Advances in Brief Gene Expression Profiling and Functional Activity of Human Dendritic Cells Induced with IFN--2b: Implications for Cancer Immunotherapy 1 Federica Moschella, Brygida Bisikirska, Antonella Maffei, Kyriakos P. Papadopoulos, Donna Skerrett, Zhuoru Liu, Charles S. Hesdorffer, and Paul E. Harris 2 The International Institute of Genetics and Biophysics, Naples, Italy [A. M.], the Department of Pathology [D. S.], the Department of Surgery [Z. L.], the Division of Medical Oncology, Department of Medicine [F. M., B. B., K. P. P., C. S. H., P. E. H.], College of Physicians and Surgeons, Columbia University, New York, New York 10033 Abstract Purpose: In this study, we have compared patterns of gene expression and functional activity of human dendritic cells (DCs) cultured under defined conditions in IFN--2b and recombinant human granulocyte macrophage colony- stimulating factor (DCA) with cells grown in granulocyte macrophage colony-stimulating factor and IL-4 (DC4) as an initial step in evaluating the clinical utility of DCA in cancer immunotherapy. Experimental Design and Results: Comparison of mRNA transcript profiles between DCA and DC4 revealed different expression patterns for cytokines, chemokines, che- mokine receptors, costimulatory molecules, and adhesion proteins. Many genes involved in antigen (Ag) processing were equally expressed in both populations; however, ex- pression of transcripts involved in Ag presentation was in- creased in DCA. DCA also showed up-regulation of Toll-like receptor 2 and 3, as well as several tumor necrosis factor family ligands. Consistent with expression profiling, func- tional assays demonstrated that DCAs were more potent stimulators of naive T-cell responses than DC4 in an inter- leukin 15 and interleukin 1-dependent manner. DCA- mediated tumor cell-directed cytotoxicity induced apoptosis in different human tumor cell lines and internalized apo- ptotic bodies to a greater extent than DC4. Lastly, in vitro priming experiments, using apoptotic cells or peptide as sources of Ag, showed that DCA drove the expansion of tumor peptide Ag-specific autologous CD8 T cells to a greater extent than DC4. Conclusions: The unique phenotype conferred by cul- turing DCs in IFN--2b may be useful in adoptive transfer regimens where the destruction of tumor cells in situ, initi- ation of T-cell responses toward tumor tissue with unknown Ags, and/or enhancement of pre-existing Ag-specific mem- ory responses are desired outcomes. Introduction IFN- is an immunoregulatory cytokine presently used clinically in a recombinant form for the treatment of tumors and chronic viral infections (1–3). Although the exact mechanism(s) by which IFN- promotes antitumor and antiviral responses is still under investigation, it is known that IFN- can act via selective toxicity toward transformed or virally infected cells, and that it modulates immune response (4). However, systemic administration of type I IFNs is associated with severe toxicity and significant side effects, thereby limiting its clinical appli- cations (5, 6). DCs 3 are central regulatory elements of both adaptive and innate immune responses by virtue of their ability to activate naı ¨ve T cells and recognize pathogen-associated molecule pat- terns (7, 8). Our current understanding of DC biology in the context of adaptive immunity suggests that the differentiation state of the DC qualitatively affects their interaction with T lymphocytes. It is well documented that, depending on the degree of maturation and direction of maturation, DCs will elaborate different profiles of cytokines and cell surface recep- tors, and express different Ag processing and presenting abilities (9 –11). Recent studies show that IFN- strongly modulates DC function and maturation (12–17). For example, treatment of peripheral blood monocytes with GM-CSF plus IFN- induces rapid maturation of these cells into potent APCs for viral epitopes (14). Other studies have found that type I IFNs induce apoptotic cell death in cultures of mature DCs (15, 16). We have developed GMP compatible culture methods for the production of DCs used in cancer immunotherapy (18). To broaden our understanding of DC phenotypes obtainable under these culture conditions, we have systematically compared gene Received 12/23/02; revised 2/18/03; accepted 2/20/03. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Supported by grants from the USPHS, NIH 1 K23 RR16078 (to K. P. P.), the Herbert Irving Cancer Center, and the Octoberwoman Foundation. 2 To whom requests for reprints should be addressed, at Medical On- cology, BB 20 –13, 650 West 168 th Street, New York, NY 10032. Phone: (212) 305-7363; Fax: (212) 305-7348; E-mail: peh1@columbia.edu. 3 The abbreviations used are: DC, dendritic cell; GM-CSF, granulocyte macrophage colony-stimulating factor; GMP, good manufacturing pro- cedure; PBMC, peripheral blood mononuclear cell; IL, interleukin; Ag, antigen; APC, antigen presenting cell; mAb, monoclonal antibody; Ab, antibody; CLIP, class II-associated invariant chain peptide; RT-PCR, reverse transcription-PCR; ELISPOT, enzyme-linked immunospot GAPDH, glyceraldehyde-3-phosphate dehydrogenase; TNF, tumor ne- crosis factor; TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; TWEAK, tumor necrosis factor-like weak inducer of apoptosis; ATC, apoptotic tumor cell. 2022 Vol. 9, 2022–2031, June 2003 Clinical Cancer Research