tailieunhanh - Báo cáo Y học: Chemical structure and immunoreactivity of the lipopolysaccharide of the deep rough mutant I-69 Rd–/b+ of Haemophilus influenzae

From the lipopolysaccharide of the deep rough mutant I-69 Rd–/b+ of Haemophilus influenzae two oligosaccharides were obtained after de-O-acylation and separation by high-performance anion exchange chromatography. Their chemical structures were determined by one- and two-dimensional 1H-, 13C- and 31P-NMR spectroscopy as aKdo-4P-(2 fi 6)-bGlcN-4P-(1 fi 6)-aGlcN-1P and aKdo-5P-(2 fi 6)-bGlcN-4P-(1 fi 6)-aGlcN-1P. | Eur. J. Biochem. 269 1237-1242 2002 FEBS 2002 Chemical structure and immunoreactivity of the lipopolysaccharide of the deep rough mutant I-69 Rd7b of Haemophilus influenzae Sven Miiller-Loennies Lore Brade and Helmut Brade Research Center Borstel Center for Medicine and Biosciences Borstel Germany From the lipopolysaccharide of the deep rough mutant I-69 Rd- b of Haemophilus influenzae two oligosaccharides were obtained after de-ơ-acylation and separation by high-performance anion exchange chromatography. Their chemical structures were determined by one- and two-dimensional H-. 13C- and 31P-NMR spectroscopy as aKdo-4P- 2 6 -bGlcN-4P- 1 6 -aGlcN-1P and aKdo-5P- 2 6 -pGlcN-4P- 1 6 -aGlcN-1P. The spe-cifcity of mAbs S42-21 and S42-16 specifc for Kdo-4P or Kdo-5P respectively Rozalski A. Brade L. Kosma P. Moxon R. Kusumoto S. Brade H. 1997 . Mol. Micro biol. 23 569-577 was confrmed with neoglycoconjugates obtained by conjugation of the isolated oligosaccharides to BSA. In addition a mAb S42-10-8 with unknown epitope specificity could be assigned using the neoglycoconjugates described herein. This mAb binds to an epitope composed of the bisphosphorylated glucosamine backbone of lipid A and Kdo-4P whereby the latter determines the specifcity strictly by the position of the phosphate group. Keywords carbohydrate antibody Kdo-phosphate neoglycoconjugate serology sugar phosphate. Haemophilus influenzae normally colonizes the human nasopharynx but may cause severe infections in particular meningitis in children. A major virulence factor of this human pathogen is the type b capsule an acidic polysaccharide composed of ribose ribitol and phosphate and which is the basis of an effective conjugate vaccine 1 . Among other virulence factors is the lipopolysaccharide LPS in which we are interested for various reasons a LPS is an essential component of the outer membrane in all Gram-negative bacteria b LPS is the endotoxin of Gramnegative bacteria c LPS is a major surface antigen .