Molecular cloning of a cDNA coding biliary glycoprotein I: primary structure of a glycoprotein immunologically crossreactive with carcinoembryonic antigen.

Hinoda, Yuji; Neumaier, Michael; Hefta, Stanley A.; Drzeniek, Zofia; Wagener, Christoph; Shively, Louise; Hefta, Laura J. F.; Shively, John E.; Paxton, Raymond J.

doi:10.1073/pnas.85.18.6959

Cited by 131 publications

(70 citation statements)

References 20 publications

(11 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As well as the different patterns of in vivo behaviour of the panel of scFv, flow cytometry has revealed differences in extent of binding to a human liver cell line: CEA6, HBB 11 and, to a very small extent, T06D 12 were partially cross-reactive with Chang cells, while the remaining affinity-matured clones were not. Although structurally related markers of the CEA superfamily are known to be expressed on liver (Hinoda et al, 1988), the binding of CEA6, HBB 11 and T06D 12 cannot be explained as cross-reactivity to such molecules, as normal human liver stained negative by immunocytochemistry for all of the scFv in the study. It is not possible to explain the cross-reactive component in these antibodies on the basis of sequence, as there is no light chain sequence consensus between CEA6, HBB 11 and T06DI2 that distinguishes them from the other clones .…”

Section: Discussionmentioning

confidence: 78%

Antigen specificity and tumour targeting efficiency of a human carcinoembryonic antigen-specific scFv and affinity-matured derivatives

Jackson

Bacon²,

Pedley³

et al. 1998

Br J Cancer

View full text Add to dashboard Cite

Summary We have examined the biological properties of CEA6, a human carcinoembryonic antigen (CEA)-specific single-chain Fv (scFv) isolated by phage display, and five related clones derived by affinity maturation and selected for improved off-rate (Kff). All clones bind strongly and specifically to CEA-positive human tumours by immunocytochemistry and show negligible cross-reactivity with normal colon. Flow cytometry of scFv on human liver cells indicates a shift in fine epitope specificity resulting from mutagenesis. All monomeric scFv have been radioiodinated, retaining effectively full binding activity. A single intravenous injection into nude mice bearing human colon tumour xenografts confirms tumour targeting in all cases. As reported in other studies, the kidney is the main route of elimination of scFv at early time points. Tumour binding of the parental antibody CEA6 consistently gives the highest tumour-blood ratios at 24 h (mean 16:1). Clone T06D11, which has a sevenfold reduced K0ff relative to CEA6, showed no difference in tumour uptake at 24 h but persisted at the tumour site for longer than CEA6. This study demonstrates a possible correlation between binding affinity and tumour residence time when examined in this model. Keywords: human scFv; carcinoembryonic antigen; affinity maturation Advances in recombinant antibody technology have allowed many of the problems encountered in antibody targeting of tumours to be overcome (Huston et al, 1993). Poor tumour penetrance of whole immunoglobulin has been addressed through the use of smaller fragments derived both enzymatically and by recombinant methods (Yokota et al, 1992;Hu et al, 1996;Rowlinson-Busza et al, 1996). Further advantages of fragments are their rapid extravasation and pharmacokinetic clearance, which are clearly contributing factors in their improved performance as imaging agents (Colcher et al, 1990;Milenic et al, 1991;Verhaar et al, 1995). Moreover, the immunogenicity of rodent monoclonal antibodies (mAbs) has been reduced either by humanization strategies or by de novo isolation of antibody fragments from combinatorial libraries of human V genes (reviewed by Johnson and Chiswell, 1993). Rapid expression in bacteria has greatly assisted the study of improved engineered antibody fragments and has avoided timeconsuming and expensive mammalian cell expression systems.The isolation and characterization of scFv from large phage display libraries has demonstrated several instances where the dissociation constant of the molecule, and more specifically the off-rate (Kodf), is the main predictive factor in performance in in vitro bioassays (Thompson et al, 1996;Schier et al, 1996; Roberts et al, in preparation). The relationship between binding kinetics and tumour targeting efficiency is rather more complicated, mainly because so many additional factors (tumour size, antigen density and rate of turnover, epitope, size and charge of antibody, dose, isotope and labelling chemistry used, presence of circulating antigen) are known to influence the...

show abstract

Section: Discussionmentioning

confidence: 78%

Antigen specificity and tumour targeting efficiency of a human carcinoembryonic antigen-specific scFv and affinity-matured derivatives

Jackson

Bacon²,

Pedley³

et al. 1998

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…It is composed of four extracellular domains and a long (CEACAM1-4L) or short (CEACAM1-4S) cytoplasmic domain which are encoded by alternative splicing (Hinoda et al, 1988;Barnett et al, 1989). The cytoplasmic domains of CEACAM1 are phosphorylated at their serine residues (Edlund et al, 1998;Fournes et al, 2001) and bind to calmodulin (Edlund et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

The CEACAM1-mediated apoptosis pathway is activated by CEA and triggers dual cleavage of CEACAM1

et al. 2008

Self Cite

View full text Add to dashboard Cite

Marked reduction in apoptosis is a hallmark of early colon tumour growth and the vast majority of these tumours exhibit a loss of expression of the glycoprotein carcinoembryonic-antigen-related cell adhesion molecule 1 (CEACAM1). We recently reported that the CEACAM1 functions as a mediator of apoptosis implicating this cell surface protein in early tumour development. However, the mechanistic involvement of CEACAM1 in cell death pathways is unclear. Here, we show that apoptosis triggers cleavage of the long form of CEACAM1 (CEACAM1-4L) at intracellular and extracellular sites in Jurkat cells and HEK293 cells. Signalling through CEACAM1 leads to caspase activation including caspase-1 and -3 and also involves non-caspase proteases. Moreover, we provide evidence that the naturally occurring CEACAM family member CEA is an inducer of CEACAM1-mediated apoptosis in HT29 colon cancer cells, an effect that depends on the abundance of CEACAM1 on the cell surface. Together, our results demonstrate that the CEACAM1-dependent cell death pathway involves dual cleavage of CEACAM1 and caspase activation and can be activated by CEA.

show abstract

“…Bgp proteins possess cytoplasmic domains of either 10 or 71 (73 in mouse) amino acids which are produced by alternative splicing of a conserved 53 bp exon (Barnett et al, 1989;NeÂ dellec et al, 1995). They are mainly expressed in epithelial cells of the gastro-intestinal tract, endothelial cells and hematopoietic cells, particularly B cells, neutrophils and macrophages (Hinoda et al, 1988;Barnett et al, 1989;O È brink, 1991;Coutelier et al, 1994;NeÂ dellec et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

Association of biliary glycoprotein with protein tyrosine phosphatase SHP-1 in malignant colon epithelial cells

et al. 1997

View full text Add to dashboard Cite

Biliary glycoprotein (Bgp) is a member of the immunoglobulin superfamily and the carcinoembryonic antigen family. Previous studies have shown that Bgp functions as an intercellular adhesion molecule and a canalicular bile salt transporter. Moreover, we and others demonstrated that Bgp can inhibit colonic and prostatic tumor cell growth in vivo, through a mechanism which depends on sequences present in its cytoplasmic domain. In this study, we have examined the possibility that the cytoplasmic domain of Bgp can interact with signal transduction molecules. We showed that tyrosine phosphorylated Bgp, expressed in mouse colon carcinoma CT51 cells, could reversibly associate with protein tyrosine phosphatase SHP-1. Mutation of either of two tyrosine residues present in the cytoplasmic domain of Bgp abrogated SHP-1 binding, suggesting that this association was mediated by both tyrosine residues. Similarly, we noted that either of the two SH2 domains of SHP-1 could bind tyrosine phosphorylated Bgp in vitro. It is therefore conceivable that some of the functions of Bgp are mediated through its ability to induce intracellular protein tyrosine dephosphorylation.

show abstract

Molecular cloning of a cDNA coding biliary glycoprotein I: primary structure of a glycoprotein immunologically crossreactive with carcinoembryonic antigen.

Cited by 131 publications

References 20 publications

Antigen specificity and tumour targeting efficiency of a human carcinoembryonic antigen-specific scFv and affinity-matured derivatives

Antigen specificity and tumour targeting efficiency of a human carcinoembryonic antigen-specific scFv and affinity-matured derivatives

The CEACAM1-mediated apoptosis pathway is activated by CEA and triggers dual cleavage of CEACAM1

Association of biliary glycoprotein with protein tyrosine phosphatase SHP-1 in malignant colon epithelial cells

Contact Info

Product

Resources

About