W

Patient's mononuclear cells

Isolate and clone activated CD8+ —

melanoma-specific CTLs §L

Tumor cells

Melanoma cell line

Coculture mononuclear cells and melanoma cells

Melanoma cell line

Identification of tumor antigens recognized by tumor-specific CTLs

Tumor cDNA library

Transfect into class I MHC+ target cell line

Coculture with CTL clone

Lysis of transfected cell line

Coculture with CTL clone

Transfect into class I MHC+ target cell line

Isolate transfected DNA and sequence ^

Gene encoding tumor antigen recognized by i ) melanoma-specific CTLv___s

FIGURE 17-3 Cloned CTL lines specific for human tumors are used to identify specific tumor antigens. A, CD8 T cells isolated from blood, lymph nodes, or tumors of patients with melanoma are propagated in culture by stimulating them with melanoma cell lines derived from the patient's tumor. Single T cells from these cultures are expanded into clonal CTL lines. B, DNA from melanoma gene libraries is transfected into class I MHC-expressing target cells. Genes that sensitize the target cells for lysis by the melanoma-specific CTL clones are analyzed to identify the melanoma protein antigens recognized by the patient's CTLs.

with cancers as well as tumor-associated antigens that may not naturally induce host immune responses but are potential targets for immunotherapy or are useful markers for clinical diagnosis and for observation of patients.

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