Measurement of antibodies to pneumococcal, meningococcal and haemophilus polysaccharides, and tetanus and diphtheria toxoids using a 19-plexed assay

The measurement of antibody responses to vaccination is useful in the assessment of immune status in suspected immune deficiency. Previous reliance on enzyme-linked immunoabsorbent assays (ELISA) has been cumbersome, time-consuming and expensive. The availability of flow cytometry systems has led to...

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Bibliographic Details
Published in:	Journal of immunological methods Vol. 377; no. 1-2; pp. 37 - 46
Main Authors:	Whitelegg, Alison M.E., Birtwistle, Jane, Richter, Alex, Campbell, John P., Turner, James E., Ahmed, Tarana M., Giles, Lynda J., Fellows, Mark, Plant, Tim, Ferraro, Alastair J., Cobbold, Mark, Drayson, Mark T., MacLennan, Calman A.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 30-03-2012
Subjects:	Adolescent Adult adults Aged antibodies Antibody antigens Bacterial bacterial vaccines blood serum Diphtheria Toxoid - immunology enzyme-linked immunosorbent assay Flow cytometry Flow Cytometry - methods Flow Cytometry - standards Haemophilus influenzae Haemophilus Vaccines - immunology Humans Immune deficiency immune response immunoglobulin G Immunoglobulin G - blood Immunoglobulin G - immunology immunoglobulin M Immunoglobulin M - blood Immunoglobulin M - immunology immunosuppression (physiological) Meningococcal Vaccines - immunology Middle Aged Multiplex Pneumococcal Vaccines - immunology polysaccharides Reproducibility of Results Sensitivity and Specificity subunit vaccines tetanus Tetanus Toxoid - immunology toxoids Vaccination Young Adult Hib Flow cytometry Bacterial PS Antibody NHSBT Multiplex Vaccination NEQAS CWPS ATCC Immune deficiency MFI Men NIBSC Pn ELISA WHO
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Summary:	The measurement of antibody responses to vaccination is useful in the assessment of immune status in suspected immune deficiency. Previous reliance on enzyme-linked immunoabsorbent assays (ELISA) has been cumbersome, time-consuming and expensive. The availability of flow cytometry systems has led to the development of multiplexed assays enabling simultaneous measurement of antibodies to several antigens. We optimized a flow cytometric bead-based assay to measure IgG and IgM concentrations in serum to 19 antigens contained in groups of bacterial subunit vaccines: pneumococcal vaccines, meningococcal vaccines, Haemophilus influenzae b (Hib), and tetanus and diphtheria toxoid vaccines. 89-SF was employed as the standard serum. The assay was used to determine specific antibody levels in serum from 193 healthy adult donors. IgG and pneumococcal IgM antibody concentrations were measurable across 3 log10 ranges encompassing the threshold protective IgG antibody levels for each antigen. There was little interference between antibody measurements by the 19-plexed assay compared with monoplexed assays, and a lack of cross-reactive IgG antibody, but evidence for cross-reacting IgM antibody for 3/19 pneumococcal antigens. 90th centile values for 15/19 IgG concentrations and 12/12 IgM concentrations of the 193 adult sera were within these ranges and percentages of sera containing protective IgG antibody levels varied from 4% to 95% depending on antigen. This multiplexed assay can simultaneously measure antibody levels to 19 bacterial vaccine antigens. It is suitable for use in standard clinical practice to assess the in vivo immune response to test vaccinations and measure absolute antibody levels to these antigens. ► Multiplex flow cytometric antibody assay optimized for use with five vaccines. ► IgG and IgM antibodies measurable across 3 log10 ranges with minimal interference. ► Antibody levels to 19 bacterial vaccine antigens measured simultaneously. ► Specific antibody levels in 193 healthy donors measured with assay.
Bibliography:	http://dx.doi.org/10.1016/j.jim.2012.01.007 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0022-1759 1872-7905
DOI:	10.1016/j.jim.2012.01.007