Biomedical subjects

Aurore Saudemont

Publications and source records attributed to Aurore Saudemont.

7 recordsLinked to original sources

PubMed2005

Targeted elimination of prostate cancer by genetically directed human T lymphocytes.

The genetic transfer of antigen receptors is a powerful approach to rapidly generate tumor-specific T lymphocytes. Unlike the physiologic T-cell receptor, chimeric antigen receptors (CARs) encompass immunoglobulin variable regions or receptor ligands as their antigen recognition moiety, thus permitting T cells to recognize tumor antigens in the absence of human leukocyte antigen expression. CARs encompassing the CD3zeta chain as their activating domain induce T-cell proliferation in vitro, but limited survival. The requirements for genetically targeted T cells to function in vivo are less well understood. We have, therefore, established animal models to assess the therapeutic efficacy of human peripheral blood T lymphocytes targeted to prostate-specific membrane antigen (PSMA), an antigen expressed in prostate cancer cells and the neovasculature of various solid tumors. In vivo specificity and antitumor activity were assessed in mice bearing established prostate adenocarcinomas, using serum prostate-secreted antigen, magnetic resonance, computed tomography, and bioluminescence imaging to investigate the response to therapy. In three tumor models, orthotopic, s.c., and pulmonary, we show that PSMA-targeted T cells effectively eliminate prostate cancer. Tumor eradication was directly proportional to the in vivo effector-to-tumor cell ratio. Serial imaging further reveals that the T cells must survive for at least 1 week to induce durable remissions. The eradication of xenogeneic tumors in a murine environment shows that the adoptively transferred T cells do not absolutely require in vivo costimulation to function. These results thus provide a strong rationale for undertaking phase I clinical studies to assess PSMA-targeted T cells in patients with metastatic prostate cancer.

Animals↗

PubMed2005

Induction of leukemia-specific CD8+ cytotoxic T cells with autologous myeloid leukemic cells maturated with a fiber-modified adenovirus encoding TNF-alpha.

Acute myeloid leukemia (AML) cells can be differentiated into dendritic cells (DCs) using appropriate combinations of cytokines but generation of autologous antileukemic cytotoxic T cells using leukemic DCs remains difficult. Transduction by adenoviral vectors has been reported to induce efficient maturation of monocyte-derived DCs but AML cells are generally resistant to adenoviral gene transfer. In this study we tested the effects of adenoviral TNF-alpha gene transfer on maturation of AML cells using the fiber-modified AdTNF.F(pK7) adenovirus. All samples expressed high and sustained levels of TNF-alpha following transduction. AdTNF.F(pK7) induced significantly greater maturation of AML cells into antigen-presenting cells (APC) than did recombinant TNF-alpha or control adenoviral vector. Maturation of leukemic cells into APCs was mediated at least partially via a PI3K/mTOR pathway, as the inhibitors LY294002, wortmannin, and rapamycin inhibited the maturation effect induced by the AdTNF.F(pK7) adenovirus. In addition, CD8+ T cells expanded with AdTNF.F(pK7)-transduced AML cells showed greater expansion and specific CD8+ CTL activity against autologous AML cells than T cells expanded by other means. Thus, fiber-modified adenoviral vectors encoding TNF-alpha are able to maturate AML cells into APCs with high efficacy and reproducibility, providing a useful tool to generate efficiently specific CD8+ CTLs against leukemic disease.

Acute Disease↗

PubMed2004

NK cells that are activated by CXCL10 can kill dormant tumor cells that resist CTL-mediated lysis and can express B7-H1 that stimulates T cells.

Tumor dormancy is a phenomenon where small numbers of tumor cells persist in the host for months or years. We previously showed in the DA1-3b/C3H mouse model of acute myeloid leukemia that dormant tumor cells resist cytotoxic T-lymphocyte (CTL)-mediated killing because they overexpress B7-H1. Here, we vaccinated mice with DA1-3b cells transduced with CXCL10. Vaccinated mice developed a strong systemic immunity that led to the cure of established leukemia without persistence of dormant tumor cells. In vivo depletion of natural killer (NK) cells from the mice abrogated the protective effect of the vaccine. Long-term persistent leukemic cells resist CTL-mediated lysis but were killed by NK cells from mice vaccinated with DA1-3b/CXCL10. These NK cells expressed B7-H1. Recombinant CXCL10, CXCL9, CXCL11, and CXCL12 chemokines induced expression of B7-H1 on mouse and human NK cells in vitro. Mouse and human B7-H1+ NK cells induced proliferation of T cells and production of interferon gamma and tumor necrosis factor alpha in vitro, and in vivo blocking of B7-H1 inhibited the protective effect of vaccination. Thus, CXCL10 induces antileukemic immunity, at least partially by stimulating NK cells to express B7-H1+. This antitumor effect is in contrast to the effect of B7-H1 when expressed on tumor cells because it stops cytotoxic lymphocytes from killing those tumor cells.

Animals↗

PubMed2004

In a model of tumor dormancy, long-term persistent leukemic cells have increased B7-H1 and B7.1 expression and resist CTL-mediated lysis.

In tumor dormancy, tumor cells persist in the host over a long period of time but do not grow. We investigated in the DA1-3b mouse model of acute myeloid leukemia how leukemic cells could persist for months in spite of an effective antileukemic immune response. Mice were immunized with irradiated interleukin 12 (IL12)- or CD154-transduced DA1-3b cells, challenged with wild-type DA1-3b cells, and randomly killed during 1-year follow-up. Quantification of residual disease 1 year after challenge showed that persistent leukemic cells represented less than 0.02% of spleen cells in most animals. These residual cells were still able to kill naive hosts, even when isolated after 1 year of persistence. Persistent leukemic cells were more resistant to specific cytotoxic T-cell (CTL)-mediated killing and had enhanced B7-H1 and B7.1 expression proportional to the time they had persisted in the host. Blocking B7-H1 or B7.1/cytotoxic T-lymphocyte-associated antigen (CTLA-4) interaction enhanced CTL-mediated killing of the persistent cells, and blocking B7-H1, B7.1, or CTLA-4 in vivo prolonged survival of naive mice injected with persistent leukemic cells. Thus, escape of leukemic cells from tumor immunity via overexpression of B7-H1 or B7.1 might represent a new mechanism of tumor dormancy in acute leukemia.

Animals↗

PubMed2004

Efficient generation of antileukemic autologous T cells by short-term culture and gamma-irradiation of myeloid leukemic cells.

Blasts from patients with acute myeloid leukemia (AML) can be differentiated in dendritic cells (DCs) using appropriate combinations of cytokines. However, generation of autologous antileukemic cytotoxic T cells using leukemic DCs remains difficult. We have previously reported that expression of costimulatory molecules in cultured AML cells could be induced by gamma-irradiation. In the present study, blasts from 21 patients with AML were cultured in vitro for 2 days, then cells were gamma-irradiated and antigen-presenting cell (APC) characteristics were assessed. gamma-Irradiation induced expression of several characteristics of APCs in AML blasts, including expression of CD80, CD86, and BDCA-4, and were stimulators of allogeneic mixed lymphocyte reactions. Autologous antileukemic cytotoxicity was induced in seven out of ten cases. This study shows that cells with APC characteristics and able to induce ex vivo stimulation of autologous antileukemic T cells can be generated from AML cells using the simple and rapid method of gamma-irradiation of cultured leukemic cells.

Acute Disease↗

PubMed2004

Short-term culture of myeloid leukemic cells allows efficient transduction by adenoviral vectors.

BACKGROUND: Ex vivo gene therapy of acute myeloid leukemia (AML) requires efficient transduction of leukemic cells. Recombinant adenovirus has been reported to be a poorly efficient vector in leukemic cells. We investigated leukemic cell culture as a possible method of improving the efficacy of this vector. METHODS: Leukemic cell lines and primary cultured AML cells were incubated with adenoviral vectors carrying GFP, LacZ, or IL-12 cDNA. Transduction efficiency was evaluated by measuring adenoviral genome copy number and transgene expression in leukemic cells. The expression of the coxsackie/adenovirus receptor (CAR), CD29, CD49e, and CD51/61 was measured, as was the effect of blocking integrin on adenoviral transduction. RESULTS: Increasing the multiplicity of infection (MOI) to 300 plaque-forming units per cell enhanced transduction of leukemic cell lines and to a lesser degree of AML cells. Analysis of adenoviral genome copy per cell showed only a partial correlation between gene transfer efficiency and transgene expression. Culture of AML cells for 3 days prior to adenoviral transduction increased both adenoviral copy number per cell and the percentage of transgene-expressing cells. CD29, CD49e, and CD51/61 but not CAR expression increased in cultured AML cells between days 0 and 3 and integrin-blocking experiments showed inhibition of transduction in two of four AML samples tested. CONCLUSIONS: Efficient ex vivo gene transfer in primary cultured AML cells can be achieved by short-term culture of leukemic cells prior to gene transfer with adenoviral vectors at a high MOI. This effect appears to be at least partially mediated by enhanced integrin expression.

Acute Disease↗

PubMed2003

Chemotherapy increases transgene expression in leukemic cells.

BACKGROUND: Patients with acute myeloid leukemia (AML) often obtain complete remission with chemotherapy but the majority of patients relapse. Combining chemotherapy and gene therapy appears to be a promising approach; however, the effects of chemotherapy on transgene expression in leukemic cells have not yet been investigated. METHODS: DA1-3b leukemic cells were transfected with pCDNA3 plasmids carrying GM-CSF or LacZ cDNA. The leukemic K562 cell line and primary cultured AML cells were transduced with an Ad5.CMV-LacZ adenoviral vector. Cells were then incubated with various concentrations of daunorubicin (DNR) and cytosine arabinoside (Ara-C), and expression of the transgene was measured. Murine DA1-3b-pCDNA3/LacZ leukemic cells were also injected into syngeneic C3H/Hej mice. RESULTS: In the cells carrying pCDNA3, DNR and Ara-C dramatically increased expression of the LacZ and GM-CSF transgenes. Over-expression depended on drug dose and was due to increased transcription. Enhancement was also observed in K562 cells and in some primary cultured AML samples transduced with the Ad5.CMV-LacZ adenovirus. Addition of N-acetyl-L-cysteine inhibited the over-expression, suggesting that reactive oxygen species were involved in activating the CMV promoter. In the A549 lung carcinoma cell line transduced with Ad5.CMV-LacZ, Ara-C had only a minor effect, and DNR had a detrimental effect, suggesting that expression depends on cell type. In vivo experiments in which mice received DA1-3b-pCDNA3/LacZ leukemic cells, and were then treated with Ara-C, also showed increased transgene expression in these leukemic cells. CONCLUSIONS: In leukemic cells, chemotherapeutic agents can induce over-expression of transgenes. This suggests a promising combined strategy for the treatment of acute leukemia.

Acute Disease↗