Heterogeneity of antibodies reactive with the dominant antigen of Actinobacillus actinomycetemcomitans
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| Published in | Infection and Immunity Vol. 65; no. 9; pp. 3794 - 3798 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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Washington, DC
American Society for Microbiology
01.09.1997
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| AbstractList | The serotype b-specific carbohydrate antigen (SbAg) of Actinobacillus actinomycetemcomitans Y4 is reported to be the O antigen of lipopolysaccharide, and the highest titers of serum antibody reactive with A. actinomycetemcomitans in early-onset periodontitis (EOP) patients bind SbAg. These high titers of serum antibody reactive with SbAg are associated with a lesser extent and severity of periodontal disease. The aim of this study was to determine if a limited number of genes code for anti-SbAg antibodies as has been shown for immunoglobulin G (IgG) reactive with the type b polysaccharide from Haemophilus influenzae. Serum IgG reactive with the SbAg was prepared from 20 high-titer EOP patients by affinity chromatography. The IgG subclass concentrations were determined, and heterogeneity was analyzed by isoelectric focusing (IEF). IgG2 was the dominant subclass (83% of total IgG) in the anti-SbAg IgG fraction and represented an average of 1.33% of total serum IgG2. The IgG2 reactive with SbAg was isolated from the affinity-purified IgG fraction by affinity chromatography with protein A and subclass-specific monoclonal antibodies. On IEF gels, only 4 to 20 bands were observed in the anti-SbAg IgG fractions, indicating limited heterogeneity. N-terminal amino acid sequence analysis of eight representative anti-SbAg IgG2 preparations indicated that variable heavy and light chains consisted largely of V(H)III and V(kappa)II, respectively. However, a significant fraction of anti-SbAg may use V(H) and V(lambda) genes with blocked N termini. In short, these findings indicate that IgG reactive with SbAg is very much like the antibody reactive with H. influenzae type b polysaccharide. Similarities include IgG2 dominance, limited bands on IEF gels, supporting an oligoclonal response, and use of genes from V(H)III and V(kappa)II regions. Services IAI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue IAI About IAI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy Connect to IAI IAI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0019-9567 Online ISSN: 1098-5522 Copyright © 2014 by the American Society for Microbiology. For an alternate route to IAI .asm.org, visit: IAI The serotype b-specific carbohydrate antigen (SbAg) of Actinobacillus actinomycetemcomitans Y4 is reported to be the O antigen of lipopolysaccharide, and the highest titers of serum antibody reactive with A. actinomycetemcomitans in early-onset periodontitis (EOP) patients bind SbAg. These high titers of serum antibody reactive with SbAg are associated with a lesser extent and severity of periodontal disease. The aim of this study was to determine if a limited number of genes code for anti-SbAg antibodies as has been shown for immunoglobulin G (IgG) reactive with the type b polysaccharide from Haemophilus influenzae. Serum IgG reactive with the SbAg was prepared from 20 high-titer EOP patients by affinity chromatography. The IgG subclass concentrations were determined, and heterogeneity was analyzed by isoelectric focusing (IEF). IgG2 was the dominant subclass (83% of total IgG) in the anti-SbAg IgG fraction and represented an average of 1.33% of total serum IgG2. The IgG2 reactive with SbAg was isolated from the affinity-purified IgG fraction by affinity chromatography with protein A and subclass-specific monoclonal antibodies. On IEF gels, only 4 to 20 bands were observed in the anti-SbAg IgG fractions, indicating limited heterogeneity. N-terminal amino acid sequence analysis of eight representative anti-SbAg IgG2 preparations indicated that variable heavy and light chains consisted largely of V sub(H)III and V sub( Kappa )II, respectively. However, a significant fraction of anti-SbAg may use V sub(H) and V sub( lambda ) genes with blocked N termini. In short, these findings indicate that IgG reactive with SbAg is very much like the antibody reactive with H. influenzae type b polysaccharide. Similarities include IgG2 dominance, limited bands on IEF gels, supporting an oligoclonal response, and use of genes from V sub(H)III and V sub( Kappa )II regions. |
| Author | K Nakashima H A Schenkein J V Califano J G Tew |
| AuthorAffiliation | Clinical Research Center for Periodontal Diseases, School of Dentistry, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298, USA |
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| Author_xml | – sequence: 1 givenname: K surname: NAKASHIMA fullname: NAKASHIMA, K organization: Clinical Research Center for Periodontal Diseases, School of Dentistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298, United States – sequence: 2 givenname: H. A surname: SCHENKEIN fullname: SCHENKEIN, H. A organization: Clinical Research Center for Periodontal Diseases, School of Dentistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298, United States – sequence: 3 givenname: J. V surname: CALIFANO fullname: CALIFANO, J. V organization: Clinical Research Center for Periodontal Diseases, School of Dentistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298, United States – sequence: 4 givenname: J. G surname: TEW fullname: TEW, J. G organization: Clinical Research Center for Periodontal Diseases, School of Dentistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298, United States |
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| Keywords | Human Immunoglobulins Immune response Antibody IgG2 Immunodominance Pasteurellaceae Antigen Immunogenetics Gene Actinobacillus actinomycetemcomitans N terminal-Sequence Bacteria Humoral immunity Polymorphism |
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StumbleUpon... The serotype b-specific carbohydrate antigen (SbAg) of Actinobacillus actinomycetemcomitans Y4 is reported to be the O antigen of lipopolysaccharide, and the... |
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| SubjectTerms | Actinobacillus actinomycetemcomitans Aggregatibacter actinomycetemcomitans - immunology Amino Acid Sequence Analysis of the immune response. Humoral and cellular immunity Antibodies, Bacterial - immunology Antibody Affinity Antibody Diversity Antibody production Biological and medical sciences Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Immunobiology Immunoglobulin G - analysis Immunoglobulin Isotypes - analysis Immunoglobulin kappa-Chains - analysis Immunoglobulin lambda-Chains - analysis Isoelectric Point Molecular Sequence Data |
| Title | Heterogeneity of antibodies reactive with the dominant antigen of Actinobacillus actinomycetemcomitans |
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