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Polyriboinosinic Polyribocytidylic Acid (Poly(I:C)) Induces Stable Maturation of Functionally Active Human Dendritic Cells¹

Department of Immunohematology and Blood Bank, Leiden University Medical Center, Leiden, The Netherlands

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
For vaccination strategies and adoptive immunotherapy purposes, immature dendritic cells (DC) can be generated from adherent monocytes using GM-CSF and IL-4. Presently, the only clinically applicable method to induce stable maturation of DC is the use of supernatants of activated monocytes (monocyte-conditioned medium (MCM)). MCM contains an undefined mixture of cytokines and is difficult to standardize. Here we report that stable maturation of DC can be simply induced by the addition of polyriboinosinic polyribocytidylic acid (poly(I:C)), a synthetic dsRNA clinically applied as an immunomodulator. Poly(I:C)-treated DC show a mature phenotype with high expression levels of HLA-DR, CD86, and the DC maturation marker CD83. This mature phenotype is retained for 48 h after cytokine withdrawal. In contrast to untreated DC, poly(I:C)-treated DC down-regulate pinocytosis, produce high levels of IL-12 and low levels of IL-10, induce strong T cell proliferation in a primary allo MLR, and effectively present peptide Ags to HLA class I-restricted CTL. In conclusion, we present a simple methodology for the preparation of clinically applicable mature DC.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
Dendritic cells (DC)³ are professional APC (1). Immature DC effectively capture and process native protein Ags (2). Mature DC express high levels of MHC and costimulatory molecules and effectively initiate primary T cell responses (3). Recently, methods to generate large numbers of DC from adherent monocytes have been developed (4). These DC generated with GM-CSF and IL-4 display an immature phenotype unless treated with additional factors. Treatment of immature DC with single cytokines or combinations of cytokines such as TNF-, IFN-ß, IFN-, IL-1ß, or PGE_2, induces reversible maturation of DC (4, 5, 6). Treatment with supernatants of activated monocytes (monocyte-conditioned medium (MCM)) induces stable maturation of DC (7). It is, however, difficult to generate standardized batches of MCM for clinical purposes.

Viral infection and inflammatory products such as LPS, bacterial DNA, and CpG-oligodeoxynucleotides trigger the maturation of DC (8, 9, 10). A synthetic dsRNA, polyriboinosinic polyribocytidylic acid (poly(I:C)), is often used in models of viral infection. Poly(I:C) is a potent IFN inducer (11, 12) and can activate monocytes to produce CSF (11), IL-1ß (11), IL-12 (12), and PGE₂ (11). Poly(I:C) has been used in various clinical trials with little or no toxicity (13, 14).

Here we describe that poly(I:C) can induce stable maturation of DC. We demonstrate that poly(I:C)-treated DC are comparable to DC treated with MCM in both their phenotypical and functional characteristics. This simple method can easily be used in closed culture systems for the generation of clinical applicable mature DC.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Culture medium

Culture medium consisted of RPMI supplemented with 100 U/ml penicillin 100 µg/ml streptomycin (both from Life Technologies, Paisley, U.K.), and 2% heat-inactivated autologous heparinized plasma or pooled human serum.

Reagents

Staphylococcus aureus Cowan I strain Pansorbin cells (SACS) were purchased from Calbiochem (La Jolla, CA). Poly(I:C) was purchased from Sigma (St. Louis, MO; lot 27H4009) and dissolved in PBS, and aliquots of 1 mg/ml were preserved at -20°C. The minor histocompatibility Ag (mHag) HA-1 peptide (15) was synthesized using a semiautomatic multiple peptide synthesizer; its purity was checked by reverse phase HPLC.

PBMC and T cell clones

PBMC were isolated by Ficoll-Hypaque density gradient separation of blood collected from healthy blood donors. Naive cord blood cells were isolated by Ficoll-Hypaque density gradient separation of cord blood. The mHag HA-1-specific HLA-A*0201-restricted CTL clone (16) was used in proliferation experiments.

Preparation of MCM

PBMC, mixed from five randomly selected healthy blood donors were allowed to adhere to petri dishes (Falcon, Lincoln Park, NJ) for 2 h at 37°C (50 x 10⁶/petridish). Nonadherent cells were removed with forceful pipetting. The adherent fraction was cultured in 8 ml of culture medium/petri dish supplemented with either SACS (1/10,000) or 50 µg/ml of poly(I:C). After 24 h the cell-free supernatants were filtered (0.22 µm pore size) and stored at -20°C.

DC culture

DC were cultured from adherent PBMC in the presence of 800 U/ml GM-CSF and 500 U/ml IL-4 as described by Romani et al. (7). On day 7, 50% MCM or different concentrations of poly(I:C) (range, 1–50 µg/ml) were added to induce DC maturation. The MCM- or poly(I:C)-treated DC as well as untreated DC were cultured for another 72 h. For determination of phenotype stability, DC were washed three times and cultured for an additional 48 h in culture medium without cytokines, MCM, or poly(I:C).

Phenotype analysis

DC were labeled using FITC- or PE-labeled specific mAb to CD14, CD40, CD80, CD86, CD83, and HLA-DR (all from Becton Dickinson, Mountain View, CA). The fluorescence intensity was measured using the FACScan (Becton Dickinson).

Pinocytosis assay

DC treated with SACS-MCM and poly(I:C)-treated and untreated DC were harvested and stored on ice in culture medium for at least 30 min before initiation of the assay. FITC-conjugated BSA (BSA-FITC; Sigma) was added at a final concentration of 1 mg/ml, and DC were incubated for 1 h at 37°C or on ice (background staining). Cells were washed twice in ice-cold medium and fixated with PBS containing 1% paraformaldehyde before FACS analysis.

Cytokine measurements

Adherent PBMC were cultured for 7 days with GM-CSF and IL-4. Cell-free culture supernatants were harvested after 24, 48, or 72 h of further culture of the DC at a concentration of 5 x 10⁵/ml with or without poly(I:C) (12.5 µg/ml). IL-12 p40 (R & D Systems, Abingdon, U.K.), IL-12 p70 (Diaclone, Besancon, France), and IL-10 (Schering Plough, Dardilly, France) release in the supernatants was measured by specific sandwich ELISA.

Primary MLR

Serial dilutions (1.5 x 10⁴ to 3 cells/well) of irradiated (30 Gy) stimulator cells were cultured in triplicate with 1.5 x 10⁵ responder cells in 96-well round-bottom plates (Costar, Corning NY). After 96 h 1 µCi/well [³H]thymidine (New England Nuclear, Boston MA) was added to the wells. The cells were harvested after 16 h, and [³H]thymidine incorporation was determined by liquid scintillation counting. The results are expressed as the mean of triplicate cultures. The SEM of the results never exceeded 15%. As responder cells, autologous PBMC or random CBC were used. As stimulator cells, PBMC, monocytes, immature DC, and DC treated with SACS-MCM or with poly(I:C) were used.

T cell proliferation assays

The HA-1-specific CTL clone was used as responder cells (1.5 x 10⁴ cells/well) and was cocultured with irradiated (30 Gy) stimulator cells (10³ cells/well) in 96-well round-bottom microtiter plates for 72 h. The mHag HA-1 peptide was added in the assay in a serial dilution of 1 ng/ml to 10 µg/ml. Sixteen hours before harvesting, the cells were labeled with 1 µCi/well [³H]thymidine. [³H]thymidine incorporation was determined by liquid scintillation counting. The results are expressed as the mean of triplicate cultures. The SEM of the results never exceeded 15%. As stimulator cells, monocytes, immature DC, and DC treated with SACS-MCM or with poly(I:C) were used.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Poly(I:C) induces mature surface phenotype of DC

We have tested whether supernatants of poly(I:C)-activated monocytes (pIC-MCM) or purified poly(I:C) alone can induce DC maturation. Untreated DC and DC treated with SACS-MCM were used as controls. Treatment of immature DC for 72 h with SACS-MCM, pIC-MCM, or purified poly(I:C) resulted in the typical DC morphology (Fig. 1) and induced significant maturation, as reflected by up-regulation of HLA-DR, CD86, and CD83 (Fig. 2). As expected, expression of the maturation marker CD83 was low or absent on untreated DC, and CD14 expression was low or absent in all DC preparations. In subsequent experiments, the maturational effect of poly(I:C)-MCM could be almost completely abrogated by RNase treatment. This demonstrates that dsRNA actually is the main active component of the MCM (data not shown). Treatment of DC with 12,5 µg/ml poly(I:C) for 72 h appeared optimal. Poly(I:C) concentrations over 25 µg/ml were toxic, whereas concentrations of poly(I:C) lower than 1 µg/ml did not induce any maturation (data not shown). Based on these results, we decided to use only purified poly(I:C) in a concentration of 12.5 µg/ml for maturation induction of the DC cultures.

View larger version (205K): [in this window] [in a new window]   FIGURE 1. Poly(I:C)-treated DC display typical dendritic morphology. DC were cultured from adherent monocytes with GM-CSF and IL-4 for 7 days followed by treatment with purified poly(I:C) at a concentration of 12,5 µg/ml for 3 days. Photomicrographs were taken on day 10 of culture with medium resolution (x100). The cells clearly show a typical dendritic morphology.

View larger version (32K): [in this window] [in a new window]   FIGURE 2. Poly(I:C) induces mature surface phenotype of DC. DC were cultured from adherent monocytes with GM-CSF and IL-4 for 7 days followed by treatment with SACS-MCM, poly(I:C)-MCM, poly(I:C) (12.5 µg/ml), or no treatment for 72 h. Filled histograms depict specific Ab staining; open histograms depict controls. DC treated with poly(I:C)-MCM, poly(I:C), or SACS-MCM express high levels of HLA DR, CD86, and the mature DC marker CD83. Untreated DC express these cell surface markers at significantly lower levels. As expected, immature DC show a very low expression of CD83. The results in this figure are representative of three independent experiments.

We investigated whether DC treated with poly(I:C) retain their mature phenotype following cytokine withdrawal for 24 and 48 h. DC treated with poly(I:C) retained stable high expression of HLA-DR, CD83, CD86, CD80, and CD40 with no expression of CD14 when cultured without IL-4 and GM-CSF for 48 h. (Fig. 3). These results were comparable to those obtained with SACS-MCM-treated DC (data not shown). Beyond 48 h of culture without cytokines the number of mature DC decreased rapidly.

View larger version (32K): [in this window] [in a new window]   FIGURE 3. Poly(I:C)-treated DC remain stable in phenotype after cytokine withdrawal. DC were cultured from adherent monocytes as described in Fig. 2. DC were washed, resuspended in medium without cytokines, and cultured for another 48 h. Cell surface marker expression was determined 0, 24, and 48 h after cytokine withdrawal. Filled histograms depict specific Ab staining; open histograms are from isotype controls. DC treated with poly(I:C) retain significantly higher expression levels of HLA-DR, CD86, CD80, CD40, and the mature DC marker CD83 compared with untreated DC 0, 24, and 48 h after cytokine withdrawal. The results in this figure are representative of three independent experiments.

Mature DC lose their pinocytic activity (17, 18). To evaluate whether the poly(I:C)-induced phenotypic maturation of DC was accompanied by down-regulation of Ag uptake, we measured BSA-FITC uptake by untreated DC, SACS-MCM-treated DC, and poly(I:C)-treated DC. In contrast to untreated DC, BSA-FITC uptake was markedly down-regulated in poly(I:C)-treated DC, comparable to that in DC cultured with SACS-MCM (Fig. 4). The uptake of BSA-FITC is a metabolically active process; metabolically inactive cells, incubated on ice, did not take up BSA-FITC (Fig. 4, open histograms).

View larger version (23K): [in this window] [in a new window]   FIGURE 4. Poly(I:C)-treated DC down-regulate pinocytic activity. DC, cultured from adherent monocytes as described in Fig. 2, were allowed to pinocytose BSA-FITC for 60 min at either 37°C or on ice. Filled histograms depict specific BSA-FITC uptake at 37°C; open histograms are from controls incubated on ice. DC treated with poly(I:C) or SACS-MCM show markedly decreased pinocytic capacity compared with untreated DC.

Mature DC can produce both IL-12 and IL-10 depending on the maturation stimulus. We tested IL-12 and IL-10 production by DC upon poly(I:C) treatment. No poly(I:C) treatment resulted in the absence of IL-12 (Fig. 5A) or IL-10 (Fig. 5B) production. Indeed, poly(I:C)-treated DC produce a high level of IL-12 p40 and p70. IL-12 secretion increased until 48 h and remained stable after 72 h. Poly(I:C)-treated DC did not produce significant amounts of IL-10 (Fig. 5B). The SACS-MCM contained high amounts of IL-10 (data not shown).

View larger version (24K): [in this window] [in a new window]   FIGURE 5. Poly(I:C)-treated DC produce high levels of IL-12 and low levels of IL-10. DC were cultured from adherent monocytes in the presence of GM-CSF and IL-4 until day 7 and matured with or without poly(I:C) (12.5 µg/ml). nd = not done. A, IL-12 p40 and IL-12 p70 production was measured by specific ELISA after 24, 48, and 72 h of culture with poly(I:C). IL-12 production increases until 48 h and remains stable at 72 h in poly(I:C)-treated DC, whereas in untreated DC no IL-12 is produced. B, IL-12 p40 and IL-10 production was measured in three individuals using specific ELISA after 48 h of culture with or without poly(I:C). Both untreated and poly(I:C)-treated DC produce insignificantly low levels of IL-10.

The Ag-presenting capacity of poly(I:C)-treated DC to naive cord blood responder T cells was assayed in a primary allo MLR. The stimulatory properties of poly(I:C)-treated DC were compared with those of PBL, monocytes, immature DC, and SACS-MCM-treated DC. As depicted in Fig. 6A, poly(I:C)- or SACS-MCM-treated DC induced comparable allo mixed lymphocyte reactions that were most markedly seen with 15,000 DC/well, in a 1:10 stimulator:responder ratio. Immature DC and monocytes were clearly inferior in stimulating capacity at all stimulator:responder ratios. Thus, poly(I:C) treatment induced DC that were potent inducers of primary allo T cell responses. The stimulation of low, but significant, T cell proliferation in an auto mixed lymphocyte reaction is a property described only for mature DC (19). Indeed, DC treated either with poly(I:C) or with SACS-MCM, but not monocytes or immature DC, were able to induce a strong auto mixed lymphocyte reaction (Fig. 6B).

View larger version (13K): [in this window] [in a new window]   FIGURE 6. Poly(I:C)-treated DC are efficient APC in primary MLR and in presentation of peptide Ags to specific CTL. DC, matured with either poly(I:C) or SACS-MCM, were used as stimulator cells. Immature DC, PBMC, and monocytes were used as control stimulator cells. Randomly selected allogeneic cord blood T cells or autologous PBMC were used as responder cells. A, Allogeneic cord blood MLR. Poly(I:C)-treated DC (filled circles) and SACS-MCM-treated DC (filled squares) produce markedly higher counts than untreated DC, monocytes, and PBMC. B, Autologous MLR. Only poly(I:C)-treated DC (filled circles) and SACS-MCM-treated DC (filled squares) induce proliferation of autologous T cells. C, Presentation of the mHag HA-1 peptide Ag by poly(I:C)-treated DC, SACS-MCM-treated DC, and immature DC to the class I HLA-A*0201-restricted mHag HA-1-specific CTL clone was tested. HA-1 peptide was added to the assay in serial dilution. Poly(I:C)-treated DC are the most efficient DC for the presentation of soluble peptide Ag.

The potential of poly(I:C)-treated DC to present peptide Ags to CTL was evaluated using the mHag HA-1 peptide. HA-1 peptide-pulsed DC from an HLA-A*0201-positive, HA-1-negative individual were analyzed with the HA-1-specific T cell clone. Peptide-pulsed poly(I:C)-treated DC are most efficient in the induction of proliferation of mHag HA-1-specific HLA-A*0201-restricted CTL clone (Fig. 6C).

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
The effects of poly(I:C) on the maturation of immature DC were investigated and compared with the effects of SACS-MCM, which is known to induce maturation of DC. Our results demonstrate that treatment of immature DC with poly(I:C) results in DC with stable and high expression of the costimulatory molecules, CD80 and CD86, and expression of the maturation marker CD83. Accordingly, the phenotypic maturation is accompanied by down-regulation of pinocytic activity and production of high levels of IL-12 and neglectable amounts of IL-10. Moreover, poly(I:C)-treated DC are potent stimulators of primary allo mixed lymphocyte reactions and adequately present peptide Ags to HLA class I-restricted mHag-specific CTL.

The stability of the mature form of DC is crucial for in vivo application of Ag-pulsed DC for immunotherapy protocols. We show that poly(I:C)-treated DC retain their stable phenotype at least 48 h after withdrawal of cytokines.

The exact mechanism by which poly(I:C) induces maturational changes is unknown. Poly(I:C) has been described to bind to scavenger receptors of macrophages (20, 21). It is not known whether this binding is accompanied by internalization or even if internalization is required for activation. Poly(I:C)-MCM can be expected to contain residual amounts of poly(I:C). RNase treatment of poly(I:C)-MCM almost completely abrogated the maturation effect. This indicates that indirect effects, such as the induction of a complex mixture of autocrine or paracrine factors, are not sufficient for maturation induction and that the presence of poly(I:C) in the DC culture is required. These findings are consistent with observations that treatment of immature DC with single cytokines or combinations of cytokines, such as TNF-, IFN-ß, IFN-, IL-1ß, or PGE_2, induces partial and reversible maturation of DC (4, 5, 6). Induction of the production of IFN, IL-1, and nitric oxide in both human endothelial cells and murine macrophages by treatment with poly(I:C) has been shown to be regulated via activation of NF-B (22, 23). It is therefore likely that the poly(I:C) treatment of DC may result in activation of the NF-B pathway.

Depending on the factors used for the maturation, the cytokine profile of DC will be determined (18). IL-12 is an important initiator of theTh1-like T cell responses (24) involved in antiviral and antitumor immunity. IL-10 is generally regarded as a Th2-type cytokine. IL-10-producing DC have been described to drive T cells in the direction of a Th2-like immune response (25). Poly(I:C)-treated DC produce high levels of IL-12 and very low levels of IL-10, which is most relevant for the immunotherapy protocols.

The potential of clinical applicable DC in anticancer therapy has been emphasized (26). DC have been used with success in antilymphoma therapy (27). We have recently developed protocols for the ex vivo generation of cytotoxic T cell lines against hemopoietic system-restricted mHag HA-1 and HA-2 (28) to be used in antileukemia therapy (29). In the latter protocol, mature DC were isolated from peripheral blood and bone marrow, loaded with mHag HA-1 or HA-2 peptide and used as APC (29). Poly(I:C)-treated DC are attractive APC for the ex vivo generation of mHag HA-1- and HA-2-specific cytotoxic T cell lines for adoptive immunotherapy of leukemia relapse as well as in other adoptive immunotherapeutic settings. This simple method can easily be adapted to closed culture systems for the generation of large amounts of clinically applicable mature DC.

Note added in proof.

While this paper was reviewed, Cella et al. (30) reported data confirming the data described in this paper.

	Acknowledgments

 
We thank Jaqueline v. Beckhoven for kindly providing cord bloods, Delphine Rea for helpful discussion and performing the IL-12 p70 ELISA, and Frits Koning for critical review of the manuscript.

	Footnotes

 
¹ This work was supported in part by grants from the Leiden University Medical Center, the Leukemia Society of America, and the J. A. Cohen Institute for Radiopathology and Radiation Protection. B.E. is a Leukemia Society of America translational research awardee.

² Address correspondence and reprint requests to Dr. Rob M. Verdijk, Department of Immunohematology and Blood Bank, Leiden University Medical Center, Building 1, E3Q, P.O. Box 9600, 2300 RC Leiden, The Netherlands. E-mail address:

³ Abbreviations used in this paper: DC, dendritic cells; MCM, monocyte-conditioned medium; poly(I:C), polyriboinosinic polyribocytidylic acid; SACS, Staphylococcus aureus Cowan I strain Pansorbin cells; mHag, minor histocompatibility Ag.

Received for publication March 16, 1999. Accepted for publication April 7, 1999.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Steinman, R. M.. 1991. The dendritic cell system and its role in immunogenicity. Annu. Rev. Immunol. 9:271.[Medline]
2. Romani, N., S. Koide, M. Crowley, M. Witmer-Pack, A. M. Livingstone, C. G. Fathman, K. Inaba, R. M. Steinman. 1989. Presentation of exogenous protein antigens by dendritic cells to T cell clones. J. Exp. Med. 169:1169.[Abstract/Free Full Text]
3. Mehta-Damani, A., S. Markowicz, E. G. Engleman. 1994. Generation of antigen specific CD8⁺ CTLs from naive precursors. J. Immunol. 153:996.[Abstract]
4. Sallusto, F., A. Lanzavecchia. 1994. Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor . J. Exp. Med. 179:1109.[Abstract/Free Full Text]
5. Bender, A., M. Sapp, G. Schuler, R. M. Steinman, N. Bhardwaj. 1996. Improved method for the generation of dendritic cells from nonproliferating progenitors in human blood. J. Immunol. Methods 196:121.[Medline]
6. Luft, T., K. C. Pang, E. Thomas, P. Hertzog, D. N. J. Hart, J. Trapani, J. Cebon. 1998. Type I IFNs enhance the terminal differentiation of dendritic cells. J. Immunol. 161:1947.[Abstract/Free Full Text]
7. Romani, N., D. Reider, M. Heuer, S. Ebner, E. Kämpgen, B. Eibl, D. Niederwieser, G. Schuler. 1996. Generation of mature dendritic cells from human blood: An improved method with special regard to clinical applicability. J. Immunol. Methods 196:137.[Medline]
8. Ridge, J. P., F. Dirosa, P. Matzinger. 1998. A conditioned dendritic cell can be a temporal bridge between a CD4(+) T-helper and a T-killer cell. Nature 393:474.[Medline]
9. Cella, M., A. Engering, V. Pinet, J. Pieters, A. Lanzavecchia. 1997. Inflammatory stimuli induce accumulation of MHC class II complexes on dendritic cells. Nature 388:782.[Medline]
10. Sparwasser, T., E. S. Koch, R. M. Vabulas, K. Heeg, G. B. Lipford, J. W. Ellwart, H. Wagner. 1998. Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells. Eur. J. Immunol. 28:2045.[Medline]
11. Akiyama, Y., G. W. Stevenson, E. Schlick, K. Matsushima, P. J. Miller, H. C. Stevenson. 1985. Differential ability of human blood monocyte subsets to release various cytokines. J. Leukocyte Biol. 37:519.[Abstract]
12. Manetti, R., F. Annunziato, L. Tomasevic, V. Gianno, P. Parronchi, S. Romagnani, E. Maggi. 1995. Polyinosinic acid:polycytidylic acid promotes T helper type 1-specific immune responses by stimulating macrophage production of interferon- and interleukin-12. Eur. J. Immunol. 25:2656.[Medline]
13. Robinson, R. A., V. T. DeVita, H. B. Levy, S. Baron, S. P. Hubbard, A. S. Levine. 1976. A phase I-II trial of multiple-dose polyriboinosinic-polyribocytidylic acid in patients with leukemia or solid tumors. J. Natl. Cancer Inst. 57:599.
14. Salazar, A. M., H. B. Levy, S. Ondra, M. Kende, B. Scherokman, D. Brown, H. Mena, N. Martin, K. Schwab, D. Donovan, et al 1996. Long-term treatment of malignant gliomas with intramuscularly administered polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose: an open pilot study. Neurosurgery 38:1096.[Medline]
15. Den Haan, J. M. M., L. M. Meadows, W. Wang, J. Pool, E. Blokland, T. L. Bishop, C. Reinhardus, J. Shabanowitz, R. Offringa, D. F. Hunt, et al 1998. The minor histocompatibility antigen HA-1: a diallelic gene with a single amino acid polymorphism. Science 279:1054.[Abstract/Free Full Text]
16. Van Els, C. A., J. D’Amaro, J. Pool, E. Blokland, A. Bakker, P. J. van Elsen, J. J. Van Rood, E. Goulmy. 1992. Immunogenetics of human minor histocompatibility antigens: their polymorphism and immunodominance. Immunogenetics 35:161.[Medline]
17. Steinman, R. M., J. Swanson. 1995. The endocytic activity of dendritic cells. J. Exp. Med. 182:283.[Free Full Text]
18. Cella, M., D. Scheidegger, K. Palmer-Lehmann, P. Lane, A. Lanzavecchia, G. Alber. 1996. Ligation of CD40 on dendritic cells triggers production of high levels of interleukin-12 and enhances T cell stimulatory capacity: T-T help via APC activation. J. Exp. Med. 184:747.[Abstract/Free Full Text]
19. Kunz-Crow, M., H. G. Kunkel. 1982. Human dendritic cells: major stimulators of the autologous and allogeneic mixed leucocyte reactions. Clin. Exp. Immunol. 49:338.[Medline]
20. Brown, M. S., J. L. Goldstein. 1983. Lipoprotein metabolism in the macrophage: implications for cholesterol deposition in atherosclerosis. Annu. Rev. Biochem. 52:223.[Medline]
21. Yoshida, I., M. Azuma, H. Kawai, H. W. Fisher, T. Suzutani. 1992. Identification of a cell membrane receptor for interferon induction by poly rI:rC. Acta Virol. 36:347.[Medline]
22. Faruqui, T. R., P. E. DiCorleto. 1997. IFN- inhibits double-stranded RNA-induced E-selectin expression in human endothelial cells. J. Immunol. 159:3989.[Abstract]
23. Heitmeier, M. R., A. L. Scarim, J. A. Corbett. 1998. Double-stranded RNA-inducible nitric oxide synthase expression and interleukin-1 release by murine macrophages requires NFB activation. J. Biol. Chem. 273:15301.[Abstract/Free Full Text]
24. Trinchieri, G.. 1998. Interleukin-12: a cytokine at the interface of inflammation and immunity. Adv. Immunol. 70:83.[Medline]
25. Kalinski, P., C. M. Hilkens, A. Snijders, F. G. Snijdewint, and M. L. Kapsenberg. 1997. Dendritic cells, obtained from peripheral blood precursors in the presence of PGE2, promote Th2 responses. Adv. Exp. Med. Biol. 417:363–7:363.
26. Schuler, G., R. M. Steinman. 1997. Dendritic cells as adjuvants for immune-mediated resistance to tumors. J. Exp. Med. 186:1183.[Free Full Text]
27. Hsu, F. J., C. Benike, F. F. Fagnoni, T. M. Liles, D. Czerwinski, B. Taidi, E. G. Engleman, R. Levy. 1996. Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat. Med. 2:52.[Medline]
28. De Bueger, M., A. Bakker, J. J. Van Rood, F. Van der Woude, E. Goulmy. 1992. Tissue distribution of human minor histocompatibility antigens. J. Immunol. 149:1788.[Abstract]
29. Mutis, T., R. Verdijk, E. Schrama, B. Esendam, A. Brand, E. Goulmy. 1999. Feasibility of immunotherapy of relapsed leukemia with ex vivo-generated cytotoxic T lymphocytes specific for hematopoietic system-restricted minor histocompatibility antigens. Blood 93:2336.[Abstract/Free Full Text]
30. Cella, M., M. Salio, Y. Sakakibara, H. Langen, I. Julkunen, A. Lanzavecchia. 1999. Maturation, activiation, and protection of dendritic cells induced by double-stranded RNA. J. Exp. Med. 189:821.[Abstract/Free Full Text]

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				A. M. Lundberg, S. K. Drexler, C. Monaco, L. M. Williams, S. M. Sacre, M. Feldmann, and B. M. Foxwell Key differences in TLR3/poly I:C signaling and cytokine induction by human primary cells: a phenomenon absent from murine cell systems Blood, November 1, 2007; 110(9): 3245 - 3252. [Abstract] [Full Text] [PDF]

				S. McBride, K. Hoebe, P. Georgel, and E. Janssen Cell-Associated Double-Stranded RNA Enhances Antitumor Activity through the Production of Type I IFN J. Immunol., November 1, 2006; 177(9): 6122 - 6128. [Abstract] [Full Text] [PDF]

				K.-M. Lee, J. P. Forman, M. E. McNerney, S. Stepp, S. Kuppireddi, D. Guzior, Y. E. Latchman, M. H. Sayegh, H. Yagita, C.-K. Park, et al. Requirement of homotypic NK-cell interactions through 2B4(CD244)/CD48 in the generation of NK effector functions Blood, April 15, 2006; 107(8): 3181 - 3188. [Abstract] [Full Text] [PDF]

				N. Hirano, M. O. Butler, Z. Xia, S. Ansen, M. S. von Bergwelt-Baildon, D. Neuberg, G. J. Freeman, and L. M. Nadler Engagement of CD83 ligand induces prolonged expansion of CD8+ T cells and preferential enrichment for antigen specificity Blood, February 15, 2006; 107(4): 1528 - 1536. [Abstract] [Full Text] [PDF]

				V. Triantis, D. E. Trancikova, M. W. G. Looman, F. C. Hartgers, R. A. J. Janssen, and G. J. Adema Identification and Characterization of DC-SCRIPT, a Novel Dendritic Cell-Expressed Member of the Zinc Finger Family of Transcriptional Regulators J. Immunol., January 15, 2006; 176(2): 1081 - 1089. [Abstract] [Full Text] [PDF]

				N. Guriec, C. Daniel, K. Le Ster, E. Hardy, and C. Berthou Cytokine-regulated expression and inhibitory function of Fc{gamma}RIIB1 and -B2 receptors in human dendritic cells J. Leukoc. Biol., January 1, 2006; 79(1): 59 - 70. [Abstract] [Full Text] [PDF]

				G. Freer, D. Matteucci, P. Mazzetti, L. Bozzacco, and M. Bendinelli Generation of Feline Dendritic Cells Derived from Peripheral Blood Monocytes for In Vivo Use Clin. Vaccine Immunol., October 1, 2005; 12(10): 1202 - 1208. [Abstract] [Full Text] [PDF]

				K. Breckpot, J. Corthals, A. Bonehill, A. Michiels, S. Tuyaerts, C. Aerts, C. Heirman, and K. Thielemans Dendritic cells differentiated in the presence of IFN-{beta} and IL-3 are potent inducers of an antigen-specific CD8+ T cell response J. Leukoc. Biol., October 1, 2005; 78(4): 898 - 908. [Abstract] [Full Text] [PDF]

				Y. Ma and A. C. Ross The anti-tetanus immune response of neonatal mice is augmented by retinoic acid combined with polyriboinosinic:polyribocytidylic acid PNAS, September 20, 2005; 102(38): 13556 - 13561. [Abstract] [Full Text] [PDF]

				S.-Y. Dai, R. Nakagawa, A. Itoh, H. Murakami, Y. Kashio, H. Abe, S. Katoh, K. Kontani, M. Kihara, S.-L. Zhang, et al. Galectin-9 Induces Maturation of Human Monocyte-Derived Dendritic Cells J. Immunol., September 1, 2005; 175(5): 2974 - 2981. [Abstract] [Full Text] [PDF]

				Y. Ma, Q. Chen, and A. C. Ross Retinoic Acid and Polyriboinosinic:Polyribocytidylic Acid Stimulate Robust Anti-Tetanus Antibody Production while Differentially Regulating Type 1/Type 2 Cytokines and Lymphocyte Populations J. Immunol., June 15, 2005; 174(12): 7961 - 7969. [Abstract] [Full Text] [PDF]

				E. Barnes, M. Salio, V. Cerundolo, J. Medlin, S. Murphy, G. Dusheiko, and P. Klenerman Impact of Alpha Interferon and Ribavirin on the Function of Maturing Dendritic Cells Antimicrob. Agents Chemother., September 1, 2004; 48(9): 3382 - 3389. [Abstract] [Full Text] [PDF]

				S. Elsen, J. Doussiere, C. L. Villiers, M. Faure, R. Berthier, A. Papaioannou, N. Grandvaux, P. N. Marche, and P. V. Vignais Cryptic O2--generating NADPH oxidase in dendritic cells J. Cell Sci., May 1, 2004; 117(11): 2215 - 2226. [Abstract] [Full Text] [PDF]

				R. Rouas, P. Lewalle, F. El Ouriaghli, B. Nowak, H. Duvillier, and P. Martiat Poly(I:C) used for human dendritic cell maturation preserves their ability to secondarily secrete bioactive IL-12 Int. Immunol., May 1, 2004; 16(5): 767 - 773. [Abstract] [Full Text] [PDF]

				L. Wen, J. Peng, Z. Li, and F. S. Wong The Effect of Innate Immunity on Autoimmune Diabetes and the Expression of Toll-Like Receptors on Pancreatic Islets J. Immunol., March 1, 2004; 172(5): 3173 - 3180. [Abstract] [Full Text] [PDF]

				C. Fujimoto, Y. Nakagawa, K. Ohara, and H. Takahashi Polyriboinosinic polyribocytidylic acid [poly(I:C)]/TLR3 signaling allows class I processing of exogenous protein and induction of HIV-specific CD8+ cytotoxic T lymphocytes Int. Immunol., January 1, 2004; 16(1): 55 - 63. [Abstract] [Full Text] [PDF]

				S. Della Bella, S. Nicola, A. Riva, M. Biasin, M. Clerici, and M. L. Villa Functional repertoire of dendritic cells generated in granulocyte macrophage-colony stimulating factor and interferon-{alpha} J. Leukoc. Biol., January 1, 2004; 75(1): 106 - 116. [Abstract] [Full Text] [PDF]

				M. Matsumoto, K. Funami, M. Tanabe, H. Oshiumi, M. Shingai, Y. Seto, A. Yamamoto, and T. Seya Subcellular Localization of Toll-Like Receptor 3 in Human Dendritic Cells J. Immunol., September 15, 2003; 171(6): 3154 - 3162. [Abstract] [Full Text] [PDF]

				T. K. Means, F. Hayashi, K. D. Smith, A. Aderem, and A. D. Luster The Toll-Like Receptor 5 Stimulus Bacterial Flagellin Induces Maturation and Chemokine Production in Human Dendritic Cells J. Immunol., May 15, 2003; 170(10): 5165 - 5175. [Abstract] [Full Text] [PDF]

				D. Stober, I. Jomantaite, R. Schirmbeck, and J. Reimann NKT Cells Provide Help for Dendritic Cell-Dependent Priming of MHC Class I-Restricted CD8+ T Cells In Vivo J. Immunol., March 1, 2003; 170(5): 2540 - 2548. [Abstract] [Full Text] [PDF]

				L. Shi, S. H. Fatemi, R. W. Sidwell, and P. H. Patterson Maternal Influenza Infection Causes Marked Behavioral and Pharmacological Changes in the Offspring J. Neurosci., January 1, 2003; 23(1): 297 - 302. [Abstract] [Full Text] [PDF]

				K. Duperrier, A. Farre, J. Bienvenu, N. Bleyzac, J. Bernaud, L. Gebuhrer, D. Rigal, and A. Eljaafari Cyclosporin A inhibits dendritic cell maturation promoted by TNF-{alpha} or LPS but not by double-stranded RNA or CD40L J. Leukoc. Biol., November 1, 2002; 72(5): 953 - 961. [Abstract] [Full Text] [PDF]

				B. Mommaas, J. Kamp, J.-W. Drijfhout, N. Beekman, F. Ossendorp, P. van Veelen, J. den Haan, E. Goulmy, and T. Mutis Identification of a Novel HLA-B60-Restricted T Cell Epitope of the Minor Histocompatibility Antigen HA-1 Locus J. Immunol., September 15, 2002; 169(6): 3131 - 3136. [Abstract] [Full Text] [PDF]

				P. Ponsaerts, G. Van den Bosch, N. Cools, A. Van Driessche, G. Nijs, M. Lenjou, F. Lardon, C. Van Broeckhoven, D. R. Van Bockstaele, Z. N. Berneman, et al. Messenger RNA Electroporation of Human Monocytes, Followed by Rapid In Vitro Differentiation, Leads to Highly Stimulatory Antigen-Loaded Mature Dendritic Cells J. Immunol., August 15, 2002; 169(4): 1669 - 1675. [Abstract] [Full Text] [PDF]

				P. Riedl, D. Stober, C. Oehninger, K. Melber, J. Reimann, and R. Schirmbeck Priming Th1 Immunity to Viral Core Particles Is Facilitated by Trace Amounts of RNA Bound to Its Arginine-Rich Domain J. Immunol., May 15, 2002; 168(10): 4951 - 4959. [Abstract] [Full Text] [PDF]

				T. R. Gibb, D. A. Norwood Jr., N. Woollen, and E. A. Henchal Viral Replication and Host Gene Expression in Alveolar Macrophages Infected with Ebola Virus (Zaire Strain) Clin. Vaccine Immunol., January 1, 2002; 9(1): 19 - 27. [Abstract] [Full Text] [PDF]

				J. A. Hobbs, S. Cho, T. J. Roberts, V. Sriram, J. Zhang, M. Xu, and R. R. Brutkiewicz Selective Loss of Natural Killer T Cells by Apoptosis following Infection with Lymphocytic Choriomeningitis Virus J. Virol., November 15, 2001; 75(22): 10746 - 10754. [Abstract] [Full Text] [PDF]

				S. Somersan, M. Larsson, J. F. Fonteneau, S. Basu, P. Srivastava, and N. Bhardwaj Primary Tumor Tissue Lysates Are Enriched in Heat Shock Proteins and Induce the Maturation of Human Dendritic Cells J. Immunol., November 1, 2001; 167(9): 4844 - 4852. [Abstract] [Full Text] [PDF]

				P.-O. Vidalain, O. Azocar, H. Yagita, C. Rabourdin-Combe, and C. Servet-Delprat Cytotoxic Activity of Human Dendritic Cells Is Differentially Regulated by Double-Stranded RNA and CD40 Ligand J. Immunol., October 1, 2001; 167(7): 3765 - 3772. [Abstract] [Full Text] [PDF]

				J. R. Bleharski, K. R. Niazi, P. A. Sieling, G. Cheng, and R. L. Modlin Signaling Lymphocytic Activation Molecule Is Expressed on CD40 Ligand-Activated Dendritic Cells and Directly Augments Production of Inflammatory Cytokines J. Immunol., September 15, 2001; 167(6): 3174 - 3181. [Abstract] [Full Text] [PDF]

				A. Th. den Boer, L. Diehl, G. J. D. van Mierlo, E. I. H. van der Voort, M. F. Fransen, P. Krimpenfort, C. J. M. Melief, R. Offringa, and R. E. M. Toes Longevity of Antigen Presentation and Activation Status of APC Are Decisive Factors in the Balance Between CTL Immunity Versus Tolerance J. Immunol., September 1, 2001; 167(5): 2522 - 2528. [Abstract] [Full Text] [PDF]

				M. Bauer, V. Redecke, J. W. Ellwart, B. Scherer, J.-P. Kremer, H. Wagner, and G. B. Lipford Bacterial CpG-DNA Triggers Activation and Maturation of Human CD11c-, CD123+ Dendritic Cells J. Immunol., April 15, 2001; 166(8): 5000 - 5007. [Abstract] [Full Text] [PDF]

				D. Rea, M. J. E. Havenga, M. van den Assem, R. P. M. Sutmuller, A. Lemckert, R. C. Hoeben, A. Bout, C. J. M. Melief, and R. Offringa Highly Efficient Transduction of Human Monocyte-Derived Dendritic Cells with Subgroup B Fiber-Modified Adenovirus Vectors Enhances Transgene-Encoded Antigen Presentation to Cytotoxic T Cells J. Immunol., April 15, 2001; 166(8): 5236 - 5244. [Abstract] [Full Text] [PDF]

				N. Kadowaki, S. Antonenko, and Y.-J. Liu Distinct CpG DNA and Polyinosinic-Polycytidylic Acid Double-Stranded RNA, Respectively, Stimulate CD11c- Type 2 Dendritic Cell Precursors and CD11c+ Dendritic Cells to Produce Type I IFN J. Immunol., February 15, 2001; 166(4): 2291 - 2295. [Abstract] [Full Text] [PDF]

				D. Weissman, H. Ni, D. Scales, A. Dude, J. Capodici, K. McGibney, A. Abdool, S. N. Isaacs, G. Cannon, and K. Kariko HIV Gag mRNA Transfection of Dendritic Cells (DC) Delivers Encoded Antigen to MHC Class I and II Molecules, Causes DC Maturation, and Induces a Potent Human In Vitro Primary Immune Response J. Immunol., October 15, 2000; 165(8): 4710 - 4717. [Abstract] [Full Text] [PDF]

				D. Yang, Q. Chen, S. Stoll, X. Chen, O. M. Z. Howard, and J. J. Oppenheim Differential Regulation of Responsiveness to fMLP and C5a Upon Dendritic Cell Maturation: Correlation with Receptor Expression J. Immunol., September 1, 2000; 165(5): 2694 - 2702. [Abstract] [Full Text] [PDF]

				M. Larsson, D. Messmer, S. Somersan, J.-F. Fonteneau, S. M. Donahoe, M. Lee, P. R. Dunbar, V. Cerundolo, I. Julkunen, D. F. Nixon, et al. Requirement of Mature Dendritic Cells for Efficient Activation of Influenza A-Specific Memory CD8+ T Cells J. Immunol., August 1, 2000; 165(3): 1182 - 1190. [Abstract] [Full Text] [PDF]

				B. Sauter, M. L. Albert, L. Francisco, M. Larsson, S. Somersan, and N. Bhardwaj Consequences of Cell Death: Exposure to Necrotic Tumor Cells, but Not Primary Tissue Cells or Apoptotic Cells, Induces the Maturation of Immunostimulatory Dendritic Cells J. Exp. Med., February 7, 2000; 191(3): 423 - 434. [Abstract] [Full Text] [PDF]

				P. Brossart, B. Spahlinger, F. Grunebach, G. Stuhler, V. L. Reichardt, L. Kanz, W. Brugger;, T. Mutis, and E. Goulmy Induction of Minor Histocompatibility Antigen HA-1-Specific Cytotoxic T Cells for the Treatment of Leukemia After Allogeneic Stem Cell Transplantation Blood, December 15, 1999; 94(12): 4374 - 4376. [Full Text] [PDF]

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