Preparation of purified lymphoma cells suitable for therapy.
BACKGROUND
Very few tumoral Ags have yet been isolated in NHL B cells. It is nevertheless possible to use whole tumor cells as a source of tumor Ags. We describe the purification of large numbers of human NHL B cells directly from lymph node or spleen biopsies, and different preparations allowing their use in a clinical setting.


METHODS
The purification procedure consists of the negative selection of tumor B cells: cells to be eliminated are opsonized by CD2 Abs, and then coupled to magnetic beads for separation by the Isolex 300 magnetic separator.


RESULTS
The mean yield of the purification was 74% for CD19+ cells, with a mean purity of 87%, dependent on the initial fraction of tumor cells in the biopsy. Using this procedure, a large number of purified tumor cells can be recovered from a biopsy in sterile conditions. We also describe treatments of B cells that can enhance their uptake by APCs, a critical step in anti-tumor immunotherapy strategies. Cells were opsonized by rituximab, or induced in apoptosis by irradiation, or necrosis by heating. Cell lysates were directly prepared from purified tumor cells.


CONCLUSIONS
These procedures were reproducible on every lymphoma cell, and treated cells were phagocytosed by APCs. The methodology described here allows the evaluation of the immunological potential of apoptotic, necrotic, opsonized lymphoma cells, or their lysates, in a clinical setting.
Lui
G
G
Department of Research and Development, Research Group on Lymphoma, EFS Rhône-Alpes Grenoble, La Tronche, France.
Manches
O
O
Chaperot
L
L
Ducrot
T
T
Molens
J-P
JP
Sotto
J-J
JJ
Bensa
J-C
JC
Plumas
J
J
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D017951:Antigen Presentation / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology; D017209:Apoptosis / Q000528:radiation effects; D001402:B-Lymphocytes / Q000276:immunology*; D001706:Biopsy; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D016393:Lymphoma, B-Cell / Q000276:immunology* / Q000473:pathology* / Q000628:therapy; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology* / Q000473:pathology* / Q000628:therapy; D009336:Necrosis; D010587:Phagocytosis / Q000276:immunology; D000069283:Rituximab; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013347:Subcellular Fractions / Q000276:immunology
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D000951:Antigens, Neoplasm; D000069283:Rituximab