[Preparation of human anti-HBs from volunteers with hepatitis B vaccine boost vaccination by modified B-cell immortalization technique].
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.
Lou
Hai-bo
HB
Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. louhaibo2008@163.com
Zhou
Yuan-ping
YP
Zhou
Fu-yuan
FY
Zhang
Ming-xia
MX
Wei
Jun-feng
JF
Zeng
Yan-li
YL
Wang
Jun-jie
JJ
Wang
Yan-jun
YJ
Wu
Ai-hua
AH
Hou
Jin-lin
JL
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
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines