Differentiation of antitumor-specific cytotoxic T lymphocytes from autologous tumor infiltrating lymphocytes in non-Hodgkin's lymphomas.
The present study describes a new culture protocol allowing the activation and proliferation of autologous tumor infiltrating T lymphocytes (TIL), and the generation of antitumor specific CTL in non-Hodgkin's lymphoma (NHL). Cells from eight patients with indolent NHL were used. We performed 3-week co-cultures of TIL with irradiated autologous malignant B cells in the presence of low doses of IL-1beta, IL-2 and IL-12. The proliferation, phenotype and cytotoxicity, and antitumor specificity of T cells recovered were studied. T-cell clonality was analyzed using TCRgamma gene rearrangement amplification by a multiplex PCR. Under these culture conditions, TIL proliferated, and the CD8+ T lymphocytes that were in a minority at the beginning of the culture increased dramatically in 6 out of 8 cases. In two cases, CD4+ T lymphocytes expanded. We showed that an oligoclonal selection of reactive T cells occurred in culture. Specific cytotoxicity developed against autologous malignant B cells in the 6 cases where there was an expansion of CD8+ T lymphocytes. Inhibition experiments performed with mAb directed against HLA class I and II molecules, CD4, CD8 and TCRgammadelta showed that the cytotoxic effector cells were CD8+ T lymphocytes probably expressing TCRalphabeta+. Cytokine secretion was analyzed in culture medium, and we detected significant levels of IFN-gamma, TNF-alpha, and IL-10 and no IL-4 (except in one case). Our results demonstrate that memory T cells from lymphoma patients can be amplified and differentiated into antitumor cytotoxic cells using a combination of the cytokines IL-1beta, IL-2, and IL-12 in association with non modified tumor cells.
Chaperot
L
L
Department of Immunology, ETS Isère-Savoie, Grenoble, France.
Delfau-Larue
M H
MH
Jacob
M C
MC
Molens
J P
JP
Roussel
B
B
Agrawal
S
S
Farcet
J P
JP
Gressin
R
R
Sotto
J J
JJ
Bensa
J C
JC
Plumas
J
J
D000368:Aged; D000911:Antibodies, Monoclonal / Q000276:immunology; D017951:Antigen Presentation; D001402:B-Lymphocytes / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002999:Clone Cells / Q000166:cytology / Q000276:immunology; D018920:Coculture Techniques; D005260:Female; D015329:Gene Rearrangement, T-Lymphocyte; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory; D016219:Immunotherapy, Adoptive; D007375:Interleukin-1 / Q000494:pharmacology*; D018664:Interleukin-12 / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology*; D016246:Lymphocytes, Tumor-Infiltrating / Q000166:cytology* / Q000187:drug effects; D008222:Lymphokines / Q000378:metabolism; D016393:Lymphoma, B-Cell / Q000276:immunology / Q000473:pathology / Q000628:therapy; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology / Q000473:pathology* / Q000628:therapy; D008297:Male; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000276:immunology; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D007375:Interleukin-1; D007376:Interleukin-2; D008222:Lymphokines; D016692:Receptors, Antigen, T-Cell, gamma-delta; D018664:Interleukin-12