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Activation of an innate immune response in the schistosome-transmitting snail Biomphalaria glabrata by specific bacterial PAMPs
Authors:John T. Sullivan  Joseph A. Belloir
Affiliation:Department of Biology, University of San Francisco, San Francisco, CA 94117, USA
Abstract:Injection of crude lipopolysaccharide (LPS) from Escherichia coli into the hemocoel of Biomphalaria glabrata stimulates cell proliferation in the amebocyte-producing organ (APO). However, it is not known if mitogenic activity resides in the lipid A or O-polysaccharide component of LPS. Moreover, the possible role of substances that commonly contaminate crude LPS and that are known to stimulate innate immune responses in mammals, e.g., peptidoglycan (PGN), protein, or bacterial DNA, is unclear. Therefore, we tested the effects of the following injected substances on the snail APO: crude LPS, ultrapurified LPS (lacking lipoprotein contamination), two forms of lipid A, (diphosphoryl lipid A and Kdo2-lipid A), O-polysaccharide, Gram negative PGN, both crude and ultrapurified (with and without endotoxin activity, respectively), Gram positive PGN, PGN components Tri-DAP and muramyl dipeptide, and bacterial DNA. Whereas crude LPS, ultrapurified LPS, and crude PGN were mitogenic, ultrapurified PGN was not. Moreover, LPS components, PGN components, and bacterial DNA were inactive. These results suggest that it is the intact LPS molecule which stimulates cell division in the APO.
Keywords:APO, amebocyte-producing organ   DMSO, dimethyl sulfoxide   EU, endotoxin unit   Kdo, keto-3-deoxyoctonoic acid   LPS, lipopolysaccharide   LPS-B4, LPS from Escherichia coli O111:B4   LPS-B4U, LPS-B4, ultrapurified   LPS-B8, LPS from Escherichia coli O127:B8   MDP, muramyl dipeptide   NAG, n-acetylglucoseamine   PAMP, pathogen associated molecular pattern   PBS, 1/3 dilution of phosphate buffered saline   PGN, peptidoglycan   PGN-BS, PGN from Bacillus subtilis   PGN-EC, PGN from E. coli O111-B4   PGN-ECU, PGN from E. coli K12, ultrapurified   PRR, pattern recognition receptor   TLR, Toll-like receptor   Tri-DAP, l-alanine-γ-d-glutamate-meso-diaminopimelate
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