INTRODUCION................................................................ 193 STUDIES ON 0 ANTIGENS...................................................... 196 Isolation and Properties of O-Specific Preparations from Bacteria.................. 196 Trichloroacetic acid extraction of the whole \"Boivin\" antigen.................... 196 Extraction of lipopolysaccharides with phenol-water, aqueous ether, and similar methods.................................................. ............. 196 Extraction ofpolysaccharides with sodium hydroxide: alkali polysaccharide ...... 197 Extraction ofpolysaccharide by acetic acid: degraded Freeman polysaccharide...... 197 Physical and biological properties of 0-specific preparations..................... 197 Sugar Constituents of the Specific Polysaccharides.............................. 198 6-Deoxyhexoses........................................................... 199 3, 6-Dideoxyhexoses ...................................................... 199 Heptoses................................................................. 200 Hexosamines.............................................................. 200 Ribose.................................................................. 200 2-Keto-3-deoxy-octonate (KDO)............................................. 200 0-phosphorylethanolamine.................................................. 201 Sugar Composition ofSpecific Polysaccharides.................................. 201 Structure of the O-Specific Side Chain.......................................... 204 Salmonella ofgroup DI ................. .................................... 204 Salmonella ofgroup B ............... ...................................... 205 Salmonella ofgroups E1 (3, 10), E2 (3, 15), E3 (3, 15, 34), and E4 (1, 3, 19) ......... 206 Salmonella of groups G (13, 22), N (30), and U (43) ...... .................... 206 Structure of 0 Factors in Specific Polysaccharides.............................. 206 Role of 3,6-dideoxyhexoses: 0 factors 2, 4, 8, 9, 35, 50........................ 207 Artificial antigens......................................................... 209 Factor 5................................................................. 209 Factors 1, 112, 19, 37....................................................... 210 Factors 3, 10, 15, and 34 .............. ..................................... 211 Factor 12........... .. .. ... .. ... .. ... ............................ 212 Location of the factors on the chain........................................ 212 Biochemical and Genetic Basis of Changes in Specificity After Phage Conversion .... 214 Appearance offactor 1..................................................... 215 Conversions in group E..................................................... 216 Appearance of factors 27A, 27B, and 27D in groups A, B, and D after conversion byphage27............................................................ 216 STUDIES ON R ANTIGENS...................................................... 219 Preparation and Composition of R (Lipo)polysaccharides........................ 221 R Mutants ofChemotype Ra (Serotype R II Mutants) ...... ...................... 221 Chemical structure ........................................................ 223 Enzymatic defects......................................................... 225 R Mutants of Chemotype Rb (Serotype R I Mutants)............................. 227 Serology.................................................................. 227 Chemical structure ........................................................ 227 Enzymatic defects......................................................... 227 0-specific polysaccharide hapten ofR I mutants ....... ......................... 228 R Mutants ofChemotype Rc (M Mutants) ........ .............................. 228 Enzymatic defects and biochemical properties.................................. 228 Chemical analysis ofchemotype Rc lipopolysaccharides......................... 229 Accumulation ofnucleoside diphospho sugars in M mutants...................... 229 R Mutants of Chemotype Rd.................................................. 230 R Mutants of Chemotype Re.................................................. 230 Salmonella T Forms......................................................... 231 Semirough Forms ........... ................................................ 231 Genetics ofR Mutants ...................................................... 232

https://mmbr.asm.org/content/mmbr/30/1/192.full.pdf

Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae.

CHAPTER 4 – Isolation and Chemical and Immunological Characterization of Bacterial Lipopolysaccharides

Publisher Summary This chapter describes the preparation and use of synthetic polypeptides, polypeptidyl proteins, and conjugates of various small molecules with synthetic polypeptides in studies of the molecular basis of immunological phenomena This line of study has already partially defined the roles of composition, accessibility, size, shape, electrical charge, optical configuration, and macromolecular conformation in immunogenicity and antigenic specificity Similar work is now underway relating these factors to the phenomena of immunological tolerance and delayed hypersensitivity The simple model antigens have afforded a better understanding of genetic variations in the immunological response that are not observable with complex natural antigens The synthesis and characterization of poly-amino acids and polypeptidyl proteins, antigenic properties of polypeptidyl proteins, reactions of synthetic polypeptides with antisera to proteins, conjugates of synthetic polypeptides, antigenic properties of natural polyglutamic acid, antigenicity of some peptides synthesized by stepwise methods, immunological unresponsiveness, and antibodies for synthetic polypeptides and polypeptidyl proteins are also discussed in the chapter

Immunological studies with synthetic polypeptides.

Binding constants of the dextran-reactive BALB/c mouse IgA myeloma proteins W3129 and QUPC 52 have been determined for each member of the isomaltose series of oligosaccharides and for methyl alphaDglucoside. Protein W3129 has maximum complementarity for isomaltopentaose (IM5) deltaf degrees = 7,180 cal/mol) with 55-60% of the total binding energy directed against methylalphaDglucoside. Protein QUPC 52 gives maximum binding with isomaltohexaose (IM6) (deltaF degrees = -5,340 cal/mol) and has about 70% of its total binding energy for isomaltotriose (IM3), but at most only 5% for isomaltose (IM2) or methyl alphaDglucoside. Protein W3129 precipitates with branched dextrans high in alpha (1 yields 6) linkages and reacts with but does not precipitate a synthetic alpha (1 yields 6)-linked linear dextran. Protein QUPC 52 precipitates both branched and linear dextrans. Thus, the immunodominant group for protein W3129 is mimicked by methyl alphaDglucoside and this protein reacts exclusively at the terminal nonreducing ends of alpha (1 yields 6)-linked dextran chains. Protein QUPC 52 has an immunodominant group which is expressed by IM3 but not smaller oligosaccharides and this protein can react at nonterminal locations along alpha (1 yields 6)-linked dextran chains. Precipitation of linear dextran seems to be a valid although not quantitative assay for antidextrans with nonterminal specificity. Quantitative precipitin reactions with branched and linear dextrans suggest that alpha (1 yields 6)-specific human antidextrans are mixtures of molecules having terminal and nonterminal specificities and that the fraction of each type can vary among individuals. Rabbit antisera against IM3 or IM6 coupled to bovine serum albumin also appear to contain antibodies with nonterminal specificity for dextran chains although a large fraction has terminal specificity. Low molecular weight clinical dextran N-150N (congruent to 60,000) reacted more like linear dextran than like its parent native-branched dextran B512. This is thought to result from an abundance of nonterminal determinants in clinical dextran N-150N but a very small number of functional terminal determinants per molecule. An appreciation of terminal and nonterminal specificities and of the different immunodominant structures in isomaltosyl chains has proven to be of a great value in understanding the immunochemical reactions of dextrans. Moreover, certain previous findings with fructosan-reactive mouse myeloma proteins and human antilevans (55, 84) also suggest terminal and nonterminal specificities for levan chains.

https://rupress.org/jem/article-pdf/142/2/435/1087142/435.pdf

Binding properties of immunoglobulin combining sites specific for terminal or nonterminal antigenic determinants in dextran.

DURING the course of our investigations on the biosynthesis of 3,6-dideoxy-hexoses, found in the endotoxins of Salmonella and Escherichia
1–3 and in the eggs of Parascaris equorum
4, the need arose for a sensitive and specific assay. A consideration of the structures of these compounds suggested to us that the malonaldehyde–thiobarbituric acid reaction, which was adapted by Waravdekar and Saslaw5 for the estimation of 2-deoxy-ribose, might equally be applicable to the quantitative determination of 3,6-dideoxy-hexoses, and indeed, of 3-deoxy sugars in general. A sample of ascarylose (3,6-dideoxy-L-arabino-hexose), which was the generous gift of Dr. E. Lederer, was used to test this hypothesis.

Estimation of 3-Deoxy Sugars by Means of the Malonaldehyde–Thiobarbituric Acid Reaction

Contribution of Deoxymethylpentoses to the Serological Specificity of Some Bacterial Polysaccharides, and the Recognition of a New Sugar, Paratose

Contribution of deoxymethylpentoses to the serological specificity of some bacterial polysaccharides, and the recognition of a new sugar, paratose.

Preparation and Immunological Properties of an Artificial Antigen with Colitose (3-Deoxy-L-Fucose) as the Determinant Group

Serological Study of O27 Factors of Salmonella.

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Contribution of deoxymethylpentoses to the serological specificity of some bacterial polysaccharides, and the recognition of a new sugar, paratose.

Preparation and Immunological Properties of an Artificial Antigen with Colitose (3-Deoxy-L-Fucose) as the Determinant Group

Serological Study of O27 Factors of Salmonella.