Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli

Induction of phenoloxidase and other immunological activities in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Lipopolysaccharide (LPS)

“…It is similar to the temporal induction kinetics by E. coli (Pang et al 2006) but later than V. alginolyticus (Pang et al 2010 (Cardenas and Dankert 1997), the mussel Mytilus edulis (Coles and Pipe 1994), and the locusts (Ratcliffe et al 1991). PO in the humoral fluids from unchallenged amphioxus comprises three bands, with a molecular mass of 72, 46, and 44 kDa, respectively, confirming our previous observation (Pang et al , 2009. Surprisingly, and interestingly, there were four bands from challenged amphioxus.…”

“…PO assay PO activity in the humoral fluids was assayed by the method of Horowitz and Shen (1952) with slight modifications (Pang et al 2004(Pang et al , 2009). The humoral fluids prepared were run on a 10 % SDS-PAGE gel with a 4 % spacer gel using the buffer system of Laemmli (1970).…”

“…After electrophoresis, the gel was cut into two parts. One part of the gel was detected for PO activity according to the method of Nellaiappan and Vinayagam (1986) with slight modifications (Pang et al , 2009). The gel was washed twice in icecold distilled water and incubated in 50 mM PBS (pH 6.5) containing 10 mM 4-methylcatechol and 0.3 % 3-methyl-2-benzothiazolinone hydrazone (MBTH) dissolved in 25 % ethanol.…”

“…The humoral immune defense system in amphioxus plays an unexceptionally pivotal role in limiting microbial invasions and mediating the clearance or killing of invading microbes. Our previous investigation has shown that phenoloxidase activity, lysozyme activity, antimicrobial activity, microbial agglutinating activity, and hemagglutinating activity in the humoral fluids from amphioxus challenged with Escherichia coli and Vibrio alginolyticus significantly increased, which probably provides some degree of protection for the animals against invading microbes (Pang et al 2006(Pang et al , 2009(Pang et al , 2010. However, it needs further testing by LPS, which is common to all Gramnegative bacteria.…”

Immune parameters in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Vibrio alginolyticus

“…Recently, induction of phenoloxidase in amphioxus challenged with microbes has also been reported [9]. However, there is still little information on the humoral immune defense system in this evolutionarily important organism.…”