Seronegative Blood Donors Research Articles

Effectiveness of Using Nucleic Acid Amplification Test to Screen Blood Donors for Hepatitis B, Hepatitis C, and HIV: A Tertiary Care Hospital Experience From Pakistan.

Background Ensuring blood safety is the primary goal of transfusion medicine. Despite extensive serological tests and strict safety measures, the risk of transfusion-transmitted infections (TTIs) still exists. As applied to blood screening, Nucleic Acid Amplification Test (NAT)offers much higher sensitivity for detecting viral infections. It is, however, currently available to a handful of centers due to the high cost. This study aims to establish the Effectiveness of NAT by assessing the NAT yield and residual risk of transmission of Hepatitis B virus (HBV), hepatitis C virus (HCV)and HIV with and without NAT testing. Material and method This prospective cross-sectional study recruited blood donors from January 2020 to November 2022. All donors underwent routine serologic screening. Only serologically negative donors were tested for HBV, HCV, and HIV by NAT. The NAT yield and residual risk (RR) per million donors were computed for viral infections in seronegative blood donors and calculated using the incidence/window period model. Result A total of 59708 donors were included during the study period. The overall prevalence of TTI's were: For HCV 1.7% (n = 1018), HBV 1.5% (n = 918), HIV 0.07% (n = 47), Syphilis 1.2% (n = 758) and malaria 0.3% (n = 218). Out of 57759 seronegative donors, thirty-four NAT-reactive samples were identified, with 3 cases of HCV, 31 cases of HBV, and Nil HIV cases. NAT yield of HBV was 1 in 1863 with an RR of 8.6 per million, followed by HCV with a NAT yield of 1 in 19253 and RR of 0.8 per million donations. NAT testing reduced RR for HBV by 48.9% and HCV by 94.5%. Conclusion Our study showed that NAT detected 34 out of 57759 cases initially missed by serological tests. The study suggests that the parallel use of serology and NAT screening of donated blood would be beneficial.

Knowledge and Practice of Hepatitis B and C Prevention among Seropositive and Seronegative Prospective Blood Donors in a Tertiary Care Hospital in Eastern Nigeria

Introduction: Hepatitis B and C infections are endemic in Nigeria and the major causes of chronic liver diseases, including hepatocellular carcinoma. One of the major routes of transmission is by transfusion of infected blood or blood products. Awareness of the disease is important, and this study seeks to compare the knowledge and practice of the disease among the seropositive and seronegative prospective blood donors. Aim: To assess the difference in the level of knowledge and preventive practice of blood borne hepatitis among hepatitis B and hepatitis C seropositive and seronegative prospective blood donors. Materials and Methods: The present study was a descriptive survey carried out between June 2019 to January 2020 in the Blood Donor’s Department of the University of Nigeria Teaching Hospital, Enugu, Nigeria. A pretested self/interviewer’s administered questionnaire was used to assess the knowledge and practice among 100 hepatitis B and hepatitis C seropositive and 305 seronegative prospective blood donors. The hepatitis B and C seropositive subjects were screened using Enzyme Immunosorbent Assay (ELISA) technique. The findings and scores were analysed using IBM Corporation 2019 Statistical Package for the Social Sciences (SPSS) software version 21.0. Results: Total of 305 respondents were seronegative while 82 and 18 were hepatitis B and C seropositive, respectively. The mean age of the study participants was 27.78±7.17 years, whereas it was 28.09±6.36 years and 27.46±7.968 years among seropositive and seronegative respondents, respectively. In the study, 98 (98%) males and 2 (2%) females were in the seropositive group while, 268 (87.9%) males and 37 (12.1%) females were in seronegative group. The overall knowledge score and practice score were 56.3% and 28.1%, respectively. In the seropositive group, the average knowledge score was 27.3% and the average practice score was 17%. While in the seronegative group, the average knowledge score was 65.7% and the average score on practice score was 31.8%. There was a significant association between seropositive and being a male (p-value=0.001), less educated (p-value=0.002), currently unmarried (p-value=0.029), knowledge above average (p-value=0.001), and practice below average (p=0.002). None of the respondents had hepatitis B vaccination. Conclusion: Based on the results obtained from the present study, it is evident that the level of knowledge and practice among the seropositive respondent is significantly low when compared with the seronegative groups. There is a need for a serious awareness campaign on the transmission and prevention of hepatitis B and C, and the possibility of extending hepatitis B immunisation coverage in the country owing to the increasing burden of the disease.

Abstract 12091: Incidence and Predictors of Progression to Chagas Cardiomyopathy: Long-Term Follow-Up of a Large Cohort of Trypanosoma cruzi- Infected Individuals

Introduction: There are few contemporary prospective cohorts of Trypanosoma cruzi ( T.cruzi )-infected individuals. This study aimed to determine the incidence of new-onset cardiomyopathy and death attributable to T. cruzi infection. Methods: We enrolled 1088 individuals: 499 T. cruzi -seropositive blood donors identified in routine donor screening, 488 age- and sex-matched T. cruzi - seronegative blood donors, and 101 patients with established Chagas cardiomyopathy recruited from a tertiary center. Subjects underwent cardiovascular evaluation with ECG and echocardiogram at the baseline (2008-2010) and follow-up (2018-2019) visits. Cardiomyopathy was diagnosed based on ECG and echocardiographic findings at baseline and follow-up. Vital status was determined by linkage with the national Brazilian mortality system. Results: Mortality was 80.9 deaths/1000 person-years (py) in established cardiomyopathy and 15.1 deaths/1000py in T. cruzi -seropositive donors with cardiomyopathy at baseline. The mortality rate among T. cruzi -seropositive blood donors without cardiomyopathy at baseline and T. cruzi -seronegative controls was similar, with mortality rates of 3.67 and 3.62 events/1000 py, respectively. In T. cruzi -seropositive donors, age (hazard ratio [HR] = 1.05 per year), anti-T.cruzi antibody level (HR = 1.40 per unit increase in serology assay signal/cut off) and the presence of cardiomyopathy at baseline (HR = 3.01) were independent predictors of mortality. The incidence of new-onset cardiomyopathy in T. cruzi -seropositive donors was 13.8 events/1000 py (95% CI 9.5-19.6) compared with 4.6 events/1000 py (95% CI 2.3-8.3) in controls, with an absolute incidence difference attributable to T. cruzi infection of 9.2 (95% CI 3.6 - 15.0) events/1000py. T. cruzi antibody level at baseline was the main independent predictor of subsequent new-onset cardiomyopathy with an adjusted odds ratio of 1.37 (95% CI 1.07-1.76) per unit increase in antibody level. Conclusions: We present a comprehensive description of the natural history of T. cruzi infection in a contemporary patient population. The results highlight the central importance of total anti- T. cruzi antibody titer as a marker of Chagas disease activity and risk of progression.

A flow cytometry-based proliferation assay for clinical evaluation of T-cell memory against SARS-CoV-2

In general, the method of choice for evaluating immunity against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is detection of antibodies against the virus in patient sera. However, this is not feasible in patients who do not produce antibodies, either due to a primary immunodeficiency or secondary to treatment with immunosuppressive drugs. Assessment of the antiviral T cell response is an alternative to serological tests, but most T cell assays are labor-intensive and unsuitable for a clinical routine laboratory.We developed a flow cytometry-based assay for T cell proliferative responses against SARS-CoV-2, based on the detection of blast transformation of activated cells. The assay was validated on previously SARS-CoV-2 infected individuals and healthy seronegative blood donors, displaying 74% sensitivity and 96% specificity for previous infection with SARS-CoV-2. The usefulness of the assay was demonstrated in a patient with common variable immunodeficiency with a history of COVID-19. The described T-cell assay is a clinically relevant complement to serology in the evaluation of cellular immunity against SARS-CoV-2, which can be emulated by any routine lab with flow cytometric competence.

Lack of neutralizing activity in nonconvalescent sera, regardless of ABO blood group and anti-A isoagglutinin titer

BackgroundSeveral ABO blood groups have been associated with the likelihood of infection, severity, and/or outcome of COVID-19 in hospitalized cohorts, raising the hypothesis that anti-A isoagglutinins in non-A-group recipients could act as neutralizing antibodies against SARS-CoV-2.Materials and methodsWe run live virus neutralization tests using sera from 58 SARS-CoV-2 seronegative blood donors (27 O-group and 31 A-group) negatives for SARS-CoV-2 IgG to investigate what degree of neutralizing activity could be detected in their sera and eventual correlation with anti-A isoagglutinin titers.ResultsWe could not find clinically relevant neutralizing activity in any blood group, regardless of anti-isoagglutinin titer,DiscussionOur findings suggest that mechanisms other than neutralization explain the differences in outcomes from COVID19 seen in different ABO blood groups.

High Frequency Occult Hepatitis B Virus Infection Detected in Non-Resolved Donations Suggests the Requirement of Anti-HBc Test in Blood Donors in Southern China.

BackgroundMost Chinese Blood Centers adopted mini pool (MP) nucleic acid testing (NAT) for HBV screening due to high cost of Individual donation (ID) NAT, and different proportions of MP-reactive but ID-non-reactive donations (MP+/ID−, defined as non-resolved donations) have been observed during daily donor screening process. Some of these non-resolved donations are occult HBV infections (OBIs), which pose potential risk of HBV transmission if they are not deferred. This study is aimed to further analyze these non-resolved donations.MethodsThe non-resolved plasma samples were further analyzed by serological tests and various HBV DNA amplification assays including quantitative PCR (qPCR) and nested PCR amplifying the basic core and pre-core promoter regions (BCP/PC; 295 base pairs) and HBsAg (S) region (496 base pairs). Molecular characterizations of HBV DNA+ non-resolved samples were determined by sequencing analysis.ResultsOf 17,226 MPs from 103,356 seronegative blood donations, 98 MPs were detected reactive for HBV. Fifty-six out of these 98 (57.1%) reactive MPs were resolved as HBV DNA+, but the remaining 42 pools (42.9%, 252 donations) were left non-resolved with a high rate (53.2%) of anti-HBc+. Surprisingly, among 42 non-resolved MPs, 17 contained one donation identified as OBIs by alternative NAT assays. Sequence analysis on HBV DNAs extracted from these OBI donations showed some key mutations in the S region that may lead to failure in HBsAg detection and vaccine escape.ConclusionA total of 53.2% of the non-resolved donations were anti-HBc+, and OBIs were identified in 40.5% of these non-resolved pools. Therefore, non-resolved donations with anti-HBc+ might pose potential risk for HBV transmission. Our present analysis indicates that anti-HBc testing in non-resolved donations should be used to identify OBIs in order to further increase blood safety in China.

Transfusion-transmitted and community-acquired cytomegalovirus infection in seronegative solid organ transplant recipients receiving seronegative donor organs.

Solid organ transplant (SOT) recipients who are cytomegalovirus (CMV) seronegative (R-) and receive seronegative donor (D-) organs have a small but currently unquantified risk of both transfusion-transmitted CMV (TT-CMV) and community-acquired CMV (CA-CMV). We retrospectively studied the incidence and clinical symptoms of TT-CMV (infection <1year posttransplant) and CA-CMV (infection >1year posttransplant) in a cohort of D-/R- adult and pediatric SOT recipients receiving leukoreduced blood products not screened for CMV seronegativity transplanted at our center between 2000 and 2011. CMV infection was defined as IgG seroconversion or detectable CMV antigenemia/DNAemia. Among 536 consecutive D-/R- recipients, 398 (81.8%) had adequate follow-up, and 231 (58%) received cellular blood products (total: 1626 red blood cell units, 470 platelet units) 30days pretransplant to 90days posttransplant. We observed no confirmed TT-CMV cases, but 14 CA-CMV cases (64% symptomatic) were seen. The estimated incidence rate of CA-CMV was higher in children (3.0/100 patient years) than adults (0.46/100 patient years, incident rate ratio of 6.52). The absence of TT-CMV over 11years suggests neither seronegative blood products nor CMV DNA blood donor screening would provide significant incremental safety when blood is already leukoreduced. D-/R- SOT recipients, particularly children, have a significantly higher and ongoing risk of CA-CMV.

Impact on mortality of being seropositive for hepatitis C virus antibodies among blood donors in Brazil: A twenty-year study

IntroductionHepatitis C virus (HCV) infection is a major health problem associated with considerable risk of mortality in different regions of the world. The purpose of this study was to investigate the contribution of HCV infection on all-cause and liver-related mortality, in a large cohort of blood donors in Brazil.MethodsThis is a retrospective cohort study of blood donors from 1994 to 2013, at Fundação Pró-Sangue—Hemocentro de São Paulo (FPS). This cohort included 2,892 and 5,784 HCV antibody seropositive and seronegative donors, respectively. Records from the FPS database and the Mortality Information System (SIM: a national database in Brazil) were linked through a probabilistic record linkage (RL). Mortality outcomes were defined based on ICD-10 (10th International Statistical Classification of Diseases and Related Health Problems) codes listed as the cause of death on the death certificate. Hazard ratios (HRs) were estimated for outcomes using Cox multiple regression models.ResultsWhen all causes of death were considered, RL identified 209 deaths (7.2%) among seropositive blood donors and 190 (3.3%) among seronegative blood donors. Donors seropositive for HCV infection had a 2.5 times higher risk of death due to all causes (95% CI: 1.76–2.62; p<0.001). When only liver-related causes of death were considered, RL identified 73 deaths among seropositive blood donors and only 6 among seronegative blood donors. Donors seropositive for HCV infection had a 23.4 times higher risk of death due to liver related causes (95% CI: 10.2–53.9; p<0.001). Donors seropositive for HCV had a 29.5 (95%CI: 3.9–221.7), 2.8 (95% CI: 1.4–5.5) and a 1.9 (95% CI: 1.2–3.0) times higher risk of death due to hepatocellular carcinoma, infection or trauma, respectively, compared to seronegative donors.ConclusionsAll-cause and liver-related mortality rate was increased among blood donors seropositive for HCV compared with the mortality rate among seronegative blood donors. Our data confirms HCV as a relevant cause of death in Brazil and also suggest that interventions directed at following patients even after access to specific drug treatment are urgent and necessary.

Chagas Disease Serological Test Performance in U.S. Blood Donor Specimens.

Chagas disease affects an estimated 300,000 individuals in the United States. Diagnosis in the chronic phase requires positive results from two different IgG serological tests. Three enzyme-linked immunosorbent assays (ELISAs) (Hemagen, Ortho, and Wiener) and one rapid test (InBios) are FDA cleared, but comparative data in U.S. populations are sparse. We evaluated 500 seropositive and 300 seronegative blood donor plasma samples. Country of birth was known for 255 seropositive specimens, which were grouped into regions as follows: Mexico (n = 94), Central America (n = 88), and South America (n = 73). Specimens were tested by the four FDA-cleared IgG serological assays. Test performance was evaluated by two comparators and latent class analysis. InBios had the highest sensitivity (97.4% to 99.3%) but the lowest specificity (87.5% to 92.3%). Hemagen had the lowest sensitivity (88.0% to 92.0%) but high specificity (99.0% to 100.0%). The level of sensitivity was intermediate for Ortho (92.4% to 96.5%) and Wiener (94.0% to 97.1%); both had high specificity (98.8% to 100.0% and 96.7% to 99.3%, respectively). The levels of antibody reactivity and clinical sensitivity were lowest in donors from Mexico, intermediate in those from Central America, and highest in those from South America. Our findings provide an initial evidence base to improve laboratory diagnosis of Chagas disease in the United States. The best current testing algorithm would employ a high-sensitivity screening test followed by a high-specificity confirmatory test.

IMPACT OF MULTIPLEX PCR IN REDUCING THE RISK OF RESIDUAL TRANSFUSION-TRANSMITTED HUMAN IMMUNODEFICIENCY AND HEPATITIS B AND C VIRUSES IN BURKINA FASO

Background and ObjectiveThe improved performance of serological tests has significantly reduced the risk of human immunodeficiency and hepatitis B and C viruses transmission by blood transfusion, but there is a persistence of residual risk. The objective of this study was to evaluate the impact of multiplex PCR in reducing the risk of residual transmission of these viruses in seronegative blood donors in Burkina Faso.MethodsThis cross-sectional study was conducted from March to September 2017. The serological tests were performed on sera using ARCHITECTSR i1000 (Abbot diagnosis, USA). Detection of viral nucleic acids was performed by multiplex PCR on mini-pools of seronegative plasma for HBV, HCV and HIV using SaCycler-96 Real Time PCR v.7.3 (Sacace Biotechnologies). Multiplex PCR-positive samples from these mini-pools were then individually tested by the same method.ResultsA total of 989 donors aged 17 to 65 were included in the present study. “Repeat donors” accounted for 44.79% (443/989). Seroprevalences for HIV, HBV, and HCV were 2.53% (25/989), 7.28% (72/989) and 2.73% (27/989), respectively. Of the 14 co-infections detected, HBV/HCV was the most common with 0.71% (7/989) of cases. Of 808 donations tested by multiplex PCR, 4.70% (38/808) were positive for HBV while no donation was positive for HIV or HCV.ConclusionOur study showed a high residual risk of HBV transmission through blood transfusion. Due to the high prevalence of blood-borne infections in Burkina Faso, we recommend the addition of multiplex PCR to serologic tests for optimal blood donation screening.

English

Changes in serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transferase (G-GT), total protein (TP), albumin (ALB), total bilirubin (T. Bil), direct bilirubin (D. Bil) are assayed to monitor liver function; while those of creatinine (CRT) and urea (UR) are for kidney functions. These two organs produce hepcidin and erythropoietins, respectively that play major roles in haemopoiesis and so may be involved in HIV- associated anemia. Anemia occurs in over 70% of HIV-infected. The objective of this study was to monitor liver and kidney derangements in HIV based on CD4+ cell levels for 6 months. This was a longitudinal descriptive study conducted at Kenyatta National Hospital Comprehensive Care Centre and involved: 184 HIV seropositive and 101 HIV seronegative blood donors as a comparative group. The comparative group demonstrated significantly higher T.Bil and D.Bil mean values in males than in females. Increases in each of: G-GT, T. Bil, D. Bil, AST, ALT and CRT above the upper limit of the control were observed. Increases in G-GT were highest in CD4+ above 200 cells /mm3; decreases in the levels of ALP and ALB were lowest in CD4+ &lt; 200 cells / mm3 group. Increases in G-GT with decreases in ALP were possibly due to cardiac-related disorders. Serum levels of AST, ALT and CRT are not affected by CD4+ levels. Decreases in the levels of ALB in CD4+ &lt; 200 cells/mm3 group were thought to be partly due to: anemia, malnutrition or hypercatabolism. Hypoalbuminemia may result in altered albumin: globulin ratio. Determinations of: albumin: globulin ratio, gender-based reference ranges for T. Bil and D. Bil, hepcidin and erythropoietin are recommended.&nbsp; &nbsp; Key words: Study, clinical values, HIV infection.

Assessment of semi-automated nucleic acid testing programme in a Regional Blood Transfusion Centre

Background: Detection of human immunodeficiency virus type-1 (HIV-1), hepatitis-C (HCV) and hepatitis-B virus (HBV) in the blood donors is crucial. An efficient form of detection is nucleic acid testing (NAT) in blood screening. We assessed the suitability of commercial NAT testing in a developing country, focusing on the Altona RealStar assay and the method of Sacace Biotechnologies.Methods: We have standardised and validated commercially available NAT kits with a semi-automated system for detection of HBV, HCV and HIV-1 in blood donations. The MP-NAT (mini-pool) assay consists of pooling of sample, virus extraction, amplification and detection with commercially available NAT kits. An internal control (IC) is incorporated in the assay to monitor the extraction, target amplification and detection process.Results: The sensitivity of the Altona RealStar assay at 10-MP for each viral target was evaluated, HBV showed amplification in all diluted positive samples of 100, 50, 25, 10 and 5 IU/ml. HIV and HCV infected samples showed amplification in all diluted positive samples of 500, 100, 50 and 30 IU/ml. For HIV, out of six diluted samples of 30 IU/ml, five were amplified. A total of 14,170 seronegative blood samples were tested by RealStar PCR kit in 10-MP and 6 (0.042%) samples/pools were positive. A total of 65,362 seronegative blood donations were also tested by kits of Sacace Biotechnologies, in 10-MP and 45 (0.075%) pools were positive. The prevalence of combined NAT yield cases among routine donors was 1 in 1559 donations tested for all the 3 viruses.Conclusion: The semi-automated combined system for NAT screening assays is robust, sensitive, reproducible, and this gives an additional layer of safety with affordable cost.

NAT positivity in seronegative voluntary blood donors from western India

Background and objectivesPrevalence and composition of Hepatitis B, Hepatitis C and HIV-1, NAT positive but seronegative voluntary blood donors from western part of India is yet to be documented. Material and methodsOver last 2 1/2 years all the seronegative voluntary blood donors were tested using 10 minipools on a semiautomated NAT testing platform. The positively tested donors were followed up for at least five months for development of seropositivity. Results79532 seronegative donations were tested by 10 minipool (MP) NAT leading to 51 positive sample (44 Hep B, 5 HIV 1 and Hep C positive). All the HIV and Hep C NAT positive donors eventually developed seropositivity and out of 44 Hep B NAT positive donors, 31 developed seropositivity within six months of follow up, following counseling of the donors. This data translate into NAT yield of 1:1559 donors for all virus taken together. NAT yield for Hep B 1:1807 donors were much higher than HIV 1 in 1:15906 and HCV yield of 1:39761. Semiautomated minipool NAT testing system was found to be cost effective way for improving blood safety. Interpretation and conclusionSeronegative NAT yield in voluntary blood donors are quiet high in western part of India and in line with rest of the country is mainly due to Hepatitis B infection. Implementation of strict donor screening, Hep B vaccination of the population and sample mutation of NAT testing should be under taken on war footing.

Prevalence of HBV DNA among 20 million seronegative blood donations in China from 2010 to 2015.

The prevalence of HBV DNA among seronegative blood donations in China has not been studied extensively on a nationwide scale. The aim of this study was to analyse the prevalence, trend, distributions, and serological characteristics of HBV DNA positive/seronegative blood donations. We collected HBV test data from all blood banks of China from 2010 to 2015 and performed supplemental serological tests and quantitative detection of HBV DNA of the seronegative/HBV DNA positive blood donations. We analysed the prevalence of HBV DNA among seronegative blood donations screened by varying nucleotide acid test (NAT) reagents. The analysis results showed that a total of 20,084,187 seronegative blood donations were screened by NAT from 2010 to 2015 in China. The average frequency of HBV DNA among seronegative blood donations was 1/1482, but there has been a steady increase from 1/1861 in 2011 to 1/1269 in 2015. The geographical distribution of seronegative and HBV DNA positive blood donations was roughly consistent with that of HBsAg. The most common serological pattern was HBeAb and HBcAb positive. In conclusion, our study offeres fundamental data of seronegative and HBV DNA positive blood donations throughout China.

HIV P24 antigen among HIV antibody seronegative blood donors in Osogbo Osun State, South Western Nigeria

Background: Efforts to curb the spread of HIV transmission through transfusion of blood and its products is still a problem because of challenge in countries using antibody-based rapid methods to detect infection during window period. Transmission of HIV through infected blood and its products accounts for approximately 10% in African region.Methods: This study analyzed true negativity of HIV infection in blood donors screened by ELISA test based on p24 core antigen detection. Four hundred and eighty (480) blood donors initially negative for HIV antibody by rapid screening kit, Determine™ HIV-1/2 (Abbott Laboratory, IL, USA) and re-screened with Immuno Comb® II HIV 1 and 2 (Bispot kit PBS Organics and Israel 2005). The samples were further tested for the presence of HIV antibody and p24 HIV core antigen using ELISA kits (Genscreen TM ULTRA HIV Ag–Ab) following manufacturer’s instructions. All donors initially tested negative for Hepatitis B virus, Hepatitis C virus.Result: Two (0.42%) of 480 blood donors tested positive for the p24 HIV core antigen. The two positive donors for the p24 antigen had multiple sexual partners and recent sexually transmitted infections.Conclusion: The association of the HIV p24 antigen with blood donation was highly significant (p = 0.000) and pose a great risk to recipients if screening of blood donor is only carried out by HIV antibody detection.

The natural history of recent hepatitis C virus infection among blood donors and injection drug users in the country of Georgia.

IntroductionHepatitis C virus (HCV) infection is a serious health problem in Georgia.MethodsWe conducted a prospective study to identify and characterize the natural history of recent HCV infection since very first days of infection. Recent HCV infection was defined as detectable plasma HCV RNA in the absence of anti-HCV antibodies.ResultsA total of 7600 HCV seronegative blood donors and 3600 HCV seronegative drug users were screened for recent HCV infection. Among them 7 (0.09 %) blood donors and 10 (0.28 %) drug users tested positive for HCV RNA and were classified as having recent HCV infection. Of these 17 patients 4 (23.5 %) spontaneously cleared the virus by the end of 24 week follow-up. Five clinical forms of recent HCV infection were identified during the follow-up. Four patients had symptomatic disease, including 3 patients with jaundice and other clinical symptoms (2 of them cleared virus) and 1 patient only had other symptoms without jaundice. All symptomatic patients had ALT elevation. Three distinct variants of asymptomatic disease were identified in 13 patients: 9 patients had ALT elevation and none cleared the virus; 2 patients developed chronic disease without ALT elevation; 2 patients cleared virus without anti-HCV seroconversion and without ALT elevation; this form can be described as transitory HCV viremia.ConclusionAdditional studies are needed to define clinical and public health implications of transitory HCV viremia. Our study suggests the need for implementing nucleic acid testing of blood donors and key populations in order to more effectively identify HCV infected persons.

Successful Treatment of Refractory CMV Chorioretinitis and Meningoencephalitis with Adoptive Transfer of Third Party CMVpp65 Specific T-Cell Lines

▪Adoptive immunotherapy with transplant donor-derived virus specific T cells is an effective strategy for the treatment of CMV viremia and disease arising after an allogeneic hematopoietic stem cell (HSCT). At our center this approach has become a standard part of the armamentarium of CMV directed therapy. However, donor-derived CMVpp65-specific cytotoxic T cells (CMV-CTLs) are not available for patients transplanted from a seronegative or cord blood donor. In addition, in the HLA disparate transplant setting the CMV-CTL line derived from non-identical donors may be restricted by non-shared HLA alleles. For these patients as we have previously reported, treatment with in vitro expanded CMV-CTLs derived from an HLA partially matched third party donors is an option.One particularly difficult clinical situation is the treatment of patients who develop CMV disease involving the sanctuary sites of the central nervous system and retina. We have treated 12 patients with primary donor derived (n=1) or third party (n=11) CMV CTLs for retinitis (7) or meningitis/encephalitis (4) or both (1). Recipients of hematopoietic stem cell transplant (HSCT) had undergone T cell depleted (5) conventional (1) or cord blood (4) transplantation. One patient was treated after solid organ transplantation and one for HIV related CMV retinitis.Third party CMV CTLS were selected from a bank of 132 lines generated under GMP conditions from normal HSCT donors specifically consented for use of their T cells in patients other than their designated transplant recipient. Third party CMV-CTLs were selected on the basis of HLA matching at a minimum of 2/8 recipient alleles and HLA restriction of the T cells by one or more HLA alleles present in the patient. A total of 10 distinct third party CMV-CTL lines and one (1) donor derived line were used. Patients received infusions of CMV-CTLs after failing a median of 146 (43-419) days of prior therapy with a median of 4 anti-viral agents.Patients received 3 weekly infusions of CMV-CTLs at doses of 1 x 10e6 T cells/kg (n=10) 2 x 10e6 T cells/kg (n=1) and 0.5 x 10e6 T-cells/kg (n=1) and were eligible to receive additional cycles of cells if they had no toxicity five weeks after the start of cellular therapy. One patient was evaluated only for toxicity as efficacy was confounded by anti-viral therapy required for treatment of varicella zoster. Of the 11 evaluable patients, 7 achieved CR, 1 PR (clinical improvement in disease and a 3 log decrease in viral load) and 1 SD. All but one of these responses were durable. One patient who had achieved a CR had recurrence of low grade viremia which was not retreated due to his overall medical deterioration. The two patients with progression of disease ultimately died of CMV.Patients were monitored for expansion of CMV-CTLs by CMV-specific interferon gamma and where appropriate tetramer analysis. Responding patients consistently had detectable expansion of CMV-specific CTL populations. In addition, in one patient, undergoing serial lumbar punctures, we were able to detect third party CMV-CTLs in the CSF. There were limited toxicities associated with CMV-CTL infusions. No patient developed de novo GvHD or a flare of prior GvHD.This study demonstrates a high response rate among patients with otherwise refractory CMV chorioretinitis or meningoencephalitis following adoptive therapy with primary donor or third party CMV-CTLs; thus demonstrating the capacity of adoptively transferred CMV-CTLs to treat disease in these sanctuary sites without toxicity. When selected based on restriction through shared alleles, third party CMV-CTLs are effective despite significant HLA disparity. The bank of CMV specific T cells can provide an immediate source of HLA partially matched appropriately restricted T cells for adoptive immunotherapy to treat CMV disease affecting the CNS. This enables treatment early in the course of disease with CMV-CTL lines previously prepared and characterized in terms of HLA restriction. This approach is anticipated to maximize the response rate as well as minimize toxicity from anti-viral therapy. DisclosuresProckop:Atara Biotherapeutics: Other: I have no financial disclosures, but Atara Biotherapeutics has exercised a licensing agreement with Memorial Sloan Kettering Cancer Center and MSKCC and some investigators at MSKCC have a financial interest in Atara.. Hasan:Atara Biotherapeutics: Research Funding. O'Reilly:Atara Biotherapeutics: Research Funding.

Occult hepatitis B viral infection among blood donors in South–Eastern Nigeria

Hepatitis B virus infection is endemic in many parts of sub-Saharan Africa including Nigeria. Occult hepatitis B virus infection (OBI) is a challenging clinical problem characterized by the absence of Hepatitis B surface Antigen (HBsAg) and low viral DNA load. We aimed at determining the prevalence of OBI among repeat blood donors in Abakaliki, south-eastern Nigeria. Of 113 informed consented repeat blood donors enrolled into the study, 12 donors (10·6%) tested positive to both serological HBsAg screening, anti-HBc total and hepatitis B virus (HBV) DNA Nested PCR tests. One donor (0·9%) tested HBsAg positive, anti-HBC positive but Nested PCR negative. All donors were negative for HIV 1 and 2 and HCV infections. Of the 100 HbsAg negative repeat blood donors, 8·0% (eight donors) were HBV DNA positive by nested PCR method and anti-HBc total positive by ELISA. The median viral load, determined by real time PCR-Taqman chemistry, in the OBI blood samples was 51 IU/ml compared to 228 IU/ml of the HBsAg screen positive donors. The observed OBI prevalence of 8·0% corroborated with high endemicity of HBV infection in Abakaliki. We therefore recommend routine HBV DNA testing by real time PCR method on all sero-negative blood donations in Abakaliki and for a similar policy to be evaluated across the sub-Saharan Africa.

Estimating window period blood donations for human immunodeficiency virus Type 1, hepatitis C virus, and hepatitis B virus by nucleic acid amplification testing in Southern Pakistan

Recently, strategic planning was initiated by the National Blood Transfusion Services Pakistan to improve its blood bank facilities. Emphasis has been placed on appropriate screening of blood products. Located in the southern region, Aga Khan University Hospital is a 700-bed tertiary care academic institute with comprehensive blood banking. Screening of blood donors has been based on verbal screening and serologic testing to date. Additionally, the need of implementing nucleic acid testing (NAT) was considered in 2011 because of an upsurge in hepatitis epidemiology. The aim of this study was to analyze the efficacy of this additional donor screening program and to evaluate the impact of NAT on the yield and residual risk of transfusion-transmissible viral infections. A total of 42,830 blood donations collected between 2011 and 2012 were screened for routine serologic assays. Only serologically negative donors (n=41,304) were tested for NAT. The frequency of viral infections was evaluated through serologic techniques and NAT yield for viral agents was estimated for computing window period donors. Residual risk per million donors was computed for viral infections in seronegative blood donors. Serologic work-up showed 1571 abnormal screening results in 1526 blood donors with the following results: hepatitis C virus antibodies (anti-HCV; n=708), hepatitis B surface antigen (n=555), human immunodeficiency virus antibodies (anti-HIV; n=29), malaria (n=30), VDRL (n=249), and coinfection (n=45). Thirty-five NAT-reactive samples were identified: HIV-1, one; HCV, 27; and hepatitis B virus (HBV), seven. Incident rates per 10(5) donors were highest for HCV (453.3) followed by HBV (171.5) and HIV (72.2). Calculated residual risk per million donors was highest at 1 in 10,900 for HBV, intermediate at 1 in 13,900 for HCV, and least at 1 in 62,600 for HIV. Incidence rates and estimated residual risk indicate that the current risk of transfusion-transmitted viral infections attributable to blood donation is relatively high in this country. The study recommends the parallel use of both serology and NAT screening of donated blood in countries that have high seroprevalence of these viral infections.

The combination of arsenic, interferon-alpha, and zidovudine restores an “immunocompetent-like” micro-environment in patients with adult T-cell leukemia lymphoma

HTLV-I associated adult T-cell leukemia/lymphoma (ATL) carries a dismal prognosis due to chemo-resistance and immuno-compromised micro-environment. The combination of zidovudine and interferon-alpha (IFN) significantly improved survival in ATL. Promising results were reported by adding arsenic trioxide to zidovudine and IFN. Here we assessed Th1/Th2/Treg cytokine gene expression profiles in 16 ATL patients before and 30 days after treatment with arsenic/IFN/zidovudine, in comparison with HTLV-I healthy carriers and sero-negative blood donors. ATL patients at diagnosis displayed a Treg/Th2 cytokine profile with significantly elevated transcript levels of Foxp3, interleukin-10 (IL-10), and IL-4 and had a reduced Th1 profile evidenced by decreased transcript levels of interferon-γ (IFN-γ) and IL-2. Most patients (15/16) responded, with CD4+CD25+ cells significantly decreasing after therapy, paralleled by decreases in Foxp3 transcript. Importantly, arsenic/IFN/zidovudine therapy sharply diminished IL-10 transcript and serum levels concomittant with decrease in IL-4 and increases in IFN-γ and IL-2 mRNA, whether or not values were adjusted to the percentage of CD4+CD25+ cells. The observed shift from a Treg/Th2 phenotype before treatment toward a Th1 phenotype after treatment with arsenic/IFN/zidovudine may play an important role in restoring an immuno-competent micro-environment, which enhances the eradication of ATL cells and the prevention of opportunistic infections.