title: [Preparation of human anti-HBs from volunteers with hepatitis B vaccine boost vaccination by modified B-cell immortalization technique].
abstract: OBJECTIVE
  To establish immortalized B lymphoblast cell lines (B-LCLs) from healthy anti-HBs antibody (anti-HBs)-positive volunteers and screen for human anti-HBs and the antibody-secreting cells.
  
  
  METHODS
  The peripheral blood mononuclear cells (PBMC) isolated from 3 healthy volunteers positive for anti-HBs with hepatitis B vaccine boost vaccination were infected with Epstein-Barr virus (EBV) and incubated in the presence of CpG DNA motifs and cyclosporin A (CyA). The anti-HBs in the culture supernatant of the immortalized B-cells was quantified by Architect anti-HBs assay with chemiluminescent microparticle technique. Immunocytochemistry was performed to identify the differentiation of the cell clones expressing anti-HBs.
  
  
  RESULTS
  Immortalized B-cell culture was successfully established from the cell clones secreting anti-HBs with EBV infection and CpG DNA stimulation. The titer of anti-HBs in the culture supernatant was at its peak at 3 weeks of cell culture and then decreased gradually. At 3 months of cell culture, the cells still retained the capacity of anti-HBs production as verified by the results of immunocytochemistry for CD20 and CD138.
  
  
  CONCLUSIONS
  Immortalized B-cell culture secreting anti-HBs from volunteers receiving boost hepatitis B vaccination has been successfully established by modified EBV immortalization technique.
authors.affiliation: Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. louhaibo2008@163.com
authors.forename: Hai-bo
authors.initials: HB
authors.lastname: Lou
authors.forename: Yuan-ping
authors.initials: YP
authors.lastname: Zhou
authors.forename: Fu-yuan
authors.initials: FY
authors.lastname: Zhou
authors.forename: Ming-xia
authors.initials: MX
authors.lastname: Zhang
authors.forename: Jun-feng
authors.initials: JF
authors.lastname: Wei
authors.forename: Yan-li
authors.initials: YL
authors.lastname: Zeng
authors.forename: Jun-jie
authors.initials: JJ
authors.lastname: Wang
authors.forename: Yan-jun
authors.initials: YJ
authors.lastname: Wang
authors.forename: Ai-hua
authors.initials: AH
authors.lastname: Wu
authors.forename: Jin-lin
authors.initials: JL
authors.lastname: Hou
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002472:Cell Transformation, Viral; D006509:Hepatitis B / Q000517:prevention & control; D006510:Hepatitis B Antibodies / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D017325:Hepatitis B Vaccines / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D007117:Immunization, Secondary; D014611:Vaccination
chemical_list: D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines