Development and applications of surface-linked single chain antibodies against T-cell antigens.
In this report the use of surface-linkage to expand the potential experimental and therapeutic applications of single chain antibody (scFv) constructs is reviewed. A strategy for the generation and functional characterization of surface-linked scFvs that bind selectively to the T-cell proteins CD3epsilon, CD28, and CD152 (CTLA-4) is described in detail. Experimental examples are provided of the use of these constructs to study the positive and negative regulation of T-cell activation and to manipulate the in vivo immunogenicity of tumor cells. In addition, a novel system for Simultaneous T-cell Activation and Retroviral Transduction (START) is described in which retroviral packaging cells are rendered mitogenic for T lymphocytes by combined expression of surface-linked scFvs. Finally, the use of random mutagenesis and yeast surface display to increase the affinity and functional efficacy of scFv constructs is demonstrated.
The Ben May Institute for Cancer Research, Dept. of Pathology and Committee on Immunology, University of Chicago, Chicago, IL, USA
M D
MD
Griffin
P O
PO
Holman
Q
Q
Tang
N
N
Ashourian
U
U
Korthäuer
D M
DM
Kranz
J A
JA
Bluestone
D000069594:Abatacept; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000276:immunology*; D018106:CD28 Antigens / Q000276:immunology*; D017252:CD3 Complex; D060908:CTLA-4 Antigen; D006801:Humans; D018796:Immunoconjugates; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D018106:CD28 Antigens; D017252:CD3 Complex; C115335:CD3E protein, human; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D018796:Immunoconjugates; D007128:Immunoglobulin Fragments; D011948:Receptors, Antigen, T-Cell; C055966:immunoglobulin Fv; D000069594:Abatacept