti-human CD16 (clone B73.1), BV650 mouse anti-human CD56 (clone B159), BV711 mouse anti-human CD11b (clone D12), BUV737 mouse anti-human CD14 (clone M5E2), PE-CF594 mouse anti-human CD64 (clone 10.1), BV605 mouse anti-human HLA-DR (clone G46-6), and PE-Cy5 mouse anti-human CD33 (clone WM-53), all antibodies were purchased from BD Bioscience.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Long-Term Clinical and Immunological Profile of Kidney Transplant Patients Given Mesenchymal Stromal Cell Immunotherapy

doi: 10.3389/fimmu.2018.01359

Figure Lengend Snippet: Histologic analysis of protocol kidney graft biopsy, profile of circulating B and T cell subsets, and immunologic functional assays of memory T cell response in patient #5, who received pretransplant infusion of mesenchymal stromal cell (MSC) in the setting of induction therapy with basiliximab/low-dose rabbit anti-thymocyte globulin (RATG). Patient #5, a 42-year-old woman with end-stage renal disease secondary to tubular interstitial nephropathy, received a kidney transplant from her sister, mismatched for two HLA alleles. She was given MSC before surgery (day −1) and induction therapy with the combination of basiliximab and low-dose RATG. A representative image of Gomori’s trichrome staining on 1-year protocol kidney graft biopsy (original magnification 100×) of MSC-treated patient #5 is shown (A) . Percentages of circulating CD45RO + RA − memory CD8 + T cells (on CD3 + CD8 + T cells) (B) , percentages of circulating regulatory T cells (Tregs) (Foxp3 + CD127 − on CD4 + CD25 high T cells) (C) , and ratio of the percentages of Tregs to CD45RO + RA − memory CD8 + T cells (D) in patient #5 (yellow circles) and in control kidney transplant recipients given basiliximab/low-dose RATG induction therapy but not MSC (white histograms) from baseline (pretransplant) to 1-year posttransplant. Number of circulating naïve IgD + CD27 − B cells (E) and CD24 high CD38 high transitional B cells (F) in patient #5 (yellow circles) and in control kidney transplant recipients given basiliximab/low-dose RATG induction therapy (white histograms) from baseline to 1-year posttransplant. ELISpot for IFNγ (G) and CD8 + T cell function by T cell-mediated lympholysis [as percentage of specific lysis at 50:1 effector-target ratio (H) ] toward donor or third-party antigens in patient #5 (yellow circles) and in control kidney transplant recipients given basiliximab/low-dose RATG induction therapy (white histograms) on peripheral blood mononuclear cells collected pretransplant and at 1 year after transplantation. Data are mean ± SEM.

Article Snippet: The mix antibodies contained BUV395 mouse anti-human CD45 (clone HI30), BV510 mouse anti-human CD3 (clone HIT3a), APC-R700 mouse anti-human CD27 (clone M-T271), BB700 mouse anti-human CD16 (clone B73.1), BV650 mouse anti-human CD56 (clone B159), BV711 mouse anti-human CD11b (clone D12), BUV737 mouse anti-human CD14 (clone M5E2), PE-CF594 mouse anti-human CD64 (clone 10.1), BV605 mouse anti-human HLA-DR (clone G46-6), and PE-Cy5 mouse anti-human CD33 (clone WM-53), all antibodies were purchased from BD Bioscience.

Techniques: Functional Assay, Staining, Enzyme-linked Immunospot, Cell Function Assay, Lysis, Transplantation Assay