title: Expansion and activation of natural killer cells from PBMC for immunotherapy of hepatocellular carcinoma.
abstract: OBJECTIVE
  To induce efficient expansion of natural killer (NK) cells from peripheral blood mononuclear cells (PBMCs) using a culture of anchorage-dependent Wilms tumor cell lines, and to provide a reliable supply for adoptive immunotherapy of hepatocellular carcinoma.
  
  
  METHODS
  Culture expansion of NK cells was achieved using PBMCs cultured with Wilms tumor cells. Cytotoxicity was measured using a standard (51)Cr release assay and crystal violet staining technique. The proportions of CD3+, CD4+, CD8+, CD16+, and CD56+ cells were determined by flow cytometry.
  
  
  RESULTS
  After PBMCs from healthy donors and hepatocellular carcinoma (HCC) were cultured with irradiated HFWT cells for 10-21 d, CD56+ CD16+ cells shared more than 50% of the cell population, and more than 80% of fresh HFWT cells were killed at an effector/target ratio of 2 over 24 h. NK-enriched lymphocyte population from HCC patients killed HCC-1 and 2 cells with sensitivities comparable to fresh TKB-17RGB cells. HCC cells proliferated 196-fold with the irradiated HFWT cells at 18 d. Stimulation by HFWT cells required intimate cell-cell interaction with PBMC. However, neither the soluble factors released from HFWT cells nor the fixed HFWT cells were effective for NK expansion. The lymphocytes expanded with IL-2 killed fresh HFWT target cells more effectively than the lymphocytes expanded with the 4-cytokine cocktail (IL-lbeta, IL-2, IL-4 and IL-6). IL-2 was the sole cytokine required for NK expansion.
  
  
  CONCLUSIONS
  Wilms tumor is sensitive to human NK cells and is highly efficient for selective expansion of NK cells from PBMCs.
authors.affiliation: Department of Hepatobiliary Surgery, First Affiliated Hospital of Sun Yat-sen University, 58 Zhongshan Road 2, Guangzhou 510080, Guangdong Province, China. pengbaogang@163.net
authors.forename: Bao-Gang
authors.initials: BG
authors.lastname: Peng
authors.forename: Li-Jian
authors.initials: LJ
authors.lastname: Liang
authors.forename: Qiang
authors.initials: Q
authors.lastname: He
authors.forename: Jie-Fu
authors.initials: JF
authors.lastname: Huang
authors.forename: Ming-De
authors.initials: MD
authors.lastname: Lu
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000628:therapy*; D002455:Cell Division; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D006801:Humans; D016219:Immunotherapy, Adoptive; D007680:Kidney Neoplasms / Q000473:pathology*; D007694:Killer Cells, Natural / Q000166:cytology* / Q000473:pathology*; D008113:Liver Neoplasms / Q000097:blood / Q000628:therapy*; D051379:Mice; D009000:Monocytes / Q000473:pathology*; D009396:Wilms Tumor / Q000473:pathology*