Journal: The Journal of Experimental Medicine

Article Title: A novel antiangiogenic and vascular normalization therapy targeted against human CD160 receptor

doi: 10.1084/jem.20100810

Figure Lengend Snippet: Expression of CD160 in tumor blood vessels but not in the blood vessels of healthy tissues. (A) Representative sections of B16 melanoma at day 16 after their subcutaneous injection (top) and healthy mouse heart (bottom), both stained with either CL1-R2 (CD160) or isolectin B4, an endothelial cell marker. Bar, 50 µm. Images are representative of four independent experiments (five mice/group). (B) Sections of human colon tumor (top) and healthy colon (bottom) stained with CL1-R2 (CD160) or a mAb against CD31, a human endothelial cell marker. Images are representative of two patient biopsies. Control IgG1 did not stain and is not shown. Bar, 50 µm.

Article Snippet: The cross-reactivity of CL1-R2 mAb with CD160 mouse proteins has been demonstrated by different means (Fig. S1): (1) using flow cytometry, we found that CL1-R2 stained mouse SVR endothelial cells as well as the BCL-1 mouse B cell chronic lymphocytic leukemia cell line (Fig. S1 A); (2) using immunoprecipitation and Western blotting, we found that the CNX46-3 anti–mouse CD160 mAb (Cedarlane laboratories) immunoprecipitated protein bands from mouse spleen or BCL-1 cells that were blotted by the CL1-R2 mAb (Fig. S1 B); and (3) five CD160 amino acids were identified by mass spectrometry in the CNX43-6–immunoprecipitated 37-kD band blotted with CL1-R2 mAb with a minimum coverage of 37.5% (Fig. S1 C).

Techniques: Expressing, Injection, Staining, Marker, Control

Journal: Blood Cancer Journal

Article Title: Minimal residual disease detection with tumor-specific CD160 correlates with event-free survival in chronic lymphocytic leukemia

doi: 10.1038/bcj.2014.92

Figure Lengend Snippet: MRD detection by CD160FCA and correlation with PCR in the pilot cohort. ( a ) The theoretical MRD expression (calculated, x axis) is plotted against the observed MRD expression (actual analyzed MRD analysis, y axis) by CD160FCA. The results are the percentage expression of CD160 of the total leukocyte population. The comparative RT-PCR results are demonstrated at the top of the graph above the corresponding dilution. The R 2 linear regression value for observed expression vs theoretical expression was R 2 =0.98. ( b ) Separation of MRD-positive and MRD-negative populations based on CD160FCA assessment ( P =0.01). ( c ) MRD assessment in paired peripheral blood and bone marrow CLL cells as determined by the CD160FCA ( n =47), irrespective of the treatment regimen (Spearman Rank R =0.87, P <0.0001).

Article Snippet: The CD160FCA incorporates CD2-FITC (Clone S5.2), CD5-APC (Clone L17F12), CD19-PerCP cy5.5 (Clone SJ25C1), CD23-APC (Clone EBVCS-5) and CD45 V500 (Clone Hi30) (BD Biosciences, Oxford, UK); CD160-PE (Clone BY55; IgM isotype; Immunotech, Beckman Coulter, Marseilles, France).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Blood Cancer Journal

Article Title: Minimal residual disease detection with tumor-specific CD160 correlates with event-free survival in chronic lymphocytic leukemia

doi: 10.1038/bcj.2014.92

Figure Lengend Snippet: MRD detection by CD160FCA vs International Standardized Protocols in the validation cohort. ( a ) Comparative assessment of CD160FCA MRD positivity as a percentage of total leucocytes against the ISA methodology ( n =48 samples, P <0.001, R =0.91). ( b ) MRD-positive and -negative cohorts based on the median fluorescence intensity (MFI) of the CD2 neg /CD5 + /CD19 + /CD23 + /CD160 + population.

Article Snippet: The CD160FCA incorporates CD2-FITC (Clone S5.2), CD5-APC (Clone L17F12), CD19-PerCP cy5.5 (Clone SJ25C1), CD23-APC (Clone EBVCS-5) and CD45 V500 (Clone Hi30) (BD Biosciences, Oxford, UK); CD160-PE (Clone BY55; IgM isotype; Immunotech, Beckman Coulter, Marseilles, France).

Techniques: Biomarker Discovery, Fluorescence

Journal: European journal of immunology

Article Title: Mycobacterium tuberculosis -specific CD8 + T cells are functionally and phenotypically different between latent infection and active disease

doi: 10.1002/eji.201243262

Figure Lengend Snippet: T-cell differentiation and exhaustion of Mtb-specific CD8+ T cells in LTBI subjects and TB patients. (A) Representative flow cytometry examples and (B) cumulative analyses of the expression of CCR7 and CD45RA on Mtb-specific CD8+ T cells from LTBI subjects and TB patients are shown. CD8+ T cells were gated as shown in Supporting Information Fig. 1A. As in all other flow cytometry analyses from this study, the gating is based on the distribution of the different markers on bulk CD8+ T cells (A; top) and it is then conserved in the analyses of Mtb-specific CD8+ T cells (A; bottom). (C) Representative flow cytometry examples and (D) cumulative analyses of the expression of PD-1, 2B4, and CD160 on Mtb-specific CD8+ T cells from LTBI subjects and TB patients are shown. (A, C) Flow cytometry profiles are gated on live CD3+ CD4− CD8+ T cells and Mtb-specific CD8+ T-cell responses were defined as IFN-γ-producing cells following stimulation with ESAT-6 and/or CFP-10 peptide pools. (B, D) For cumulative analyses, all the possible combinations of the different markers are shown on the x-axis whereas the percentages of the distinct T-cell subsets within Mtb-specific CD8+ T cells are shown on the y-axis. The pie charts summarize the data, and each slice corresponds to the mean proportion of Mtb-specific CD8+ T cells positive for a certain combination of markers. (B, D) Comparisons of markers distribution were performed using a Student’s t-test and a partial permutation test as described [50].

Article Snippet: Flow cytometry analyses The following antihuman monoclonal antibodies were used in various combinations: CD3, CD4, CD8, IFN-γ, TNF-α, IL-2, granzyme B, PD-1, and CD45RA were purchased from BD; granzyme A and CD160 from Lucerna Chem; CD4 and 2B4 from BioLegend; granulysin from Bender MedSystems GmbH; Perforin from Biotest AG and CCR7 from R&D System.

Techniques: Cell Differentiation, Flow Cytometry, Expressing