Monday, December 10, 2001 - 1:12 PM
0308

Characterization of the effects of different lipopolysaccharides on the cellular immune system of tobacco hornworm, Manduca sexta

Jon C. Bedick1, R. L. Pardy2, and David W. Stanley1. (1) University of Nebraska-Lincoln, Insect Biochemical Physiology Laboratory, 311 Plant Industry Bldg, University of Nebraska, Lincoln, NE, (2) University of Nebraska-Lincoln, Laboratory for the Biology of Lipopolysaccharides, 246 Manter Hall, University of Nebraska, Lincoln, NE

Nodulation is the predominant insect cellular reaction to bacterial infections. Nodulation begins with formation of microaggregates of circulating hemocytes and entrapped bacterial cells. Microaggregates grow with addition of hemocytes and their attached bacteria, and the process is completed with addition of a final layer of plasmatocytes. The mature nodules are darkened by melanization. We reported that nodulation, along with other cellular defense reactions, is mediated by eicosanoids, which are signal moieties formed by enzymatic oxygenation of certain polyunsaturated fatty acids. Most recently we reported that cellular defense reactions are stimulated by treating insects with the lipopolysaccharide (LPS) components purified from the gram-negative bacterium Serratia marcescens. In this paper we report on the effects of LPS purified from four gram-negative bacteria (Halomonas elongata, Klebesella, Provedentia 2, Escherichia coli), three strains of Prototheca portoricensis algae, and two Lipid A moieties (a component of LPS) on nodulation reactions in Manduca sexta. We observed different degrees of nodule formation to challenges with the LPS from the different sources. At the low end, larvae treated with H. elongata stimulated no more nodules than controls while those treated with E. coli had very high numbers of nodules. Larvae treated with three different strains of P. portoricensis produced similar numbers of nodules and treatment with the two synthetic Lipid As did not depart from controls. In a second series of experiments E. coli and Provedentia 2 were incubated with Polymyxin B (PMB) before injection. PMB binds specifically to the Lipid A portion of LPS. In both cases larvae treated PMB-bound LPS had lower numbers of nodules than controls treated with unbound LPS. This research supports the hypothesis that the Lipid A is a necessary, but not sufficient component of LPS to stimulate insect nodulation reactions.

Species 1: Lepidoptera Sphingidae Manduca sexta (tobacco hornworm)
Keywords: bacteria, algae

The ESA 2001 Annual Meeting - 2001: An Entomological Odyssey of ESA