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J Biol Chem, Vol. 274, Issue 17, 11727-11735, April 23, 1999
From the European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany
Cell lines from the malaria vector
Anopheles gambiae have been established as a tool for the
study of the mosquito innate immune system in vitro. Here,
we describe the first continuous insect cell line that produces
prophenoloxidase (PPO). This cell line (4a-3B) expresses constitutively
six PPO genes, three of which are novel (PPO4, PPO5, and
PPO6). The PPO genes show distinct temporal expression
profiles in the intact mosquito, spanning stages from the embryo to the
adult in an overlapping manner. Transient induction of larva-specific
PPO genes in blood-fed adult females suggests that the developmental
hormone 20-hydroxyecdysone may be involved in PPO gene regulation.
Indeed, exposure of 4a-3B cells to 20-hydroxyecdysone in culture
results in induction of those PPO genes that are mainly expressed in
early developmental stages, and repression of PPO5, which
is preferentially expressed at the adult stage. The cell line shows
bacteria-induced immune transcripts that encode defensin and
Gram-negative bacteria-binding protein, but no induction of PPO
transcripts. This cell line most likely derives from a hemocyte
lineage, and represents an appropriate in vitro model for
the study of the humoral and cellular immune defenses of A. gambiae.
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