Risk Of Transfusion-transmitted Infections Research Articles

Microelimination of hepatitis C in patients with chronic hemolytic anemias: a single-center experience

BackgroundPatients with chronic hemolytic anemias (CHA) are at a high risk for transfusion-transmitted infections. Various studies in Egypt have shown a prevalence of hepatitis C virus (HCV) infection in 24–37% of those patients. Elimination of hepatitis C virus (HCV) in patients with CHA would prevent early progression of liver disease. In this study, we aimed to assess the efficacy, safety, and tolerability of sofosbuvir (SOF) and daclatasvir (DAC) in the special population of HCV-infected patients with CHA. In this prospective study, 21 consenting hepatitis C patients were recruited and treated using ribavirin-free SOF/DAC regimen for either 12 or 24 weeks according to categorization of patients into easy or hard-to-treat in accordance with the national protocols. Sustained virological response was assessed by RT-PCR for HCV-RNA at 12 weeks post-treatment (SVR12). Any treatment-related adverse events were noted.ResultsAll patients were adherent to treatment with no discontinuation of therapy. SVR12 was achieved in 19 out of 21 patients (90.5%). There was a significant improvement in levels of ALT (p<0.009) after completion of therapy. On the other hand, the hemoglobin, total bilirubin, and ferritin levels showed a non-significant difference (p<0.501, p<0.542, and p<0.339, respectively). Moderate adverse events were observed in 2 out of 21 patients (9.5%), including sickling crisis and hepatic decompensation.ConclusionThe results of this study substantiate the favorable efficacy, safety, and tolerability of ribavirin-free direct-acting antivirals (DAAs) in the special population of HCV-infected patients with CHA. Micro-elimination of HCV in special patient populations allows for pragmatic delivery of care to patients with co-morbid conditions who are in most need for treatment and allows for achievement of global elimination of HCV worldwide.

Pandemic blood donor demographics – Do changes impact blood safety?

BackgroundCOVID‐19 safety measures and possibly SARS‐CoV‐2 antibody testing may alter blood donor demography, which has the potential to alter blood safety. We characterized pre‐pandemic and pandemic rates of donor infectious disease marker (IDM) reactivity which reflect the residual risk of transfusion‐transmitted infections (TTIs) undetectable by current testing.MethodsThis cross‐sectional analysis of allogeneic blood donor presentations and successful donations in a large national US blood collector identifies changes in self‐reported behavioral risk factors and IDM reactivity. Data on allogeneic blood donor presentations and successful donations from March 1 through August 31, 2020 and the same period in 2019 were retrieved from the blood center's computer system. Donor demographics and deferrals for reported behavioral risk factors and confirmed‐positive IDMs were compared in pre‐pandemic and pandemic periods.ResultsWith increasing mobile blood drive cancellations, pandemic donors were more likely than 2019 donors to be female, over age 30, non‐Hispanic Whites, and have a post‐secondary degree. First‐time donations (at highest risk for confirmed‐positive IDMs) did not substantially increase. Pandemic donors reported fewer behavioral risks and IDMs declined among these donors. Mid‐pandemic introduction of screening for SARS‐CoV‐2 antibodies did not affect IDM rates.ConclusionsUnlike disasters, which tend to bring out more first‐time donors with increased IDM reactivity and TTI residual risk, COVID‐19 donors had lower IDM rates which were not affected by SARS‐CoV‐2 antibody testing. Already‐low TTI residual risk is likely to have declined as a result.

Seropositivity of Transfusion Transmitted Infections among Blood Donors in Hilly Region of Karnataka, India

Introduction: One of the major problem associated with blood transfusions are Transfusion Transmitted Infections (TTIs). Accurate estimates of risk of TTIs are essential for monitoring the safety of blood supply. National AIDS Control Organisation (NACO) guidelines state that all donor’s blood samples should mandatorily be tested for Human Immunodeficiency Virus (HIV), Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Syphilis and Malaria. Aim: To estimate the seropositivity of HIV, Hepatitis B surface Antigen (HBsAg), HCV, Syphilis and Malaria among blood donors for tertiary care hospital of hilly region in Karnataka, India. Materials and Methods: This cross-sectional study was done from January 2012 to March 2018 in District Hospital Blood bank, Department of Pathology, Kodagu Institute of Medical Sciences, Madikeri, Karnataka, India. The blood donors were screened with detailed history and physical examination according to Standard Operating Procedures (SOP) of Blood bank as per Guidelines for Blood donor selection and referral. Donor blood sample was subjected to screening investigations for TTIs diseases: HIV, HBV, HCV, Syphilis and Malaria. Data collected was tabulated in Microscoft Excel and results were expressed as percentage. Results: The study included total 9599 blood donors. The total number of male donors was 8985 and female donors were 614. There were 74 seropositive donors of which 71 (96%) were males and 3 (4.0%) were females. The total seropositivity rate of TTIs was 0.77% among which the incidence of seropositivity was higher in males as compared to females. There were 55 donors (0.58%) positive for HBV, eight donors (0.08%) positive for Syphilis, five donors (0.05%) positive for HIV, four donors (0.04%) positive for Malaria and two donors (0.02%) positive for HCV. Conclusion: Screening and better selection of donors by testing for TTIs are necessary to improve blood safety in the blood transfusion centre of hospital.

Directed Donation: Special Considerations and Review for Contemporary Clinical Practices.

Background: Directed blood donation is defined as the donation of blood or its components for the purpose of transfusion into a specified individual. Directed blood donation holds historic significance, and although practices as of 2021 encourage voluntary, nonrenumerated blood donations, public interest in directed donation remains. Requests to discuss the risks and benefits of directed donations are a common inquiry for transfusion medicine, transplant, and hematology/oncology professionals. This narrative review discusses the history of directed donation and summarizes directed donation considerations in the context of modern transfusion practices.Methods: We conducted a systematic search of PubMed for published literature on the topic of directed blood donation and gathered information about its benefits and potential harms with respect to the variety of products used in transfusion medicine.Results: The drawbacks of directed donation include transfusion-transmitted infection risk, alloimmunization risk, increased transfusion-associated graft vs host disease risk, decreased expediency in treatment, and increased administrative burdens. However, a role remains for directed blood donation in specific patient populations, such as individuals with rare blood types or immunoglobulin A deficiencies, because of the difficulties in finding compatible blood for transfusion.Conclusion: Clinicians should consider the risks and benefits when discussing directed blood donations with patients and family members.

Efficacy of COVID-19 Pathogen Inactivated Convalescent Plasma for Patients with Moderate to Severe Acute COVID-19: A Case Matched Control Study

Background. COVID-19, caused by the SARS-CoV-2 virus, is a pandemic disease with high morbidity and mortality. Currently, available therapeutic options for COVID-19 are limited. Prior experience in epidemics with convalescent plasma (CP) containing antibodies to viruses has demonstrated variable indications of therapeutic efficacy for: Influenza, Argentine Hemorrhagic Fever, and SARS. Characterizing antibody titers to viruses has indicated correlation with therapeutic efficacy. Convalescent COVID-19 patients with potent SARS-CoV-2 antibody responses can serve as plasma donors for immune therapy. However, antibody responses are variable, many donors are first-time higher risk blood donors, and rapid assays to select optimal CP immune efficacy are limited. Pathogen inactivation (PI) of CP can reduce the risk of transfusion-transmitted infection by unrecognized pathogens. Objectives. This study characterized COVID-19 PI-CP activity; and evaluated efficacy and safety of PI CP transfusion in a case matched controlled cohort of acute COVID-19 patients. Methods. COVID-19 apheresis CP (650 - 1300 mL) was collected from nasopharyngeal PCR + outpatients following 2 PCR negative tests or 28 days after symptom resolution. Amotosalen-UVA PI of CP (INTERCEPT Blood System for Plasma) was performed, and antibody efficacy before and after PI was characterized by: VSV reporter pseudo-virus plaque neutralization (RVPN) NT-50 titer (Vitalant Research Institute, San Francisco), antibody to S and N virus proteins by agglutination-dependent antibody PCR (ADAP, Enable Biosciences, San Francisco), virus ACE-2 soluble receptor neutralization assay (Enable Biosciences), and SARS-CoV-2 antibody profile by coronavirus microarray (University of California, Irvine). Patient inclusion criteria were: confirmed SARS-CoV-2 infection, hospitalization, pulmonary infiltrates, availability of ABO compatible CP, and informed consent. CP patients were matched with control patients (CTRL) for disease severity at diagnosis by standardized clinical risk score (W. Liang et al JAMA Intern Med 2020) and concomitant Tocilizumab use. CP Patients received a total of 400 mL of PI CP from 2 donors over 48 hours and standard therapy. CTRL patients received standard COVID-19 therapy without CP. The primary outcome was in-hospital death to day 28. Secondary outcomes included: progression to intubation, admission to ICU, time to discharge, serious adverse events, NP viral clearance, plasma viral clearance, and humoral immune responses. Differences between CP and CTRL patients were assessed by the Mann-Whitney test for continuous variables, and by Fisher’s exact test for categorical variables. Progression to ICU and intubation were analyzed as odds ratios calculated by conditional logistic regression. Results. 15 CP and 30 CTRL patients were enrolled. One CP patient was admitted in cardiogenic shock. Only 2 of 15 CP cohort patients had detectable IgG antibody to SARS CoV-2 S1 antigen at study entry. 3 of 15 PI CP donors had negligible SARS CoV-2 IgG antibodies to all antigens, and demonstrated poor neutralization efficacy. 12/15 CP had effective RVPN titers (> 1:80), RVPN titers were correlated with ACE-2 neutralization antibody titers (r2 = 0.83), and had significant activity specific for S and RBD antigens by microarray profiling (Figure 1). SARS CoV-2 antibody levels were variable between CP donors, but not impacted by PI (Figure 1). Baseline characteristics of CP and matched CTRL patients were similar (Table 1). Sensitivity analysis was performed assessing mortality after exclusion of one CTRL patient admitted in cardiogenic shock and the 2 respective controls. In-hospital 28-day mortality was lower in the CP cohort (0/14) compared to 5/28 CTRL, p = 0.151, 2-sided Fisher’s exact test. Progression to intubation, ICU admission, and days in hospital were not significantly different (Table 1). There was a trend toward decreased inflammatory response (CRP normalization) in CP patients. Conclusions. In hospital mortality of COVID-19 patients was lower in the PI-CP cohort, but not statistically significant. 15% of CP had ineffective antibody by multiple assays. However, PI did not impact CP anti-SARS-CoV-2 activity. PI of plasma provides reduced risk of transfusion transmitted infection from COVID-19 CP donors. In this study, PI CP was safe, and may be effective for early treatment of hospitalized COVID-19 patients.Disclosuresvon Goetz:Cerus Corporation: Current Employment, Current equity holder in publicly-traded company. Khan:Nanommune Inc.: Current equity holder in private company. Tsai:Enable Biosciences: Current Employment. Robinson:Enable Biosciences: Current Employment. Seftel:Enable Biosciences: Current Employment, Current equity holder in private company. Bagri:Cerus Corporation: Current Employment, Current equity holder in publicly-traded company. Corash:Cerus Corporation: Current Employment, Current equity holder in publicly-traded company.

Risk for Hepatitis E Virus Transmission by Solvent/Detergent-Treated Plasma.

Hepatitis E has emerged as a major transfusion-transmitted infectious risk. Two recipients of plasma from 2 lots (A and B) of pooled solvent/detergent–treated plasma were found to be infected by hepatitis E virus (HEV) that was determined to have been transmitted by the solvent/detergent–treated plasma. HEV RNA viral loads were 433 IU in lot A and 55 IU in lot B. Retrospective studies found that 100% (13/13) of evaluable lot A recipients versus 18% (3/17) of evaluable lot B recipients had been infected by HEV (p<0.001), albeit not necessarily at time of transfusion. Among evaluable recipients, 86% with a transfused HEV RNA load >50,000 IU were infected, most likely by the HEV-containing solvent/detergent–treated plasma, versus only 7% with a transfused HEV RNA load <50,000 IU (p<0.001). Overall, solvent/detergent–treated plasma might harbor HEV. Such an occurrence might result in a dose-dependent risk for transfusion-transmitted hepatitis E.

Trends and epidemiological analysis of hepatitis B virus, hepatitis C virus, human immunodeficiency virus, and human T-cell lymphotropic virus among Iranian blood donors: strategies for improving blood safety

BackgroundBlood transfusion is associated with potential risks of transfusion-transmitted infections (TTIs). Different strategies are needed to monitor blood safety and screen the donors’ efficacy, such as evaluation of the prevalence and trends of TTIs. This study was conducted to evaluate the prevalence and trends of TTIs, including hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and human T-cell lymphotropic virus (HTLV 1/2), and the impact of the donors’ characteristics such as age, sex, and donor status on the prevalence of TTIs in blood donors in seven large provinces of Iran from 2010 to 2018.MethodsThis study was conducted on the data collected from all blood donations in seven Iranian Blood Transfusion Centers including Ardabil, Alborz, Guilan, West Azarbaijan, North, Razavi, and South Khorasan from April 2010 to March 2018. Demographic characteristics, number of donations, donor status, and screening and confirmatory serological results of all blood donations were collected from Iranian Blood Transfusion Organizations (IBTO) national database. The prevalence and trend of HBV, HCV, HIV, and HTLV 1/2 infections were reported according to the donation year and donor’s characteristics.ResultsThe analysis of the prevalence and trend of TTIs in 3,622,860 blood donors showed a significant decreasing trend in first-time and regular donors. Additionally, compared to first- time donors, regular donors made safer blood donations with lower risks of HBV, HIV, HCV and HTLV 1/2 (P < 0.0001). Although the prevalence of HTLV 1/2 and HBV was higher in females, TTIs had a significant decreasing trend in males and females. Finally, it was found that the prevalence of HBV and HTLV 1/2 increased with age up to 40–49 years and then decreased thereafter.ConclusionsThe decreasing trends of TTIs in Iranian donors during 9 years may indicate that the various strategies implemented by IBTO have been effective in recent years. Other factors such as a decrease in the prevalence of specific TTIs in the general population might have also contributed to these declines.

Hepatitis B Virus Nucleic Acid Amplification Testing Assay in Detecting Window Period and Occult Hepatitis B Virus Infections in Blood Donors

SummaryTo reduce the residual risk of transfusion-transmitted infections, nucleic acid amplification testing (NAT) of donated blood with higher sensitivity for HBV, HCV, and HIV 1/2 was implemented in Bulgaria at the end of 2019. This study aimed to assess the clinical sensitivity of HBsAg testing and NAT testing of donated blood to detect all forms of HBV infection. A total of 9498 consecutive blood donations collected for six months, from February 10 to July 17, 2020, from first-time and repeat donors at the Regional Center of Transfusion Hematology Pleven, Bulgaria, were screened for HBsAg and HBV DNA. The detection of HBsAg was performed by enzyme-linked immunoassay and chemiluminescent immunoassay. Detection of HBV DNA was performed using the HIV1/2 /HCV / HBV multiplex Procleix Ultrio Elite assay in a fully automated and integrated Procleix Panther System. The overall HBsAg prevalence was 0.05%. HBV DNA was detected in 25 blood units (0.26%), but only 12 (0.13%) were found positive after repeat testing and were confirmed by a discriminatory test. The other 13 units were false positive, with the initial reactive result and negative results after repeat testing. HBV DNA’s overall incidence was significantly higher in HBsAg-positive donors than the HBsAg- negative (Fisher exact p=0.0063). In our study, blood donations were not tested for anti-HBc and anti-HBs, so it is difficult to determine whether HBV DNA-positive/HBsAg-negative results were associated with the early phase of infection or persistent occult infection. There was no statistical difference in the incidence of HBV DNA between repeat-donors (0.16%) and first-time donors (0.06%) (Fisher exact test p=0.239 NS), and also between the incidence in female donors (0.12%) and male donors (0.13%) (Fisher exact test p=1.0 NS). The results of this study showed a low rate of detection of the hepatitis B virus in donated blood. NAT testing demonstrates higher sensitivity for the detection of HBV, as compare to HBsAg screening.

Pathogen reduction of double-dose platelet concentrates from pools of eight buffy coats: Product quality, safety, and economic aspects.

BackgroundPathogen reduction (PR) of platelet concentrates (PCs) contributes to the safety of platelet (PLT) transfusion by reducing the risk of transfusion‐transmitted infections and transfusion‐associated graft‐versus‐host disease. In vitro quality of pathogen‐reduced double‐dose PC (PR‐PC) made of eight whole blood (WB)‐derived buffy coats (BCs) were evaluated.MethodsEight small‐volume WB BCs from donors with at least 200 × 109 PLT/L were pooled with an additive solution to produce double‐dose PCs (DD‐PCs), which were treated with amotosalen/ultraviolet A light in a dual storage processing set, yielding 2 units of PR‐PC. Quality controls were undertaken as per European Directive for the Quality of Medicines (EDQM) guidelines. PLT recovery rates were measured. Production costs and savings were compared over the 3 years before and after PR implementation.ResultsIn the pre‐PR period, 19 666 PCs were produced, compared to 17 307 PCs in the PR period. Single BC in the PR period had 41 ± 2 mL, hematocrit 0.39 ± 0.04 and 1.06 ± 0.18 × 1011 PLTs, and showed a recovery of 91% ± 8%. After pooling, separation, PR treatment of DD‐PC, and splitting, each single PC had 189 ± 6 mL with 2.52 ± 0.34 × 1011 PLTs, compared to 2.48 ± 0.40 in the pre‐PR period. The PLT recovery rate after PR was 87% ± 14%. EDQM requirements were met. An increase of about €12 (+7.5%) per PC from the pre‐PR to the PR period was identified.ConclusionA new production method resulting in two PR‐PCs made from pools of 8 BCs with use of one PR set was successfully introduced, and our experience of nearly 3 years demonstrated the high efficacy and in vitro quality of the PR‐PCs obtained.

Seroprevalence of HBV, HCV and HIV-1 and Correlation with Molecular Markers among Multi-Transfused Thalassemia Patients in Western India.

BackgroundMultitransfused β-thalassemia major patients are always at high risk of having Transfusion Transmitted Infections (TTIs). This study was aimed to determine the seroprevalence of HBsAg, Anti-HIV-1/2, and Anti-HCV among these patients and to correlate the same with NAT testing.MethodsA total of 196 patients with β-thalassemia were included in the study. Patients were screened for the presence of viral markers by third-generation ELISA test as well as for viral DNA/RNA by NAT test.ResultsAmong 196 multi-transfused Beta-thalassemia patients, the seroprevalence of anti-HCV was very high 100 (51.1%), however, anti-HIV1/2 was 6 (3.1%), and HBsAg were 3 (1.5%). Surprisingly similar patterns were observed in the prevalence of molecular markers, as HCV-RNA were 66 (33.7%) of the patients along with HIV-1 RNA were 8 (4.1%), and HBV-DNA were 5 (2.5%) patients. Overall eight (4.1%) patients were found to have coinfections, where two were positive for HBsAg/anti-HCV by ELISA along with 3 (1.5%) were positive for HBV-DNA/ HCV-RNA, 1 (0.5%) was positive for HIV-RNA/HBV-DNA, and 2 (1%) had coinfection of HIV-RNA/ HCV RNA by NAT testingConclusionThe prevalence of HCV infection among multi-transfused β-thalassemia patients is significantly higher than that of the HBV and HIV infections. This scenario should be controlled and monitored by doing regular follow-up testing schedules of such patients and also the administration of the booster dose of the HBV vaccine along with HCV treatment with antiviral DAAs.

Generation and characterization of HLA-universal platelets derived from induced pluripotent stem cells

Platelet demand has increased around the world. However, the inadequacy of donors, the risk of transfusion-transmitted infections and associated reactions, and the refractory nature of platelet transfusions are among the limitations of allogeneic platelet transfusions. To alleviate these problems, we propose generating platelets in a laboratory that do not induce alloimmunity to human leukocyte antigen (HLA) class I, which is a major cause of immune reaction in platelet transfusion refractoriness. Induced pluripotent stem cells (iPSCs) were generated from peripheral blood mononuclear cells (PBMCs) of a healthy Thai woman. We then knocked out the β2-microglobulin (β2m) gene in the cells using paired CRISPR/Cas9 nickases and sequentially differentiated the cells into haematopoietic stem cells (HSCs), megakaryocytes (MKs) and platelets. Silencing of HLA class I expression was observed on the cell surface of β2m-knockout iPSCs, iPSC-derived HSCs, MKs and platelets. The HLA-universal iPSC-derived platelets were shown to be activated, and they aggregated after stimulation. In addition, our in vivo platelet survival experiments demonstrated that human platelets were detectable at 2 and 24 hours after injecting the β2m-KO MKs. In summary, we successfully generated functional iPSC-derived platelets in vitro without HLA class I expression by knocking out the β2m gene using paired CRISPR/Cas9 nickases.

High‐yield production of hematopoietic growth factors by tobacco BY‐2 cells for ex vivo production of red blood cells

Ex vivo generation of clinically available red blood cells (RBCs) from hematopoietic stem cells (HSCs) represents a promising approach for overcoming the limitations associated with the use of donor’s blood, such as shortage of blood and risk of transfusion‐transmitted infections. Both expansion and differentiation of stem cells are highly reliant on growth and differentiation factors, with the activity and purity of these molecules being critical to successful research and clinical applications. The overarching goal of this project is to develop a novel plant cell‐based bioproduction platform for high‐quality (i.e. animal‐free and endotoxin‐free) erythropoietic growth factors (eGFs) that facilitate mass production of RBCs from HSCs at significantly reduced cost. In addressing the low productivity bottleneck of plant cell culture technology, this proposal aims to leverage a unique, plant‐specific, O‐glycosylation process called “hydroxyproline (Hyp)‐O‐glycosylation” for de novo design and engineering of Hyp‐O‐ glycosylated peptides (HypGPs”) that presumably function as a molecular carrier in boosting secretion of conjoined eGFs from cultured plant cells. This will dramatically increase the secreted yields of plant cell‐derived eGFs and simplify the product purification process. Three key eGFs essential for HSCs expansion and differentiation: erythropoietin (EPO), stem cell factor (SCF) and interleukin 3 (IL‐3), are expressed in tobacco BY‐2 cells as fusion with a strategically designed HypGP tag. The secreted protein yields, molecular structure, and Hyp‐O‐glycosylation of the HypGP‐tagged eGFs are characterized. The bioactivities of individual HypGP‐tagged eGF in stimulating proliferation and differentiation of relevant hematopoietic cell lines, includingTF1 erythroleukemic cell and umbilical cord blood‐derived CD34+ cell, are evaluated to establish a suitable design of HypGP tag (size and orientation) for each eGF. A scalable cell culture system fortified sorely with plant cell‐produced eGFs is developed for production of RBCs from CD34+ HSC cells. This project provides a proof‐of‐concept for ex vivo manufacturing of stem cell‐derived RBCs with plant cell‐produced eGFs.Support or Funding InformationNational Science Foundation Directorate for Engineering [1605564]; National Institutes of Health‐Arkansas INBRE [P20GM103429]

Platelet Pathogen Reduction Technologies Alter the MicroRNA Profile of Platelet-Derived Microparticles.

Despite improvements in donor screening and increasing efforts to avoid contamination and the spread of pathogens in clinical platelet concentrates (PCs), the risks of transfusion-transmitted infections remain important. Relying on an ultraviolet photo activation system, pathogen reduction technologies (PRTs), such as Intercept and Mirasol, utilize amotosalen, and riboflavin (vitamin B2), respectively, to mediate inactivation of pathogen nucleic acids. Although they are expected to increase the safety and prolong the shelf life of clinical PCs, these PRTs might affect the quality and function of platelets, as recently reported. Upon activation, platelets release microparticles (MPs), which are involved in intercellular communications and regulation of gene expression, thereby mediating critical cellular functions. Here, we have used small RNA sequencing (RNA-Seq) to document the effect of PRT treatment on the microRNA profiles of platelets and derived MPs. PRT treatment did not affect the microRNA profile of platelets. However, we observed a specific loading of certain microRNAs into platelet MPs, which was impaired by treatment with Intercept or its Additive solution (SSP+). Whereas, Intercept had an impact on the microRNA profile of platelet-derived MPs, Mirasol did not impact the microRNA profile of platelets and derived MPs, compared to non-treated control. Considering that platelet MPs are able to transfer their microRNA content to recipient cells, and that this content may exert biological activities, those findings suggest that PRT treatment of clinical PCs may modify the bioactivity of the platelets and MPs to be transfused and argue for further investigations into PRT-induced changes in clinical PC content and function.

Inactivation of yellow fever virus with amotosalen and ultraviolet A light pathogen-reduction technology.

BACKGROUNDThe reemergence of yellow fever virus (YFV) in Africa and Brazil, and massive vaccine campaigns triggered to contain the outbreaks, have raised concerns over blood transfusion safety and availability with increased risk of YFV transfusion‐transmitted infections (TTIs) by native and vaccine‐acquired YFV. Blood donor deferral for 2 to 4 weeks following live attenuated YFV vaccination, and deferral for travel to endemic/epidemic areas, may result in blood donor loss and impact platelet component (PC) stocks. This study investigated the efficacy of INTERCEPT Blood System pathogen reduction (PR) with use of amotosalen and ultraviolet A (UVA) light to inactivate high levels of YFV in PCs.MATERIALSFour units of apheresis platelets prepared in 35% plasma/65% platelet additive solution (PC‐PAS) and 4 units of PC in 100% human plasma (PC‐Plasma) were spiked with high infectious titers of YFV (YFV‐17D vaccine strain). YFV‐17D infectious titers were measured by plaque assay and expressed as plaque‐forming units (PFU) before and after amotosalen/UVA treatment to determine log reduction.RESULTSThe mean YFV‐17D infectious titers in PC before inactivation were 5.5 ± 0.1 log PFU/mL in PC‐PAS and 5.3 ± 0.1 log PFU/mL in PC‐Plasma. No infectivity was detected immediately after amotosalen/UVA treatment.CONCLUSIONThe amotosalen/UVA PR system inactivated high titers of infectious YFV‐17D in PC. This PR technology could reduce the risk of YFV TTI and help secure PC supplies in areas experiencing YFV outbreaks where massive vaccination campaigns are required.

Impact of Amotosalen/UVA Pathogen Inactivated Platelet Components on Outcomes after Allogeneic Hematopoetic Stem Cell Transplantation: A Single Center Analysis

Background: Platelet component (PC) transfusion is required for allogeneic hematopoietic stem cell transplantation (HCT) recipients. Contamination with infectious pathogens (bacteria, viruses, or protozoa) and T-cells is a risk factor for transfusion-transmitted infection (TTI) and transfusion associated graft-versus-host disease (TA-GVHD). Pathogen inactivation (PI) treatment of PC with amotosalen-UVA (PI-PC, INTERCEPT Blood System, Cerus Corp) in platelet additive solution (PAS) without bacterial screening, gamma irradiation, CMV serology, and with 7-day storage has been the standard of care in Switzerland since 2011 to manage risk of TTI and TA-GVHD. PI-PC have replaced conventional PC (C-PC) prepared in PAS with gamma irradiation and 5 day storage. We previously reported platelet usage in two consecutive five year periods at the University Hospital of Basel. Mean PI-PC dose was higher (3.0 vs. 2.8 x 1011, p=0.001) and mean storage duration longer (4.2 vs. 3.4 days: p=0.001) than with C-PC. PC expiration wastage was reduced with 7-day PI-PC storage vs. 5-day storage (1.5% vs. 8.7%). For HCT recipients, days of PC support; PC use per patient; and RBC use per patient were similar, despite 24.3% lower corrected count increments (CCI) with PI-PC. Now, we report the impact of these observations on treatment related mortality (TRM) and overall survival (OS) 100 days after HCT. Patients and Methods: A two-period retrospective cohort study was conducted to evaluate PI-PC impact on outcomes of consecutive first allogeneic HCT recipients from January 2006 to December 2010 (Period 1, P1), when gamma-irradiated apheresis C-PC were used, and Period 2 (P2) from January 2011 to December 2017, when apheresis and whole blood-derived PI-PC were used. The review utilized 100-day OS and 100-day TRM to determine the impact of PI-PC on HCT outcomes. Descriptive statistics were used for continuous variables and log-rank analysis for survival outcomes. Univariate analysis was performed using Pearson χ2 statistics. Multivariate Cox regression modelling analyses included: PC period (P1, P2), donor match (HLA identical/twin, matched related, matched unrelated), disease state (early, intermediate, late), and conditioning regimen (reduced intensity, myeloablative) with TRM as the outcome. This was an IRB approved single-center analysis. Results: In P1 and P2, 256 and 557 consecutive first-time allogeneic HCT recipients were included, respectively. By univariate analysis, the distribution of European Group for Bone Marrow Transplantation (EBMT) risk scores (grouped 0-2, 3-4, 5-7) and mean patient age were higher during P2 (p = 0.001 and p &lt;0.001, respectively). Primary disease status (p = 0.039); stem cell source (p &lt;0.001); GVHD prophylaxis with ATG (p &lt;0.001); total body irradiation (p &lt;0.001); and conditioning regimen (p &lt;0.001) were different between P1 and P2. Donor match (p=0.084) and disease status (p = 0.628) were similar in P1 and P2. TRM at day 100 post HCT was significantly less (31/557, 5.5%) for PI-PC recipients in P2 vs. C-PC recipients in P1 (37/256, 14.5%, p&lt;0.001). Overall proportion of survivors at day 100 post HCT was significantly greater for PI-PC recipients (507/557, 91.0 %) compared to C-PC recipients (209/256, 81.6%, p &lt;0.001). By multivariate Cox regression analysis, P2 with PI-PC component support was associated with improved TRM (p = 0.001; adjusted hazard ratio 0.433; 95% confidence interval: 0.262, 0.716). Donor match (p = 0.019), disease state (p = 0.022), and myeloablative conditioning (p = 0.034) were associated with significantly poorer TRM (Table). Stem cell source was not significant (p=0.157) in the model. Hemorrhage was reported as cause of death in 1/50 (2.0%) patients during P2 with PI-PC and 4/47 (8.5%) patients during P1 with C-PCs. Conclusions: Universal implementation of PI-PC in routine with extended storage to 7 days in P2 was associated with reduced TRM and better overall survival 100 days post HCT, despite transplantation of older patients with higher EBMT risk scores. Multivariate analysis revealed an adjusted hazard ratio of 0.433 (95% C.I. 0.262, 0.716) for TRM by 100 days, suggesting better outcomes in P2. This retrospective analysis at a single site indicated that PI-PC treated with amotosalen /UVA stored up to 7 days did not have a negative impact on TRM and OS in HCT recipients, and was an integral part of improving clinical outcomes at our institution. . Table. Disclosures Heim: Novartis: Research Funding. Irsch:Cerus Corporation: Employment, Equity Ownership. Lin:Cerus Corporation: Employment, Equity Ownership. Benjamin:Cerus Corporation: Employment, Equity Ownership. Corash:Cerus Corporation: Employment, Equity Ownership.

Neonatal and pediatric platelet transfusions: current concepts and controversies.

In this review, we focus on three specific concepts related to platelet transfusion in the neonatal and pediatric population: choice of transfusion threshold; use of ABO-mismatched platelets; transfusion of pathogen-reduced or inactivated platelets. Recent trials support the use of lower platelet transfusion thresholds (25 000/μl) in preterm neonates, although data is limited to guide transfusion among more mature neonates. In children, there is low-level evidence as to what the prophylactic platelet transfusion threshold should be in many situations of thrombocytopenia, revealing major variability in platelet transfusion practices. Most pediatric guidelines are extrapolated from adult studies with the most evidence in treatment-associated hypoproliferative thrombocytopenia varying between a platelet transfusion threshold of 10 000/μl to 20 000/μl. Although pathogen-reduced platelets may lower the risks of transfusion-transmitted infection, the effects on platelet refractoriness and transfusion burden in this population warrant additional study. Our review highlights recent advances in neonatal and pediatric platelet transfusion and also emphasizes the urgent need for better evidence to guide practice given recent studies showing the potential harms of platelet transfusion, particularly with liberal use.

Phased implementation of pathogen-reduced platelets in a health system facilitates increased manufacturing at the blood center.

BACKGROUNDPathogen reduction treatment (PRT) reduces the risk of transfusion‐transmitted infections from established and emerging organisms. Manufacturing, however, is complex. In our university health system, we phased in pathogen‐reduced platelets (PR PLTs) by patient population. We then assessed the implementation strategy and investigated factors in the supply chain that prevented us from meeting the goal of providing greater than 90% PR PLTs within 6 months.STUDY DESIGN AND METHODSIn Phase 1, PR PLTs were provided in the outpatient cancer center. Phase 2 added inpatients undergoing bone marrow transplantation, and Phase 3 included all patients. In Phase 4, the blood center implemented manufacturing optimization strategies. Product supply and usage during the first 23 months after implementation were evaluated. Investigation of the supply chain included analysis of (1) the number of in‐state hospitals receiving PR PLTs; (2) the fraction of products eligible for PRT before and after manufacturing improvements.RESULTSDuring Phases 1 and 2, PR products comprised 44% and 53% of PLTs transfused in the phased‐in areas. At 6 months, 41% of PLTs were PR, and at 23 months, 92%. The fraction of PR PLTs transfused in our system correlated logarithmically with the number of in‐state hospitals receiving them (R2 = 0.71) and the number of PR PLTs sold to those hospitals (R2 = 0.80).CONCLUSIONPhased implementation is a practical and ethical way to introduce PR PLTs in a health system and facilitates scalability at the blood center. Widespread availability of PR products may require collective action and can be increased by optimization strategies during manufacturing.

Bacterial Contamination of Platelet Units is an Under-Recognized Contributor to Healthcare-associated infections (HAIs)

BACKGROUND HAIs create a major logistical, financial and patient safety concerns within the healthcare system. Bacterial contamination of platelet components (PCs) is the most common, under recognized, infectious risk of transfusion and contributor to HAIs. Contaminated PCs can lead to central line-associated bloodstream infections, bacteremia, and sepsis, with major long term complications and/or death. While US blood centers employ primary bacterial culture screening (pBCS) today, FDA's Draft Guidance and Blood Products Advisory Committee recommend additional approaches to reduce continued risk including FDA-approved pathogen reduction technology (PRT). Our objective is to determine whether PRT is an effective means to reduce the infectious risk of PCs. METHODS We compiled national hemovigilance (HV) data (2006-2016) from the UK, France, Switzerland, Belgium and the US to examine use of pBCS and PRT with amotosalen/UVA and their associated reports of PC bacterial contamination and sepsis. RESULTS Introduction of pBCS decreased HV reports of PC bacterial contamination in the US by 72.5% (p 700,000 PRT PCs over the same time period. CONCLUSIONS pBCS does not eliminate the risk of bacterial contamination; In its Draft Guidance, FDA acknowledges this shortcoming and recommends further measures. In contrast, PRT inactivates a broad spectrum of pathogens present in PCs, regardless of low level contamination, eliminating the need for pBCS. PRT provides clinicians and patients with transfusion-ready PCs, mitigating the risk of transfusion-transmitted infections and representing an approach to increase hospital safety while reducing the financial challenges of transfusion-related HAIs.

Plant Cell‐Secreted Growth Factors for ex vivo Mass Production of Red Blood Cells

Ex vivo generation of clinically available red blood cells (RBCs) from hematopoietic stem cells (HSCs) represents a promising approach for overcoming the limitations associated with the use of donor's blood, such as shortage of blood and risk of transfusion-transmitted infections. Both expansion and differentiation of stem cells are highly reliant on growth and differentiation factors, with the activity and purity of these molecules being critical to successful research and clinical applications. The overarching goal of this project is to develop a novel plant cell-based bioproduction platform for high-quality (i.e. animal-free and endotoxin-free) erythropoietic growth factors (eGFs) that facilitate mass production of RBCs from HSCs at significantly reduced cost. In addressing the low productivity bottleneck of plant cell culture technology, this proposal aims to leverage a unique, plant-specific, O-glycosylation process called “hydroxyproline (Hyp)-O-glycosylation” for de novo design and engineering of Hyp-O-glycosylated peptides (HypGPs”) that presumably function as a molecular carrier in boosting secretion of conjoined eGFs from cultured plant cells. This will dramatically increase the secreted yields of plant cell-derived eGFs and simplify the product purification process. Three key eGFs essential for HSCs expansion and differentiation: erythropoietin (EPO), stem cell factor (SCF) and interleukin 3 (IL-3), are expressed in tobacco BY-2 cells as fusion with a strategically designed HypGP tag. The secreted protein yields, molecular structure, and Hyp-O-glycosylation of the HypGP-tagged eGFs are characterized. The bioactivities of individual HypGP-tagged eGF in stimulating proliferation and differentiation of relevant hematopoietic cell lines, including TF1 erythroleukemic cell and umbilical cord blood-derived CD34+ cell, are evaluated to establish a suitable design of HypGP tag (size and orientation) for each eGF. A scalable cell culture system fortified sorely with plant cell-produced eGFs will be finally developed for mass production of RBCs from CD34+ HSC cells. This project provides a new cost-effective bioproduction platform for eGFs, facilitating manufacturing of HSCs-derived RBCs at large scale for clinical applications. Support or Funding Information Arkansas INBRE-Research Development Grant (RDG) under Grant No. P20GM103429; National Science Foundation under Grant No. 1605564; Arkansas Biosciences Institute Schematic of the proposed research for mass production of red blood cells (RBCs) ex vivo from hematopoietic stem cells (HSCs) with plant produced eGFs (EPO, SCF, IL-3 and IGF-1) All the eGFs will be engineered in plant cells (tobacco BY-2 cell) with a Hyp-O-glycosylated peptide (HypGP) tag that function as a molecular carrier in boosting the secretion of conjoined erythropoietic growth factors (eGFs). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Transfusion of pathogen-reduced platelet components without leukoreduction.

BACKGROUNDLeukoreduction (LR) of platelet concentrate (PC) has evolved as the standard to mitigate risks of alloimmunization, clinical refractoriness, acute transfusion reactions (ATRs), and cytomegalovirus infection, but does not prevent transfusion‐associated graft‐versus‐host disease (TA‐GVHD). Amotosalen–ultraviolet A pathogen reduction (A‐PR) of PC reduces risk of transfusion‐transmitted infection and TA‐GVHD. In vitro data indicate that A‐PR effectively inactivates WBCs and infectious pathogens.STUDY DESIGN AND METHODSA sequential cohort study evaluated A‐PR without LR, gamma irradiation, and bacterial screening in hematopoietic stem cell transplant (HSCT) recipients. The first cohort received conventional PC (control) processed without LR, but with gamma irradiation and bacterial screening. The second cohort received A‐PR PC (test) processed without: LR, bacterial screening, or gamma irradiation. The primary efficacy outcome was the 1‐hour corrected count increment. The primary safety outcome was treatment‐emergent ATR. Secondary outcomes included clinical refractoriness, and 100‐day status for engraftment, TA‐GVHD, HSCT‐GVHD, infections, and mortality.RESULTSMean corrected count increment (× 103) of 33 test PC recipients was similar (18.9 ± 8.8 vs. 16.6 ± 8.4; p = 0.296) to that of 31 control PC recipients. Test recipients had a reduced, but nonsignificant, incidence of ATR (test = 9.1%, Control = 19.4%; p = 0.296). The frequencies of clinical refractoriness (0 of 33 vs. 4 of 31 patients) and refractory transfusions (6.6% vs. 19.3%) were lower in the test cohort (p = 0.05 and 0.02), respectively. No patient in either cohort had TA‐GVHD. Day 100 engraftment, HSCT‐GVHD, mortality, and infectious disease complications were similar between cohorts.CONCLUSIONSThis study indicated that A‐PR PC without LR, gamma irradiation, or bacterial screening is feasible for support of HSCT.