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In vitro anti-tumor immune response induced by dendritic cells transfected with recombinant adenovirus carrying mutant k-ras genes

  • Zhao Feng
  • Zhou Qinghua
  • Lu Yanrong
  • Qin Yang
  • Zhang Jie
  • Li Jinsong
  • Wang Jianjun
Article
  • 31 Downloads

Summary

The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) transfected with recombinant adenovirus carrying mutant k-ras genes was investigated. DCs were generated from mouse bone marrow in the presence of rmGM-CSF (3.3 ng/mL) and rmIL-4 (1.3 ng/mL) and detected by FACS, and then transfected with the recombinant adenovirus encoding mutant k-ras gene. The efficacy of transfection and T cell stimulating activity of DCs were detected. CTL activity of the mice vaccinated with DCs was observed. The results showed that DCs had dendritic veiled morphology. BmDCs highly expressed B7-1 (80%), B7-2 (77%), MHC II (70%), CD11c (65%), CD40 (70%) and CD54 (96%) with FACS, and no significant difference in the expression was observed before and after the transfection (P>0.05). The DCs transfected by mutant k-ras gene could significantly stimulate lymphocytes proliferation as compared with those transfected by Ad-c or non-modified DCs (P<0.05). DC vaccine transfected by mutant k-ras gene could induce CTL activity against Lewis lung cancer, but not against B16. The specific cytotoxicity against Lewis lung cancer in Ad-k-ras/12-transduced DC group was significantly higher than those in the control, vector and non-transfected DCs groups (P<0.05). It was concluded that special antitumor response could be induced by DCs transfected with recombinant adenovirus carrying mutant k-ras genes.

Key words

adenovirus vector mutant k-ras gene dendritic cell T lymphocyte 

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References

  1. 1.
    Abrams S I, Stanziale S F, Lunin S Det al. Identification of overlapping epitopes in mutant ras oncogene peptides that activate CD4+ and CD8+ T cell responses. Eur J Immunol. 1996,26:435PubMedCrossRefGoogle Scholar
  2. 2.
    Bergmann-Leitner E S, Kantor J A, Lesley Shupert Wet al. Identification of a human CD8+ T lymphocyte neo-epitope created by a ras codon 12 mutation which is restricted by the HLA-A2 allele. Celluar Immunology, 1998,187:103CrossRefGoogle Scholar
  3. 3.
    Bos J L. ras oncogenes in human cancer: a review. Cancer Res, 1989,49:4682PubMedGoogle Scholar
  4. 4.
    Ramakrishna G, Sithanandam G, Cheng R Yet al. K-ras p21 expression and activity in lung and lung tumors. Exp Lung Res, 2000,26:659PubMedCrossRefGoogle Scholar
  5. 5.
    Abrams S I, Dobrzanski M J, Wells D Tet al. Peptide-specific activation of cytolytic CD4 + T lymphocytes against tumor cells bearing mutated epitopes of K-ras p21. Eur J Immunol, 1995,25:2588PubMedCrossRefGoogle Scholar
  6. 6.
    Banchereau J, Steinman R M. Dendritic cells and the control of immunity. Nature, 1998,392:245PubMedCrossRefGoogle Scholar
  7. 7.
    Zhao F, Zhou Q H, Qing Yet al. Construction of mutant k-ras gene recombinant adenovirus. Chin J Lung Cancer (Chinese), 2002,5(1):14Google Scholar
  8. 8.
    Jackson J H, Cochrane C G, Bourne J Ret al. Farnesol modification of Kirsten-ras exon 4B protein is essential for transformation. Proc Natl Acad Sci U S A, 1990,87:3042PubMedCrossRefGoogle Scholar
  9. 9.
    He T C, Zhou S, Costa L Tet al. A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci U S A, 1998,95:2509PubMedCrossRefGoogle Scholar
  10. 10.
    Inaba K, Inaba M, Romani Net al. Generation of large number of dendritic cells from mouse bone marrow cultures supplement with GM-CSF. J Exp Med, 1992, 176(6):1693PubMedCrossRefGoogle Scholar
  11. 11.
    Hunger R E, Brand C U, Streit Met al. Successful induction of immune responses against mutant ras in melanoma patients using intradermal injection of peptides and GM-CSF as adjuvant. Exp Dermatol, 2001,10(3): 161PubMedCrossRefGoogle Scholar
  12. 12.
    Linard B, Bezieau S, Benlalam Het al. A ras-mutated peptide targeted by CTL infiltrating a human melanoma lesion. J Immunol, 2002, 168:4802PubMedGoogle Scholar
  13. 13.
    Zhong L, Granelli-Piperno A, Choi Yet al. Recombinant adenovirus is an efficient and nonperturbing genetic vector for human dendritic cells. Eur J Immunol, 1999,29:964PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Zhao Feng
    • 1
  • Zhou Qinghua
    • 2
  • Lu Yanrong
    • 2
  • Qin Yang
    • 2
  • Zhang Jie
    • 2
  • Li Jinsong
    • 1
  • Wang Jianjun
    • 1
  1. 1.Department of Thoracic Surgery, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Cancer Center of West China HospitalWest China Medical School, Sichuan UniversityChengduChina

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