Table I.

Patient characteristics and cloning efficiency

PatientMM14MM18L02DECHMM15
SexFMFFF
Age4451653959
Stage (AJCC)IIIIIIIVIIIIV
MetastasesLymph nodesLymph nodesSkinLymph nodesSkin
LungSkinLung
Adrenals
TherapySurgerySurgerySurgerySurgerySurgery
DacarbazineDacarbazine
1,3-Bis(2-chloroethyl)-1-nitrosourea
Cisplatin
Tamoxifen
IFN-γ
Tumor AgaMelan-AMelan-AMelan-AMAGE3Melan-A
TyrosinaseTyrosinaseTyrosinase
Tissue sampleTILNTILNPBLPBLSkin
Metastasis
Tissue cultureIL-7, IL-2IL-7, IL-2NilPeptidePHA
IL-7, IL-2IL-2
Tetramer in sortA2/melan-AA2/tyrosinaseA2/melan-AA2/melan-AA2/MAGE3A2/melan-A
Clones sorted60631756011080
Clones expandedb8 (13%)1 (2%)11 (6%)7 (12%)2 (2%)7 (9%)
Specific clonesc8111726
Source population per specific cloned25,00078,75053,03014,290183,3301,875
  • a Tumors were typed for expression of melan-A and tyrosinase by immunohistochemistry and for MAGE3 by RT-PCR.

  • b Number (and percent) of clones that expanded to at least 5 × 106 cells.

  • c Expanded clones that subsequently stained strongly with the appropriate tetramer or were specific for the appropriate Ag in chromium release or enzyme-linked immunospot assay.

  • d Number of starting lymphocytes needed to generate each melanoma-specific clone, derived by dividing the number of cells plated per specific expanded clone by the percentage of the starting population that fell within the cloning gate.