Raisa Balbuena-Merle1, Manjula Santhanakrishnan1, Lesley Devine1, David R Gibb2, Christopher A Tormey3, Alexa J Siddon4, Susanna A Curtis5, Patrick G Gallagher6, Jason S Weinstein7, Jeanne E Hendrickson8. 1. Yale University, Department of Laboratory Medicine, New Haven, CT, United States. 2. Cedars-Sinai Medical Center, Department of Pathology and Laboratory Medicine, Los Angeles, CA, United States. 3. Yale University, Department of Laboratory Medicine, New Haven, CT, United States; VA Connecticut, Pathology and Laboratory Medicine Service, West Haven, CT, United States. 4. Yale University, Department of Laboratory Medicine, New Haven, CT, United States; VA Connecticut, Pathology and Laboratory Medicine Service, West Haven, CT, United States; Yale University, Department of Pathology, New Haven, CT, United States. 5. Yale University, Division of Hematology/Oncology, New Haven, CT, United States. 6. Yale University, Department of Pathology, New Haven, CT, United States; Yale University, Department of Pediatrics, New Haven, CT, United States; Yale University, Department of Genetics, New Haven, CT, United States. 7. Rutgers New Jersey Medical School, Center for Immunity and Inflammation, Newark, NJ, United States. 8. Yale University, Department of Laboratory Medicine, New Haven, CT, United States; Yale University, Department of Pediatrics, New Haven, CT, United States. Electronic address: jeanne.hendrickson@yale.edu.
Abstract
BACKGROUND: People living with sickle cell disease (SCD) are prone to red blood cell (RBC) alloimmunization. We hypothesized that subjects with alloantibodies (responders) would have differences in circulating T-follicular helper (Tfh)-like cells compared to subjects without alloantibodies (non-responders). MATERIALS AND METHODS: Peripheral blood mononuclear cells were collected from 28 subjects, including those with SCD and controls. Circulating CD4 T-cell subsets were first evaluated at baseline. CD4 T-cell subsets were also evaluated after naïve CD4 T-cells were differentiated into Tfh-like cells following in vitro culture with CD3/CD28 beads, IL-7, IL-12, and Activin A. Transfusion and alloantibody histories were extracted from the electronic medical record. RESULTS: Non-responders had a lower percentage of CD45RA negative Tmemory cells than responders or controls (p<0.05). Notably, there were no differences in circulating Tfh-like cells between any group. However, naïve CD4 T-cells from subjects with SCD were more likely to express CXCR5 after in vitro culture than cells from controls. After culture, CXCR5 expressing cells from responders were more likely to express PD1 and ICOS (16.43 %, sd. 20.23) compared to non-responders (3.69 %, s.d. 3.09) or controls (2.78 %, s.d. 2.04). DISCUSSION: The tendency for naïve CD4 T-cells from responders to differentiate into Tfh-like cells after in vitro culture may suggest these cells are prepared to assist B-cells with antibody production regardless of antigen specificity. Further studies are needed, but it is possible that these results may explain why some responders form RBC alloantibodies with multiple specificities, in addition to RBC autoantibodies and HLA alloantibodies.
BACKGROUND:People living with sickle cell disease (SCD) are prone to red blood cell (RBC) alloimmunization. We hypothesized that subjects with alloantibodies (responders) would have differences in circulating T-follicular helper (Tfh)-like cells compared to subjects without alloantibodies (non-responders). MATERIALS AND METHODS: Peripheral blood mononuclear cells were collected from 28 subjects, including those with SCD and controls. Circulating CD4 T-cell subsets were first evaluated at baseline. CD4 T-cell subsets were also evaluated after naïve CD4 T-cells were differentiated into Tfh-like cells following in vitro culture with CD3/CD28 beads, IL-7, IL-12, and Activin A. Transfusion and alloantibody histories were extracted from the electronic medical record. RESULTS: Non-responders had a lower percentage of CD45RA negative Tmemory cells than responders or controls (p<0.05). Notably, there were no differences in circulating Tfh-like cells between any group. However, naïve CD4 T-cells from subjects with SCD were more likely to express CXCR5 after in vitro culture than cells from controls. After culture, CXCR5 expressing cells from responders were more likely to express PD1 and ICOS (16.43 %, sd. 20.23) compared to non-responders (3.69 %, s.d. 3.09) or controls (2.78 %, s.d. 2.04). DISCUSSION: The tendency for naïve CD4 T-cells from responders to differentiate into Tfh-like cells after in vitro culture may suggest these cells are prepared to assist B-cells with antibody production regardless of antigen specificity. Further studies are needed, but it is possible that these results may explain why some responders form RBC alloantibodies with multiple specificities, in addition to RBC autoantibodies and HLA alloantibodies.
Authors: Marilyn J Telen; Araba Afenyi-Annan; Melanie E Garrett; Martha R Combs; Eugene P Orringer; Allison E Ashley-Koch Journal: Transfusion Date: 2014-12-01 Impact factor: 3.157
Authors: Stella T Chou; Tannoa Jackson; Sunitha Vege; Kim Smith-Whitley; David F Friedman; Connie M Westhoff Journal: Blood Date: 2013-05-30 Impact factor: 22.113