and the level of ADCC was quantified as the percentage of NK cells that were positive for CD107a staining by flow cytometry .Neutrophils were isolated from freshly collected venous blood using the EasySep Direct Human Neutrophil Isolation Kit according to the manufacturer’s instructions and used on the same day as collection.
CSP was coated on 96 well, white, flat bottom plates at 4 μg/mL in PBS and left overnight wrapped in parafilm in a damp box at 4°C. The liquid was then removed, and plates were washed with PBS followed by blocking with PBS 0.1% BSA for 1 h at room temperature.
Given the non-normal distribution of the data and presence of zero-values, generalised linear mixed modelling was used to estimate latent growth-curve models exploring the subject-specific nature of the association between time and the magnitude and functions of antibodies. Generalised latent growth-curve models were estimated for each IgG subclass and functional antibody parameter using linear splines ; this was used to model the functional form of the effect of time .
In order to estimate the longitudinal associations between the magnitude and functions of antibodies, latent growth-curve models were extended to include covariates in addition to linear splines , with unadjusted and adjusted effects estimated. RR and ARR were estimated and represent the percent change in the magnitude of an antibody parameter outcome for each unit increase in covariate parameter .
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