DETECTION OF TOTAL AND ROTAVIRUS SPECIFIC ANTIBODY SECRETING CELLS IN HEALTHY ADULTS
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Antibodies (Ab) play a critical role in the immune response against rotavirus (RV). Protector Ab, are produced by antibody secreting cells (ASC). The capacity to detect ASC becomes fundamental in process like antiviral response analysis and vaccination testing. Here, we analyzed the frequency and isotype of total and RV-specific ASC by ELISPOT (functional assay) and flow cytometry (FC) in healthy adults using CD38
and CD27 like ASC markers. In each million of peripherals blood mononuclear cells (PBMC), approximately 2,550 totals ASC were detected. IgA was the isotype preferentially expressed for circulating ASC followed by IgG and IgM with 63%, 29.4% and 7.6% of total ASC respectively. Paired analysis of immunomagnetic sorting, ELISPOT and FC shown that 85% and 90% of total ASC expressed CD38 and CD27 respectively. CD38 and CD27 ASC enrichment population were used to detect RV-specific ASC. None to 40 ASC per million of PBMC were detected by ELISPOT and FC and more interesting yet, a high correlation were found in the frequency of RV-specific ASC detected by both methods. This mixture assay can be used in the evaluation of new RV vaccines.
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