title: Immunotherapeutic potential of dendritic cells generated in long-term stroma-dependent cultures.
abstract: Long term cultures (LTC) producing dendritic cells (DC) have been established from spleen. A well developed stromal cell layer supported production of DC in numbers suitable for experimentation. Cells had obvious membrane pseudopodia and could be collected from culture every 2-3 days. Large cells produced in LTC stained with fluorescently labelled monoclonal antibodies specific for DC such as 33D1, and M1/70 which is specific for DC and myeloid cells. These staining patterns confirmed the presence of DC within the LTC population. LTC-DC were tested and shown capable of migration in vivo in B10.A(2R) mice following footpad inoculation. Most cells entered the spleen and a small number entered popliteal lymph node. LTC-DC have migratory capacity comparable with control spleen lymphocytes. LTC-DC were tested for capacity to induce an anti-tumour immune response after exposing cells to tumour cell membranes. LTC-DC pulsed with BL/VL3 tumour antigens were able to induce a BL/VL3-specific primary cytotoxic T lymphocyte (CTL) response detectable in popliteal lymph nodes and spleen of C57BL/6J mice within 6 days of priming. BL/VL3 tumour cells grew in sublethally irradiated C57BL/6J mice giving 100% mortality. Adoptive transfer of spleen cells from mice given BL/VL3 antigen-pulsed LTC-DC, two weeks previously, significantly slowed the growth of BL/VL3 tumour cells in mice. DC produced in LTC can function as antigen presenting cells (APC) when adoptively transferred into animals. Their capacity to migrate effectively, to induce a CTL response and to reduce tumour load suggests that DC grown using this in vitro system may have valuable clinical potential in humans.
authors.affiliation: Division of Biochemistry & Molecular Biology, School of Life Sciences, Australian National University, Canberra ACT, Australia. Helen.Oneill@anu.edu.au
authors.forename: H C
authors.initials: HC
authors.lastname: O'Neill
authors.forename: N
authors.initials: N
authors.lastname: Jonas
authors.forename: H
authors.initials: H
authors.lastname: Wilson
authors.forename: K
authors.initials: K
authors.lastname: Ni
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005260:Female; D007942:Leukemia, Experimental / Q000276:immunology / Q000628:therapy; D007953:Leukemia, Radiation-Induced / Q000276:immunology / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D009374:Neoplasms, Experimental / Q000276:immunology* / Q000628:therapy*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D017154:Stromal Cells / Q000166:cytology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D013945:Thymoma / Q000276:immunology / Q000628:therapy; D013953:Thymus Neoplasms / Q000276:immunology / Q000628:therapy