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Characterization of Lipopolysaccharide of Haemophilus influenzae

Lipopolysaccharide from strains of Haemophilus influenzae was extracted and isolated by the hot phenol-water procedure. The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosph...

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Published in:The Journal of infectious diseases 1978-12, Vol.138 (6), p.719-730
Main Authors: Flesher, Alan R., Insel, Richard A.
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Language:English
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Insel, Richard A.
description Lipopolysaccharide from strains of Haemophilus influenzae was extracted and isolated by the hot phenol-water procedure. The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosphate (4.7%); protein content was
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The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosphate (4.7%); protein content was &lt;1 %. Thin-layer chromatography, gas-liquid chromatography, and colorimetric assays detected glucose, galactose, glucosamine, heptose, and a 2-keto-3-deoxyoctonate- like molecule &lt;1 %). Neither methylpentose nor dideoxyhexose was detected. The lipid portion was composed of fatty acids common to lipopolysaccharide of Salmonella. The preparations provoked positive dermal Shwartzman reactions and biphasic febrile responses in rabbits, responses typical of endotoxic activity. The 50% lethal dose for mice was decreased from 16.5 μg/g to 0.015 μg/g by concomitant administration of actinomycin D. The preparations were shown to be polyclonal activators of bone marrow-derived (B) cells. Limulus lysate gelation was seen with 8.0 ng of lipopolysaccharide. Preliminary hemagglutination data suggested at least three different antigenic factors associated with the lipopolysaccharide of H. influenzae type b. 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The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosphate (4.7%); protein content was &lt;1 %. Thin-layer chromatography, gas-liquid chromatography, and colorimetric assays detected glucose, galactose, glucosamine, heptose, and a 2-keto-3-deoxyoctonate- like molecule &lt;1 %). Neither methylpentose nor dideoxyhexose was detected. The lipid portion was composed of fatty acids common to lipopolysaccharide of Salmonella. The preparations provoked positive dermal Shwartzman reactions and biphasic febrile responses in rabbits, responses typical of endotoxic activity. The 50% lethal dose for mice was decreased from 16.5 μg/g to 0.015 μg/g by concomitant administration of actinomycin D. The preparations were shown to be polyclonal activators of bone marrow-derived (B) cells. Limulus lysate gelation was seen with 8.0 ng of lipopolysaccharide. Preliminary hemagglutination data suggested at least three different antigenic factors associated with the lipopolysaccharide of H. influenzae type b. 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purification</subject><subject>Male</subject><subject>Mice</subject><subject>Phosphates - isolation &amp; purification</subject><subject>Polymyxins</subject><subject>Rabbits</subject><subject>Salmonella</subject><subject>Scientific Articles</subject><subject>Shwartzman Phenomenon - etiology</subject><subject>Solubility</subject><subject>Spleen cells</subject><issn>0022-1899</issn><issn>1537-6613</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1978</creationdate><recordtype>article</recordtype><recordid>eNpFkD1PwzAQhi3EV_nYGUDqxJb2HDu2s0EroEgVDBSEWCzHcYQhrYOdSJRfj6tUMJ10z917ugehMwwjDDkZ21VV2jDGRIzYiON8Bw1wRnjCGCa7aACQpgkWeX6IjkL4AABKGD9A-wSDyLIBupq-K690a7z9Ua11q6GrhnPbuMbV66C0jtiWZtOdKbN0zbutuzCMZ-vOrH6UOUF7laqDOd3WY_R8e7OYzpL549399Hqe6JSJNhGVAKbLkmpu0pwWAJkWtBSVzonOgBQlTpmimmpTFJBCBqpUqaBcFYIozMkxuuxzG---OhNaubRBm7pWK-O6IDklJI1Px0HoB7V3IXhTycbbpfJriUFunMnemYzOJJPRWVw532Z3xdKUfwu9pIgvevwRWuf_KcQ0IJuLSc9taM33H1f-UzJOeCZnr2_y7WUxofzhSU7ILwu0gpg</recordid><startdate>197812</startdate><enddate>197812</enddate><creator>Flesher, Alan R.</creator><creator>Insel, Richard A.</creator><general>The University of Chicago Press</general><general>University of Chicago Press</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>197812</creationdate><title>Characterization of Lipopolysaccharide of Haemophilus influenzae</title><author>Flesher, Alan R. ; 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purification</topic><topic>Male</topic><topic>Mice</topic><topic>Phosphates - isolation &amp; purification</topic><topic>Polymyxins</topic><topic>Rabbits</topic><topic>Salmonella</topic><topic>Scientific Articles</topic><topic>Shwartzman Phenomenon - etiology</topic><topic>Solubility</topic><topic>Spleen cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Flesher, Alan R.</creatorcontrib><creatorcontrib>Insel, Richard A.</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of infectious diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Flesher, Alan R.</au><au>Insel, Richard A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of Lipopolysaccharide of Haemophilus influenzae</atitle><jtitle>The Journal of infectious diseases</jtitle><addtitle>J Infect Dis</addtitle><date>1978-12</date><risdate>1978</risdate><volume>138</volume><issue>6</issue><spage>719</spage><epage>730</epage><pages>719-730</pages><issn>0022-1899</issn><eissn>1537-6613</eissn><abstract>Lipopolysaccharide from strains of Haemophilus influenzae was extracted and isolated by the hot phenol-water procedure. The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosphate (4.7%); protein content was &lt;1 %. Thin-layer chromatography, gas-liquid chromatography, and colorimetric assays detected glucose, galactose, glucosamine, heptose, and a 2-keto-3-deoxyoctonate- like molecule &lt;1 %). Neither methylpentose nor dideoxyhexose was detected. The lipid portion was composed of fatty acids common to lipopolysaccharide of Salmonella. The preparations provoked positive dermal Shwartzman reactions and biphasic febrile responses in rabbits, responses typical of endotoxic activity. The 50% lethal dose for mice was decreased from 16.5 μg/g to 0.015 μg/g by concomitant administration of actinomycin D. The preparations were shown to be polyclonal activators of bone marrow-derived (B) cells. Limulus lysate gelation was seen with 8.0 ng of lipopolysaccharide. Preliminary hemagglutination data suggested at least three different antigenic factors associated with the lipopolysaccharide of H. influenzae type b. The H. influenzae lipopolysaccharide appeared biologically similar to that of enterobacteria but chemically different.</abstract><cop>United States</cop><pub>The University of Chicago Press</pub><pmid>310855</pmid><doi>10.1093/infdis/138.6.719</doi><tpages>12</tpages></addata></record>
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source Oxford University Press:Jisc Collections:Oxford Journal Archive: Access period 2024-2025
subjects Animals
Antigens, Bacterial - isolation & purification
Antiserum
B lymphocytes
Bacterial Proteins - isolation & purification
Endotoxins - immunology
Fatty acids
Fatty Acids - isolation & purification
Galactose - isolation & purification
Glucosamine - isolation & purification
Glucose - isolation & purification
Haemophilus influenzae - analysis
Haemophilus influenzae type b
Heptoses
Heptoses - isolation & purification
Ketoses - isolation & purification
Lipids
Lipopolysaccharides
Lipopolysaccharides - analysis
Lipopolysaccharides - immunology
Lipopolysaccharides - isolation & purification
Male
Mice
Phosphates - isolation & purification
Polymyxins
Rabbits
Salmonella
Scientific Articles
Shwartzman Phenomenon - etiology
Solubility
Spleen cells
title Characterization of Lipopolysaccharide of Haemophilus influenzae
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