Serum bactericidal antibody assays – The role of complement in infection and immunity

Vaccine ◽  
2015 ◽  
Vol 33 (36) ◽  
pp. 4414-4421 ◽  
Author(s):  
E.D.G. McIntosh ◽  
M. Bröker ◽  
J. Wassil ◽  
J.A. Welsch ◽  
R. Borrow
2010 ◽  
Vol 18 (2) ◽  
pp. 243-247 ◽  
Author(s):  
Nadine G. Rouphael ◽  
Sarah Satola ◽  
Monica M. Farley ◽  
Karen Rudolph ◽  
Daniel S. Schmidt ◽  
...  

ABSTRACTHaemophilus influenzaetype a (Hia) is an important pathogen for some American Indian, Alaskan native, and Northern Canada aboriginal populations. Assays to measure serum bactericidal activity (SBA) to Hia have not been developed or validated. Here, we describe two methods for the measurement of SBA: SBA with a viability endpoint (CFU counts) and SBA with a fluorometric endpoint using alamarBlue as the metabolic indicator. Both SBA assays measure Hia-specific functional antibody and correlate with anti-Hia IgG enzyme-linked immunosorbent assay (ELISA) concentration of naturally acquired antibodies.


Author(s):  
Jay Lucidarme ◽  
Jennifer Louth ◽  
Kelly Townsend-Payne ◽  
Ray Borrow

Author(s):  
Sian E. Faustini ◽  
Sian E. Jossi ◽  
Marisol Perez-Toledo ◽  
Adrian M. Shields ◽  
Joel D. Allen ◽  
...  

AbstractBackgroundDetecting antibody responses during and after SARS-CoV-2 infection is essential in determining the seroepidemiology of the virus and the potential role of antibody in disease. Scalable, sensitive and specific serological assays are essential to this process. The detection of antibody in hospitalized patients with severe disease has proven straightforward; detecting responses in subjects with mild disease and asymptomatic infections has proven less reliable. We hypothesized that the suboptimal sensitivity of antibody assays and the compartmentalization of the antibody response may contribute to this effect.MethodsWe systemically developed an ELISA assay, optimising different antigens and amplification steps, in serum and saliva from symptomatic and asymptomatic SARS-CoV-2-infected subjects.ResultsUsing trimeric spike glycoprotein, rather than nucleocapsid enabled detection of responses in individuals with low antibody responses. IgG1 and IgG3 predominate to both antigens, but more anti-spike IgG1 than IgG3 was detectable. All antigens were effective for detecting responses in hospitalized patients. Anti-spike, but not nucleocapsid, IgG, IgA and IgM antibody responses were readily detectable in saliva from non-hospitalized symptomatic and asymptomatic individuals. Antibody responses in saliva and serum were largely independent of each other and symptom reporting.ConclusionsDetecting antibody responses in both saliva and serum is optimal for determining virus exposure and understanding immune responses after SARS-CoV-2 infection.FundingThis work was funded by the University of Birmingham, the National Institute for Health Research (UK), the NIH National Institute for Allergy and Infectious Diseases, the Bill and Melinda Gates Foundation and the University of Southampton.


1971 ◽  
Vol 262 (6) ◽  
pp. 338-346 ◽  
Author(s):  
RALPH D. FEIGIN ◽  
DENNIS RICHMOND ◽  
MARK W. HOSLER ◽  
PENELOPE G. SHACKELFORD

2011 ◽  
Vol 79 (8) ◽  
pp. 3188-3194 ◽  
Author(s):  
Janet C. Lindow ◽  
Kelly A. Fimlaid ◽  
Janice Y. Bunn ◽  
Beth D. Kirkpatrick

ABSTRACTAlthough vaccines have been available for over a century, a correlate of protection for typhoid fever has yet to be identified. Antibodies are produced in response to typhoid infection and vaccination and are generally used as the gold standard for determining vaccine immunogenicity, even though their role in clearance ofSalmonella entericaserovar Typhi infections is poorly defined. Here, we describe the first functional characterization ofS.Typhi-specific antibodies following vaccination with a new vaccine, M01ZH09 (Ty2 ΔaroCΔssaV). We determined that postvaccination sera increased the uptake of wild-typeS.Typhi by human macrophages up to 2.3-fold relative to prevaccination (day 0) or placebo samples. These results were recapitulated using immunoglobulins purified from postvaccination serum, demonstrating that antibodies were largely responsible for increases in uptake. Imaging verified that macrophages internalized 2- to 9.5-fold moreS.Typhi when the bacteria were opsonized with postvaccination sera than when the bacteria were opsonized with day 0 or placebo sera. Once inside macrophages, the survival ofS.Typhi was reduced as much as 50% when opsonized with postvaccination sera relative to day 0 or placebo serum samples. Lastly, bactericidal assays indicated that antibodies generated postvaccination were recognized by complement factors and assisted in killingS.Typhi: mean postvaccination bactericidal antibody titers were higher at all time points than placebo and day 0 titers. These data clearly demonstrate that there are at least two mechanisms by which antibodies facilitate killing ofS.Typhi. Future work could lead to improved immunogenicity tests associated with vaccine efficacy and the identification of correlates of protection against typhoid fever.


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