Immune activation of apolipophorin-III and its distribution in hemocyte from Hyphantria cunea

Hong Ja Kim, Hyun Jeong Je, Shin Yong Park, In Hee Lee, Byung Rae Jin, Hwa Kyung Yun, Chi Young Yun, Yeon Soo Han, Young Jin Kang, Sook Jae Seo
Insect Biochemistry and Molecular Biology 2004, 34 (10): 1011-23
Apolipophorin-III (apoLp-III) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium. Recently, apoLp-III in Galleria mellonella larvae was shown to play an unexpected role in insect immune activation. We identified the cDNA sequence of Hyphantria cunea apoLp-III by oligonucleotide-primed amplification, and 5'- and 3'-RACE PCR. Since H. cunea has an unusually low level of apoLp-III in the hemolymph, a recombinant apoLp-III was overexpressed using a baculovirus expression system to investigate its biological activity. Recombinant apoLp-III and/or Escherichia coli were injected into the hemocoel of last instar larvae, and the expression of antimicrobial peptide from fat body was determined by Northern blot. Injection of apoLp-III as well as E. coli induced slight up-regulation of its transcription rate in fat body, whereas the expression of antimicrobial peptide was dramatically induced by the injection of apoLp-III and E. coli. H. cunea hemocytes had apoLp-III in the granules and expressed its transcript, albeit at a much lower level than in the fat body. Upon bacterial injection, a subpopulation of hemocytes showed degranulation and degradation. Local discharge of apoLp-III from hemocytes caused by the injection of E. coli might be related to the immune response through an unknown mechanism.

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