Transgenic expression of CD40L and interleukin-2 induces an autologous antitumor immune response in patients with non-Hodgkin's lymphoma.
The malignant B cells of non-Hodgkin's lymphoma (B-NHL cells) express peptides derived from tumor-specific antigens such as immunoglobulin idiotypes, and also express major histocompatibility complex antigens. However, they do not express co-stimulatory molecules, which likely contributes to their protection from host antitumor immunity. To stimulate NHL-specific immune responses, we attempted to transfer the human CD40 ligand (hCD40L) gene to B-NHL cells and enhance their co-stimulatory potential. We found that an adenoviral vector encoding human CD40L (AdhCD40L) was ineffective at transducing B-NHL cells because these cells lack the coxsackievirus B-adenovirus receptor and alpha(v) integrins. However, preculture of the B-NHL cells with the human embryonic lung fibroblast line, MRC-5, significantly up-regulated expression of integrin alpha(v)beta 3 and markedly increased their susceptibility to adenoviral vector transduction. After prestimulation, transduction with AdhCD40L increased CD40L expression on B-NHL cells from 1.3+/-0.2% to 40.8+/-11.9%. Transduction of control adenoviral vector had no effect. Expression of transgenic human CD40L on these CD40-positive cells was in turn associated with up-regulation of other co-stimulatory molecules including B7-1/-2. Transduced B-NHL cells were now able to stimulate DNA synthesis of autologous T cells. However, the stimulated T cells were unable to recognize unmodified lymphoma cells, a requirement for an effective tumor vaccine. Based on previous results in an animal model, we determined the effects of combined use of B-NHL cells transduced with AdhCD40L and AdhIL2 vectors. The combination enhanced initial T-cell activation and generated autologous T cells capable of specifically recognizing and killing parental (unmodified) B-NHL cells via major histocompatibility complex--restricted cytotoxic T lymphocytes. These findings suggest that the combination of CD40L and IL2 gene-modified B-NHL cells will induce a cytotoxic immune response in vivo directed against unmodified tumor cells.
Takahashi
S
S
Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.
Yotnda
P
P
Rousseau
R F
RF
Mei
Z
Z
Smith
S
S
Rill
D
D
Younes
A
A
Brenner
M K
MK
D000256:Adenoviridae / Q000235:genetics*; D000911:Antibodies, Monoclonal; D001324:Autoantigens / Q000276:immunology*; D023201:CD40 Ligand / Q000235:genetics* / Q000276:immunology; D018920:Coculture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry; D006801:Humans; D016023:Integrins / Q000378:metabolism; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics* / Q000276:immunology; D008212:Lymphocyte Depletion; D016393:Lymphoma, B-Cell / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D011991:Receptors, Virus / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D019076:Transgenes / Q000235:genetics*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D016023:Integrins; D007376:Interleukin-2; D011991:Receptors, Virus; D023201:CD40 Ligand; D007371:Interferon-gamma