Stimulation of Autologous Antitumor T-Cell Responses Against Medullary Thyroid Carcinoma Using Tumor Lysate-Pulsed Dendritic Cells

Abstract Effective presentation of tumor antigens is fundamental to strategies aimed at enrolling the immune system in eradication of residual disease after conventional treatments. Myeloid malignancies provide a unique opportunity to derive dendritic cells (DCs), functioning antigen-presenting cells, from the malignant cells themselves. These may then co-express leukemic antigens together with appropriate secondary signals and be used to generate a specific, antileukemic immune response. In this study, blasts from 40 patients with acute myeloid leukemia (AML) were cultured with combinations of granulocyte-macrophage colony-stimulating factor, interleukin 4, and tumor necrosis factor α, and development to DCs was assessed. After culture, cells from 24 samples exhibited morphological and immunophenotypic features of DCs, including expression of major histocompatibility complex class II, CD1a, CD83, and CD86, and were potent stimulators in an allogeneic mixed lymphocyte reaction (MLR). Stimulation of autologous T-cell responses was assessed by the proliferative response of autologous T cells to the leukemic DCs and by demonstration of the induction of specific, autologous, antileukemic cytotoxicity. Of 17 samples, 11 were effective stimulators in the autologous MLR, and low, but consistent, autologous, antileukemic cytotoxicity was induced in 8 of 11 cases (mean, 27%; range, 17%-37%). This study indicates that cells with enhanced antigen-presenting ability can be generated from AML blasts, that these cells can effectively prime autologous cytotoxic T cells in vitro, and that they may be used as potential vaccines in the immunotherapy of AML.

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Enhanced in vitro stimulation of rhesus macaque dendritic cells for activation of SIV-specific T cell responses

Journal of Immunological Methods ◽

10.1016/s0022-1759(01)00544-0 ◽

2002 ◽

Vol 260 (1-2) ◽

pp. 219-234 ◽

Cited By ~ 52

Author(s):

Erin Mehlhop ◽

Loreley A. Villamide ◽

Ines Frank ◽

Agegnehu Gettie ◽

Christine Santisteban ◽

...

Keyword(s):

Dendritic Cells ◽

T Cell ◽

Rhesus Macaque ◽

T Cell Responses ◽

Cell Responses ◽

Stimulation Of

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Superior Acute Myeloid Leukemia-Specific T Cell Responses Using Dendritic Cells Pulsed with Apoptotic Bodies, vs.Tumor Lysates or mRNA.

Blood ◽

10.1182/blood.v106.11.295.295 ◽

2005 ◽

Vol 106 (11) ◽

pp. 295-295

Author(s):

Dongxia Xing ◽

William K. Decker ◽

Sufang Li ◽

Richard E. Champlin ◽

John D. MacMannis ◽

...

Keyword(s):

Dendritic Cells ◽

T Cell ◽

T Lymphocytes ◽

T Cell Responses ◽

Myelogenous Leukemia ◽

Apoptotic Bodies ◽

Apoptotic Body ◽

Tumor Lysate ◽

Antigen Uptake ◽

Cell Responses

Abstract Dendritic cells (DCs), which hold promise for cancer immunotherapy have two distinct functions. Immature DCs internalize antigens; while mature DCs trigger native T cell activation. To date, no consensus has been reached concerning the optimal source of tumor antigen nor the mode of antigen uptake for efficient cross-priming of tumor specific response. Reports have demonstrated that human DCs can acquire relevant antigens and stimulate MHC class I-restricted cytotoxic T lymphocytes (CTLs) by phagocytosing apoptotic tumor cells. Apoptosis was established to be the critical trigger of cross-priming in this process. Apoptotic bodies form when cells undergo programmed cell death. In this study, three acute myelogenous leukemia in (AML) antigen preparations were compared in an autologous setting. Apoptotic bodies, Tumor lysate Tumor mRNA Methods: DCs were generated by CD14 magnetic selection of peripheral blood progenitor cells (PBPCs) and six days of culture in GM-CSF and IL-4. Tumor lysate was made by repeated freezing and thawing of patient AML cells (leukodepletion product >95% blasts); mRNA was extracted using a kit (Qiagen); apoptotic bodies were generated with UVB irradiation of a fraction of the same AML product. The rate of apoptosis was detected using Annexin V FITC flow cytometry. The DCs were loaded with mRNA by electroporation. The DCs were loaded with lysate and apoptotic bodies by incubation. Autologous T cells stimulated with DCs loaded with the three differing antigen preparations were compared for interferon(INF)-g production in ELISPOT and proliferation assays. Results: The DCs pulsed with apoptotic bodies; lysate and mRNA expressed similar levels of the costimulatory molecules CD80, CD86, and theDC maturation marker CD83. The internalization rate of apoptotic bodies was about 20% detected by DAPI (label the apoptotic bodies) and DC83 double staining. The lysate pulsed DC and mRNA pulsed DCs were above 90% detected by flow of trace FITC- albumin and tracer GFP mRNA from previous experiments. Yet IFN-g secretion of T lymphocytes was statistically significant higher (P<0.05), using apoptotic body-loaded DC as compared to total lysate or mRNA. The T cell proliferation stimulated by apoptotic body-loaded DC was also better than that of lysate or mRNA loaded DCs (p<0.05). Conclusions: These preliminary data demonstrate that apoptotic body-loaded DCs induced better Th1 T cell responses than DCs loaded with AML lysate or mRNA. One potential advantage conferred by the use of apoptotic body loaded DCs is that presentation of intracellular proteins in tumor lysates and mRNA that are irrelevant to the induction of anti-tumor responses are eliminated, which may lead to better antigen presentation and consequently an improved Th1 response.

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Pilot trial of autologous dendritic cells loaded with tumor lysate(s) from allogeneic tumor cell lines in patients with metastatic medullary thyroid carcinoma

Oncology Reports ◽

10.3892/or_00000391 ◽

2009 ◽

Vol 21 (6) ◽

Cited By ~ 34

Author(s):

Bachleitner-Hofmann

Keyword(s):

Dendritic Cells ◽

Thyroid Carcinoma ◽

Tumor Cell ◽

Medullary Thyroid Carcinoma ◽

Cell Lines ◽

Pilot Trial ◽

Tumor Cell Lines ◽

Tumor Lysate ◽

Medullary Thyroid ◽

Allogeneic Tumor Cell

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Combined immunotherapy with whole tumor lysate–pulsed interleukin-15–activated dendritic cells and cucurbitacin I promotes strong CD8+ T-cell responses and cures highly aggressive lymphoma

Cytotherapy ◽

10.1016/j.jcyt.2015.01.006 ◽

2015 ◽

Vol 17 (5) ◽

pp. 647-664 ◽

Cited By ~ 5

Author(s):

S. Kumar Hira ◽

Indrani Mondal ◽

Partha P. Manna

Keyword(s):

Dendritic Cells ◽

T Cell ◽

T Cell Responses ◽

Cd8 T Cell ◽

Aggressive Lymphoma ◽

Tumor Lysate ◽

Cell Responses ◽

Interleukin 15 ◽

Combined Immunotherapy

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Inhibition of invariant chain expression in dendritic cells presenting endogenous antigens stimulates CD4+ T-cell responses and tumor immunity

Blood ◽

10.1182/blood-2003-06-1867 ◽

2003 ◽

Vol 102 (12) ◽

pp. 4137-4142 ◽

Cited By ~ 23

Author(s):

Yangbing Zhao ◽

David Boczkowski ◽

Smita K. Nair ◽

Eli Gilboa

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Cd4 T Cells ◽

T Cell Responses ◽

Cd4 T Cell ◽

Cytotoxic T Cell ◽

Invariant Chain ◽

Cell Responses ◽

Stimulation Of

Abstract Induction of potent and sustained antiviral or antitumor immunity is dependent on the efficient activation of CD8+ and CD4+ T cells. While dendritic cells constitute a powerful platform for stimulating cellular immunity, presentation of endogenous antigens by dendritic cells transfected with nucleic acid-encoded antigens favors the stimulation of CD8+ T cells over that of CD4+ T cells. A short incubation of mRNA-transfected dendritic cells with antisense oligonucleotides directed against the invariant chain enhances the presentation of mRNA-encoded class II epitopes and activation of CD4+ T-cell responses in vitro and in vivo. Immunization of mice with the antisense oligonucleotide-treated dendritic cells stimulates a more potent and longer lasting CD8+ cytotoxic T-cell (CTL) response and enhances the antitumor efficacy of dendritic cell-based tumor vaccination protocols. Transient inhibition of invariant chain expression represents a simple and general method to enhance the stimulation of CD4+ T-cell responses from endogenous antigens. (Blood. 2003;102:4137-4142)

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