Activation of mouse B lymphocyte by proteins containing hexahistidine.
Many recombinant proteins are produced as a fusion protein tagging hexahistidine, which has been used for studying their biological function both in vitro and in vivo. Unexpectedly, we observed activation of BALB/c mouse splenocytes when treated with hexahistidine-tagged recombinant proteins. This activation was hexahistidine-specific since the anti-pentahistidine antibody completely neutralized the effect, and the bovine serum albumin conjugated with the hexahistidine peptides also showed a similar activation effect. The B cells seemed to be the activated splenocytes, since the cell population stained with the anti-immunoglobulin antibody and anti-CD80 antibody was increased after the treatment. However, the activation signal by hexahistidine was insufficient to fully differentiate the B lymphocytes. This result suggests that caution must be taken in the use of hexahistidine-tagged recombinant proteins, due to their nonspecific activation of B lymphocyte.
Kim
K H
KH
Laboratory of Endocrinology, Institute for Medical Sciences, Ajou University School of Medicine, Suwon, Korea.
Kim
K S
KS
Choi
S E
SE
Kim
J K
JK
Park
J H
JH
Lee
J H
JH
Kang
Y
Y
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006639:Histidine; D008213:Lymphocyte Activation / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D013154:Spleen / Q000166:cytology; D014759:Viral Envelope Proteins / Q000276:immunology
D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D006639:Histidine