Placenta Derived Adherent Cells (PDACs) Suppress Tumor Cells of Diverse Origin.

Abstract The placenta is a readily available and ethically non-controversial source of large amounts of therapeutic stem cells. We isolated adherent cells from the Umbilical Cord (UC) and the Amnion Chorion (AC) of term placentas. These Placenta Derived Adherent Cells (PDACs) displayed a cell surface phenotype of CD29+, CD44+, CD73+, CD90+, CD105+, CD200+, CD34−, and constitutively secreted IL-6, IL-8 and Monocyte Chemoattractant protein-1 (MCP-1). PDACs demonstrated in vitro pluripotency and suppressed T cell proliferation in both the allogeneic mixed leukocyte reaction, and the autologous EBV regression assay. Because progenitor cells have been described to inhibit tumor growth in some systems, the role of PDACs in tumor suppression was investigated. EBV transformed tumor cells were cultured either alone, or with AC or UC PDACs. After 6 days, live (7-AAD− Annexin-V−) tumor cells were counted on a flow cytometer. Free growing tumor cells numbered 40,000. When tumor cells were co-cultured with AC or UC PDACs, the growth was suppressed to 10,000 cells, a suppression of 75%. We designed and tested a panel of tumor cell lines that included retinoblastoma (RB), histiocytic lymphoma (HL), chronic myelogenous leukemia (CML), and colon adenocarcinoma (CAC). The tissue origin of the selected tumor cell lines included neuronal tissue (retinoblastoma), epithelium (carcinomas) and the hematopoietic lineage (lymphomas and CML). Co-culture experiments showed that the growth of all tumor lines was suppressed more than 40%. When a transwell was introduced separating PDACs and tumor cells, contact dependency of suppression was 12% for HL, 22% for CML, 42% for CAC, and 51% for EBV transformed tumor cells. These results indicate that PDACs may have therapeutic value with respect to certain cancers.

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Synthesis and Characterization of New Palladium(II) Complexes with Ligands Derived from Furan-2-carbaldehyde and Benzaldehyde Thiosemicarbazone and their in vitro Cytotoxic Activities against Various Human Tumor Cell Lines

Zeitschrift für Naturforschung B ◽

10.1515/znb-2010-1015 ◽

2010 ◽

Vol 65 (10) ◽

pp. 1271-1278 ◽

Cited By ~ 4

Author(s):

Wilfredo Hernández ◽

Juan Paz ◽

Fernando Carrasco ◽

Abraham Vaisberg ◽

Jorge Manzur ◽

...

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Human Tumor ◽

Myelogenous Leukemia ◽

Cytotoxic Activities ◽

Spectroscopic Techniques ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines

With the ligands 4-phenyl-1-(furan-2-carbaldehyde)thiosemicarbazone, HTSC1, (1), 4-phenyl-1- (5´-phenyl-furan-2-carbaldehyde)thiosemicarbazone, HTSC2 (2), o-methoxy-benzaldehydethiosemicarbazone, HTSC3 (3), and o-cyano-benzaldehydethiosemicarbazone, HTSC4 (4), the corresponding palladium(II) complexes, Pd(TSC1)2 (5), Pd(TSC2)2 (6), Pd(TSC3)2 (7), and Pd(TSC4)2 (8) were synthesized and characterized by elemental analysis and spectroscopic techniques. The crystal structure of Pd(TSC3)2 (7) was determined by single-crystal X-ray diffraction. Complex 7 shows a squareplanar geometry, where two deprotonated ligands are coordinated to the PdII center through the nitrogen and sulfur atoms in a trans arrangement. In vitro antitumor studies against different human tumor cell lines have revealed that the palladium(II) complexes 5- 8 are more cytotoxic (IC50 values in the range of 0.21 - 3.79 μM) than their corresponding ligands (1 - 4) (> 60 μM). These results indicate that the antiproliferative activity is enhanced when thiosemicarbazone ligands are coordinated to the metal. Among the studied palladium(II) complexes, 8 exhibits high antitumor activity on K562 chronic myelogenous leukemia cells with a low value of the inhibitory concentration (IC50 = 0.21 μM).

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Breast Tumor Cell Lines From Pleural Effusions2

JNCI Journal of the National Cancer Institute ◽

10.1093/jnci/53.3.661 ◽

1974 ◽

Vol 53 (3) ◽

pp. 661-674 ◽

Cited By ~ 562

Author(s):

R. Cailleau ◽

R. Young ◽

M. Olivé ◽

W. J. Reeves

Keyword(s):

Chromosome Number ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Breast Tumor ◽

Tumor Cell Lines ◽

Breast Cancer Patients ◽

Pleural Effusions ◽

Epithelial Tumor

Summary During 1973, 4 new epithelial tumor cell lines were isolated from pleural effusions from breast cancer patients. We describe 3 of these lines: MDA-MB-134, with a mean chromosome number of 43; MDA-MB-175, with a mean chromosome number of 49; and MDA-MB-231, with a mean chromosome number between 65 and 69. We isolated the same cell type from 4 of 10 effusions from MDA-MB-134 and from 6 of 8 effusions from MDA-MB-175. We found that pleural effusions as a source of breast tumor cells to be cultured and studied in vitro have the following advantages: 1) large amounts of material and the possibility of obtaining sequential samples from the same patient; 2) high viability of tumor cells; 3) scarcity or absence of fibroblasts; and 4) the possibility of separating the tumor cells from other “contaminating” cell types by differences in their speed or degree of attachment to the flask. All lines from different patients differed, as seen grossly and microscopically. All lines from sequential pleural effusions from the same patient were apparently alike. No viruses or mycoplasmas were detected in any line.

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Antiproliferative and Antitumor Activity of the 2-Cyanoaziridine Compound Imexon on Tumor Cell Lines and Fresh Tumor Cells In Vitro

JNCI Journal of the National Cancer Institute ◽

10.1093/jnci/84.16.1238 ◽

1992 ◽

Vol 84 (16) ◽

pp. 1238-1244 ◽

Cited By ~ 20

Author(s):

E. M. Hersh ◽

C. R. Gschwind ◽

C. W. Taylor ◽

R. T. Dorr ◽

R. Taetle ◽

...

Keyword(s):

Antitumor Activity ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Tumor Cell Lines

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High affinity Fc receptor binding and potent induction of antibody-dependent cellular cytotoxicity (ADCC) in vitro by anti-epidermal growth factor receptor antibody cetuximab

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.3041 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 3041-3041 ◽

Cited By ~ 4

Author(s):

X. Kang ◽

D. Patel ◽

S. Ng ◽

M. Melchior ◽

D. Ludwig ◽

...

Keyword(s):

Tumor Cell ◽

Nk Cells ◽

Tumor Cells ◽

Cell Lines ◽

Growth Factor Receptor ◽

Tumor Cell Lines ◽

Antibody Dependent Cellular Cytotoxicity ◽

Effector Cells ◽

Epidermal Growth

3041 Background: Cetuximab is an IgG1 monoclonal antibody specific for human epidermal growth factor receptor (EGFR). Cetuximab acts as a functional antagonist by blocking ligand (EGF and TGFa) binding to EGFR and, therefore, inhibits EGFR activation and downstream signaling in tumor cells. Methods: In the present study, we analyzed the binding activity of cetuximab to human Fc receptors and tested whether cetuximab can elicit antibody-dependent cellular cytotoxicity (ADCC) in vitro using tumor cell lines expressing various levels of EGFR. Results: Cetuximab bound to human FcRI and FcRIII with high affinity (EC50 of 0.13 nM and 6 nM, respectively), whereas an aglycosylated form of cetuximab or an IgG2 antibody to EGFR (panitumumab) did not bind to FcRI and FcRIII. A panel of head and neck, colon, and breast tumor cell lines were tested for cetuximab-mediated ADCC using human peripheral blood mononuclear cells (PBMC) from normal donors as effector cells. We found that cetuximab-induced ADCC to these tumor cell lines ranging from 20–90% specific killing. Interestingly, the extent of ADCC induced by cetuximab correlated with the level of EGFR expression on cell surface. The aglycosylated form of cetuximab or panitumumab, did not elicit ADCC of EGFR expressing tumor cells, even though cetuximab and panitumumab bind to EGFR with similar affinity (Kd=87 pM and 83 pM, respectively). To identify the immune cell population responsible for ADCC, NK cells were isolated from PBMC and tested in an ADCC assay. Purified NK cells elicited high levels of specific killing of cetuximab bound tumor cells, suggesting that NK cells are one of effector cell populations in human PBMCs responsible for the observed ADCC activity. Conclusions: Thus, cetuximab can effectively link effector cells to EGFR expressing tumor cells and mediate potent ADCC against EGFR-expressing human tumors. [Table: see text]

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Human Placental CD34 +-Derived Natural Killer Cells with High Affinity and Cleavage Resistant CD16 (CYNK-101) in Combination with Daratumumab for Immunotherapy Against CD38 Expressing Hematological Malignancies

Blood ◽

10.1182/blood-2021-148869 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 2773-2773

Author(s):

Irene Raitman ◽

Joseph Gleason ◽

Salvatore Rotondo ◽

Shuyang He ◽

Valentina Rousseva ◽

...

Keyword(s):

B Cells ◽

Tumor Cell ◽

Nk Cells ◽

Tumor Cells ◽

Cell Lines ◽

Xenograft Model ◽

Tumor Cell Lines ◽

Publicly Traded

Abstract Natural Killer (NK) cells are key mediators of antibody dependent cellular cytotoxicity (ADCC) via the CD16 Fc receptor. NK cellular therapies can effectively be targeted to tumor antigens when combined with tumor specific antibodies. Celularity Inc. is developing human placental CD34 +-derived, cryopreserved, off-the-shelf, allogenic NK cells (CYNK-101) with a high IgG binding affinity and proteinase cleavage resistant CD16 variant (CD16VP) for cancer treatment. We hypothesize that expressing CD16VP augments anti-tumor ADCC activity. Reported here are the in vitro and in vivo results of evaluating CYNK-101 cytotoxicity against CD38 expressing multiple myeloma (MM) and lymphoma tumor cell lines when in combination with daratumumab, an anti-CD38 monoclonal antibody. Human placental CD34 + cells were transduced with lentivirus expressing CD16VP and cultured in the presence of cytokines to generate CYNK-101 cells. The in vitro cytotoxic activity of CYNK-101 against CD38 + MM (MOLP-8, LP-1, MM.1S) and lymphoma (Daudi) tumor cell lines, and normal B-cells was assessed in combination with daratumumab via flow cytometry based ADCC assays and cytokine secretion was assessed via multiplex Luminex analysis. In vivo ADCC activity of CYNK-101 was assessed using a disseminated B-cell lymphoma xenograft model in B-NDG-hIL15 mice. B-NDG-hIL15 mice lack T, B, and NK cells and are transgenic for human IL-15 to support CYNK-101 persistence and maturation. Luciferase expressing Daudi cells (3×10 6) were intravenously (IV) injected on Day 0 three days after preconditioning with a myeloablative dose of busulfan to allow for better tumor cell engraftment. CYNK-101 cells (1x10 7) and/or daratumumab (0.05 mg/kg) were IV injected on Days 7, 14 and 21. Tumor burden was assessed weekly by bioluminescence imaging (BLI) and the mice were followed for assessment of their survival (n=5 mice per group). In vitro ADCC studies indicate enhanced cytolysis of CYNK-101 in combination with daratumumab against both MM and lymphoma tumor cells compared to that of IgG control. At 24h at the effector to target (E:T) ratio of 5:1, CYNK-101 (n=5 donors) demonstrated a cytolysis of 87.6 ± 6.3% with daratumumab vs. 37.3 ± 9.5% with IgG control against MOLP-8 (p<0.001), 73.9 ± 2.5% vs. 32.1 ± 7.2% against LP-1 (p<0.001), 77.2 ± 11.5% vs. 67.4 ± 10.7% against MM.1S (p<0.001), and 54.7 ± 24.0% vs. 4.3 ± 2.6% against Daudi (p<0.01) tumor cells. Secretion of GM-CSF, IFN-γ, and TNF-α was increased in CYNK-101 co-cultured with the tumor cell lines in the presence of daratumumab for 24h (n=5 donors, p<0.05). When cocultured with mixed LP-1 and CD38 + normal B-cells, CYNK-101 in combination with daratumumab displayed specific cytotoxicity against LP-1, while sparing CD38 + normal B-cells even at an E:T ratio up to 100:1, demonstrating that CYNK-101 can distinguish CD38 + tumor cells from CD38 + normal cells. Additionally, despite expression of CD38 on CYNK-101 there was no NK fratricide observed when CYNK-101 were in combination with daratumumab. In vivo studies in the lymphoma xenograft model revealed a significant decrease in tumor burden as evidenced from bioluminescence imaging at day 28 (1 week after last CYNK-101 injection) for mice that received CYNK-101 in combination with daratumumab compared to vehicle control (p<0.001), CYNK-101 (p<0.05) and daratumumab (p<0.05). Furthermore, CYNK-101 in combination with daratumumab demonstrated an enhanced survival benefit with a median survival of 35 days versus a median survival of 28 days for the vehicle treated group (p<0.005). In summary, our results demonstrate enhanced in vitro and in vivo ADCC activities of CYNK-101 in combination with daratumumab against CD38 + hematological tumors and warrant further development of this combination therapy for these cancers. Disclosures Raitman: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Gleason: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Rotondo: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. He: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Rousseva: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Guo: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Rana: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. van der Touw: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Ye: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Kang: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Hariri: Celularity Inc.: Current Employment, Current equity holder in publicly-traded company. Zhang: Celularity Inc.: Current equity holder in publicly-traded company, Ended employment in the past 24 months.

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Nanobody Armed T Cells Endow CAR-T Cells the Ability to Against Lymphoma Cells

10.21203/rs.3.rs-546282/v1 ◽

2021 ◽

Author(s):

Hongxia Wang ◽

Liyan Wang ◽

Yanning Li ◽

Guangqi Li ◽

Xiaochun Zhang ◽

...

Keyword(s):

T Cells ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Phage Display Library ◽

Target Cells ◽

Tumor Cell Lines ◽

Car T Cells ◽

Car T

Abstract BackgroundTaking advantages of nanobody (Nb) in immunotherapy, here we investigate the cytotoxicity of Nb based Chimeric antigen receptor T cells (Nb CAR-T) against Lymphoma cells.MethodsCD19 Nb CAR-T, CD20 Nb CAR-T, and Bispecific Nb CAR-T cells were generated by panning anti-human CD19, CD20 specific nanobodies sequences from naive phage display library, then integrating Nb genes with lentiviral cassette that included other CARs elements, and finally transducing T cells that were expanded under optimization system with above prepared CARs lentiviruses. Prepared Nb CAR-T cells were co-cultured with tumor cell lines or primary tumor cells for 24 hours or 5 days to evaluate the biological function. ResultsObtained several Nb sequences specific to CD19 and CD20. Optimized culture conditions of T cells that expand 87.5 folds after 7 days of activation. Generated Nb CAR-T cells that could recognize Burkitt lymphoma cell lines (Raji and Daudi), induce activation, proliferation, and therefore kill target cells specifically. Furthermore, same results were also obtained from patient samples with cytotoxicity about 60%. ConclusionsOur study demonstrated that nanobody based single and bispecific CAR-T cells have certain killing ability against both tumor cell lines and patient-derived tumor cells in vitro.

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Function and Therapeutic Potential of In Vitro-Expanded Natural Killer T (NKT) Cells from Patients with Hematopoietic Malignancies.

Blood ◽

10.1182/blood.v110.11.2742.2742 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2742-2742 ◽

Cited By ~ 1

Author(s):

Paul R. Rogers ◽

Melissa Emerson ◽

Esme Nguyen ◽

Edward Ball ◽

Asad Bashey ◽

...

Keyword(s):

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Nkt Cells ◽

Inkt Cell ◽

Tumor Cell Lines ◽

Inkt Cells ◽

Nkt Cell ◽

Hematopoietic Malignancies

Abstract Injection of glycolipid KRN7000 (α-galactosylceramide) into mice induces activation of a specific subpopulation of T cells called invariant NKT (iNKT) cells which are important for tumor cell eradication. NKT cell activation is antigen-specific and dependent on presentation of agonist by the non-polymorphic class I-like molecule CD1d on antigen-presenting cells. In contrast to mouse, injection of KRN7000 into cancer patients did not induce any clinical responses in a published phase I study. This lack of response in part reflects the very low frequency of iNKT (Vα24/Vβ11 T cell receptor) cells in humans and especially in some cancer patients. Therefore, therapies which can increase iNKT cell numbers in patients may enhance the chances of tumor cell killing. Adoptive transfer of in vitro-expanded iNKT cells in combination with KRN7000 may be one therapeutic option. Our group and others have shown that CD1d-positive tumor cell lines are very sensitive to killing by human NKT cells. In contrast, killing of CD1d-negative tumor cells is not directly mediated by NKT cells but can be influenced through cytokines secreted by NKT cells. Based on these results and the reduced frequency of iNKT cells in patients, we hypothesized that NKT cell adoptive therapy could be most beneficial for treatment of patients with CD1d-positive cancers. Previous reports have shown that multiple myeloma (MM), acute myeloid leukemia (AML)-M4, and T-acute lymphoblastic leukemia (ALL) tumor cells express CD1d and may be susceptible to NKT cell killing in vitro. In this proof-of-concept study, we analyzed iNKT cell frequency, expansion, and function from six patients with hematopoietic malignancies (MM and AML). Although the frequency of iNKT cells in patients was extremely low (<0.01% in 5 of 6 patients), in vitro culture of PBMCs with KRN7000 and rhIL-2 induced tremendous NKT cell expansion. After the initial 12–14 day culture, iNKT cells expanded from 300-fold to greater than 2000-fold in 5 of 6 patient samples. Purified iNKT cell lines were generated from 5 of 6 patients and then restimulated every 8–12 days. Within one month, it was possible to obtain more than 100 million NKT cells (4 of 6 donors) from a starting culture of 10 million PBMCs (∼100–6,000 NKT cells). The sorted and expanded iNKT cell lines were then tested for cytokine secretion and cytotoxic activity. Upon activation, patient iNKT cell lines produced relatively normal levels of IFN-γ, IL-4, IL-5, and GM-CSF but slightly reduced amounts of IL-2 and TNFα compared to normal donor iNKT lines. Importantly, patient iNKT cell lines showed potent and dose-dependent killing activity against B, T and monocytic tumor cell lines in a 4h chromium release assay. The cytotoxic activity was both CD1d-restricted and antigen-specific. Near maximal killing of tumor cells was seen at an effector to target cell ratio of 5:1, but killing was seen even at 1:1 (E:T). Three of four patient iNKT cell lines showed strong killing of Jurkat (57–66% killing), MOLT-4 (36–57% killing), CEM (30–46%killing), U-937 (47–49% killing), and CD1d-transfected B cell (54–78% killing) targets at a 5:1 NKT:tumor cell ratio. Based on this data and results of the preliminary toxicology study in non-human primate, we believe that adoptive transfer of expanded iNKT cells plus KRN7000 is safe and may be particularly effective against CD1d-positive tumor cells in vivo.

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Interaction of Human Tumor Cells with Human Platelets and the Coagulation System

Thrombosis and Haemostasis ◽

10.1055/s-0038-1665296 ◽

1983 ◽

Vol 50 (03) ◽

pp. 726-730 ◽

Cited By ~ 11

Author(s):

Hamid Al-Mondhiry ◽

Virginia McGarvey ◽

Kim Leitzel

Keyword(s):

Tumor Cell ◽

Tumor Cells ◽

Cell Lines ◽

Human Tumor ◽

Human Platelets ◽

Factor Vii ◽

Coagulation System ◽

Breast Adenocarcinoma ◽

Activate Factor ◽

Tumor Cell Lines

SummaryThis paper reports studies on the interaction between human platelets, the plasma coagulation system, and two human tumor cell lines grown in tissue culture: Melanoma and breast adenocarcinoma. The interaction was monitored through the use of 125I- labelled fibrinogen, which measures both thrombin activity generated by cell-plasma interaction and fibrin/fibrinogen binding to platelets and tumor cells. Each tumor cell line activates both the platelets and the coagulation system simultaneously resulting in the generation of thrombin or thrombin-like activity. The melanoma cells activate the coagulation system through “the extrinsic pathway” with a tissue factor-like effect on factor VII, but the breast tumor seems to activate factor X directly. Both tumor cell lines activate platelets to “make available” a platelet- derived procoagulant material necessary for the conversion of prothrombin to thrombin. The tumor-derived procoagulant activity and the platelet aggregating potential of cells do not seem to be inter-related, and they are not specific to malignant cells.

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Synthesis and Cytotoxicity of New Mannich Bases of 6-[3-(3,4,5-Trimetoxyphenyl)-2- propenoyl]-2(3H)-Benzoxazolone

Letters in Drug Design & Discovery ◽

10.2174/1570180816666190730164952 ◽

2020 ◽

Vol 17 (4) ◽

pp. 512-517

Author(s):

Ognyan Ivanov Petrov ◽

Yordanka Borisova Ivanova ◽

Mariana Stefanova Gerova ◽

Georgi Tsvetanov Momekov

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Biological Activities ◽

Human Tumor ◽

Mannich Bases ◽

In Vitro Cytotoxicity ◽

Tumor Cell Lines ◽

Human Tumor Cell ◽

Human Tumor Cell Lines

Background: Chemotherapy is one of the mainstays of cancer treatment, despite the serious side effects of the clinically available anticancer drugs. In recent years increasing attention has been directed towards novel agents with improved efficacy and selectivity. Compounds with chalcone backbone have been reported to possess various biological activities such as anticancer, antimicrobial, anti-inflammatory, analgesic, antioxidant, etc. It was reported that aminomethylation of hydroxy chalcones to the corresponding Mannich bases increased their cytotoxicity. In this context, our interest has been focused on the design and synthesis of the so-called multi-target molecules, containing two or more pharmacophore fragments. Methods: A series of Mannich bases were synthesized by the reaction between 6-[3-(3,4,5- trimethoxyphenyl)-2-propenoyl]-2(3Н)-benzoxazolone, formaldehyde, and a secondary amine. The structures of the compounds were confirmed by elemental analysis, IR and NMR spectra. The new Mannich bases were evaluated for their in vitro cytotoxicity against a panel of human tumor cell lines, including BV-173, SKW-3, K-562, HL-60, HD-MY-Z and MDA-MB-231. The effects of selected compounds on the cellular levels of glutathione (GSH) were determined. Results: The new compounds 4a-e exhibited concentration-dependent cytotoxic effects at micromolar concentrations in MTT-dye reduction assay against a panel of human tumor cell lines, similar to those of starting chalcone 3. The tested agents led to concentration - dependent depletion of cellular GSH levels, whereby the effects of the chalcone prototype 3 and its Mannich base-derivatives were comparable. Conclusion: The highest chemosensitivity to the tested compounds was observed in BV- 173followed by SKW-3 and HL-60 cell lines.

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