title: Specific cellular immune response elicited by the necrotic tumor cell-stimulated macrophages.
abstract: OBJECTIVE
  To determine whether the necrotic tumor cell-stimulated macrophages (NTCSM) could elicit specific immune response.
  
  
  METHODS
  Mice were immunized with the necrotic H22 tumor cell lysate-stimulated macrophages and the specific immune responses against the same tumor challenge were examined. The morphologic characteristics were observed with the transmission electron microscope and scanning electron microscopy. The expression of CD14, CD68, CD80 and CD86 were detected with the flow cytometer. The cytotoxicity and cytokine production of splenocytes were measured with the MTT assay and ELISA assay respectively.
  
  
  RESULTS
  Our research results reveal that NTCSMs are larger cells which generally generate spherical and elongated protrusions, folding membrane, and vesicles on their surface. Also, abundant lysosomes, secondary lysosomes, phagosomes, rough endoplasmic reticulum, and lipid bodies were found in their cytoplasm. The flow cytometry results show that the necrotic H22 tumor cell lysate could enhance the expression of CD14 and CD86 molecules and the NTCSM was characterized by the expression of CD14+/-CD68+CD80-CD86+. After the mice were vaccinated with NTCSMs, the tumor forming rate, tumor volume and weight of the NTCSM-vaccinated group were significantly lower than those of the sterile saline-injected group and untreated macrophage-vaccinated group (p<0.05). The cytotoxicity to H22 tumor cells of the splenocytes obtained from the NTCSM-immunized group was higher than that of the sterile saline-injected group and untreated macrophage-vaccinated group (p<0.05). Meanwhile, the levels of IL-2 and IFN-γ in the culture supernatant of the NTCSM-immunized group were higher significantly than those of the saline-injected group and untreated macrophage-vaccinated group. The level of IL-4 of the NTCSM-immunized group was significantly lower than those of the other two groups.
  
  
  CONCLUSIONS
  Our results indicated that NTCSMs could elicit specific cellular immune responses in vivo.
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China. Electronic address: xiaolinyuandoc@yeah.net.
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Yuan
authors.affiliation: The Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine, Harbin 150001, PR China.
authors.forename: Weina
authors.initials: W
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Yifen
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhan
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Xiaohuan
authors.initials: X
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Shengfan
authors.initials: S
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Qinglin
authors.initials: Q
authors.lastname: Guan
authors.affiliation: Affiliated Hospital of Dalian University, Dalian 116001, PR China.
authors.forename: Xiuyan
authors.initials: X
authors.lastname: Sun
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D002454:Cell Differentiation; D002457:Cell Extracts / Q000276:immunology; D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D007111:Immunity, Cellular; D008099:Liver / Q000473:pathology*; D008113:Liver Neoplasms / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology* / Q000637:transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D009336:Necrosis; D009368:Neoplasm Transplantation; D047368:Tumor Burden
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D002457:Cell Extracts; D016207:Cytokines
keywords: Cellular immunity; Cytotoxicity; Immune response; Necrotic tumor cell; Stimulated macrophages