1. Endogenous Tumor Necrosis Factor as a Predictor of Doxorubicin Sensitivity in Leukemic Patients
by Daisuke Kobayashi, Naoki Watanabe, Naofumi Yamauchi, Naoki Tsuji, Tsutomu Sato, and Yoshiro Niitsu
Blood
Volume 89(7):
April 1, 1997
©1997 by American Society of Hematology

2. Linear representation of recombinant retrovirus vector used for antisense TNF expression. pRXhTNFanti vector was derived from pRX.
Linear representation of recombinant retrovirus vector used for antisense TNF expression. pRXhTNFanti vector was derived from pRX. Antisense TNF is encoded by the subgenomic mRNA expressed from the upstream CMVIEΔLTR. CMVIE, cytomegalovirus immediate early enhancer; LTR, long terminal repeat; SD, splice donor; SA, splice acceptor.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

3. Changes in TNF/DOX sensitivity and intracellular resistant factors in a patient (case 4) who developed resistance to DOX. Leukemic cells were obtained at the pretreatment points (January 18, 1995) and posttreatment points (March 30, 1995) of chemotherapy wi...
Changes in TNF/DOX sensitivity and intracellular resistant factors in a patient (case 4) who developed resistance to DOX. Leukemic cells were obtained at the pretreatment points (January 18, 1995) and posttreatment points (March 30, 1995) of chemotherapy with the regimen which includes DOX, and used for each assay as described in Materials and Methods. enTNF expression is represented as the relative value compared with that of KG-1 cells. The results shown are the mean (±SD) of three independent experiments. *P < .01; **P < .02; ***P < .05 by Student's t-test.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

4. Expression of p-glycoprotein in case 4.
Expression of p-glycoprotein in case 4. Immunofluorescent flow cytometric analysis of p-glycoprotein on leukemic cells from the patient (case 4) was performed. Cells were stained with anti–p-glycoprotein monoclonal antibody JSB-1 ( — ) followed by PE-conjugated goat anti-mouse IgG as a secondary antibody. As a negative control ( ), only secondary antibody was used.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

5. Analysis of MRP gene expression in case 4.
Analysis of MRP gene expression in case 4. DNA molecular size marker (lane 1); MRP gene expression in cells from time points indicated in Fig 2 (lanes 2 and 3); β-actin band in each point (lanes 4 and 5). RT-PCR was performed as described in Materials and Methods.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

6. Correlation between enTNF expression and chemosensitivity.
Correlation between enTNF expression and chemosensitivity. Values of enTNF expression for 14 cases treated with anthracyclins and treatment outcome are plotted. enTNF expression is represented as the relative value compared with that of KG-1 cells. NR, no response; PR, partial remission; CR, complete remission.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

7. enTNF expression in KG-1 cells, pSV2neo transfected clones, and pTNFΔpro transfected clones. enTNF expression is represented as the relative value compared with that of KG-1 cells.
enTNF expression in KG-1 cells, pSV2neo transfected clones, and pTNFΔpro transfected clones. enTNF expression is represented as the relative value compared with that of KG-1 cells. The data represent the mean (±SD) of three independent experiments. KP1 and KP2 cells were transfected with pSV2neo alone and KD1-KD11 cells were cotransfected with pTNFΔpro and pSV2neo.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

8. Analysis of TNF gene expression.
Analysis of TNF gene expression. KG-1 cells (lane 1); KG-1 (pSV2neo) KP2 cells (lane 2); KG-1 (pTNFΔpro) KD3 cells (lane 3); KG-1 (pTNFΔpro) KD5 cells (lane 4); KG-1 (pTNFΔpro) KD10 cells (lane 5). RT-PCR was performed as described in Materials and Methods.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology

9. Reversal of DOX resistance by transfection of antisense TNF cDNA with retroviral vector pRXhTNFanti into leukemic cells.
Reversal of DOX resistance by transfection of antisense TNF cDNA with retroviral vector pRXhTNFanti into leukemic cells. The leukemic cells obtained freshly from five patients were used (cases 2, 5, 6, 10, and 11) for the cytotoxic assay as described in Materials and Methods. The data represent the mean (±SD) of three independent experiments and statistical significance was determined by Student's t-test. *P < .01; **P < .02; ***P < .05.
Daisuke Kobayashi et al. Blood 1997;89:
©1997 by American Society of Hematology