In Vitro Propagated Dendritic Cells from Patients with Human-Papilloma Virus-Associated Preneoplastic Lesions of the Uterine Cervix: Use of Flt3 Ligand
[en] Dendritic cells (DC) are the most efficient antigen presenting cells. The clinical use of DC as vectors for antitumor and anti-infectious disease immunotherapy has been limited by their low level and accessibility in normal tissue. Substantial numbers of DC can be generated from peripheral blood cultured in the presence of interleukin-4 (IL-4) and granulocyte/macrophage-colony-stimulating factor (GM-CSF). We showed in this study that substantial numbers of DC can be obtained from the peripheral blood of patients with (pre)neoplastic lesions of the uterine cervix. The procedure required relatively small blood samples (10 ml) and the presence of 100 U/ml IL-4 and 800 U/ml GM-CSF in the culture medium. There was no significant difference in the morphology, yield, phenotype and function of generated DC between patients with cervical (pre)neoplastic lesions and healthy individuals. When the hematopoietic factor Flt3 ligand (Flt3L, 40 ng;ml) was added, there was an average increase in the DC population of 26% compared to cultures with GM-CSF and IL-4 alone. Approximately 1.2 x 10(6) cells with the characteristics of dendritic cells could be obtained when Flt3L was included in the medium. The addition of Flt3L did not modify the phenotypic profile of DC (HLA-DR+, CD1a+, CD4+, CD54+, CD80+, CD86+. CD40+, CD3- and CD14-). In addition, Flt3L generated functional DC capable of stimulating the proliferation of alloreactive T cells. These results suggest that Flt3L, in association with GM-CSF and IL-4, provides an advantageous tool for the large-scale generation of DC and that an immunotherapy based on the use of DC generated in vitro is possible in patients with (pre)neoplastic lesions of the uterine cervix.
Disciplines :
Reproductive medicine (gynecology, andrology, obstetrics)
Author, co-author :
Hubert, Pascale ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Greimers, Roland ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Franzen-Detrooz, E.
Sele, Jocelyne ; Centre Hospitalier Universitaire de Liège - CHU > Pneumologie-Allergologie
Delanaye, Pierre ; Centre Hospitalier Universitaire de Liège - CHU > Néphrologie
Boniver, Jacques ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Delvenne, Philippe ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Language :
English
Title :
In Vitro Propagated Dendritic Cells from Patients with Human-Papilloma Virus-Associated Preneoplastic Lesions of the Uterine Cervix: Use of Flt3 Ligand
Al-Salch W, Delvenne P, Arrese Estrada J, Nikkels AF, Pierard JE, Boniver J (1995) Inverse modulation of intraepithelial Langerhans' cell and stromal macrophage dendrocyte populations in human papillomavirus-associated squamous intraepithelial lesions of the cervix. Virchows Arch 427:41
Al-Saleh W, Giannini S, Jacobs N, Moutschen M, Doyen J, Boniver J, Delvenne P (1998) Correlation of T helper secretory differentiation and types of antigen-presenting cells in squamous intraepithelial lesions of the uterine cervix. J Pathol 184:283
Bernhard H, Disis ML, Heimfeld S, Hand S, Gralow JR, Cheever MA (1995) Generation of immunostimulatory dendritic cells from human CD34+ hematopoietic progenitor cells of the hone marrow and peripheral blood. Cancer Res 55:1099
Bhardwaj N, Young JW, Nisanian AJ, Baggers J, Steinman RM (1993) Small amounts of superantigen, when presented on dendritic cells, are sufficient to initiate T cell responses. J Exp Med 178:633
Castello G, Esposito G, Giovanni S, Mora LD, Abate G, Germano A (1986) Immunological abnormalities in patients with cervical carcinoma. Gynecol Oncol 25:61
Caux C, Vanbervliet B, Massacrier C, Azuma M, Okumura K, Lanier L, Banchereau J (1994) B70/B7-2 is identical to CD86 and is the major functional ligand for CD28 expressed on human dendritic cells. J Exp Med 180:1841
Caux C, Liu YJ, Banchereau J (1995) Recent advances in the study of dendritic cells and follicular dendritic cells. Immunol Today 16:2
Di Girolama W, Goni J, Laguens RP (1985) Langerhans' cells in squamous metaplasia of the human uterine cervix. Gynecol Obstet Invest 19:38
Evans EM, Man S, Evans AS, Borysiewicz LK (1997) Infiltration of cervical cancer tissue with human papillomavirus-specific cytotoxic T-lymphocytes. Cancer Res 57:2943
Freudenthal PS, Steinman RM (1990) The distinct surface of human blood dendritic cells, as observed alter an improved isolation method. Proc Natl Acad Sci USA 87:7698
Gabbianelli M, Pelosi E, Montesoro E, Valtieri M, Luchetti L, Samoggia P, Vitelli L, Barberi T, Testa U, Lyman S, Peschle C (1995) Multi-level of flt3 ligand on human hematopoiesis: expansion of putative stem cells and proliferation of granulomonocytic progenitors monocytic precursors. Blood 86: 1661
Gabrilovich D, Corak J, Ciernik IF, Kavanaugh D, Carbone DP (1997) Decreased antigen presentation by dendritic cells in patients with breast cancer. Clin Cancer Res 3:483
Giannini SL, Al-Saleh W, Piron H, Jacobs N, Doyen J, Boniver J, Delvenne P (in press) Cytokine expression in squamous intraepithelial lesions of the uterine cervix: implications for the generation of local immunosuppression. Clin Exp Immunol
Heufler C, Koch F, Schuler G (1988) Granulocyte/macrophage colony-stimulating factor and interleukin 1 mediate the maturation of murine epidermal Langerhans cells into potent immuno-stimulatory dendritic cells. J Exp Med 167:700
Hsu FJ, Benike C, Fagnoni F, Liles TM, Czerwinski D, Taidi B, Engelman EG, Levy R (1996) Vaccination of patients wilh B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nature Med 2:52
Hughes RG, Norval M, Howie SEM (1988) Expression of major histocompatibility class II antigens by Langerhan's cells in cervical intraepithelial neoplasia. J Clin Pathol 41:253
Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, Meramatsu S, Steinamn RM (1992) Generation of large numbers of dendritic cells from mouse hone marrow cultures supplemented with granulocyte macrophage colony-stimulating factor. J Exp Med 176:1693
Jacobs N, Giannini SL, Doyen J, Baptista A, Moutschen M, Boniver J, Delvenue P (1998) Inverse modulation of IL-10 and IL-12 in the blood of women with preneoplastic lesions of the uterine cervix. Clin Exp Immunol 111:219
Jansen JH, Wientjens G-JHM, Fibbe WE, Willemze R, Kluin-Nelemans HC (1989) Inhibition of human macrophage colony formation by interleukin 4. J Exp Med 170:577
Lehtinen M, Rantala I, Toivonen A, Luoto H, Aine R, Lauslahti K, Ylä-Outinen A, Romppanen U, Paavonen J (1993) Depletion of Langerhans cells in cervical HPV infection is associated with replication of the virus. APMIS 101:833
Lorincz AT, Temple GF, Kurman RJ, Jenson AB, Lancaster WD (1987) Oncogenic association of specific human papillomavirus types with cervical neoplasia. J Natl Cancer Inst 79:671
Lyman SD (1966) Biology of Flt3 ligand and receptor. Int J Hematol 62:63
Lyman SD, James L, VandenBos T, de Vries P, Brasel K, Gliniak B, Hollingsworth LT, Picha KS, McKenna HJ, Splett RR, Fletcher FF, Maraskovsky E, Farrah T, Foxworthe D, Williams DE, Beckmann MP (1933) Molecular cloning of a ligand for the flt3/flk-2 tyronise kinase receptor:a proliferative factor for primitive hematopoietic cells. Cell 75:1157
Maraskovsky F, Brasel K, Teepe M, Roux ER, Lyman SD, Shortman K, McKenna HJ (1996) Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice:multiple dendritic cells subpopulations identified. J Exp Med 184:1953
Mayordomo JI, Zorina T, Storkus WJ, Zitvonel L, Celluzzi C, Falo LD, Melief CJ, Ildstad ST, Kast WM, Deleo AB, Lotze MT (1995) Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity. Nat Med 1:1297
McArdle JP, Muller HK (1986) Quantitative assessement of Langerhans' cells in human cervical intraepithelial neoplasia and wart virus infection. Am J Obstet Gynecol 154:509
Moore TA, Zlotnik A (1997) Differential effects of Flk-2/Flt-3 ligand and stem cell factor on murine thymic progenitor cells. J Immunol 158:4187
Morelli A, Larregina A, Dipaola G, Fainboim L (1991) Intraepithelial CD1 positive Langerhans cell precursor in transformation zone squamous metaplasia. Cervix 9:69
Morelli AE, Sananes C, Di Paola G, Paredes A, Fainboim L (1993) Relationship between types of human papillomavirus and Langerhans' cells m cervical condyloma and intraepithelial neoplasia. Am J Clin Pathol 99:200
Nuovo GJ, Friedman D (1990) In situ hybridization analysis of HPV DNA segregaton patterns in lesions of the female genital tract. Gynecol Oncol 36:256
Peters JH, Gieseler R, Thiele B, Steinbach F (1996) Dendritic cells:from ontogenetic orphans to myelomonocytic descendants. Immunol Today 17:273
Radmayr C, Böck G, Hobisch A, Klocker H, Bartsch G, Thurnher M (1995) Dendritic antigen-presenting cells from the peripheral blood of renal-cell-carcinoma patients. Int J Cancer 63:627
Reid CDL, Fryer PR, Clifford C, Kirk A, Tikerpae J, Knight SC (1990) Identification of hematopoietic progenitors of macrophages and dendritic Langerhans cells (DL-CFU) in human bone marrow and peripheral blood. Blood 76:1139
Reid CDL, Stackpoole A, Meager A, Tikerpae J (1992) Interactions of tumor necrosis factor with granulocyte-macrophage colony-stimulating factor and other cytokines in the regulation of dendritic cell growth in vitro from early bipotent CD34 progenitors in human bone marrow. J Immunol 149:2681
Romani N, Schuler G (1992) The immunologie properties of epidermal Lungerhans cells as a part of the dendritic cell system. Springer Semin Immunopathol 13:265
Romani N, Gruner S, Brang D, Kämpgen E, Lenz A, Trockenbacher B, Konwalinka G, Fritsh PO, Steinman RM, Schuler G (1994) Proliferating dendritic-cell progenitors in human blood. J Exp Med 180:83
Rosini S, Caltagirone S, Tallini G, Lattanzio G, Demopoulos R, Piantelli M, Musiani P (1996) Depletion of stromal and intraepithelial antigen-presenting cells in cervical neoplasi in human immunodeficiency virus infection. Hum Pathol 27:834
Sallusto F, Lanzavecchia A (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 alpha. J Exp Med 179:1109
Santiago-Schwarz F, Belilos E, Diamond B, Carsons SE (1992) TNF in combination with GM-CSF enhances the differentiation of neonatal cord blood stem cells into dendritic cells and macrophages. J Leukoc Biol 52:274
Shurin MR, Pandharipande PP, Zorina TD, Haluszczak C, Subbotin VM, Hunter O, Brumfield A, Storkus WJ, Maraskovsky E, Lotze MT (1997) FLT3 ligand induces the generation of functionally active dendritic cells in mice. Cell Immunol 179:174
Siena S, Di Nicola M, Bregni M, Mortarini R, Anichini A, Lombardi L, Ravagnani F, Parmiani G, Gianni AM (1995) Massive ex vivo generation of functional dendritic cells from mobilized CD34 blood progenitors for anticancer therapy. Exp Hematol 23:1463
Small D, Levenstein M, Kim E, Carow C, Amin S, Rockwell P, Witte L, Burrow C, Ratajczak MZ, Gewirtz AM, Civin CI (1994) STK-I, the human homolog of flk2 flt3, is selectively expressed in CD34+ human hone marrow cells and is involved in the proliferation of early progenitor stem cells. Proc Natl Acad Sci USA 91:459
Smolle J, Soyer HP, Ehall R, Bartenstein S, Kerl H (1986) Langerhans cell density in epithelial differentiation hut not with the peritumoral infiltrate. J Invest Dermatol 87:477
Sornasse T, Flamand V, Becker GD, Bazin H, Tielemans F, Thielemans K, Urbain J, Leo O, Moser M (1992) Antigen-pulsed dendritic cells can efficiently induce an antibody response in vivo. J Exp Med 175:15
Steinman RM, Witmer MC (1978) Lymphoid dendritic cells are potent stimulators of the primary mixed leukocyte reaction in mice. Proc Natl Acad Sci USA 75:5132
Tang A, Amagai M, Granger LG, Stanley JR, Udey MC (1993) Adhesion of epidermal Langerhans cells to keratinoeytes mediated by E-cadherin. Nature 361:82
Tay SK, Jenkins D, Maddox P, Campion M, Singer A (1987) Lymphocyte phenotypes in cervical intraepithelial neoplasia and human papillomavirus infection. Br J Obstet Gynaecol 94:16
Thivolet J, Viac J, Staquet MG (1982) Cell mediated immunity in wart virus infection. Int J Dermatol 21:94
Young JW, Inaba K (1996) Dendritic cells as adjuvants for class I major histocompatibility complex-restricted antitumour immunity. J Exp Med 183:7
Zitvogel L, Mayordomo JI, Tjandrawan T, Deleo AB, Clarke MR, Lotze MT, Storkus WJ (1996) Therapy of murine tumors with tumor peptide-pulsed dendritic cells:dependenee on T cells, B7 costimulation, and T helper cell I associated cytokines. J Exp Med 183:87
