Treatment of colon and lung cancer patients with ex vivo heat shock protein 70-peptide-activated, autologous natural killer cells: a clinical phase i trial

Clin Cancer Res. 2004 Jun 1;10(11):3699-707. doi: 10.1158/1078-0432.CCR-03-0683.


Purpose: The 14 amino acid sequence (aa(450-463)) TKDNNLLGRFELSG (TKD) of heat shock protein 70 (Hsp70) was identified as a tumor-selective recognition structure for natural killer (NK) cells. Incubation of peripheral blood lymphocyte cells with TKD plus low-dose interleukin 2 (IL-2) enhances the cytolytic activity of NK cells against Hsp70 membrane-positive tumors, in vitro and in vivo. These data encouraged us to test tolerability, feasibility, and safety of TKD-activated NK cells in a clinical Phase I trial.

Experimental design: Patients with metastatic colorectal cancer (n = 11) and non-small cell lung cancer (n = 1) who had failed standard therapies were enrolled. After ex vivo stimulation of autologous peripheral blood lymphocytes with Hsp70-peptide TKD (2 microg/ml) plus low-dose IL-2 (100 units/ml), TKD was removed by extensive washing, and activated cells were reinfused i.v. The procedure was repeated for up to six cycles, applying a dose escalation schedule in 4 patients.

Results: The percentage of activated NK cells in the reinfused leukapheresis products ranged between 8 and 20% of total lymphocytes, corresponding to total NK cell counts of 0.1 up to 1.5 x 10(9). Apart from restless feeling in 1 patient and itching in 2 patients, no negative side effects were observed. Concomitant with an enhanced CD94 cell surface density, the cytolytic activity of NK cells against Hsp70 membrane-positive colon carcinoma cells was enhanced after TKD/IL-2 stimulation in 10 of 12 patients. Concerning tumor response, 1 patient was in stable disease during therapy by formal staging criteria and another patient showed stable disease in one metastases and progression in another.

Conclusions: Reinfusion of Hsp70-activated autologous NK cells is safe. Immunological results warrant additional studies in patients with lower tumor burden.

Publication types

  • Clinical Trial
  • Clinical Trial, Phase I
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Antigens, CD / biosynthesis
  • Cell Membrane / metabolism
  • Cell Movement
  • Colonic Neoplasms / therapy*
  • Cytotoxicity, Immunologic
  • Disease Progression
  • Female
  • Follow-Up Studies
  • Granzymes
  • HSP70 Heat-Shock Proteins / chemistry*
  • Humans
  • Interferon-gamma / blood
  • Interleukin-2 / chemistry
  • Killer Cells, Natural / cytology*
  • Lectins, C-Type / biosynthesis
  • Lung Neoplasms / therapy*
  • Male
  • Middle Aged
  • NK Cell Lectin-Like Receptor Subfamily D
  • Neoplasm Metastasis
  • Phenotype
  • Serine Endopeptidases / blood
  • Time Factors
  • Treatment Outcome
  • Tumor Necrosis Factor-alpha / biosynthesis


  • Antigens, CD
  • HSP70 Heat-Shock Proteins
  • Interleukin-2
  • KLRD1 protein, human
  • Lectins, C-Type
  • NK Cell Lectin-Like Receptor Subfamily D
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma
  • GZMB protein, human
  • Granzymes
  • Serine Endopeptidases