科研之友
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摘要
p38MAPK is a subclass of mitogen activated protein kinases (MAPK), which plays an important role in the signal transduction of immune response in higher vertebrates. It was found that there are two isoforms of p38 MAPK in Lampetra japonica, and they were identified as p38 α (Lja-mapkl4) and p38 β (Lja-mapk11) through cloning their open reading frames and doing homologous sequences alignment and phylogenetic analysis. In order to examine whether Lja-mapk14 and lja-mapk11 are involved in lamprey immune response, mixed antigens are used to stimulate lampreys and their protein samples are detected by Western blotting. It was found that Lja-mapkl4 expression reached highest levels in peripheral blood lymphocytes, gill tissues and supraneural myeloid bodies after 36 h, 24 h and 24 h stimulation, respectively. The relative expression levels of Lja-mapkl4 were 2.9, 2. 1 and 2.6 folds higher than those of the control group, respectively. Lja-mapk11 expression levels all reach highest in these tissues after 36 h stimulation, and the relative expression levels of Lja-mapkll were 2. 2, 2. 5 and 6. 3 folds of those in the control group. The protein expressional patterns of Lja-mapk14 and Lja-mapk11 were further verified at transcriptional levels by real-time quantitative PCR (qPCR) methods in several tissues isolated from animals stimulated by mixed antigens. The qPCR results showed that the relative expression levels of Lja-mapk14 mRNA were up-regulated 2. 3, 1. 5 and 3. 4 folds in lymphocytes, gill tissues and supraneural myeloid bodies compared to their control group after 36 h immunization, respectively, while diose of Lja-mapk11 mRNA were up-regulated by 1.3, 2.6 and 1.6 folds in lymphocytes, gill tissues and hearts, respectively. The above results proved that Lja-mapk14 and Lja-mapk11 were involved in lamprey immune response at mRNA and protein levels. In order to investigate which sub-group of lymphocytes is related to Lja-mapk14 and Lja-mapk11, the lipopolysaccharide (LPS) and phytohemagglutinin (PHA) were used to stimulate lampreys. The results of Western blotting showed that Lja-mapkl4 and Lja-mapkll were up-regulated 1.3-4.1 folds in lymphocytes, gill tissues and supraneural myeloid bodies after 36 h of LPS immunization, while the expression levels of Lja-mapkl4 and Lja-mapkll were not significantly changed in the above tissues after 36 h of PHA immunization. These results suggest that Lja-mapkl4 and Lja-mapk11 may be involved in the B-cell mitogen LPS mediated immune response of VLRB+lymphocyte subset.
