title: Identification and characterisation of the immune response properties of Lampetra japonica BLNK.
abstract: B cell linker protein (BLNK) is a central linker protein involved in B cell signal transduction in jawed vertebrates. In a previous study, we have reported the identification of a BLNK homolog named Lj-BLNK in lampreys. In this study, a 336 bp cDNA fragment encoding the Lj-BLNK Src homology 2 (SH2) domain was cloned into the vector pET-28a(+) and overexpressed in Escherichia coli BL21. The recombinant fragment of Lj-BLNK (rLj-BLNK) was purifiedby His-Bind affinity chromatography, and polyclonal antibodies against rLj-BLNK were raised in male New Zealand rabbits. Fluorescenceactivated cell sorting (FACS) analysisrevealed that Lj-BLNK was expressed in approximately 48% of the lymphocyte-like cells of control lampreys, and a significant increase in Lj-BLNK expression was observed in lampreys stimulated with lipopolysaccharide (LPS). Western blotting analysis showed that variable lymphocyte receptor B (VLRB) and Lj-BLNKwere distributed in the same immune-relevant tissues, and the levels of both were upregulated in supraneural myeloid bodies and lymphocyte-like cells after LPS stimulation. Immunofluorescence demonstrated that Lj-BLNK was localized in VLRB(+) lymphocyte-like cells. These results indicate that the Lj-BLNK protein identified in lampreys might play an important role in the VLRB-mediated adaptive immune response.
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Yinglun
authors.initials: Y
authors.lastname: Han
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Biyue
authors.initials: B
authors.lastname: Shi
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Xiao
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Gou
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Life Science, Liaoning Normal University, Dalian 116029, China.|Lamprey Research Center, Liaoning Normal University, Dalian 116029, China.
authors.forename: Qingwei
authors.initials: Q
authors.lastname: Li
mesh_terms: D056704:Adaptive Immunity; D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D007798:Lampreys / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology; D011817:Rabbits; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D008070:Lipopolysaccharides; D011947:Receptors, Antigen, B-Cell; D011994:Recombinant Proteins