scholarly journals A retroviral vector system ‘STITCH’ in combination with an optimized single chain antibody chimeric receptor gene structure allows efficient gene transduction and expression in human T lymphocytes

Gene Therapy ◽  
1998 ◽  
Vol 5 (9) ◽  
pp. 1195-1203 ◽  
Author(s):  
MEM Weijtens ◽  
RA Willemsen ◽  
EH Hart ◽  
RLH Bolhuis
Keyword(s):  
T Lymphocytes ◽  
Gene Structure ◽  
Receptor Gene ◽  
Retroviral Vector ◽  
Vector System ◽  
Chimeric Receptor ◽  
Gene Transduction ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Efficient Gene

scholarly journals Antigen-dependent CD28 Signaling Selectively Enhances Survival and Proliferation in Genetically Modified Activated Human Primary T Lymphocytes

1998 ◽  
Vol 188 (4) ◽  
pp. 619-626 ◽  
Author(s):  
Anja Krause ◽  
Hong-Fen Guo ◽  
Jean-Baptiste Latouche ◽  
Cuiwen Tan ◽  
Nai-Kong  V. Cheung ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Tumor Cells ◽  
Small Cell Lung Carcinoma ◽  
Interleukin 2 ◽  
Culture Conditions ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Cell Lung Carcinoma ◽  
Antiidiotypic Antibody ◽  
Antigen Presenting

Most tumor cells function poorly as antigen-presenting cells in part because they do not express costimulatory molecules. To provide costimulation to T lymphocytes that recognize tumor cells, we constructed a CD28-like receptor specific for GD2, a ganglioside overexpressed on the surface of neuroblastoma, small-cell lung carcinoma, melanoma, and other human tumors. Recognition of GD2 was provided by a single-chain antibody derived from the GD2-specific monoclonal antibody 3G6. We demonstrate that the chimeric receptor 3G6-CD28 provides CD28 signaling upon specific recognition of the GD2 antigen on tumor cells. Human primary T lymphocytes retrovirally transduced with 3G6-CD28 secrete interleukin 2, survive proapoptotic culture conditions, and selectively undergo clonal expansion in the presence of an antiidiotypic antibody specific for 3G6-CD28. Polyclonal CD8+ lymphocytes expressing 3G6-CD28 are selectively expanded when cultured with cells expressing allogeneic major histocompatibility complex class I together with GD2. Primary T cells given such an antigen-dependent survival advantage should be very useful to augment immune responses against tumor cells.

scholarly journals Lysis of ovarian cancer cells by human lymphocytes redirected with a chimeric gene composed of an antibody variable region and the Fc receptor gamma chain.

1993 ◽  
Vol 178 (1) ◽  
pp. 361-366 ◽  
Author(s):  
P Hwu ◽  
G E Shafer ◽  
J Treisman ◽  
D G Schindler ◽  
G Gross ◽  
...  
Keyword(s):  
Ovarian Carcinoma ◽  
Human Lymphocytes ◽  
Human Tumor ◽  
Ovarian Cancer Cells ◽  
Chimeric Receptor ◽  
Single Chain ◽  
Gamma Chain ◽  
Single Chain Antibody ◽  
Ovarian Carcinoma Cell Line ◽  
Variable Regions

To expand the spectrum of recognition of effector lymphocytes and to redirect them towards predefined targets, we have altered the specificity of human tumor-infiltrating lymphocytes (TIL) through stable modification with chimeric receptor genes consisting of single-chain antibody variable regions linked to the gamma subunit common to the immunoglobulin (Ig)G and IgE Fc receptors. Using either hapten or ovarian carcinoma-specific monoclonal antibodies, we constructed chimeric receptor genes and retrovirally introduced them into CD8+ TIL. Redirected TIL specifically lysed trinitrophenyl-labeled Daudi or a human ovarian carcinoma cell line (IGROV-1), and secreted granulocyte/macrophage colony-stimulating factor upon stimulation with the appropriate antigen. This strategy may allow new approaches towards the adoptive immunotherapy of cancer in humans.

Abstract 1735: Targeted Application of Interleukin-2 Activates T-Lymphocytes and Reduces Plaque Formation

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Kristof Graf ◽  
Thore Dietrich ◽  
Christiane Schneemann ◽  
Kirstin Atrott ◽  
Philipp Stawowy ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Fusion Protein ◽  
Thoracic Aorta ◽  
Aortic Root ◽  
Cell Cancer ◽  
Interleukin 2 ◽  
Single Chain ◽  
Lesion Formation ◽  
Cellular Mechanisms ◽  
Single Chain Antibody

A newly developed fusion protein (L19-IL2) consisting of an anti-ED-B fibronectin single chain antibody and the cytokine interleukin-2 (IL2), is a potent antitumor agent for renal cell cancer. ED-B fibronectin (ED-B) is expressed in inflammatory plaque lesions. Preliminary studies suggested that delivering IL2 locally to atherosclerotic tissue seem to reduce plaque progression. Therefore we investigated the application of L19-Il2 on lesion formation in apoE-deficient mice (apoE −/− ). 6-month old apoE −/− , fed with high fat diet (n = 5/group) were injected intravenously L19-IL2 (4290IU/g bodyweight), with L19 (antibody alone), with the control fusion protein D1-IL2 (4290IU/g bodyweight) or NaCL (control) at day 1, 3 and 5 and were sacrificed at day 7. Plaque lesion formation was analyzed by histology, immunohisto-chemistry (Mac3 = macrophages, CD31; actin, human IL2, ED-B, CD25, CD4), morphometry of the aortic root and by Sudan stain of the thoracic aorta followed by densitometry. Macrophage, ED-B, CD4, CD25 (IL-2 receptor subunit) content was analysed after immunohistochemistry. Results: Treatment with L19-IL2, L19, and D1-IL2 did not affect bodyweight, survival of mice compared to control (NaCL). Serum levels for blood glucose, cholesterol, liver and cardiac enzymes, and troponin were not different between L19-IL2, L19, and NaCL. Cardiac histology was also unaffected. Human IL2 was only detectable in plaque lesions after L19-IL2, but not after L19 or D1-IL2 treatment. L19-IL2 application significantly reduced plaque lesion size in the aortic root (L19-Il2: 23.2 ± 1.7%, L19: 35.2 ± 1.2%, D1-IL2: 31.6 ± 3.0%, NaCL: 31.0 ± 3.2%, p < 0.01), plaque extension in the thoracic aorta (L19-Il2: 5.3 ± 0.7%, L19: 8.2 ± 0.4%, D1-IL2: 9.8 ± 1.0%, NaCL: 8.9 ± 1.2%, p < 0.01). Mac3 immunoreactivity was reduced under L19-IL2, whereas CD4 and CD25 were strongly increased compared to L19, D1-IL2 and NaCL treated groups. Conclusion: A fusion protein, which delivers human IL-2 into plaque lesions, induces T-lymphocyte activation, reduction of macrophages and a reduction in plaque growth. The present study suggests that local application of IL2 activates cellular mechanisms involving T-lymphocytes leading to a reduction of atherosclerotic disease.

Identification of a murine CD28 dileucine motif that suppresses single-chain chimeric T-cell receptor expression and function

Blood ◽  
2003 ◽  
Vol 102 (13) ◽  
pp. 4320-4325 ◽  
Author(s):  
Phuong Nguyen ◽  
Ioana Moisini ◽  
Terrence L. Geiger
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
T Cell Receptor ◽  
Therapeutic Potential ◽  
Cell Receptor ◽  
Receptor Expression ◽  
Chimeric Receptor ◽  
Single Chain ◽  
Dileucine Motif

AbstractRecent preclinical and clinical trials have demonstrated the therapeutic potential of T lymphocytes redirected with genetically engineered T-cell receptor (TCR) surrogates against infected, cancerous, or autoreactive cells. These surrogate TCRs link a ligand-recognition domain to signaling regions from the TCR. We previously compared the function of surrogate TCRs that include TCR or TCR and CD28 signaling regions. We found that primary murine T cells modified to specifically target Kb-restricted CD8+ T cells using either Kb-ζ or Kb-CD28-ζ receptors had similar functional activities, although the CD28-ζ receptor showed a 2-fold to 4-fold decreased expression. We have now identified a previously unrecognized dileucine motif in the murine CD28 signaling domain that accounts for this reduced expression. Inactivation of this motif increased chimeric receptor surface expression 2- to 5-fold. T cells expressing the dileucine-mutated CD28-ζ chimeric receptor demonstrated enhanced proliferation, cytokine production, and cytolytic activities. Further, cells expressing this dileucine-mutated receptor were highly effective in eliminating antigen-specific CD8+ T lymphocytes in vivo. These results therefore identify a critical motif limiting the function of receptor-modified T lymphocytes, demonstrate that inactivation of this motif enhances chimeric receptor function, and illustrate a potential novel application of receptor-modified T lymphocytes in the induction of immune tolerance. (Blood. 2003;102:4320-4325)

Retroviral Vector Targeting to Melanoma Cells by Single-Chain Antibody Incorporation in Envelope

1998 ◽  
Vol 9 (5) ◽  
pp. 737-746 ◽  
Author(s):  
F. Martin ◽  
J. Kupsch ◽  
Y. Takeuchi ◽  
S. Russell ◽  
F.-L. Cosset ◽  
...  
Keyword(s):  
Retroviral Vector ◽  
Melanoma Cells ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Vector Targeting
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