关键词：乳腺癌;疾病载体;T淋巴细胞;解剖模型
摘 要：The overall objective of this project is to develop targeted therapy tools directed against the breast cancer vasculature. We aim to bypass many obstacles in the field by developing a treatment based on the targeting of tumor blood vessels with T cells bearing chimeric immune receptors (CIRs). We have constructed and tested first generation CIR lentiviral vectors targeting hTEM1, which were derived from a panel of scFvs of varying affinity. These constructs signal through CD3 but can be augmented by the addition of costimulatory molecules (CD28 and 4-1BB). In vitro, only T cells bearing the scFv78 CIR were activated upon co-culture with recombinant hTEM1 protein. However, no activation was observed when scFv78 CIR bearing T cells were co- cultured with cells expressing hTEM1 on the surface. Additional strategies, including hinge elongation and the utilization of loadable CIR bearing T cells, also failed to elicit a response against the hTEM1 bearing target cells. We engineered a new yeast-display scFv library from patients with ovarian cancer to isolate a second generation of anti-TEM1 scFvs. We validated a panel of multivalent antibodies for binding to recombinant proteins by ELISA assays. We have generated CIRs against PSMA using scFv (PZ1) and T cells bearing PZ1 and CD28 produce a robust polyfunctional repertoire of cytokines. A tumor vascular mouse model expressing human TEM1 was developed and PET imaging with (124I)-MORAb-004 indicated TEM1-specific accumulation of antibody. We have established optimized HSV-tk as extra safety and tracer for PET imaging.