Daudi cell stroma: An alternative to dithiothreitol to resolve daratumumab interference in pretransfusion testing

Transfusion ◽  
2020 ◽  
Author(s):  
Tony Tremblay ◽  
Donald R. Branch ◽  
Lionel Loubaki
Keyword(s):  
Daudi Cell

scholarly journals Association of the H-Y male antigen with beta2-microglobulin on human lymphoid and differentiated mouse teratocarcinoma cell lines.

1978 ◽  
Vol 148 (1) ◽  
pp. 58-70 ◽  
Author(s):  
M Fellous ◽  
E Günther ◽  
R Kemler ◽  
J Wiels ◽  
R Berger ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Hla Antigens ◽  
Lymphoid Cell Line ◽  
Hla Antigen ◽  
Teratocarcinoma Cell ◽  
Daudi Cell ◽  
Human Male ◽  
Lymphoid Lines ◽  
Mouse Teratocarcinoma

The expression of the H-Y antigen has been tested on several human lymphoid lines and mouse teratocarcinoma cell lines during differentiation. The human male lymphoid cell line Raji is a very useful target for studies of the H-Y antigen by lymphocytotoxicity test with rat anti-H-Y sera. With a few exceptions, all cells carrying the Y chromosome were H-Y positive. One of the exceptions is the human Daudi cell line which, besides lacking H-Y antigen, also lacks beta2-microglobulin. We have studied a possible association between the H-Y antigen, beta2-microglobulin, and HLA antigen with redistribution experiments. The results strongly suggest that H-Y antigen is not associated with HLA antigens but with beta2-microglobulin.

Targeting xIAP Is More Important Than Bcl-Xl in Lymphomas with Constitutive Activation of NF-kB.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2390-2390
Author(s):  
Yanjuan He ◽  
Joan Cain ◽  
Lee Ratner ◽  
Leon Bernal-Mizrachi
Keyword(s):  
Cell Lines ◽  
Annexin V ◽  
Fold Increase ◽  
Daudi Cell ◽  
Xiap Expression ◽  
Apoptotic Effect ◽  
Effective Doses ◽  
Induction Of Apoptosis ◽  
Apoptotic Pathways

Abstract Pathways resulting in resistance to apoptosis are essential to the process of lymphomagenesis. One such pathway, the nuclear factor-kB (NFkB), has been shown to be a key element in coordinating the anti-apoptotic effect of these malignancies. However the mechanisms used by which NFkB prevents apoptosis are not well understood. It has been suggested that NFkB inhibits activation of the intrinsic, extrinsic and common apoptotic pathways. Previous work in our lab using two different virally mediated lymphoma models (Tax/HTLV1 and LMP1/EBV driven tumors) has identified two candidates that could explain these results: X chromosome-linked inhibitor of apoptosis (xIAP) and BCL-xL. Although the current literature extensively demonstrates the role of BCL-xL in lymphomas, little is known about the importance of xIAP in these malignancies. To answer this question we tested the apoptotic effect of etoposide or tumor necrosis factor (TNF) after knocking down bcl-xL and xIAP expression in our lymphoma models (SC and Daudi cell lines) using a lentivirus expressing siRNAs. After 24 hours of treatment with etoposide and TNF, we measured apoptosis by flow cytometry using double staining with Annexin V-Alexa Fluorescense and propidium iodide. Interestingly, xIAP siRNA-expressing cell lines demonstrated 2–4 fold increase in the induction of apoptosis after treatment with etoposide as compared to a nearly 2 fold increase in those expressing Bcl-xL siRNA (see Table below). No synergism was seen after treatment with TNF. Based on this finding, we then tested a novel small molecule, homolog smac, (SHC, kindly provided by Dr. PG Harren) to determine the possible therapeutic effect of xIAP inhibitors. After titration, the two most effective doses were selected (25 μM and 50 μM) to treat Daudi cell lines for 24hrs, with either etoposide or TNF. At doses of 25 μM , we observed a 2 fold increase in the induction of apoptosis produced by etoposide compared to that seen in control (DMSO + etoposide) or SHC alone and no synergism with TNF confirming the siRNA data. More importantly, at doses of 50 μM, SHC alone demonstrated activity with a 5 fold increase in apoptosis and a nearly 10 fold increase as compared to control (DMSO) when etoposide was added. Overall, we have demonstrated that xIAP and bcl-xL are important in mediating NFkB-resistance to apoptosis. However, our findings suggested that xIAP is a more potent anti-apoptotic signal and opens the door for further drug development aimed at testing xIAP-inhibitors in lymphomas. Induction of Apoptosis in xIAP or Bcl-xL siRNA expressing cell lines siRNA/Compound Etoposide TNF Untreated xIAP 43.1 ± 17.6 17.04 ± 1.4 14.3 ± 2 SC Bcl-xL 18.39± 3.7 9.4 ± 0.22 12.5 ± 2.7 Luc/DMSO 14.9 ± 1.8 14.4 ± 5.6 14.03 ± 1.25 xIAP 9.2 ± 3.2 4.7 ± 0.48 4.6 ± 0.44 Bcl-xL 8.9 ± 0.5 5.3 ± 1.7 4.16 ± 0.4 Daudi Luc/DMSO 5.49 ± 1.71 4.28 ± 0.5 6.2 ± 0.9 SHC 25 μM 20.07 ± 4.8 12.8 ± 3.9 12.1 ± 3.2 SHC 50 μM 47.7 ± 14.55 38.3 ± 0.99 32.7 ± 8.99

scholarly journals Cytotoxicity of a cell-reactive immunotoxin containing ricin A chain is potentiated by an anti-immunotoxin containing ricin B chain.

1984 ◽  
Vol 160 (1) ◽  
pp. 341-346 ◽  
Author(s):  
E S Vitetta ◽  
R J Fulton ◽  
J W Uhr
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Ricin A Chain ◽  
Daudi Cell ◽  
A Cell ◽  
A Chain ◽  
Ricin A

In vitro killing of the human Daudi cell line by either univalent [F(ab')] or divalent (IgG) forms of rabbit anti-human Ig (RAHIg) coupled to ricin A chain can be specifically potentiated by a "piggyback" treatment with ricin B chain coupled to goat anti-rabbit Ig (GARIg). When cells are treated with univalent immunotoxin (IT) [F(ab') RAHIg-A] and then cultured, IT can be detected on the cell surface for at least 5 h, since GARIg-B can still enhance killing at this time. These results provide a strategy for in vivo use of A chain- and B chain-containing IT.

hIgD promotes human Burkitt lymphoma Daudi cell proliferation by accelerated G1/S transition via IgD receptor activity

2016 ◽  
Vol 64 (4) ◽  
pp. 978-987 ◽  
Author(s):  
Xing Dai ◽  
YuJing Wu ◽  
XiaoYi Jia ◽  
Yan Chang ◽  
HuaXun Wu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Burkitt Lymphoma ◽  
Receptor Activity ◽  
Daudi Cell

Retargeting of Activated T Cells (ATC) with Gemtuzumab (anti-CD33) Ozogamicin x Anti-CD3 Bispecific Antibody (CD33-OBi) Enhances Cytotoxicity Directed at K562.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4094-4094
Author(s):  
Abhinav Deol ◽  
Archana Thakur ◽  
Lawrence G. Lum
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
Cell Line ◽  
K562 Cells ◽  
Resistant Cell ◽  
Leukemia Stem Cells ◽  
Activated T Cells ◽  
Daudi Cell ◽  
Specific Cytotoxicity ◽  
Daudi Cells

Abstract Abstract 4094 Poster Board III-1029 Background CD33 is expressed in about 90% of blasts in acute myeloid leukemia but the response rate of gemtuzumab ozogamicin (GO) is about 30%. GO is anti-CD33 hP67.6 linked to calicheamicin. The mechanism of resistance of blasts to GO is not clearly understood. Our previous studies show the anti-CD3 activated T cells (ATC) can be redirected with bispecific antibodies (BiAb) to Her2/neu, CD20, and EGFR. We produced CD33-OBi by chemically heteroconjugating GO with OKT3 (anti-CD3). We determined whether ATC armed with CD33-OBi can enhance the killing of a low expressing GO-resistant cell line K562. Methods ATC were produced by stimulating peripheral blood mononuclear cells (PBMC) with anti-CD3 in the presence of IL-2. K562 cell line, expressing 67% CD33 expression was used as a target given the relative resistance of this cell line to GO. Daudi cell line expressing no CD33 was used as control. Cytotoxicity to K562 and Daudi cell lines was performed using 51Cr labeled cells as targets with different concentrations of GO alone, CD33-OBi, ATC alone and CD33-OBi armed ATC. Armed ATC were washed free of arming BiAb unless otherwise indicated. Various arming doses of the CD33-OBi for ATC were also tested, ADCC with CD33-OBi utilizing fresh PBMC were tested. ADCC with GO was not tested as the mechanism of action for GO is through internalization and DNA damage from the cleaved calicheamicin. Results In antibody doses of 5, 50, 500, and 5000 ng/ml of GO or CD33-OBi alone, specific cytotoxicity was 5,7,14, and 14% for GO and was essentially null at all concentrations for CD33-OBi. At arming doses of 0.5, 5, 50 and 500 ng of CD33-OBi/106 ATC, the plateau of mean specific cytotoxicity directed at K562 cells was 38%, achieved between arming doses of 50 and 500 ng/106ATC at an effector to target ratio (E/T) of 25:1. Daudi CD33-negative control cells were not lysed by either GO or CD33-OBi alone. Subsequently, ATC from 3 normal donors (n=3) armed with CD33-OBi with 50 ng/106 cells at E/T of 25:1, 12.5:1, 6.25:1 and 3.125:1 showed % mean [±SEM] specific cytotoxicity of 34.3[±3.2], 28.5[±3.2], 22[±2.8] and 16.2[±2.1]%, respectively to K562. On the other hand, ATC alone at similar E/T showed % mean specific cytotoxicity of 25[±4.6], 16.3[±3.6], 10.3[±2.4] and 5.3[±1.3].There were significant differences between CD33-OBi-ATC and ATC with p values (Wilcoxan signed rank test) at E/T of 25:1, 12.5:1, 6.25:1 and 3.125:1 that were 0.043, 0.033, 0.014 and 0.012, respectively. There was no specific cytotoxicity directed at Daudi cells under similar conditions. ADCC was performed with CD33-OBi added to co-culture of PBMC and labeled K562 and Daudi cells for 24 hrs which showed background specific cytotoxicity. Conclusions ATC armed with CD33-OBi at 50 ng/106 cells showed enhanced cytotoxicity directed at K562 cells which historically is relatively resistant to GO. The unarmed ATC have been shown to have non MHC restricted cytotoxicity and arming them with the CD33-OBi retargets and enhances the cytotoxicity of ATC to targets that express CD33. Given that patients with CD33 positive blasts may contain a population of CD33 positive leukemia stem cells, this approach may provide a non-toxic strategy to target leukemia stem cells. Testing additional GO susceptible and resistant cell lines under conditions in combination with chemotherapy or hematopoietic stem cell transplant may provide proof of principle for developing this approach. Disclosures: Lum: Transtarget Corporation: Founder.

Cellular and Molecular Effects of NVP-BKM120 in Lymphoblastic Leukemia Cells

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2241-2241
Author(s):  
João Kleber Novais Pereira ◽  
João Agostinho Machado-Neto ◽  
Matheus Rodrigues Lopes ◽  
Fabiola Traina ◽  
Fernando Ferreira Costa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Lymphoblastic Leukemia ◽  
Cyclin B1 ◽  
Pyrimidine Derivative ◽  
Leukemia Cells ◽  
Dependent Manner ◽  
Daudi Cell ◽  
Induction Of Apoptosis ◽  
Orally Bioavailable ◽  
Dose Dependent

Abstract Background: PI3K/AKT/mTOR signaling controls most hallmarks of cancer. Constitutive activation of PIK3 pathway in T cells acute lymphoblastic leukemia (T-ALL) has been reported; in a mouse model, PI3K activation, together with MYC, cooperates in Burkitt lymphoma (BL) pathogenesis. NVP-BKM120 is an orally bioavailable 2,6-dimorpholine pyrimidine derivative, and considered a highly selective pan-class I PI3K inhibitor. In preclinical studies, it has shown efficacy in a variety of malignancies and is currently being investigated in phase I/II/III clinical testing, mainly for advanced solid tumors (clinicaltrials.gov). Aims: Here, we described the effects of the pan-PI3K/AKT/mTOR inhibitor NVP-BKM120 on T-ALL and BL cell lines. Methods: T-ALLcell lines, Jurkat and MOLT-4, and BL cell lines, NAMALWA and Daudi, were obtained from ATCC. NVP-BKM120 was kindly provided by Novartis, and was prepared as a 10mM stock solution in DMSO. Different concentrations of the drug were used as indicated, where cells treated only with DMSO served as control. Cell viability was measured by MTT. Colony formation was carried out in semisolid methyl cellulose medium. The induction of apoptosis was assessed by annexin-V-APC/PI and by caspase cleavage. The cell cycle was analyzed by a PI-staining method. Western blot analysis was performed by standard methods. Vital staining and flow cytometry analysis with acridine orange was performed for the detection and quantification of acidic vesicular organelles (AVOs). Comparisons between the two groups were performed by the t test. Pvalue <0.05 was considered statistically significant. Results: Cell viability decreased in a concentration-dependent manner, with an IC50 range of 7-8 μM and the clonogenic growth was significantly decreased at the concentration of 1μM and at 10µM the colony formation was completely inhibited in all cells tested. After 6 hours of NVP-BKM120 treatment, we observed an increase in apoptotic cells, as well as an increase in the cleavage of procaspase 3, 8 and 9 in Jurkat, MOLT-4 and NAMALWA cells. Compared with DMSO control, NVP-BKM120 does not have any effect during apoptosis induction in the Daudi cell line. NVP-BKM120 treatment also resulted in G2/M arrest, associated with a decrease in the G1population and a decrease in Cyclin B1 protein levels. Immunoblotting analysis of cells treated with the drug revealed decreased phosphorylation, in a dose-dependent manner, of AKT, P70SK6 and 4EBP1, with stable total proteins levels. Additionally, we observed a dose-dependent decrease in BAD phosphorylation, followed by an increase in BAX:BCL2 ratio. Quantification of AVOs showed a dose-dependent increase of AVOs in all cells tested, after NVP-BKM120 treatment. Conclusions: NVP-BKM120 induced apoptosis in a dose-dependent manner in Jurkat, MOLT-4 and NAMALWA cells, while effects of the drug in the Daudi cell line were mainly cytostatic. In those cells, the induction of apoptosis suggested that the death receptor and mitochondrial pathways were activated after drug treatment. In our study, we found that NVP-BKM120 decreased the phosphorylation levels of BAD, which is linked to a pro-apoptotic activity, and up-regulated the BAX:BCL2 ratio. These results are consistent with the activation of caspase-9 and 3, related to the mitochondrial apoptosis. The accumulation of leukemia cells in the G2/M phase of the cell cycle has been associated with enhanced apoptosis. Our results suggest that decreased Cyclin B1 protein expression might be the molecular mechanism through which NVP-BKM120 induces G2/M arrest. The effects of NVP-BKM120 on the PI3K pathway indicate that NVP-BKM120 treatment may overcome rapamycin-induced AKT activation. P70SK6 and 4EBP1 are the two best-characterized substrates of mTOR1. Hence, the decrease in the phosphorylation levels of P70SK6 and 4EBP1 results in impaired oncogenic protein synthesis. Moreover, P70SK6 is one of the kinases whose phosphorylation by mTOR1 results in opposing autophagy. Accordingly, NVP-BKM120 resulted in increased AVOs, which is a characteristic feature of cells engaged in autophagy. In summary, our present study establishes that NVP-BKM120 effectively presents an antitumor activity against T-ALL and BL cell lines. The reduction of proliferation is possibly by down-regulation of Cyclin B1 and the increased BAX:BCL2 ratio is one of the mechanisms involved in the induction of apoptosis. Disclosures Off Label Use: NVP-BKM120 is an orally bioavailable 2,6-dimorpholino pyrimidine derivative, and considered a highly selective pan-class I PI3K inhibitor. .

A protein from Daudi cell line conditioned medium induces ovarian dysgenesis inDrosophila melanogaster

1992 ◽  
Vol 21 (2) ◽  
pp. 77-89 ◽  
Author(s):  
Noëlle Richard-Mercier ◽  
Michèle Thomas-Orillard ◽  
Madeleine Morinière ◽  
Patrick Porcheron ◽  
Daniel Soyez ◽  
...  
Keyword(s):  
Cell Line ◽  
Conditioned Medium ◽  
Daudi Cell ◽  
Ovarian Dysgenesis
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