Yonsei Med J.  2011 Nov;52(6):990-998. 10.3349/ymj.2011.52.6.990.

Immunotherapy of Malignant Melanoma with Tumor Lysate-Pulsed Autologous Monocyte-Derived Dendritic Cells

Affiliations
  • 1Department of Dermatology and Cutaneous Biology Research Institute, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea. mglee@yuhs.ac
  • 2Department of Dermatology, Bundang CHA Medical Center, CHA University, Seongnam, Korea.
  • 3Department of Laboratory Medicine, Yonsei Cell Therapy Center, Yonsei University College of Medicine, Seoul, Korea.
  • 4Clinical Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
  • 5Division of Nuclear Medicine, Department of Diagnostic Radiology, Yonsei University College of Medicine, Seoul, Korea.

Abstract

PURPOSE
Dendritic cell (DC) vaccination for melanoma was introduced because melanoma carries distinct tumor-associated antigens. The purpose of this study was to investigate the efficacy and safety of DC vaccination for melanoma in Korea.
MATERIALS AND METHODS
Five patients with stage IV and one with stage II were enrolled. Autologous monocyte-derived DCs (MoDCs) were cultured and pulsed with tumor-lysate, keyhole limpet hemocyanin, and cytokine cocktail for mature antigen-loaded DC. DC vaccination was repeated four times at 2-week intervals and 2-4x107 DC were injected each time.
RESULTS
Reduced tumor volume was observed by PET-CT in three patients after DC vaccination. Delayed type hypersensitivity responses against tumor antigen were induced in five patients. Tumor antigen-specific IFN-gamma-producing peripheral blood mononuclear cells were detected with enzyme-linked immunosorbent spot in two patients. However, the overall clinical outcome showed disease progression in all patients.
CONCLUSION
In this study, DC vaccination using tumor antigen-loaded, mature MoDCs led to tumor regression in individual melanoma patients. Further standardization of DC vaccination protocol is required to determine which parameters lead to better anti-tumor responses and clinical outcomes.

Keyword

Dendritic cell; immunotherapy; malignant melanoma

MeSH Terms

Dendritic Cells/*cytology
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Humans
Immunotherapy/*methods
Melanoma/*therapy
Monocytes/*cytology
Treatment Outcome

Figure

  • Fig. 1 Patients had multiple cutaneous metastases and recurrent melanoma in the tonsil. Because the whole tumor-lysate was used for antigen pulsing, it was important to have sufficient tumor volume.

  • Fig. 2 The immunophenotypic profiles of enriched monocytes (A); immature DC on day 6 of culture with GM-CSF and IL-4 (B); and mature DC cultured after an additional 2 days with tumor lysate, KLH and cytokine cocktail (TNF-a, IL-1b, IL-6, and PGE2) (C). Each of the cells was stained with monoclonal antibodies specific to CD1a, CD14, CD45, CD80, CD83, CD86, HLA-ABC and HLA-DR. Mature DCs have a higher expression of HLA-DR, CD86, and CD80 than immature DCs. Only mature DCs express CD83. The data shown are from one representative experiment. DC, dendritic cell; KLH, keyhole limpet hemocyanin.

  • Fig. 3 Tumor volume reduction in PET-CT. In the case of patient 3, reduction of tumor volume was noted in PET-CT after the 4th vaccination. The lymph nodes at the Lt. supraclavicular fossa, bilateral retroperitoneal lymph node chain, Rt. inguinal area and upper part of right axillary lymph nodes were pointed out as sites of improvement.

  • Fig. 4 Patient 3 showing positive reactions to the DTH test. DTH was examined before and then 2 weeks after the 4th vaccination. Patients received i.d. injections of 100 µL of normal saline, 100 µg tumor lysate, 50 µg KLH, 100 µg tumor lysate, plus 50 µg KLH, at separate sites on the forearm. Forty-eight hours later, DTH was scored positive if the areas of erythema and induration were greater than 5 mm. DTH, delayed type hypersensitivity; KLH, keyhole limpet hemocyanin.

  • Fig. 5 ELISPOT for the detection of Ag-specific IFN-γ producing PBMCs in patient 2, 3. Both patients showed positive results. ELISPOT, enzyme-linked immunosorbent spot.


Cited by  1 articles

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