title: The Application of Magnetic Bead Selection to Investigate Interactions between the Oral Microbiota and Salivary Immunoglobulins.
abstract: The effect of humoral immunity on the composition of the oral microbiota is less intensively investigated than hygiene and diet, in part due to a lack of simple and robust systems for investigating interactions between salivary immunoglobulins and oral bacteria. Here we report the application of an ex situ method to investigate the specificity of salivary immunoglobulins for salivary bacteria. Saliva collected from six volunteers was separated into immunoglobulin and microbial fractions, and the microbial fractions were then directly exposed to salivary immunoglobulins of "self" and "non-self" origin. Antibody-selected bacteria were separated from their congeners using a magnetic bead system, selective for IgA or IgG isotypes. The positively selected fractions were then characterized using gel-based eubacterial-specific DNA profiling. The eubacterial profiles of positively selected fractions diverged significantly from profiles of whole salivary consortia based on volunteer (P≤ 0.001%) and immunoglobulin origin (P≤ 0.001%), but not immunoglobulin isotype (P = 0.2). DNA profiles of separated microbial fractions were significantly (p≤ 0.05) less diverse than whole salivary consortia and included oral and environmental bacteria. Consortia selected using self immunoglobulins were generally less diverse than those selected with immunoglobulins of non-self origin. Magnetic bead separation facilitated the testing of interactions between salivary antibodies and oral bacteria, showing that these interactions are specific and may reflect differences in recognition by self and non-self immunoglobulins. Further development of this system could improve understanding of the relationship between the oral microbiota and the host immune system and of mechanisms underlying the compositional stability of the oral microbiota.
authors.affiliation: Manchester Pharmacy School, The University of Manchester, Manchester, M13 9PT, United Kingdom.
authors.forename: Tejal
authors.initials: T
authors.lastname: Madhwani
authors.affiliation: Manchester Pharmacy School, The University of Manchester, Manchester, M13 9PT, United Kingdom.
authors.forename: Andrew J
authors.identifier: http://orcid.org/0000-0002-5255-5425
authors.initials: AJ
authors.lastname: McBain
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000276:immunology*; D001419:Bacteria / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D007136:Immunoglobulins / Q000276:immunology*; D018189:Immunomagnetic Separation; D064307:Microbiota; D010802:Phylogeny; D012463:Saliva / Q000276:immunology* / Q000382:microbiology*
chemical_list: D000907:Antibodies, Bacterial; D004269:DNA, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007136:Immunoglobulins