Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A

Janice N. Cormier, Michael L. Salgaller, Tracy Prevette, Kathleen C. Barracchini, Licia Rivoltini, Nicholas P. Restifo, Steven A. Rosenberg, Francesco M. Marincola

Research output: Contribution to journalArticle

Abstract

PURPOSE: In this study, we tested the effectiveness of a melanomaassociated antigen-derived peptide, MART-127-35, in eliciting cellular immune responses in vivo in the context of a phase I active immunization protocol. This peptide (AAGIGILTV) corresponds to residues 27- 35 from the nonmutated melanoma-associated antigen MART-1/Melan A and is recognized by most melanoma-specific, HLA-A*0201-restricted, tumor- infiltrating lymphocytes. To test the in vivo induction of cytotoxic T lymphocyte (CTL) sensitization, we compared CTL reactivity in vitro from peripheral blood mononuclear cell (PBMC) pools obtained before and after vaccination. PATIENTS AND METHODS: MART-127-35 was administered to HLA- A*0201 melanoma patients subcutaneously in an emulsification with incomplete Freund's adjuvant. A vaccination course included four inoculations of peptide at 3-week intervals. PBMC collected by leukapheresis and separated by Ficoll- Hypaque gradient before and after vaccination were analyzed in 18 patients by in vitro sensitization with MART-127-35. To induce MART-127-35- specific CTL, PBMC were incubated with 1 μM peptide (on day 0) and interleukin-2 (IL-2) (300 IU/mL, on days 1 and 4 after each stimulation). At weekly intervals, cells were harvested and an aliquot was cryopreserved for later analysis. The remaining cells were replated and restimulated using irradiated autologous PBMC pulsed with 1 μM of relevant peptide. After three restimulations, all samples from one patient were tested simultaneously for HLA-A*0201-restricted anti-MART-127-35 reactivity by microcytotoxicity and cytokine (IFN-γ) release assays. RESULTS: Toxicities were minimal and consisted of local irritation at the site of vaccine administration. None of the patents sustained a clinical response. The first eight patients were monitored by inducing CTL reactivity from PBMC obtained preimmunization and after two and four vaccinations. Only two prevaccination cultures were reactive to MART-1, compared with five and seven cultures from PBMC obtained after two and four vaccinations, respectively. Thus, an enhancement in cytotoxic activity could be detected in postvaccination CTL cultures, and serial vaccine administrations appeared to boost the detectability of cytotoxicity in vitro. For completeness, the analysis compared prevaccination with postvaccination PBMC cultures. Specific anti-MART 127-35 cytotoxicity (≤ 10 lytic units) could be detected in two prevaccination and 12 postvaccination cultures after two in vitro stimulations. In 15 postvaccination CTL cultures, a more than threefold increase in specific release of IFN-γ was noted, compared with prevaccination. DISCUSSION: In vivo administration of a melanoma-associated antigen peptide, emulsified in incomplete Freund's adjuvant, could safely augment CTL reactivity against epitopes commonly expressed by melanoma cells. Although the enhancement of CTL reactivity did not achieve tumor regression, it is possible that the use of recombinant immunogens with increased immunomodulatory capabilities in future clinical trials could reach the threshold of CTL activation necessary for tumor regression.

Original languageEnglish (US)
Pages (from-to)37-44
Number of pages8
JournalCancer Journal from Scientific American
Volume3
Issue number1
StatePublished - 1997
Externally publishedYes

Fingerprint

MART-1 Antigen
Cytotoxic T-Lymphocytes
Cellular Immunity
Melanoma
Blood Cells
Peptides
Vaccination
Melanoma-Specific Antigens
Vaccines
Diatrizoate
Leukapheresis
Tumor-Infiltrating Lymphocytes
Ficoll
Patents
Lymphocyte Activation
Interleukin-2
Epitopes
Neoplasms
Cell Culture Techniques
Clinical Trials

Keywords

  • immunization
  • MART-1
  • melanoma

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Cormier, J. N., Salgaller, M. L., Prevette, T., Barracchini, K. C., Rivoltini, L., Restifo, N. P., ... Marincola, F. M. (1997). Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A. Cancer Journal from Scientific American, 3(1), 37-44.

Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A. / Cormier, Janice N.; Salgaller, Michael L.; Prevette, Tracy; Barracchini, Kathleen C.; Rivoltini, Licia; Restifo, Nicholas P.; Rosenberg, Steven A.; Marincola, Francesco M.

In: Cancer Journal from Scientific American, Vol. 3, No. 1, 1997, p. 37-44.

Research output: Contribution to journalArticle

Cormier, JN, Salgaller, ML, Prevette, T, Barracchini, KC, Rivoltini, L, Restifo, NP, Rosenberg, SA & Marincola, FM 1997, 'Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A', Cancer Journal from Scientific American, vol. 3, no. 1, pp. 37-44.
Cormier JN, Salgaller ML, Prevette T, Barracchini KC, Rivoltini L, Restifo NP et al. Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A. Cancer Journal from Scientific American. 1997;3(1):37-44.
Cormier, Janice N. ; Salgaller, Michael L. ; Prevette, Tracy ; Barracchini, Kathleen C. ; Rivoltini, Licia ; Restifo, Nicholas P. ; Rosenberg, Steven A. ; Marincola, Francesco M. / Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A. In: Cancer Journal from Scientific American. 1997 ; Vol. 3, No. 1. pp. 37-44.
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author = "Cormier, {Janice N.} and Salgaller, {Michael L.} and Tracy Prevette and Barracchini, {Kathleen C.} and Licia Rivoltini and Restifo, {Nicholas P.} and Rosenberg, {Steven A.} and Marincola, {Francesco M.}",
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TY - JOUR

T1 - Enhancement of cellular immunity in melanoma patients immunized with a peptide from MART-1/Melan A

AU - Cormier, Janice N.

AU - Salgaller, Michael L.

AU - Prevette, Tracy

AU - Barracchini, Kathleen C.

AU - Rivoltini, Licia

AU - Restifo, Nicholas P.

AU - Rosenberg, Steven A.

AU - Marincola, Francesco M.

PY - 1997

Y1 - 1997

N2 - PURPOSE: In this study, we tested the effectiveness of a melanomaassociated antigen-derived peptide, MART-127-35, in eliciting cellular immune responses in vivo in the context of a phase I active immunization protocol. This peptide (AAGIGILTV) corresponds to residues 27- 35 from the nonmutated melanoma-associated antigen MART-1/Melan A and is recognized by most melanoma-specific, HLA-A*0201-restricted, tumor- infiltrating lymphocytes. To test the in vivo induction of cytotoxic T lymphocyte (CTL) sensitization, we compared CTL reactivity in vitro from peripheral blood mononuclear cell (PBMC) pools obtained before and after vaccination. PATIENTS AND METHODS: MART-127-35 was administered to HLA- A*0201 melanoma patients subcutaneously in an emulsification with incomplete Freund's adjuvant. A vaccination course included four inoculations of peptide at 3-week intervals. PBMC collected by leukapheresis and separated by Ficoll- Hypaque gradient before and after vaccination were analyzed in 18 patients by in vitro sensitization with MART-127-35. To induce MART-127-35- specific CTL, PBMC were incubated with 1 μM peptide (on day 0) and interleukin-2 (IL-2) (300 IU/mL, on days 1 and 4 after each stimulation). At weekly intervals, cells were harvested and an aliquot was cryopreserved for later analysis. The remaining cells were replated and restimulated using irradiated autologous PBMC pulsed with 1 μM of relevant peptide. After three restimulations, all samples from one patient were tested simultaneously for HLA-A*0201-restricted anti-MART-127-35 reactivity by microcytotoxicity and cytokine (IFN-γ) release assays. RESULTS: Toxicities were minimal and consisted of local irritation at the site of vaccine administration. None of the patents sustained a clinical response. The first eight patients were monitored by inducing CTL reactivity from PBMC obtained preimmunization and after two and four vaccinations. Only two prevaccination cultures were reactive to MART-1, compared with five and seven cultures from PBMC obtained after two and four vaccinations, respectively. Thus, an enhancement in cytotoxic activity could be detected in postvaccination CTL cultures, and serial vaccine administrations appeared to boost the detectability of cytotoxicity in vitro. For completeness, the analysis compared prevaccination with postvaccination PBMC cultures. Specific anti-MART 127-35 cytotoxicity (≤ 10 lytic units) could be detected in two prevaccination and 12 postvaccination cultures after two in vitro stimulations. In 15 postvaccination CTL cultures, a more than threefold increase in specific release of IFN-γ was noted, compared with prevaccination. DISCUSSION: In vivo administration of a melanoma-associated antigen peptide, emulsified in incomplete Freund's adjuvant, could safely augment CTL reactivity against epitopes commonly expressed by melanoma cells. Although the enhancement of CTL reactivity did not achieve tumor regression, it is possible that the use of recombinant immunogens with increased immunomodulatory capabilities in future clinical trials could reach the threshold of CTL activation necessary for tumor regression.

AB - PURPOSE: In this study, we tested the effectiveness of a melanomaassociated antigen-derived peptide, MART-127-35, in eliciting cellular immune responses in vivo in the context of a phase I active immunization protocol. This peptide (AAGIGILTV) corresponds to residues 27- 35 from the nonmutated melanoma-associated antigen MART-1/Melan A and is recognized by most melanoma-specific, HLA-A*0201-restricted, tumor- infiltrating lymphocytes. To test the in vivo induction of cytotoxic T lymphocyte (CTL) sensitization, we compared CTL reactivity in vitro from peripheral blood mononuclear cell (PBMC) pools obtained before and after vaccination. PATIENTS AND METHODS: MART-127-35 was administered to HLA- A*0201 melanoma patients subcutaneously in an emulsification with incomplete Freund's adjuvant. A vaccination course included four inoculations of peptide at 3-week intervals. PBMC collected by leukapheresis and separated by Ficoll- Hypaque gradient before and after vaccination were analyzed in 18 patients by in vitro sensitization with MART-127-35. To induce MART-127-35- specific CTL, PBMC were incubated with 1 μM peptide (on day 0) and interleukin-2 (IL-2) (300 IU/mL, on days 1 and 4 after each stimulation). At weekly intervals, cells were harvested and an aliquot was cryopreserved for later analysis. The remaining cells were replated and restimulated using irradiated autologous PBMC pulsed with 1 μM of relevant peptide. After three restimulations, all samples from one patient were tested simultaneously for HLA-A*0201-restricted anti-MART-127-35 reactivity by microcytotoxicity and cytokine (IFN-γ) release assays. RESULTS: Toxicities were minimal and consisted of local irritation at the site of vaccine administration. None of the patents sustained a clinical response. The first eight patients were monitored by inducing CTL reactivity from PBMC obtained preimmunization and after two and four vaccinations. Only two prevaccination cultures were reactive to MART-1, compared with five and seven cultures from PBMC obtained after two and four vaccinations, respectively. Thus, an enhancement in cytotoxic activity could be detected in postvaccination CTL cultures, and serial vaccine administrations appeared to boost the detectability of cytotoxicity in vitro. For completeness, the analysis compared prevaccination with postvaccination PBMC cultures. Specific anti-MART 127-35 cytotoxicity (≤ 10 lytic units) could be detected in two prevaccination and 12 postvaccination cultures after two in vitro stimulations. In 15 postvaccination CTL cultures, a more than threefold increase in specific release of IFN-γ was noted, compared with prevaccination. DISCUSSION: In vivo administration of a melanoma-associated antigen peptide, emulsified in incomplete Freund's adjuvant, could safely augment CTL reactivity against epitopes commonly expressed by melanoma cells. Although the enhancement of CTL reactivity did not achieve tumor regression, it is possible that the use of recombinant immunogens with increased immunomodulatory capabilities in future clinical trials could reach the threshold of CTL activation necessary for tumor regression.

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KW - MART-1

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