A Human Melanoma Cell Line Transduced with an Interleukin-4 Gene by a Retroviral Vector Releases Biologically Active IL-4 and Maintains the Original Tumor Antigenic Phenotype

Cecilia Melani, Josep Sulé-Suso, Flavio Arienti, Cristina Maccalli, Flavio Passerini, Mario P. Colombo, Giorgio Parmiani

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Experimental models of vaccination with tumor cells engineered to produce interleukin-4 (IL-4) have shown that the local release of this cytokine is associated with the development of antitumor immunity that may induce regression of established cancer. The aim of this study was to transduce a human melanoma cell line with the gene coding for human IL-4, and to analyze cytokine production, phenotypic characteristics, and antigen expression after transduction. A retroviral vector, constructed by inserting IL-4 cDNA into the LXSN vector, was used to infect the human melanoma cell line Me14932, known to express the MHC class I HLA-A2 and the melanoma-associated antigen Melan-A/MART-1, recognized by HLA-A2-restricted T-cells. The confluence of all G418-resistant cells (Me14932/IL-4) was then analyzed for proviral integration and IL-4 mRNA expression. Substantially stable IL-4 release was detected by ELISA in the supernatant of transduced cells, ranging from 1.6 to 4.6 ng/ml per 105 cells per 24 hr; such a cytokine displayed a specific biologic activity, as revealed by the stimulation of blast cell proliferation and the inhibition of lymphokine activated killer cell (LAK) induction by IL-2. After 200 Gy irradiation, IL-4 release remained detectable for 5 weeks, whereas cell proliferation ceased within 7 days. Morphology and immunophenotypic characteristics of the parental cell line (expression of MHC classes I and II, ICAM-1, LFA 3, melanoma-associated antigens, etc.) were retained by the IL-4 gene-transduced melanoma as assayed by microscopy and immunofluorescence; likewise, susceptibility to lysis by LAK cells as well as a T-cell clone recognizing the Melan-A/MART-1 antigen did not change. These results, together with the lack of replication-competent retrovirus, suggest that the Me14932/IL-4 cell line displays suitable characteristics for its use in the treatment of HLA-matched melanoma patients.

Original languageEnglish
Pages (from-to)1427-1436
Number of pages10
JournalHuman Gene Therapy
Volume6
Issue number11
DOIs
Publication statusPublished - 1 Nov 1995
Externally publishedYes

Fingerprint

Interleukin-4
Melanoma
Phenotype
Cell Line
MART-1 Antigen
Genes
Neoplasms
Melanoma-Specific Antigens
HLA-A2 Antigen
Lymphokine-Activated Killer Cells
Cytokines
CD58 Antigens
Cell Proliferation
T-Lymphocytes
Retroviridae
Intercellular Adhesion Molecule-1
Fluorescence Microscopy
Interleukin-2
Immunity
Vaccination

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

A Human Melanoma Cell Line Transduced with an Interleukin-4 Gene by a Retroviral Vector Releases Biologically Active IL-4 and Maintains the Original Tumor Antigenic Phenotype. / Melani, Cecilia; Sulé-Suso, Josep; Arienti, Flavio; Maccalli, Cristina; Passerini, Flavio; Colombo, Mario P.; Parmiani, Giorgio.

In: Human Gene Therapy, Vol. 6, No. 11, 01.11.1995, p. 1427-1436.

Research output: Contribution to journalArticle

Melani, Cecilia ; Sulé-Suso, Josep ; Arienti, Flavio ; Maccalli, Cristina ; Passerini, Flavio ; Colombo, Mario P. ; Parmiani, Giorgio. / A Human Melanoma Cell Line Transduced with an Interleukin-4 Gene by a Retroviral Vector Releases Biologically Active IL-4 and Maintains the Original Tumor Antigenic Phenotype. In: Human Gene Therapy. 1995 ; Vol. 6, No. 11. pp. 1427-1436.
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abstract = "Experimental models of vaccination with tumor cells engineered to produce interleukin-4 (IL-4) have shown that the local release of this cytokine is associated with the development of antitumor immunity that may induce regression of established cancer. The aim of this study was to transduce a human melanoma cell line with the gene coding for human IL-4, and to analyze cytokine production, phenotypic characteristics, and antigen expression after transduction. A retroviral vector, constructed by inserting IL-4 cDNA into the LXSN vector, was used to infect the human melanoma cell line Me14932, known to express the MHC class I HLA-A2 and the melanoma-associated antigen Melan-A/MART-1, recognized by HLA-A2-restricted T-cells. The confluence of all G418-resistant cells (Me14932/IL-4) was then analyzed for proviral integration and IL-4 mRNA expression. Substantially stable IL-4 release was detected by ELISA in the supernatant of transduced cells, ranging from 1.6 to 4.6 ng/ml per 105 cells per 24 hr; such a cytokine displayed a specific biologic activity, as revealed by the stimulation of blast cell proliferation and the inhibition of lymphokine activated killer cell (LAK) induction by IL-2. After 200 Gy irradiation, IL-4 release remained detectable for 5 weeks, whereas cell proliferation ceased within 7 days. Morphology and immunophenotypic characteristics of the parental cell line (expression of MHC classes I and II, ICAM-1, LFA 3, melanoma-associated antigens, etc.) were retained by the IL-4 gene-transduced melanoma as assayed by microscopy and immunofluorescence; likewise, susceptibility to lysis by LAK cells as well as a T-cell clone recognizing the Melan-A/MART-1 antigen did not change. These results, together with the lack of replication-competent retrovirus, suggest that the Me14932/IL-4 cell line displays suitable characteristics for its use in the treatment of HLA-matched melanoma patients.",
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AU - Melani, Cecilia

AU - Sulé-Suso, Josep

AU - Arienti, Flavio

AU - Maccalli, Cristina

AU - Passerini, Flavio

AU - Colombo, Mario P.

AU - Parmiani, Giorgio

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