title: 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.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
authors.affiliation: Department of Hematology/Oncology, Institute for Clinical Chemistry, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Krause
authors.forename: Robert
authors.initials: R
authors.lastname: Gastpar
authors.forename: Reinhard
authors.initials: R
authors.lastname: Andreesen
authors.forename: Catharina
authors.initials: C
authors.lastname: Gross
authors.forename: Heidrun
authors.initials: H
authors.lastname: Ullrich
authors.forename: Gerald
authors.initials: G
authors.lastname: Thonigs
authors.forename: Karin
authors.initials: K
authors.lastname: Pfister
authors.forename: Gabriele
authors.initials: G
authors.lastname: Multhoff
mesh_terms: D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000096:biosynthesis; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D003110:Colonic Neoplasms / Q000628:therapy*; D003602:Cytotoxicity, Immunologic; D018450:Disease Progression; D005260:Female; D005500:Follow-Up Studies; D053804:Granzymes; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D007371:Interferon-gamma / Q000097:blood; D007376:Interleukin-2 / Q000737:chemistry; D007694:Killer Cells, Natural / Q000166:cytology*; D037181:Lectins, C-Type / Q000096:biosynthesis; D008175:Lung Neoplasms / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D051920:NK Cell Lectin-Like Receptor Subfamily D; D009362:Neoplasm Metastasis; D010641:Phenotype; D012697:Serine Endopeptidases / Q000097:blood; D013997:Time Factors; D016896:Treatment Outcome; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D015703:Antigens, CD; D018840:HSP70 Heat-Shock Proteins; D007376:Interleukin-2; C497869:KLRD1 protein, human; D037181:Lectins, C-Type; D051920:NK Cell Lectin-Like Receptor Subfamily D; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases