Peptide-specific CTL in tumor infiltrating lymphocytes from metastatic melanomas expressing MART-1/Melan-A, gp100 and Tyrosinase genes: a study in an unselected group of HLA-A2.1-positive patients

Int J Cancer. 1995 Oct 20;64(5):309-15. doi: 10.1002/ijc.2910640505.

Abstract

Peptide specificity of cultured tumor-infiltrating lymphocytes (TIL) was systematically investigated in a group of HLA-A2.1+ metastatic melanoma patients consecutively referred to our department for surgical treatment. Seven samples from 6 patients were studied. All surgical specimens showed evidence of gp 100, MART-1/Melan-A and Tyrosinase gene expression as detectable by reverse PCR (rPCR). Cultured TIL from 2 patients displayed cytotoxic activity against autologous or HLA-matched EBV-transformed cells previously pulsed with MART-1/Melan-A27-35 peptide. In contrast, no CTL activity against gp100(280-288) or tyrosinase1-9 peptides could be observed. TIL were then repeatedly stimulated in vitro with the same peptides. After 6 restimulation courses at weekly intervals, specific recognition of gp100(280-288) and MART-1/Melan-A peptides was detectable in 3 and 5 TIL populations, respectively. In one case Tyrosinase1-9-specific CTL could be demonstrated. Two TIL populations from metastases resected from a melanoma patient at 6 months' distance showed a different peptide specificity pattern, and no specific CTL could be generated from simultaneously sampled peripheral blood mononuclear cells (PBMC). All peptide-specific CTL populations also displayed significant cytotoxic activity against HLA-A2.1 matched melanoma cell lines expressing the antigens under investigation. Our data indicate that CTL specific for MART-Melan-A27-35, gp100(280-288) or Tyrosinase1-9 peptides could be expanded with varying frequency from TIL derived from 4 out of 6 HLA-A2.1+ patients whose tumors expressed the genes encoding these tumor-associated antigens (TAA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Alleles
  • Amino Acid Sequence
  • Antigens, Neoplasm / biosynthesis
  • Antigens, Neoplasm / genetics*
  • Base Sequence
  • Cell Transformation, Viral
  • Cytotoxicity, Immunologic
  • Epitopes
  • Female
  • Granulocytes
  • HLA-A2 Antigen / immunology*
  • Herpesvirus 4, Human
  • Humans
  • Isoantigens / biosynthesis
  • Isoantigens / genetics
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / immunology
  • Lymphocytes, Tumor-Infiltrating / immunology
  • Lymphocytes, Tumor-Infiltrating / metabolism
  • Lymphocytes, Tumor-Infiltrating / physiology*
  • Male
  • Melanoma / genetics*
  • Melanoma / immunology*
  • Melanoma / metabolism
  • Membrane Glycoproteins / biosynthesis
  • Membrane Glycoproteins / genetics
  • Middle Aged
  • Molecular Sequence Data
  • Monophenol Monooxygenase / biosynthesis
  • Monophenol Monooxygenase / genetics
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics*
  • Peptides
  • Phenotype
  • Polymerase Chain Reaction
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / metabolism
  • T-Lymphocytes, Cytotoxic / physiology*
  • gp100 Melanoma Antigen

Substances

  • Antigens, Neoplasm
  • Epitopes
  • HLA-A2 Antigen
  • HNA-4A antigen
  • Isoantigens
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • PMEL protein, human
  • Peptides
  • gp100 Melanoma Antigen
  • gp100(280-288) melanoma antigen peptide
  • Monophenol Monooxygenase