Generation and functional characterisation of dendritic cells from patients with pancreatic carcinoma with special regard to clinical applicability.
BACKGROUND
We studied dendritic cell (DC) function in patients affected by pancreatic carcinoma, and the possibility of obtaining DC adequate for immunological treatment modalities.


METHODS
Leucocytes were isolated from buffy coats obtained by autotransfusion of six patients undergoing pancreatico-duodenectomy. The leucocytes were cryopreserved and, after thawing, were purified by density gradient and/or plastic adhesion. They were then cultured in vitro in cytokine-enriched medium (granulocyte/macrophage-colony-stimulating factor + interleukin-4) with different sources of serum: 10% fetal calf serum (FCS), 2% autologous human serum or 2% pooled human AB serum.


RESULTS
The DC obtained were identical to those from healthy donors in terms of phenotype, antigen uptake capacity, capacity for antigen presentation and their capacity to mature after exposure to stimuli like CD40L. DC differentiated in human serum demonstrated more mature behaviour than did DC cultured in FCS but, after exposure to CD40L, this difference disappeared. In one patient soluble factors in serum were able to inhibit the capacity of DC to stimulate T cells.


CONCLUSIONS
It's possible to obtain DC from autotransfusion of patients with pancreatic carcinoma: these cells do not show evident quantitative or qualitative alterations, are able to present soluble antigen even when cultured in the presence of human serum and may be used in immunological tumour treatments.
Piemonti
L
L
Surgical Department , S Raffaele Scientific Institute, Milan, Italy. piemonti.lorenzo@hsr.it
Monti
P
P
Zerbi
A
A
Balzano
G
G
Allavena
P
P
Di Carlo
V
V
D000230:Adenocarcinoma / Q000097:blood / Q000276:immunology / Q000473:pathology*; D000818:Animals; D017951:Antigen Presentation / Q000187:drug effects; D015703:Antigens, CD / Q000032:analysis; D018122:B7-1 Antigen / Q000032:analysis; D051940:B7-2 Antigen; D001685:Biological Factors / Q000097:blood / Q000494:pharmacology; D001790:Blood Physiological Phenomena; D023201:CD40 Ligand / Q000494:pharmacology; D002417:Cattle; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured / Q000187:drug effects; D018920:Coculture Techniques; D004396:Coloring Agents / Q000378:metabolism; D015925:Cryopreservation; D003470:Culture Media / Q000494:pharmacology; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology*; D003911:Dextrans / Q000378:metabolism; D004705:Endocytosis; D005312:Fetal Blood / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006680:HLA Antigens / Q000032:analysis; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive; D015847:Interleukin-4 / Q000494:pharmacology; D007546:Isoquinolines / Q000378:metabolism; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000032:analysis; D010190:Pancreatic Neoplasms / Q000097:blood / Q000276:immunology / Q000473:pathology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013045:Species Specificity; D013601:T-Lymphocytes / Q000276:immunology; D013744:Tetanus Toxin / Q000276:immunology
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D001685:Biological Factors; C497964:CD86 protein, human; D004396:Coloring Agents; D003470:Culture Media; D003911:Dextrans; D006680:HLA Antigens; D007546:Isoquinolines; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D013744:Tetanus Toxin; C015219:fluorescein isothiocyanate dextran; D023201:CD40 Ligand; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C017475:lucifer yellow; D016650:Fluorescein-5-isothiocyanate