title: Insoluble fraction of tumor cell homogenate is a useful material for eliciting cytotoxic T lymphocytes: a unique method for protein solubilization.
abstract: OBJECTIVE
  Dendritic cell (DC)-based cancer immunotherapy using tumor homogenate has been evaluated. In all previously reported cases, DCs have been pulsed with a soluble fraction (lysate) of the tumor homogenate. The aim of this study was the evaluation of DCs pulsed with solubilized insoluble fraction of tumor cells.
  
  
  METHODS
  Solubilized recombinant murine TRP-2 and solubilized-insoluble fraction of B16 melanoma was prepared by a novel method using nucleotides. Bone marrow-derived DCs were electroloaded with the solubilized proteins.
  
  
  RESULTS
  Cytotoxic T lymphocytes were elicited in the splenocytes of C57BL/6 mice immunized with electroloaded DCs with solubilized rTRP-2. CD8(+) T-cells derived from immunized mice with electroloaded DCs using the solubilized insoluble fraction of B16 melanoma had specific killing activity. The effects were augmented when DCs were electroloaded with both the soluble and insoluble fractions of B16 homogenate.
  
  
  CONCLUSIONS
  Insoluble fraction of tumor cells is a useful material for cancer immunotherapy.
authors.affiliation: Medinet Medical Institute, MEDINET Co., Ltd., 2-2-8 Tamagawadai, Setagayaku, Tokyo 158-0096, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Kuwada
authors.forename: Kaori
authors.initials: K
authors.lastname: Kambara
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Tadaki
authors.forename: Katsuo
authors.initials: K
authors.lastname: Noguchi
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology; D001671:Biochemistry / Q000379:methods*; D002457:Cell Extracts / Q000737:chemistry*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007114:Immunization; D019746:Intramolecular Oxidoreductases / Q000378:metabolism; D058965:MART-1 Antigen / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000378:metabolism*; D009369:Neoplasms / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D009711:Nucleotides / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility / Q000187:drug effects; D013154:Spleen / Q000166:cytology; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; D002457:Cell Extracts; D058965:MART-1 Antigen; D009363:Neoplasm Proteins; D009711:Nucleotides; D011994:Recombinant Proteins; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase