Assessment of immune response to meningococcal disease: comparison of a whole-blood assay and the serum bactericidal assay

Ison, C; Anwar, Natasha; Cole, Michelle; Galassini, Rachel; Heyderman, Robert S.; Klein, Nigel; West, John T.; Pollard, Andrew J.; Morley, Sarah L.; Levin, Michael; Group, Meningococcal Research

doi:10.1006/mpat.1999.0296

Cited by 40 publications

(29 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…As previously reported, the WBA is more sensitive than the SBA assay (22,23,32), with killing being detected in subjects with no SBA activity.…”

Section: Discussionsupporting

confidence: 63%

“…In addition, a surface labeling assay, which detects antibody binding to meningococci, has also been developed (1,2,16), but further data are required to determine whether these assays may add to our knowledge of correlates of protection. The whole-blood assay (WBA), which measures the bactericidal activity of blood, including phagocytosis, in conjunction with complement-mediated lysis has been postulated as being more sensitive than the SBA assay (22,23,32). However, the WBA requires large volumes of fresh blood, is difficult to standardize and control, and may therefore not be suitable for vaccine trials.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Comparison and Correlation of Neisseria meningitidis Serogroup B Immunologic Assay Results and Human Antibody Responses following Three Doses of the Norwegian Meningococcal Outer Membrane Vesicle Vaccine MenBvac

Findlow

Taylor²,

Aase

et al. 2006

Infect Immun

View full text Add to dashboard Cite

The prediction of efficacy of Neisseria meningitidis serogroup B (MenB) vaccines is currently hindered due to the lack of an appropriate correlate of protection. For outer membrane vesicle (OMV) vaccines, immunogenicity has primarily been determined by the serum bactericidal antibody (SBA) assay and OMV enzyme-linked immunosorbent assay (ELISA). However, the opsonophagocytic assay (OPA), surface labeling assay, whole blood assay (WBA), and salivary antibody ELISA have been developed although correlation with protection is presently undetermined. Therefore, the aim of the study was to investigate further the usefulness of, and relationships between, MenB immunologic assays. A phase II trial of the OMV vaccine, MenBvac, with proven efficacy was initiated to compare immunologic assays incorporating the vaccine and six heterologous strains. Correlations were achieved between the SBA assay, OMV ELISA, and OPA using human polymorphonuclear leukocytes and human complement but not between an OPA using HL60 phagocytic cells and baby rabbit complement. Correlations between the surface labeling assay, the SBA assay, and the OMV ELISA were promising, although target strain dependent. Correlations between the salivary antibody ELISA and other assays were poor. Correlations to the WBA were prevented since many samples had results greater than the range of the assay. The study confirmed the immunogenicity and benefit of a third dose of MenBvac against the homologous vaccine strain using a variety of immunologic assays. These results emphasize the need for standardized methodologies that would allow a more robust comparison of assays between laboratories and promote their further evaluation as correlates of protection against MenB disease.

show abstract

“…As previously reported, the WBA is more sensitive than the SBA assay (22,23,32), with killing being detected in subjects with no SBA activity.…”

Section: Discussionsupporting

confidence: 63%

mentioning

confidence: 99%

Comparison and Correlation of Neisseria meningitidis Serogroup B Immunologic Assay Results and Human Antibody Responses following Three Doses of the Norwegian Meningococcal Outer Membrane Vesicle Vaccine MenBvac

Findlow

Taylor²,

Aase

et al. 2006

Infect Immun

View full text Add to dashboard Cite

show abstract

“…The whole-blood assay (WBA) was performed as described previously (15). Briefly, N. meningitidis was grown to mid-log phase by inoculation from an overnight culture into fresh GC (Difco Laboratories, Moseley, Surrey, United Kingdom) medium supplemented with IsoViteliX and incubated for 4 h at 37°C in 6% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Vaccination with Attenuated Neisseria meningitidis Strains Protects against Challenge with Live Meningococci

Sun

Ison

et al. 2004

Infect Immun

View full text Add to dashboard Cite

Meningococcal disease is a life-threatening infection caused by Neisseria meningitidis. Currently, there are no vaccines to prevent infection with serogroup B N. meningitidis strains, the leading cause of meningococcal meningitis in Europe and North America. Here we describe the construction and characterization of two attenuated serogroup B N. meningitidis strains, YH102 (MC58⌬sia ⌬rfaF) and YH103 (MC58⌬sia ⌬metH). Both strains are markedly attenuated in their capacity to cause bacteremia in rodent models and have a reduced ability to survive in a human whole-blood assay. Immunization of adult mice with these strains leads to the development of bactericidal antibodies and confers sterilizing protection against challenge with homologous live bacteria. Furthermore, we show that the strains confer protection against infection by other serogroups. Use of the attenuated strains in animals with gene knockouts or after depletion of immunological effectors could be used to define the basis of protection, and human volunteer studies could be undertaken to examine the immune response following exposure to this important human pathogen.

show abstract

“…The amount of highaffinity antimeningococcal antibodies detected by an affinity ELISA (8,10) has been shown to correlate with SBA (10), and previous ELISA studies using meningococcal serogroup C polysaccharide have also been shown to correlate with SBA results (27). However, a number of other assays have recently been developed to reveal the functional characteristics of antimeningococcal antibodies, including a whole-blood assay (17), an opsonophagocytic killing assay (34,39), and chemiluminescence-and flow cytometry (FCM)-based species-and antigenspecific opsonophagocytosis assays (OPAs) (11,14,20,23,24,38). The traditional SBA is highly dependent on both the complement source (44) and the target strain used and is not ideal since the contribution of these variables to the end point, bacterial killing, cannot be easily distinguished from that of functional antibodies.…”

mentioning

confidence: 99%

Functional Opsonic Activity of Human Serum Antibodies to Inner Core Lipopolysaccharide (galE) of Serogroup B Meningococci Measured by Flow Cytometry

et al. 2001

View full text Add to dashboard Cite

A recently described flow cytometric opsonophagocytic assay (OPA) was adapted to quantify the functional activity of serum antibodies specifically directed against serogroup B inner core lipopolysaccharide (LPS) of Neisseria meningitidis. The percentage of human peripheral polymorphonuclear leukocytes and monocytes (PMNms) ingesting fluorescently labeled, ethanol-fixed N. meningitidis organisms (phagocytic activity) in the presence of human sera was measured to reflect the serum opsonic activity against the bacterium. The contribution to opsonophagocytic activity of antibodies to inner core LPS was estimated by comparing the opsonic activities of adult and infant sera before and after adsorbing anti-LPS antibodies from the sera using purified LPS extracted from an LPS mutant (galE) of N. meningitidis strain MC58 (B:15:P1.7,16:L3). The specificity of the assay was further investigated using monoclonal antibody (MAb) B5, which binds to an inner core LPS epitope of N. meningitidis. A dose-dependent decrease in phagocytic activity was observed when MAb B5 was incubated with LPS from an inner core LPS (galE) mutant. Similarly, the number of PMNms ingesting fluorescently labeled polystyrene beads coated with inner core (galE) LPS decreased in a dose-dependent fashion when MAb B5 was incubated with various concentrations of the homologous inner core LPS. Strong correlations were found between the concentration of serum antibodies to inner core LPS (galE) versus the phagocytic activity using healthy adult sera (r 2 ‫؍‬ 0.89). There was a correlation between phagocytic ingestion and initiation of intracellular oxidative burst (r 2 ‫؍‬ 0.99) using polystyrene beads coated with inner core LPS and opsonized with the same sera using the oxidative burst indicator system dihydrorhodamine123/rhodamine 123. OPA results were also found to correlate closely with the results of the serum bactericidal assay using MAb B5 against the N. meningitidis MC58 galE mutant in the presence of human complement (r 2 ‫؍‬ 0.994, P ‫؍‬ 0.003, two-tailed test). These studies demonstrate that functional antibodies are produced in humans against meningococcal inner core LPS and that the OPA is a useful approach to study the opsonic activity of antibodies to inner core LPS in health and disease.

show abstract

Assessment of immune response to meningococcal disease: comparison of a whole-blood assay and the serum bactericidal assay

Cited by 40 publications

References 26 publications

Comparison and Correlation of Neisseria meningitidis Serogroup B Immunologic Assay Results and Human Antibody Responses following Three Doses of the Norwegian Meningococcal Outer Membrane Vesicle Vaccine MenBvac

Comparison and Correlation of Neisseria meningitidis Serogroup B Immunologic Assay Results and Human Antibody Responses following Three Doses of the Norwegian Meningococcal Outer Membrane Vesicle Vaccine MenBvac

Vaccination with Attenuated Neisseria meningitidis Strains Protects against Challenge with Live Meningococci

Functional Opsonic Activity of Human Serum Antibodies to Inner Core Lipopolysaccharide (galE) of Serogroup B Meningococci Measured by Flow Cytometry

Contact Info

Product

Resources

About