![]() We mixed the phage library with sufficient volume of serum to provide 2 μg of IgG (between 2 and 8 μL) in each of 2 replicates per sample. 15 We first quantified total IgG from each sample using the Human IgG ELISA Quantitation Set (Bethyl Laboratories, Inc, Montgomery, TX). VirScan was carried out according to the previously published method with the following minor modifications. We amplified the VirScan T7 bacteriophage library from the original library (Elledge Laboratory) according to the manufacturer’s suggested protocol (EMD Millipore) and sequenced the product to quantify the frequency of targets in our input library. 7-11 Much less is known about the reconstitution of immunity toward other viruses, 12 the role of other viruses in post-HCT outcomes, or how other factors associated with altered B-cell responses, including graft-versus-host disease (GVHD) or CMV infection, might influence reconstitution of humoral antiviral immunity. ![]() 3-6 Much is known about how cytomegalovirus (CMV) impacts clinical outcomes, including death, following HCT, and recent data demonstrate the importance of humoral immunity and far-reaching immunologic effects of this virus. Following HCT, preformed antibodies from the recipient continue to circulate, with an average half-life of ∼26 days, and are thought to be slowly replaced by donor IgG following establishment of the donor B cell and plasma cell population much later. 1, 2 Human viral pathogens number in the hundreds, with thousands of possible strains, and clinically used proxies for risk of viral infection, such as absolute lymphocyte count (ALC) or total immunoglobulin levels (immunoglobulin G ), do not strongly predict protection against specific viral pathogens. This work builds a foundation to test whether such systematic profiling could serve as a biomarker of immune reconstitution, predict clinical events after HCT, or help refine selection of optimal donors.įollowing myeloablative allogeneic hematopoietic cell transplantation (HCT), full lymphocyte reconstitution and resolution of hypogammaglobulinemia may be protracted. We used VirScan to highlight contributions of donor and recipient to antiviral humoral immunity and evaluate longitudinal changes. Gain or loss of epitopes to common viruses over the year post-HCT differed by donor and recipient pre-HCT serostatus, with highest gains in naive donors to seropositive recipients for several human herpesviruses and adenoviruses. The recipient repertoire was most similar (pairwise β-diversity) to that of donor at day 100, but more similar to pre-HCT self by day 365. Other clinical characteristics, including pre-HCT treatment and conditioning, did not affect antiviral repertoire metrics. Donor age and donor/recipient cytomegalovirus (CMV) serostatus and receipt systemic glucocorticoids were most strongly associated with VirScan metrics at day 100. ![]() We applied ecologic metrics (α- and β-diversity) and evaluated predictors of metrics and changes over time. We performed VirScan on cryopreserved serum from pre-HCT and 30, 100, and 365 days after myeloablative HCT from 37 donor-recipient pairs. We interrogated the viral antibody repertoire before and after HCT using a novel serosurvey (VirScan) that detects immunoglobulin G responses to 206 viruses. Currently, estimation of humoral immune recovery is inferred from lymphocyte counts or immunoglobulin levels and does not address vulnerability to specific viral infections. Further insight into humoral viral immunity after hematopoietic cell transplantation (HCT) could have potential impact on donor selection or monitoring of patients.
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