Leukemia Specific Complement Dependent Cytotoxicity Contributes to Anti-Leukemia Immunity Induced by Leukemia Cell-Derived HSP70 Immunization in Mice.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3910-3910
Author(s):  
Junko Jimbo ◽  
Kazuya Sato ◽  
Takaaki Hosoki ◽  
Motohiro Shindo ◽  
Katsuya Ikuta ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Target Cells ◽  
Antigenic Peptides ◽  
Humoral Immune ◽  
Complement Dependent Cytotoxicity ◽  
Igg Level

Abstract Background: Tumor-derived heat shock proteins (HSPs) can be used for the vaccination against cancer because tumor-derived HSPs bind the tumor specific antigenic peptides and these peptides are carried onto MHC molecules by HSPs. We previously reported that immunotherapy using leukemia cell-derived HSPs against minimal residual leukemia after syngeneic bone marrow transplantation in mice prolonged survival by inducing leukemia-specific CD8 + cytotoxic T lymphocyte (Sato et al. Blood, 2001). It is known that induction of cell-mediated immune response plays a main role of tumor rejection in the HSP-based immunotherapy. However we also showed that CD4+ as well as CD8+ T cell is necessary for survival prolongation of HSP-immunized mice, suggesting that humoral immune response though CD4+ T cell contributes to the eradication of leukemia cells in our mouse model. To investigate the contribution of humoral immune responses in anti-leukemia immunity induced by HSP-based immunotherapy, we tried to detect the anti-leukemia cell antibody and analyzed cytotoxicities by the antibody in the leukemia cell-derived HSP70 immunization mice model. Methods: Balb/c mice and syngeneic A20 B-cell leukemia cell line were used in this study. HSP70 was purified from A20 cells or liver tissue from healthy mice. Three weeks after subcutaneous administration of A20-derived HSP70 (A20-HSP70), liver-derived HSP70 (liver-HSP70) or PBS to the healthy mice, the sera were harvested to perform following experiments. To detect anti-A20 antibodies, mean fluorescence intensity (MFI) of A20 cells with immunized mice sera and FITC-conjugated anti-mouse-IgG was analyzed by flowcytometry. Additionally, the sera were subjected to ELISA using HRP labeled anti-mouse-IgG to detect specific antibody against A20-HSP70. Complement dependent cytotoxicity (CDC) activities were determined by trypan blue uptake of mouse target cells (A20, YAC1: lymphoma, or T27A: myeloid leukemia) after incubation with mice sera and rabbit complement. Results: MIF of A20 with the sera from A20-HSP70 immunized mice was significantly higher than that from PBS or liver-HSP70 immunized mice. In addition, anti-A20-HSP70 IgG level by ELISA was significantly increased in the A20-HSP70 immunized mice. However, anti-A20-HSP70 IgG level in liver-HSP70 mice was not elevated, suggesting that A20-HSP70 immunization induced the specific IgG against not HSP70 itself but peptides bound for A20-HSP70. The sera of mice immunized with A20-HSP70 showed no cytotoxic activity without complement but showed significantly high CDC activity against A20 in vitro. This CDC activity was not observed against YAC-1 or T27A. Conclusions: Leukemia specific CDC by the antibodies against the peptides bound for leukemia cell-derived HSP70 is suggested to be one of the mechanisms of anti-leukemia immunity induced by immunization with leukemia cell-derived HSP70 in mice. This would be an important finding to eradicate leukemia cells effectively in HSP-based immunotherapy for leukemia patients.

Leukemia Specific Humoral Immune Responses in Immunotherapy with Leukemia Cell-Derived HSP70 in Mice.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5192-5192
Author(s):  
Junko Jimbo ◽  
Kazuya Sato ◽  
Takaaki Hosoki ◽  
Motohiro Shindo ◽  
Katsuya Ikuta ◽  
...  
Keyword(s):  
T Cell ◽  
Immune Responses ◽  
Leukemia Cell ◽  
Leukemia Cells ◽  
Specific Antibodies ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Mice Liver ◽  
Igg Level

Abstract Background: Tumor-derived heat shock proteins (HSPs), which bind the tumor specific antigenic peptides and carry them onto MHC molecules, are used for the vaccination against cancers. We previously reported that immunotherapy using leukemia cell-derived HSPs against minimal residual leukemia in mice prolonged survival by leukemia-specific CD8 + cytotoxic T lymphocyte induction. In addition, we showed that CD4+ as well as CD8+ T cell is indispensable for survival prolongation (Sato et al. Blood, 2001), which suggests that humoral immune response by CD4+ T cell is also important to eradicate leukemia cells. Contributions of humoral immune responses in anti-leukemia immunity induced by HSP-based immunotherapy remain unknown and are important information to induce effective anti-leukemia immunity. In this study, we investigated the humoral immune responses and cytotoxic activities against leukemia cells in the leukemia cell-derived HSP70 immunization mice model in vitro. Methods: Balb/c mice and syngeneic A20 B-cell leukemia cell line were used in this study. HSP70 was purified from A20 cells or healthy mice liver tissue. After subcutaneous administration of A20-derived HSP70 (A20-HSP), liver-derived HSP70 (liver-HSP) to the healthy mice, the sera were harvested to perform following experiments. To detect anti-A20 antibodies, mean fluorescence intensity (MFI) of A20 cells with mice sera and FITC-conjugated anti-mouse-IgG was analyzed by flowcytometry. The sera were subjected to ELISA to detect the specific IgG against A20-HSP, or IgG secreted by A20 cells (A20 Ig) as putative A20-specific antigen. Complement dependent cytotoxicity (CDC) activities were determined by trypan blue uptake of mouse target cells (A20, YAC1: lymphoma, or T27A: myeloid leukemia) after incubation with mice sera and rabbit complement. Results: MIF of A20 with the sera from A20-HSP immunized mice (A20-HSP mice) was significantly higher than that from liver-HSP immunized mice (liver-HSP mice). IgG level against A20-HSP by ELISA was significantly increased in the A20-HSP mice compared with liver-HSP mice. The reactivities of A20-HSP mice sera against A20-HSP were completely lost by ATP treatment, which treatment dissociates the antigenic peptide from A20-HSP. In addition, IgG level against A20 Ig in the A20-HSP mice was significantly higher than that in the liver-HSP mice, and this reactivity against A20 Ig were inhibited by preincubation of sera with A20 Ig-idiotipe-derived peptide (A20 IP) DYWGQGTEL, which is known as the A20-specific peptide. The sera from A20-HSP mice showed no cytotoxic activity itself but showed significantly high CDC activity with complement against A20 but not to YAC-1 or T27A in vitro. Conclusions: Immunization with leukemia cell-derived HSP70 induces the leukemia specific antibodies against peptides bound with leukemia cell-derived HSP70, including an idiotipic peptide of IgG secreted by leukemia cells. In addition, CDC by these leukemia specific antibodies is though to be one of the mechanisms of anti-leukemia immunity induced by leukemia cell-derived HSP70 immunization. These findings enable the effective monitoring of therapeutic effects on the HSP-based immunotherapy for patients with leukemia by using the leukemia specific antibodies, and might develop a new strategy to enhance the leukemia specific CDC activities induced by HSP70-immunization.

A Novel Cytotoxic Mechanism by Leukemia-Specific Antibody in Immunotherapy with Leukemia Cell-Derived Heat Shock Protein 70.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2302-2302
Author(s):  
Kazuya Sato ◽  
Junko Jimbo ◽  
Takaaki Hosoki ◽  
Motohiro Shindo ◽  
Katsuya Ikuta ◽  
...  
Keyword(s):  
T Cell ◽  
Immune Responses ◽  
Leukemia Cell ◽  
Cd4 T Cell ◽  
Leukemia Cells ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
B Cell Leukemia ◽  
Mice Liver ◽  
Igg Level

Abstract Introduction: Tumor-derived heat shock proteins (HSPs), which bind the tumor-specific antigenic peptides, are good application for cancer vaccine. We previously reported that immunotherapy using leukemia cell-derived HSPs against leukemia cell in mice prolonged survival by leukemia-specific cellular immune responses through CD8 + cytotoxic T-cell (Sato et al. Blood, 2001; Iuchi et al. Int J Hematol, 2006). We also indicated that CD4+ as well as CD8+ T-cell is indispensable for the survival prolongation (Sato et al. Blood, 2001), suggesting that humoral immune response by CD4+ T-cell also contributes to eradicate leukemia cells. Contributions of humoral immune responses, including tumor-specific antibodies or cytotoxic activities, in anti-tumor immunity induced by tumor-derived HSP-based immunotherapy remain unclear. We therefore investigated humoral immune responses against leukemia cells in the leukemia cell-derived HSP70-immunized mouse model. Methods: Balb/c mice and syngeneic A20 B-cell leukemia cell line were used in this study. HSP70 was purified from A20 cells or healthy mice liver tissue. After subcutaneous administration of A20-derived HSP70 (A20-HSP), liver-derived HSP70 (liver-HSP; control) to the healthy mice, the sera were harvested to perform following experiments. To detect anti-A20 antibodies, mean fluorescence intensity (MFI) of A20 cells with mice sera and FITC-conjugated anti-mouse-IgG was analyzed by flowcytometry. The sera were subjected to ELISA to detect the specific IgG against A20-HSP, or A20 secreted IgG (A20-Ig) as an A20-specific antigen. To investigate a contribution of A20-HSP70 specific CD4+ T-cell, expression of intracellular Th2-cytokine IL4 in the A20-HSP70 stimulated CD4+ T-cell in the HSP70-immunizaed mice was measured by flowcytometry. Complement dependent cytotoxicity (CDC) activities were determined by trypan blue uptake of mouse target cells (A20, YAC1: lymphoma, or T27A: myeloid leukemia) after incubation with mice sera and complement. Results: MIF of A20 with the sera from A20-HSP immunized mice (A20-HSP mice) was significantly higher than that from liver-HSP immunized mice (liver-HSP mice). IgG level against A20-HSP by ELISA was significantly increased in the A20-HSP mice compared with liver-HSP mice. The reactivities of A20-HSP mice sera against A20-HSP were completely lost by dissociation of the antigenic peptide from A20-HSP after ATP-treatment. Additionally, IgG level against A20-Ig in the A20-HSP mice was significantly higher than that in the liver-HSP mice, and this reactivity was blocked by preincubation of the sera with A20-idiotype derived peptide (A20-IP), which is the A20-specific peptide. A20-HSP70-reactive IL4-producing CD4 + T-cells in the A20-HSP mice are extremely more than those in the liver-HSP mice. The sera from A20-HSP mice showed no cytotoxic activity itself but showed significantly high CDC activity with complement against A20 but not to YAC-1 or T27A in vitro. Conclusions: Immunization with leukemia cell-derived HSP70 induces the leukemia-specific antibodies against peptides binding to leukemia cell-derived HSP70, including B-cell leukemia idiotypic peptide, via activation of the leukemia-specific CD4+ T-cell. In addition, leukemia-specific antibody-mediated CDC contributes to the eradication of leukemia cells. To utilize the leukemia-specific CDC activities induced by HSP-based immunotherapy would be a novel therapeutic strategy to eradicate leukemia cells in the patients with leukemia.

Establishment of a NOD/SCID/γc−/−-Dependent Lymphoid Leukemia Cell Line, D593, with Novel RUNX1-LAF4 Fusion Gene.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4276-4276
Author(s):  
Akihiro Abe ◽  
Manabu Ninomiya ◽  
Shizuka Imagama ◽  
Momoko Suzuki ◽  
Fumihiko Hayakawa ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Fusion Gene ◽  
Lymphoblastic Leukemia ◽  
Leukemia Cell ◽  
Cell Receptor ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Pcr Analysis ◽  
Lymphoid Leukemia

Abstract We established a NOD/SCID/γc−/−(NOG mouse)-dependent human lymphoid leukemia cell line, D593, by repeated xenotransplantation of pediatric T-cell acute lymphoblastic leukemia cells with the translocation t(2;21). The cell line, D-593, could be serially transplanted from mouse to mouse over a 2-year period. D593 had the same immuno-phenotype as the original leukemia cells: positive for CD2, 5, 7, 14, and 34, and negative for CD3, 4, 8, 19, and 41a. Cytoplasmic CD3 was positive and the rearrangement of T-cell receptor was detected by Southern blot analysis. A previously unreported translocation of t(2;21)(q11;q22) was observed in both the original patient sample and D593. The split signal of RUNX1 was detected by fluorescence in site hybridization in D593 indicating the involvement of RUNX1. Using 3′-RACE and RT-PCR analysis, we identified novel chimeric transcripts of RUNX1-LAF4 joining exon 7 of RUNX1 to exon 4 of LAF4. In the transplanted NOG mice, D593 homed into the trabecular endosteal region of bone marrow (BM), and proliferated from the endosteum to medulla. At the late stage of engraftment, the BM was filled with human lymphoblasts and metastases into the trabecular of the spleen and Glisson’s sheath of the liver were also observed. These findings suggest that D593 is a useful cell line to study not only the leukemia-related biology of RUNX1-LAF4 but also the novel therapeutic model against core-binding factor (CBF) leukemia.

Pretreatment of Low-Dose Decitabine to Leukemia Cells Markedly Enhances the Cytotoxicity of Gemtuzumab Ozogamicin

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5000-5000
Author(s):  
Miwa Kurimoto ◽  
Hiroshi Matsuoka ◽  
Nobuyoshi Hanaoka ◽  
Shima Uneda ◽  
Takashi Sonoki ◽  
...  
Keyword(s):  
Cell Line ◽  
Low Dose ◽  
Leukemia Cell ◽  
Gemtuzumab Ozogamicin ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Target Cells ◽  
Enhancing Effect ◽  
Concurrent Use ◽  
Refractory Aml

Abstract Abstract 5000 Prognosis of relapsed/refractory acute myeloid leukemia (AML) patients has been poor due to its drug resistance. Although gemtuzumab ozogamicin (GO) targets CD33 antigen, which is overexpressed on most AML cells, the efficacy of GO alone for relapsed/refractory patients remains to be unsatisfied. Therefore we aimed to improve GO effects by combination with the other drugs. GO needs target cells intact to complete several steps for its cytotoxicity: GO binding to CD33 antigen, internalization of GO with CD33 antigen, fusion of the endosome to lysosome, digestion of the linker by lysozyme and release of calicheamicin, and intercalation of activated calicheamicin to DNA. GO could exert its cytotoxicity only when these cellular steps of target cells are intact. This paradoxical mechanism prompted us to study decitabine (DAC), azacitidine (AZA) and, valproic acid (VPA), which modify epigenetic alterations in leukemia cells with modest cytotoxicity. Low-dose DAC, AZA, or VPA have been reported to differentiate AML cells, with the increased expression of myeloperoxidase and lysozyme. Differentiation of AML cells by some agents, for example G-CSF, has been reported to reduce the expression of P-glycoprotein. Thus, we hypothesized that DAC, AZA or VPA could activate the cellular functions of target cells and enhance the cytotoxicity of GO. As refractory leukemic cells, we used SKK-1 and SKM-1 cells derived from patients with leukemia progressed from myelodysplastic syndrome, and SKNO-1, KO52, and Kasumi3 cells with complex karyotypes. We also used an acute promyelocytic leukemia cell line (NB4) and an acute myelomonocytic leukemia cell line (U937). Low-dose of DAC (100nM), AZA (2μM), or VPA (1mM) showed negligible cytotoxic effects. Simultaneous administration of GO (2.5 μg/ml) and DAC did not enhance cytotoxicity of GO. In contrast, GO cytotoxicity was enhanced only when cells were pretreated with DAC prior to GO. Pretreatment with DAC for 48 hours showed maximum enhancing effects. AZA also required pretreatment for enhancing effect. It was suggested that DAC and AZA induced some cellular response which enhance the cytotoxic steps of GO during pretreatment. VPA enhanced the cytotoxicity added either simultaneously or antecedently. The enhancing effect varied in each cell line. Pretreatment with DAC showed the most prominent enhancing effects among the three agents. DAC treatment changed neither the expression of CD33 antigens nor internalization of GO. DAC enhanced GO cytotoxicity partly by both promoting intercalation of calicheamicin to DNA and suppressing the activity of a drug-efflux pump, MRP-1. DAC-enhanced GO cytotoxicity in vitro was also observed in primary leukemic blasts of four (29%) among 14 relapsed/refractory AML patients. A large phase III study about GO, S0106 trial conducted by SWOG, revealed no significant improvement either in CR rate or in LFS for de novo AML patients, whereas significant increase of early complication death, in concurrent use of chemotherapy and GO. The result of SWOG trial and the mechanism of GO cytotoxicity might suggest that concurrent use of strongly-cytotoxic agents is not the best way of combination for GO. In contrast to SWOG trial, the present study targeted relapsed/refractory AML cells, employed pretreatment of low-dose DAC, and demonstrated preclinical possibility that the combination of DAC and GO would be useful for relapsed/refractory AML. Disclosures: No relevant conflicts of interest to declare.

Effects of hydroxylated resveratrol analogs on oxidative stress and cancer cells death in human acute T cell leukemia cell line

2014 ◽  
Vol 209 ◽  
pp. 96-110 ◽  
Author(s):  
Malgorzata Kucinska ◽  
Hanna Piotrowska ◽  
Michał W. Luczak ◽  
Justyna Mikula-Pietrasik ◽  
Krzysztof Ksiazek ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
T Cell ◽  
Cancer Cells ◽  
Cell Line ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
T Cell Leukemia ◽  
Cell Leukemia ◽  
Resveratrol Analogs ◽  
Cells Death

scholarly journals Epithelium-Specific Ets-Like Transcription Factor 1, ESE-1, Regulates ICAM-1 Expression in Cultured Lung Epithelial Cell Lines

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Zhiqi Yu ◽  
Jun Xu ◽  
Jinbao Liu ◽  
Jing Wu ◽  
Chan Mi Lee ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Airway Inflammation ◽  
Cell Lines ◽  
Leukemia Cell ◽  
A549 Cells ◽  
Leukemia Cell Line ◽  
Intercellular Adhesion Molecule ◽  
Cytokine Signaling ◽  
Target Cells ◽  
Transcription Factor 1

Cystic fibrosis (CF) patients suffer from chronic airway inflammation with excessive neutrophil infiltration. Migration of neutrophils to the lung requires chemokine and cytokine signaling as well as cell adhesion molecules, such as intercellular adhesion molecule-1 (ICAM-1), which plays an important role in mediating adhesive interactions between effector and target cells in the immune system. In this study, we investigated the relationship betweenICAM-1and epithelium-specific ETS-like transcription factor 1 (ESE-1) and found thatICAM-1expression is upregulated in cell lines of CF (IB3-1) as well as non-CF (BEAS-2B and A549) epithelial origin in response to inflammatory cytokine stimulation. SinceESE-1is highly expressed in A549 cells without stimulation, we examined the effect ofESE-1knockdown onICAM-1expression in these cells. We found thatICAM-1expression was downregulated whenESE-1was knocked down in A549 cells. We also tested the effect ofESE-1knockdown on cell-cell interactions and demonstrate that the knocking downESE-1in A549 cells reduce their interactions with HL-60 cells (human promyelocytic leukemia cell line). These results suggest thatESE-1may play a role in regulating airway inflammation by regulatingICAM-1expression.

scholarly journals A combination of a T cell-derived lymphokine differentiation-inducing activity and a physiologic concentration of retinoic acid induces HL-60 to differentiate to cells with functional chemotactic peptide receptors

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1273-1280
Author(s):  
M Imaizumi ◽  
TR Breitman
Keyword(s):  
Retinoic Acid ◽  
T Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Chemotactic Peptide ◽  
Phagocytic Cells ◽  
Peptide Receptors ◽  
Morphologic Characteristics ◽  
O2 Production

The human acute promyelocytic leukemia cell line HL-60 is induced by retinoic acid (RA) and N,N-dimethylformamide (DMF) to differentiate into cells having many of the functional and morphologic characteristics of mature granulocytes. With normal human phagocytic cells there is both superoxide anion (O2-) production and chemotaxis in response to chemoattractants such as N-formyl-methionyl-leucyl- phenylalanine (FMLP). We have now found that although HL-60 cells induced with RA alone produce O2- in response to 12–0-tetradecanoyl- phorbol-13-acetate (TPA) they are deficient in FMLP-stimulated O2- production and chemotaxis. In contrast, HL-60 induced either with DMF or with a combination of 10 nmol/L RA and a T cell-derived lymphokine, differentiation-inducing activity (DIA), produce O2- and exhibit chemotaxis in response to FMLP. The basis for these results appears to be the concentration of cell surface chemotactic peptide receptors. Thus, untreated HL-60 and HL-60 induced with either RA alone or DIA alone do not have measurable levels of FMLP receptors, whereas HL-60 induced with a combination of RA and DIA has 5,400 receptors per cell. HL-60 induced with RA and DIA plus 1 mumol/L dexamethasone have 25,000 receptors per cell and have greater chemotactic activity than HL-60 induced with the combination of RA and DIA. Thus, differentiation of HL- 60 to cells with many properties of normal phagocytes can be induced in vitro by physiologic substances.

Sign in / Sign up

Export Citation Format

Share Document