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Bacterial capsular polysaccharides are important vaccine immunogens. However, the study of polysaccharide-specific immune responses has been hindered by technical restrictions. Here, we developed and validated a high-throughput method to analyse antigen-specific B cells using combinatorial staining with fluorescently-labelled capsular polysaccharide multimers. Concurrent staining of 25 cellular markers further enables the in-depth characterization of polysaccharide-specific cells. We used this assay to simultaneously analyse 14 Streptococcus pneumoniae or 5 Streptococcus agalactiae serotype-specific B cell populations. The phenotype of polysaccharide-specific B cells was associated with serotype specificity, vaccination history and donor population. For example, we observed a link between non-class switched (IgM+) memory B cells and vaccine-inefficient S. pneumoniae serotypes 1 and 3. Moreover, B cells had increased activation in donors from South Africa, which has high-incidence of S. agalactiae invasive disease, compared to Dutch donors. This assay allows for the characterization of heterogeneity in B cell immunity that may underlie immunization efficacy. A high-throughput method is developed for simultaneous quantification and in-depth characterization of bacterial polysaccharide antigen-specific B cells. [ABSTRACT FROM AUTHOR]

The point-of-care urine based strip test for the detection of circulating cathodic antigen (POC-CCA) in schistosome infections is a frequently used tool for diagnosis and mapping of Schistosoma mansoni in school-aged children. Because of its ease of use, the test is increasingly applied to adults and preschool-aged children (PSAC), but its performance has not been specifically evaluated in these target groups. Recent observations have raised concerns about possible reduced specificity, in particular in pregnant women (PW) and PSAC. We thus explored specificity of the POC-CCA urine strip test (Rapid Medical Diagnostics, Pretoria, South Africa) in a non-endemic, nonexposed population of 47 healthy nonpregnant adults (NPAs), 52 PW, and 58 PSAC. A total of 157 urines were tested with POC-CCA, of which five (10.6%) NPAs, 17 (32.7%) PW, and 27 (46.5%) PSAC were positive. The highest scores were found in the youngest babies, with an infant of 9 months being the oldest positive case. On measuring pH, it appeared that all POC-CCA strongly positive urines were acidic (pH range 5-5.5), whereas addition of pH-neutral buffer to a subsample reversed the false positivity. We conclude that the POC-CCA test has reduced specificity in PW and infants younger than 9 months, but that the false positivity might be eliminated by modifications in the buffers used in the test.