Identification and characterisation of the immune response properties of Lampetra japonica BLNK
Han, Y.; Liu, X.; Shi, B.; Xiao, R.; Gou, M.; Wang, H.; Li, Q. (2016). Identification and characterisation of the immune response properties of Lampetra japonica BLNK. NPG Scientific Reports 6(25308): 8 pp. http://dx.doi.org/10.1038/srep25308 In: Scientific Reports (Nature Publishing Group). Nature Publishing Group: London. ISSN 2045-2322; e-ISSN 2045-2322, more |   |
| Keywords | Lampetra japonica (Martens, 1868) [WoRMS]
Marine/Coastal |
| Authors | | Top | - Han, Y.
- Liu, X.
- Shi, B.
- Xiao, R.
| - Gou, M.
- Wang, H., more
- Li, Q.
| |
| 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) analysis revealed 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 |
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