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Development and evaluation of a rapid neutralizing antibody test for

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Development and evaluation of a rapid neutralizing antibody test for Powered By Docstoc
					Resveratrol exhibits a strong cytotoxic
activity in cultured cells and has an
antiviral action against polyomavirus:
potential clinical use

Author: Valerio Berardi, Francesca Ricci, Mauro Castelli,
Gaspare Galati, Gianfranco Risuleo

Source: Journal of Experimental & Clinical Cancer
Research 2009, 28:96

Speaker:陳彥安
Polyomavirus
 Papovaviridae
 dsDNA
 circular form
 icosahedral capsid
 non-envelope
Polyomavirus
 包含了六個基因
 其中三個基因是在感染初期產生的T-
  antigens,可分為small T, middle T和large
  T,其主要的作用為引發並維持腫瘤的
  形成
 另外三個基因為感染後期產生的蛋白質
  ,分別為VP1、VP2及VP3,為結構性蛋
  白質,會形成病毒外殼
Resveratrol
 一種存在於植物和水果中的非類黃酮多
  酚化合物,特別是葡萄含有高濃度的
  Resveratrol
 其他作者的研究顯示此種天然化合物可
  抑制不同病毒的增殖,例如單純泡疹病
  毒、水痘帶狀皰疹病毒和A型流感病毒
Resveratrol
 苦楝油(neem oil)經由甲醇萃取所獲得
 苦楝油(neem oil)是由苦楝樹(neem
  tree,Azadirachta indica)的種子製備而
  成
Neem tree(Azadirachta indica)
Methods
Cell cultures
 3T6(mouse fibroblast line)
 HL60(human promyelocytic leukemia cell
  line)
 High glucose DMEM
   newborn bovine serum (10%)
   glutamine (50 mM)
   penicillin-streptomycin (10000 U/mL).
 37 °C, 5% CO2,incubator
    3T6,HL60

             treated with 20uM
             40uM of resveratrol

trypan blue staining    Vital cell count




Viral infect 3T6 HL60
             treated with
             20uM of resveratrol

                        Assessment of the
trypan blue staining
                        cell mortality rate




    agarose gel
                          Viral DNA yield
  electrophoresis
Results
  Vital cell count
 Vital cell count after trypan blue staining of cells treated with
 resveratrol (20 μM) for 24 hours




the tumor cells HL60:The results clarly show that RV can significantly
inhibit the cell growth already at a concentration of 25 μM
Cytotoxicity of resveratrol assay
             3T6 cells
Cytotoxicity of resveratrol assay
              HL60 cells
 Assessment of the cell mortality rate
Assessment of the cell mortality rate due to Py proliferation
        Viral DNA yield
      Viral DNA yield obtained at 24 hours post-infection
              20   40




5kb


                                                     20 μM   40 μM
       Viral DNA yield
the culture medium was changed to fresh drug-free medium at the
following times post-infection. The incubation was continued for 24
hours

      4h 8h 12h16h




                                                4h 8h 12h16h
Discussion
   Discussion
                Resveratrol


  3T6(mouse               HL60(human
fibroblast line)         promyelocytic
                       leukemia cell line)
    cytotoxic
                              cytotoxic
Discussion
 Resveratrol
  dose-dependent
  time-dependent
 Resveratrol
  16h,24h
  significant effect of viral DNA replication
Conclusion
 The target of Resveratrol is the
  replication of viral DNA
 further studies are required for the full
 elucidation of the inhibitory mechanism
 mediated by RV leading to the abrogation
 of the viral DNA synthesis
 the possibility that RV may find a potential
  clinical use for the control of proliferative
  pathologies or as an antiviral drug

				
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