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Antibodies generated by the linear or cyclic SV loop peptides were not bactericidal, however, even when rabbit serum was used as a complement source to by-pass the host-restricted serum resistance of this strain.
Baby rabbit serum was used as a complement source.
Our in vitro data in this report rely only on direct apoptosis, as we do not add human serum as a complement source nor effector cells.
This process can be measured in vitro using the opsonophagocytic antibody assay (OPA), which is a complex assay composed of live S. aureus bacteria, a complement source, phagocytic effector cells such as differentiated HL-60 cells, and test serum.
These assays are designed to quantify functional antibodies in human serum samples that enable phagocytic effector cells (such as polymorphonuclear leukocytes or differentiated HL-60 cells) to recognize and destroy bacteria in the presence of a complement source.
These bactericidal titers were low even though we used a complement source (baby rabbit serum) that is associated with higher SBA titers than the human complement used in Goldschneider et al.'s study in the 1960s, which found considerably higher levels of protection (2 ).
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To further investigate this host restriction, we also compared NHS with BBS as a nonhuman complement source.
Later on, the SBA was improved using rabbit serum (rSAB) as an exogenous complement source.
An in vitro opsonic reaction is initiated when bacterial cells, human serum (heat inactivated to abrogate endogenous complement activity) containing functional antibodies to the bacteria, and an exogenous complement source are mixed together.
Antibody titers were measured using an rSBA assay, due to the practical advantages of a commercially available heterologous complement source.
The cut off of the rSBA assay was a 1 8 dilution and was considered indicative of seroprotection., Rabbit complement source (rather than human complement) was used because of its wider availability, and because a cut-off for a population based protective titer (≥ 1 8) has been estimated post-surveillance data in the UK after licensure of meningococcal serogroup C conjugate.
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