Phage libraries for generation of clinically useful antibodies

Chester, Kerry; Begent, R. H. J.; Robson, L; Keep, P. A.; Pedley, RB; LIBiol, J.A. Boden; Boxer, G. M.; Green, A.; Winter, Greg; Cochet, Olivier; Hawkins, Robert E.

doi:10.1016/s0140-6736(94)92695-6

Cited by 163 publications

(70 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our study, the chimeric receptor consisted of an extracellular antigen-binding domain (scFv) specific for CEA (Chester et al, 1994;Begent et al, 1996), attached to the CD3z component of the T-cell receptor complex. This CIR has already been demonstrated to be effective in initiating cytokine secretion and the cytotoxic activities of normal donor T lymphocytes .…”

Section: Discussionmentioning

confidence: 99%

“…The CIRs used in this study were either an anti-CEA-specific scFv, MFE23 (derived by phage display technology) (Chester et al, 1994;Begent et al, 1996) or a control scFv (anti-neural cell adhesion molecule (NCAM)) (D29) (Whittington et al, 2001) fused to the human CD3z protein. Both receptors have been shown to be active when introduced into primary human T lymphocytes derived from healthy donors .…”

mentioning

confidence: 99%

See 1 more Smart Citation

T lymphocytes isolated from patients with advanced colorectal cancer are suitable for gene immunotherapy approaches

et al. 2003

View full text Add to dashboard Cite

Despite improvements in treatment, the 5-year survival for metastatic colorectal cancer remains poor. Novel approaches such as gene immunotherapy are being investigated to improve treatment. Retroviral gene transfer methods have been shown to transduce primary human T lymphocytes effectively resulting in the expression of therapeutic genes. However, a number of defects have been identified in T lymphocytes isolated from patients bearing tumour, which may have critical implications for the development of genetargeted T cells as an anticancer therapy. To address this issue, primary T lymphocytes were isolated from patients with advanced colorectal cancer and tested for their ability to be transduced and to express subsequently a chimeric immune receptor consisting of a single-chain antibody fragment antigen-binding moiety specific for carcinoembryonic antigen (CEA) fused to the T cell receptor (TCR) CD3z chain. In 10 out of 10 patients, T lymphocytes were transduced, expanded in the absence of selection and tested for functional activity against CEA-expressing tumour cells. In each case, functional-specific cytotoxic activity was observed. Negligible activity was found in control cultures. This study highlights the feasibility of patient-derived T lymphocytes as a source of immune cells for autologous gene immunotherapy approaches.

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

T lymphocytes isolated from patients with advanced colorectal cancer are suitable for gene immunotherapy approaches

et al. 2003

View full text Add to dashboard Cite

show abstract

“…A 767-bp fragment of DNA coding for scFv of anti-CEA and a marker epitope from c-myc were amplified by PCR from the MFE-23 vector (28) and subcloned into XbaI/BstEII-digested pRSVscFv␥R (a kind gift from Z. Eshhar, Weizmann Institute, Rehovot, Israel). The chimeric gene constructs were composed of the scFv-anti-CEA mAb, a membrane proximal hinge region of human CD8, and one of the transmembrane and cytoplasmic regions of the human TCR-chain (scFv-anti-CEA-), the transmembrane and cytoplasmic regions of the mouse CD28 signaling chain (scFvanti-CEA-CD28), or the transmembrane and cytoplasmic regions of the mouse CD28 signaling chain fused to the cytoplasmic region of TCR-(scFv-anti-CEA-CD28-) (Fig.…”

Section: Chimeric Receptor Gene Constructionmentioning

confidence: 99%

Rejection of Syngeneic Colon Carcinoma by CTLs Expressing Single-Chain Antibody Receptors Codelivering CD28 Costimulation

Haynes

Trapani

Teng

et al. 2002

The Journal of Immunology

View full text Add to dashboard Cite

A new strategy to improve the therapeutic utility of redirected T cells for cancer involves the development of novel Ag-specific chimeric receptors capable of stimulating optimal and sustained T cell antitumor activity in vivo. Given that T cells require both primary and costimulatory signals for optimal activation and that many tumors do not express critical costimulatory ligands, modified single-chain Ab receptors have been engineered to codeliver CD28 costimulation. In this study, we have compared the antitumor potency of primary T lymphocytes expressing carcinoembryonic Ag (CEA)-reactive chimeric receptors that incorporate either TCR-ζ or CD28/TCR-ζ signaling. Although both receptor-transduced T cell effector populations demonstrated cytolysis of CEA+ tumors in vitro, T cells expressing the single-chain variable fragment of Ig (scFv)-CD28-ζ chimera had a far greater capacity to control the growth of CEA+ xenogeneic and syngeneic colon carcinomas in vivo. The observed enhanced antitumor activity of T cells expressing the scFv-CD28-ζ receptor was critically dependent on perforin and the production of IFN-γ. Overall, this study has illustrated the ability of a chimeric scFv receptor capable of harnessing the signaling machinery of both TCR-ζ and CD28 to augment T cell immunity against tumors that have lost expression of both MHC/peptide and costimulatory ligands in vivo.

show abstract

“…Originally identified by phage library selection, this scFv was later humanized by resurfacing and engineered in yeast for greater stability and solubility (shMFE) as well as affinity (sm3E) (31)(32)(33). Both of these engineered molecules are well expressed in yeast and have K d values of ϳ7 nM and ϳ30 pM, respectively.…”

mentioning

confidence: 99%

Convergent Potency of Internalized Gelonin Immunotoxins across Varied Cell Lines, Antigens, and Targeting Moieties

Pirie¹,

Hackel²,

Rosenblum

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Gelonin-based immunotoxins vary widely in their cytotoxic. Results were matched with cytotoxicity measurements made at equivalent concentration and exposures. Unexpectedly, when matched internalization and cytotoxicity data were combined, a conserved internalized cytotoxicity curve was generated that was common across experimental conditions. Considerable variations in antigen expression, trafficking kinetics, extracellular immunotoxin concentration, and exposure time were all found to collapse to a single potency curve on the basis of internalized immunotoxin. Fifty percent cytotoxicity occurred when ϳ5 ؋ 10 6 toxin molecules were internalized regardless of the mechanism of uptake. Cytotoxicity observed at a threshold internalization was consistent with the hypothesis that endosomal escape is a common, highly inefficient, rate-limiting step following internalization by any means tested. Methods designed to enhance endosomal escape might be utilized to improve the potency of gelonin-based immunotoxins.Immunotoxins are a promising approach to the targeted delivery of highly potent, cancer-specific, cytotoxic agents. Immunotoxins are frequently composed of a targeting moiety (derived from antibodies or other cell-binding proteins) either chemically conjugated or genetically fused to highly cytotoxic plant or bacterial protein toxins. Clinical success for immunotoxins has been mostly limited to hematological malignancies due to transport limitations in solid tumors (1). Such limitations have been extensively studied experimentally (2) and with several computational models (3, 4).The potency of a particular immunotoxin is dependent on the ability to deliver the toxin to the cytoplasm, which is commonly considered to be the rate-limiting step. For some native toxins such as ricin, intracellular delivery is achieved through lectin binding, followed by internalization and toxin release with membrane fusion or retrograde trafficking (5). Immunotoxins attempt to recreate this scenario by replacing the indiscriminate lectin binding with cancer-specific antigen binding as a means of targeting and internalization (6). Subsequent intracellular trafficking, release, and endosomal escape are often achieved using existing toxin characteristics, translocation domains, protease cleavage sites, disulfide bonds, and/or signaling peptides (7-10). However, the inclusion of toxins with domains facilitating cytoplasmic access can also lead to increased nonspecific toxicity in vivo (11,12).Gelonin is a plant toxin and classified as a type I ribosomeinactivating protein because it lacks any cell-binding or cytoplasmic delivery domains. Recombinant gelonin (rGel) 2 is an ϳ30-kDa N-glycosidase with activity similar to the ricin A chain but exhibiting better stability and lower immunogenicity (13,14). The use of rGel in tumor-targeted cytotoxic agents has been well studied (15, 16). Furthermore, rGel has been shown to be active without cleavage from the binding domain and without negative impact on the targeting agent's pharmacokinetics (...

show abstract

Phage libraries for generation of clinically useful antibodies

Cited by 163 publications

References 4 publications

T lymphocytes isolated from patients with advanced colorectal cancer are suitable for gene immunotherapy approaches

T lymphocytes isolated from patients with advanced colorectal cancer are suitable for gene immunotherapy approaches

Rejection of Syngeneic Colon Carcinoma by CTLs Expressing Single-Chain Antibody Receptors Codelivering CD28 Costimulation

Convergent Potency of Internalized Gelonin Immunotoxins across Varied Cell Lines, Antigens, and Targeting Moieties

Contact Info

Product

Resources

About