Journal: Med (New York, N.y.)

Article Title: Early T cell and binding antibody responses are associated with COVID-19 RNA vaccine efficacy onset

doi: 10.1016/j.medj.2021.04.003

Figure Lengend Snippet:

Article Snippet: No-GFP-CD16.NK-92 cell line (High affinity 176V) , ATCC , PTA-6967.

Techniques: Virus, Recombinant, Staining, Luminex, Neutralization, Generated, Software

Journal: Transplantation and cellular therapy

Article Title: NK Cell Adoptive Immunotherapy of Cancer: Evaluating Recognition Strategies and Overcoming Limitations

doi: 10.1016/j.bbmt.2020.09.030

Figure Lengend Snippet: Sample List of Active Trials Involving Allogeneic NK Cells

Article Snippet: Anderson Cancer Center; National Cancer Institute CB-derived NK cells Rituximab Stem cell transplant Recurrent or refractory B cell non-Hodgkin lymphoma NCT03027128 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Metastatic of locally advanced solid tumors NCT03068819 Washington University School of Medicine Cytokine-induced memory-like NK cells CD3 + T cells donor lymphocyte infusion Relapsed AML NCT03136406 NantKwest, Inc. NK92 cell line ALT-803 Other immune modulators Pancreatic cancer NCT03167164 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Merkel cell carcinoma NCT03167177 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Melanoma NCT03169738 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Non-small-cell lung cancer NCT03169764 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Head and neck squamous cell carcinoma NCT03169777 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Colorectal cancer NCT03169790 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Non-Hodgkin lymphoma NCT03175666 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Triple-negative breast cancer NCT03197571 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Urothelial carcinoma NCT03197584 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Ovarian cancer NCT03300492 University Hospital, Basel, Switzerland Haploidentical NK cells Acute myeloid leukemia; myelodysplastic syndromes NCT03329248 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Pancreatic cancer NCT03383978 Johann Wolfgang Goethe University Hospital NK92 cell line gene modified to express anti-Her2 CAR-expressing CD28 and ζ Glioblastoma NCT03387085 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Triple-negative breast cancer NCT03387098 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Pancreatic cancer NCT03387111 NantKwest, Inc. NK92 cell line modified to express IL-2 and CD16 (haNK) Other immune modulators Squamous cell carcinoma NCT03415100 The Third Affiliated Hospital of Guangzhou Medical University Autologous or allogeneic NK cells transfected with mRNA expressing NKG2D-directed CAR Metastatic solid tumors NCT03420963 M.D.

Techniques: Transplantation Assay, Modification, Transfection, Expressing, Derivative Assay

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: (A) Schematic representation of recombinant lentiviral vector encoding third generation CD4CAR, driven by spleen focus-forming virus (SFFV) promoter. The construct contains a leader sequence, anti-CD4 scFv, hinge domain (H), transmembrane (TM) and signaling domains CD28, 4-1BB, and CD3 zeta. (B) HEK293FT cells were transfected with GFP vector control (lane 1) and CD4CAR (lane 2) lentiviral plasmids. Forty-eight hours after transfection, cells were removed and subsequently used for Western blot analysis with mouse anti-human CD3z antibody. (C) Illustration of third-generation CAR NK-92 cells targeting CD4 expressing cells.

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Recombinant, Plasmid Preparation, Virus, Construct, Sequencing, Transfection, Control, Western Blot, Expressing

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: ( A , upper panel) CD4CAR expression levels on NK-92 cells prior to being sorted by FACS (N=3); ( A , lower panel) CD4CAR expression on NK-92 cells after sorting and expansion, prior to co-culture experiments (N=3).

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Expressing, Co-Culture Assay

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: Co-culture experiments were performed at an effector to target ratio of 2:1 for 24 hours and were directly analyzed by flow cytometry for CD56 and CD4 (panels A and B). Each assay consists of target cells alone control (left), and target cells incubated with NK-92 cells transduced with vector control (center) or CD4CAR (right) lentiviral supernatant. (A) Top row: KARPAS-299 (N=3). Middle row: HL-60 T-cells (N=2). Bottom row: CCRF-CEM cells (N=2). (B) CD4CAR NK-92 cells eliminated primary T-cell leukemia cells from a patient with CD4 + T-cell lymphoma/ Sézary syndrome (N=2) and CD4 expressing pediatric T-cell ALL (N=2). (C) Bar graph summarizing co-culture assay results for both 2:1 and 5:1 E:T ratios.

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Co-Culture Assay, Flow Cytometry, Control, Incubation, Transduction, Plasmid Preparation, Expressing, Co-culture Assay

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: (A) Co-culture assays were performed at an effector to target ratio of 2:1 for 24 hours, after which, cells were stained with mouse anti-human CD56 and CD4 antibodies. Target cells were incubated alone as a control (left). NK-92 cells were transduced with either vector control (center) or CD4CAR (right) lentiviral supernatant and incubated with CD4 + T-cells obtained from human cord blood. (N=2) (B) CD4CAR NK-92 cells were incubated at co-culture effector:target ratios of 2:1 and 5:1 respectively with 500 CD34+ cord blood cells for 24 hours in NK cell media supplemented with IL-2. Experimental controls used were CD34+ cells alone, and non-transduced NK-92 cells were co-cultured at respective 2:1 and 5:1 effector:target ratios with CD34+ CB cells. Hematopoietic compartment output was assessed via formation of erythroid burst-forming units (BFU-E) and number of granulocyte/monocyte colony-forming units (CFU-GM) at Day 16. CFU statistical analysis was performed via 2-way ANOVA with alpha set at 0.05.

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Co-Culture Assay, Staining, Incubation, Control, Transduction, Plasmid Preparation, Cell Culture

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: CD4CAR NK-92 cells lyse CD4-expressing leukemic cell lines in a dose-dependent and specific manner. CD4CAR NK and vector control cells were incubated with an equal ratio of CFSE-stained “on-target” (KARPAS-299 or CCRF-CEM) cells and CMTMR-stained “off target” MOLT4 cells at 0.25 to 1, 0.5 to 2, and 1 to 1 effector to target ratios. After 24 hours, 7-AAD dye was added and remaining live cells were analyzed by flow cytometry. Percent killing of target cells was measured by comparing CD4 + KARPAS-299 or CCRF-CEM cell survival in CD4CAR NK-92 cell co-cultures relative to survival in vector control NK-92 cell co-cultures.

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Expressing, Plasmid Preparation, Control, Incubation, Staining, Flow Cytometry

Journal: Oncotarget

Article Title: Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells

doi: 10.18632/oncotarget.22626

Figure Lengend Snippet: NSG mice were sublethally irradiated and intradermally injected with luciferase-expressing KARPAS-299 cells (Day 0) to induce measurable tumor formation. On day 1 and every 5 days for a total of 6 courses, mice were intravenously injected with 5 × 10 6 CD4CAR NK-92 cells or vector control NK-92 cells. (A) On days 7, 14, and 21, mice were injected subcutaneously with RediJect D-Luciferin and subjected to IVIS imaging. (B) Average light intensity measured for the CD4CAR NK-92 injected mice was compared to that of vector control NK-92 injected mice. (C) On day 1, and every other day after, tumor size area was measured and the average tumor size between the two groups was compared. (D) Percent survival of mice was measured and compared between the two groups.

Article Snippet: CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4 + human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo .

Techniques: Irradiation, Injection, Luciferase, Expressing, Plasmid Preparation, Control, Imaging