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. 2017 Feb 6;24(2):e00412-16. doi: 10.1128/CVI.00412-16

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Copyright © 2017 Shimanovich et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 1 — Shigella-specific serum bactericidal antibody (SBA) and opsonophagocytic killing antibody (OPKA) assay optimization. The percentages of organisms killed were determined using different amounts of baby rabbit complement (BRC) (A), DMF-differentiated HL60 cells (B), and serum dilutions (C and D) tested during assay optimization. Paired individual serum samples from two volunteers obtained before (nonimmune) and 28 days after (immune) immunization with Shigella vaccine strain CVD 1208S were used for assay optimization. Each assay included a negative control containing all reagents except serum. Curves show SBA (E) and OPKA (F) activity using the optimized assays; data represent mean percent killing ± SEM of four independent assays. The intersection of dashed lines exemplifies SBA and OPKA titers, calculated as the reciprocal of the serum dilution that produces 50% Shigella killing using Reed-Muench regression analysis.