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Diffuse large B-cell lymphoma (DLBCL) is a high-grade non-Hodgkin lymphoma with increased incidence among people living with HIV-infection (PLWH). Although its frequency is reportedly attenuated by antiretroviral therapy (ART), we have previously shown a similar rate of DLBCL in the post-ART era (2017) in Johannesburg, South Africa compared with that observed when ART had only limited availability in the South Africa state-sector (2007). Here, we present a more detailed analysis of DLBCL in the pre-and post-ART eras in Johannesburg.All cases of DLBCL diagnosed in the state-sector hospitals of Johannesburg in 2007 and 2017 were extracted from the laboratory information system, and factors of interest compared. Most (gt;85%) were observed among PLWH at both time-points; ART-coverage was significantly higher in 2017 compared with 2007, but with failed immunological recovery in 50% of cases. The immunohistochemically-defined cell of origin differed according to HIV-status; the germinal center (GC) and non-GC subtypes predominating in the PLWH and the HIV-negative group, respectively. MYC-gene rearrangement was more common than is reported elsewhere (22.1%), whereas BCL6 and BCL2 gene rearrangements were less so (14.6% and 0%, respectively). Slight improvement in survival was noted in the post-ART era, but remained poor, with bone marrow involvement and albumin levels ≤30 g/L independently associated with mortality.Although the frequency of DLBCL in Johannesburg has not dropped significantly in the post-ART era, a slight improvement in survival is observed. However, outcomes remain poor, indicating a need for further improvements in care.

Introduction Extended differential parameters (EDPs) are generated with the automated differential count by Sysmex XN-series automated hematology analysers, and include the immature granulocyte count (IG%), the neutrophil fluorescent light intensity (NE-SFL) and the neutrophil fluorescent light distribution width (NE-WY). These have been proposed as early biomarkers of bacteremia. This study aimed to evaluate the NE-SFL, NE-WY and IG% in comparison to neutrophil CD64 (nCD64) expression (as a high quality sepsis biomarker) among patients with suspected bacterial sepsis at the Chris Hani Baragwanath Academic Hospital in Johannesburg, South Africa. Methods A daily search of the laboratory information system identified samples submitted for a blood culture (BC) and a concurrent full blood count (FBC). Automated differential counts using a Sysmex XN-9000 haematology analyser and neutrophil CD64 expression by flow cytometry were assessed on the residual FBC samples. Results A total of 151 samples were collected, of which 83 were excluded due to equivocal results with regards to the presence of bacterial infection. The remaining 68 samples included 23 with bacteremia, 28 with evidence of non-bacteremic bacterial infection, 13 with no evidence of bacterial infection and 4 with Tuberculosis. HIV status was documented in 90 of the patients, with a seropositivity rate of 57.8%. The EDPs were all significantly higher among patients with bacteremia as compared to those without bacterial infection, but on ROC curve analyses, only the NE-SFL showed good performance (AUC>0.8) for discriminating cases with bacteremia from those without bacterial infection at a cut-off value of 49.75. In comparison to the nCD64, the NE-SFL showed moderate agreement (kappa = 0.5). On stratification of the ROC analysis by HIV status, the NE-SFL showed superior performance among persons with HIV infection (AUC = 1), while the automated IG% showed better performance among the patients without HIV infection (AUC = 0.9). Conclusion In this study, EDPs showed differential performance as biomarkers for bacteremia according to HIV-status in the South African setting, with the most promising results seen with the NE-SFL and IG% parameters among people with and without HIV infection, respectively. Further assessment of these parameters without pre-selection of patients likely to have infection is required to further determine their clinical utility, particularly among patients with underlying inflammatory conditions or malignancy.

Background: B-cell lymphoblastic leukaemia (B-ALL) is a malignancy of immature B-cells with several described recurrent genetic abnormalities. These have distinct clinico-pathological associations and show regional variation in prevalence. In all previously reported series, the translocation t(1;19) is uncommon, comprising 10% of all cases. The genetic composition of B-ALL in Africa is unknown. Aim: The aim of this study was to assess the genetic landscape of B-ALL in Johannesburg, South Africa. Setting: The Johannesburg state-sector. Methods: All cases of B-ALL diagnosed by flow cytometry in the state-sector hospitals of Johannesburg over 36 months between 2016 and 2019 were identified and pertinent data were recorded from the laboratory information system. Results: A total of 108 patients with B-ALL were identified, 82 (75.9%) of whom were children or adolescents. The translocation t(1;19)(q23;p13) was the most common genetic abnormality identified (23.7% of cases), predominating in young patients. The translocation t(9;22)(q34;q11) was the next most common aberration (17.5%) occurring predominantly in adults 40 years of age, but also in 8.1% of children. Crude survival rates were overall poor (44.6% overall and 57.4% in patients 18 years of age). On survival analysis, older age, KMT2A-rearrangement and t(1;19) were independently associated with relapse-related death. The t(9;22) was not associated with mortality independently from age. Conclusion: B-ALL shows a distinct pattern of lymphoblastic leukaemia-associated chromosomal translocations in Johannesburg.

BACKGROUND South Africa has a high HIV prevalence, which associates with an increased risk of lymphoma. Antiretroviral therapy (ART) became accessible in 2004, but the program has substantially expanded. Changes in lymphoma patterns are documented in high-income countries after wide-scale ART including declining high-grade B-cell non-Hodgkin lymphomas (HG B-NHLs), particularly diffuse large B-cell lymphoma, and increased Hodgkin lymphoma (HL). There are limited data from Africa. This study aimed to compare HG B-NHL characteristics in the early (2007) and later (2017) ART era. METHODS All incident lymphomas at the National Health Laboratory Service, Johannesburg, were identified using the laboratory information system, and data were collected for each patient. RESULTS The total number of lymphoma cases increased from 397 (2007) to 582 (2017). This was associated with improved lymphoma classification and patient referral for oncological care. HG B-NHL remained the most diagnosed lymphoma subtype in 2017 comprising 70% of HIV-associated lymphomas, followed by HL (24%). Diffuse large B-cell lymphoma comprised 65% of all HG B-NHLs and 45% of all lymphomas in people with HIV in 2017. Significantly more patients were on ART in 2017, with improvements in virological control documented. Despite this, 47.6% of patients were not virologically suppressed, and 37.5% of patients were ART-naive at time of diagnosis in 2017. Immunological reconstitution was suboptimal, which may reflect late initiation of ART. CONCLUSION Public health initiatives to initiate ART as early as possible and to retain patients in ART programs may assist in decreasing the number of HIV-associated lymphomas in our setting.

From its early origins, COVID-19 has spread extensively and was declared a global pandemic by the World Health Organization in March of 2020. Although initially thought to be predominantly a respiratory infection, more recent evidence points to a multisystem systemic disease which is associated with numerous haematological and immunological disturbances in addition to its other effects. Here we review the current knowledge on the haematological effects of COVID-19.

OBJECTIVES Plasmablastic lymphoma (PBL) is a clinically aggressive lymphoma which has a predilection for extranodal sites and is frequently HIV-associated. The incidence of non-Hodgkin lymphoma is thought to be reduced by widescale antiretroviral therapy (ART) coverage, but the literature is sparse as regards the impact of ART on the incidence of PBL and its outcomes in South Africa (SA). This study aimed to compare factors of interest in cases of PBL diagnosed before and after the widespread availability of ART in Johannesburg, SA. METHODS All cases of PBL diagnosed in the state sector hospitals of Johannesburg in 2007 and 2017 (before and after the widespread availability of ART, respectively) were extracted from the laboratory information system, and factors of interest compared. RESULTS The majority (> 95%) of cases of PBL were seen among people with HIV infection (PWH) at both time-points, and the proportion of patients on ART and with virological suppression (VS) increased significantly in 2017. However, the number of cases of PBL did not differ significantly between the two years assessed, comprising 46/397 (11.6%) and 53/582 (9.6%) of all lymphomas in 2007 and 2017, respectively (P = 0.23). Ongoing risk for PBL among PWH with virological control and immunological recovery was evident in 2017, as 18.9% of the patients had both VS and CD4 counts > 200 cells/μL at diagnosis. Inferior survival times were associated with elevated lactate dehydrogenase (LDH) levels and Epstein Barr virus (EBV) negativity, but were not influenced by the presence of AIDS, ART or VS. EBV negativity was significantly associated with VS, and appeared to flag a particularly aggressive form of the disease. CONCLUSIONS Widescale ART coverage has not reduced the incidence of PBL in Johannesburg, and an ongoing risk for this disease among PWH with adequate virological control and immunological recovery persists.

Background The incidence of HIV-associated Hodgkin lymphoma (HIV-HL) has not dropped in the era of widespread antiretroviral therapy (ART), and there have reportedly been shifts in the most prevalent variants encountered. In this study, factors of interest in cases of HIV-HL diagnosed before and after the widespread availability of ART in Johannesburg, South Africa, were compared. Methods All cases of HIV-HL diagnosed in 2007 and 2017 were extracted from the laboratory information system, and pertinent factors compared. Results The number of cases of HL increased significantly over the period assessed, but without a clear increase in the incidence of HIV-HL. As has been reported previously, the proportion of HIV-HL subclassified as the Nodular Sclerosis and Mixed Cellularity subtypes increased and decreased respectively over the period. The number of unclassifiable cases also increased significantly largely because of more frequent diagnosis in bone marrow (BM). BM involvement was highly prevalent at both timepoints (51.7% in 2007 vs 66.2% in 2017; P = 0.18), but was more frequently associated with multiple cytopenias in 2017. Despite significant ART upscaling, the median CD4 count was significantly lower in 2017 (242.5 cells/μL in 2007 vs 85.5 in 2017; P = 0.002). This particularly affected patients with BM involvement, and the median survival time was significantly shorter among BM+ patients diagnosed in 2017 as compared to those diagnosed in 2007. Notably, 40.8% of the patients with BM involvement diagnosed in 2017 died before the diagnosis was established. Conclusion HIV-HL with BM involvement identifies a very high-risk subpopulation in the post-ART era.

Thromboembolic conditions are a leading cause of mortality, estimated to account for 1 in 4 deaths worldwide in 2010. Over time, the incidence and mortality rates of these conditions have declined in developed countries, but are increasing in developing countries. A delicate balance exists between procoagulant and anticoagulant factors within the vascular system. Numerous acquired or inherited conditions may tip the balance either way, i.e. towards a prothrombotic or prohaemorrhagic state. Acquired thrombotic disorders are the subject of discussion in this issue, the second of a 2-part series on thrombophilia.

A peripheral blood smear review is a useful but labour intensive adjunct to the full blood count and differential count. In this study, we retrospectively evaluated the performance of the International Consensus Group for Haematology smear review rules for detection of malaria in 153 samples with Plasmodium falciparumparasitaemia. Review rules were triggered in 132 (86.3%) samples, including all patients with severe malaria. Of the 21 false negative samples, 14 (66.7%) had haemoglobin values ≥ 10g/dl and platelets > 120x109/l.

Hepatosplenic T-cell lymphoma (HSTCL) is a rare type of Non-Hodgkin Lymphoma (NHL), grouped under the mature or peripheral T-cell lymphomas. It is characterised by extranodal infiltration and proliferation of malignant T-cells within the sinusoids of the liver, sinuses and red pulp of the spleen, and the bone marrow. The tumour cells express CD2 and CD3, but are CD4, CD5 and CD8 negative and express a clonally restricted gamma–delta (or less commonly alpha–beta) T-cell receptor. The disease has an aggressive clinical course associated with a poor prognosis. We highlight and report three patients from South Africa with HSTCL, all of whom had hepatosplenomegaly and cytopaenias, and despite being HIV seronegative and immunocompetent, had a poor outcome, with a mean survival of 7.5 months in the two evaluable patients. This rare entity has not previously been reported from South Africa and as yet needs to be adequately characterised in a population where lymphoma is the most common haematological malignancy in adults, and where approximately two thirds of the adult lymphoma population are HIV seropositive. Keywords: Hepatosplenic T-cell lymphoma, Hepatosplenomegaly, Cytopaenias, Aggressive course, Poor prognosis, South Africa

Introduction South Africa has a high prevalence of Human immunodeficiency virus (HIV) infection with an estimated 7 million people with HIV (PWH). Significant progress has been made in implementing anti-retroviral therapy (ART) with 4 million patients on ART in 2017 compared with 400 000 in 2007. HIV-associated lymphomas remain a public health concern in our setting. Patterns of changes in lymphoma are well documented in high-income countries following wide-scale ART, however, there are limited published data available from our region. This study aimed to compare epidemiological characteristics of high-grade B-cell lymphomas (HG BLPDs) in PWH presenting to the academic hospitals in Johannesburg, South Africa, prior to and post implementation of a wide-scale ART programme. Methods This study was approved by the Human Research Ethics committee of the University of the Witwatersrand (protocol number: M130280). All patients with newly diagnosed lymphoma (B- and T-cell non-Hodgkin and Hodgkin lymphoma) during 2007 and 2017 in the Johannesburg academic complex of hospitals (South Africa) were identified via a comprehensive search of the laboratory information system (LIS) utilizing SNOMED codes and a detailed pathology record review. Limited demographic and clinical data were collected for each patient. Statistical analysis was performed using GraphPad Prism software, version 7.0 (San Diego, California, United States). Continuous data are presented as the median (interquartile range (IQR)) and categorical data as frequencies and percentages. The Mann-Whitney and Fisher's exact or Chi-square test were used to compare continuous and categorical variables respectively. Results and Discussion The total number of cases of lymphoma diagnosed in the Johannesburg state-sector hospitals showed a marked increase from 397 in 2007 to 582 in 2017. This may represent an improvement in access to health-care, more extensive diagnostic work-up and significantly increased HIV testing in lymphoma patients (p There were significantly more PWH presenting with HG BLPD on ART in 2017 (p PWH presenting with lymphoma showed significant changes in virologic control as expected, with a significant decline in the median HIV viral load (HIVVL) (p=0.0032), increase in the proportion of patients with an HIVVL Conclusions: A marked increase in the number of lymphoma cases diagnosed in the state-sector hospitals in Johannesburg was noted over the study period with HG BLPD remaining the most commonly diagnosed subtype in PWH despite significant improvements in virological control. This suggests that virological control alone may not be protective and that delayed or incomplete immunologic recovery as a consequence of late ART initiation may be contributing. Public health initiatives to promote early access to ARV therapy at higher CD4 counts may assist in decreasing the incidence of HG BLPDs in our setting. Disclosures No relevant conflicts of interest to declare.

Thrombocytopenia is common in HIV-infected individuals and often requires a diagnostic bone marrow examination. Interpretation may, however, be limited due to the multifactorial nature of HIV-associated thrombocytopenia and the difficulty in assessing megakaryocyte function morphologically. The immature platelet fraction (IPF) is a parameter which reportedly reflects megakaryocyte activity, with an IPF7.7% suggesting increased platelet production. The aim of this study was to correlate the IPF with the bone marrow findings as well as other clinical variables of interest in South African patients with HIV-associated thrombocytopenia.Seventy-eight HIV-positive patients with thrombocytopenia were enrolled from the Charlotte Maxeke Johannesburg Academic Hospital. The IPF levels were measured using a Sysmex XE-5000 haematology analyzer and were correlated with bone marrow and other findings.The median IPF was 7.6%, ranging from 1.3 to 44%. It was raised in 78% of patients with immune thrombocytopenia (ITP) (median = 16.3%) and low in 79% of patients with hypocellular marrow (median = 6.5%). Surprisingly, it was highly variable among patients with malignant marrow infiltration and mycobacterial infection of the bone marrow (BMTB) (median = 8.4 and 7.1%, respectively). Multivariate linear regression analysis confirmed a significant independent inverse relationship between the IPF and hypocellular marrow (P0.0001), a marginally significant positive association with ITP (P = 0.059), and the absence of any relationship with malignant infiltration or BMTB. The IPF had a significant inverse association with the platelet count (P = 0.0006), but was unrelated to the CD4 count and exposure to anti-retroviral therapy. Unexpectedly, it showed a significant positive association with the HIV viral load (P = 0.005). We speculate this to reflect increased megakaryocyte activity in compensation for accelerated platelet clearance due to HIV-driven platelet activation.This study investigates the role of the IPF in HIV-associated thrombocytopenia, and emphasizes the limitations of morphological analysis in determining megakaryocyte function.

Acute erythroid leukemia is rare, with isolated reports on presentation as an extramedullary tumor mass (myeloid sarcoma). We describe a case of pure erythroid leukemia presenting as an orbital mass in a 1-year, 9-month-old girl. This is only the second case described in a child. Tissue biopsy of the tumor mass showed medium-sized cells that were glycophorin A positive and negative with conventional myeloid markers. Flow cytometry, bone marrow aspirate, and trephine confirmed the diagnosis of pure erythroid leukemia.

Introduction: The CellaVision™ DM96 (DM96) is a digital microscopy system which performs well in developed countries. However, to date it has not been evaluated in Africa, where the pathology spectrum encountered is very different. In particular, its utility in a setting with high HIV prevalence has not been assessed, which is of interest because of the morphological aberrations often seen in HIV-positive patients. Objectives: This study aimed to evaluate the accuracy of the DM96 in a South African laboratory, with emphasis on its performance in samples collected from HIV-positive patients. Methods: A total of 149 samples submitted for a routine differential white cell count in 2012 and 2013 at the Chris Hani Baragwanath Academic Hospital in Johannesburg, South Africa were included, of which 79 (53.0%) were collected from HIV-positive patients. Results of DM96 analysis pre- and post-classification were compared with a manual differential white cell count and the impact of HIV infection and other variables of interest were assessed. Results: Pre- and post-classification accuracies were similar to those reported in developed countries. Reclassification was required in 16% of cells, with particularly high misclassification rates for eosinophils (31.7%), blasts (33.7%) and basophils (93.5%). Multivariate analysis revealed a significant relationship between the number of misclassified cells and both the white cell count ( p = 0.035) and the presence of malignant cells in the blood ( p = 0.049), but not with any other variables analysed, including HIV status. Conclusion: The DM96 exhibited acceptable accuracy in this South African laboratory, which was not impacted by HIV infection. However, as it does not eliminate the need for experienced morphologists, its cost may be unjustifiable in a resource-constrained setting.

Acute adult T-cell leukaemia (AATL) is a clinical subtype of adult T-cell leukaemia/lymphoma, a rare type of peripheral T cell lymphoma. It is associated with infection by the retrovirus Human T-Lymphotropic Virus Type-1 (HTLV-1), and is seen most commonly in HTLV-1 endemic areas. The tumour cells have polylobulated nuclei morphologically and are CD3+, CD4+ or CD8+, CD7-, express T-cell receptor , and are CD25+. The disease has an aggressive clinical course associated with marked immunosuppression caused by HTLV-1, and a poor prognosis. We report a rare case of a young male patient who presented with AATL complicated by multiple opportunistic infections including pulmonary tuberculosis and Norwegian scabies. He also had chronic hepatitis B. This constellation of illnesses has not previously been reported from South Africa in a population where another retrovirus, Human Immunodeficiency Virus (HIV), is a far more common occurrence, as are the HIV-related lymphomas.

SummaryIntroduction Peripheral blood smear review (PBSR) is a labour-intensive test, and skilled morphologists are in short supply. It is therefore helpful for laboratories to establish rules for PBSR to improve laboratory efficiency. Previously published guidelines in this regard are useful, but make few recommendations specific to neonates. Neonatal blood is characterized by several peculiarities which would be considered pathological if present in adults. Consequently, smear review rules (SRR) are often triggered in neonates without significant value being added on review. This study aimed to assess and fine-tune the SRR triggered in neonatal samples in order to improve laboratory efficiency. Methods Full blood counts collected from 188 neonatal inpatients of the Chris Hani Baragwanath Academic Hospital in South Africa were retrospectively reviewed, the triggered rules documented, and the value added on PBSR determined. Results Smear review rules were triggered in 148 (78.7%) samples, with significant morphological abnormalities identified in 84 (54.4%), and a false-positive rate of 34.0%. In patients with unhelpful review, the commonest rules triggered were the flags querying the presence of abnormal lymphocytes, blasts or nucleated red blood cells. When one or more of these flags were triggered in the absence of any other SRR, PBSR was always noncontributory. Disregarding these flags in the current cohort would reduce both the review and the false-positive rates by >20% without increasing the false-negative rate. Conclusion False-positive smear review is common in neonates, and minor modifications to SRR can substantially reduce the smear review rate without increasing the false-negative rate.

Introduction: Primary plasma cell leukaemia is a rare and highly aggressive disease that is commonly diagnosed a decade earlier than multiple myeloma, at a median age of 55 years. However, it has also been described in younger patients, as documented in this case report. It often presents with hepatosplenomegaly and lymphadenopathy, whilst the presence of bony lesions are less-commonly seen when compared to multiple myeloma. Case presentation: This report describes the case of a young woman who presented with symptoms of anaemia and a history of menorrhagia. On further careful examination, she was found to have additional signs and symptoms and was later diagnosed with primary plasma cell leukaemia. Management and outcome: On admission, the patient received supportive care measures, including blood products. At diagnosis, a specific chemotherapy regimen was commenced; however, this failed to induce remission. The decision to continue with supportive care only was made and the patient died seven months later. Discussion: This case study is presented because of its rarity, the young age of the patient at presentation and the unusual clinical and laboratory findings. Persistent anaemia unresponsive to standard treatment should raise the index of suspicion and further investigations directed to exclude malignancies should be considered. (Full article to follow)

Background. The HIV epidemic in South Africa (SA) has had a substantial impact on laboratory services, at least partially owing to the well-described propensity to cytopenias in HIV-positive patients. Objectives. ( i ) To formally gauge the impact of HIV infection on the state sector haematology services in SA by determining the HIV seropositivity rate among full blood counts (FBCs) performed at a large academic state sector laboratory; and ( ii ) to document the prevalence of cytopenias among HIV-positive patients in this setting. Methods. Randomly selected FBCs submitted to the National Health Laboratory Service laboratory at Chris Hani Baragwanath Academic Hospital, Johannesburg, were extracted from the laboratory information system (LIS) and retrospectively reviewed. HIV test results and other pertinent information in the LIS were documented, as was the presence of any cytopenias. Results. HIV status was documented in 561 of 1 006 samples (55.8%), with 307 (54.7%) of these being HIV-positive. Of the HIV-positive patients, 63.2% had one or more cytopenia/s. Anaemia was present in 183/307 (59.6%) of the HIV-positive patients, and was severe (haemoglobin

Causes of thrombocytopenia range from laboratory errors to life-threatening pathological conditions. To establish the cause, appropriate laboratory investigation is required.To determine the prevalence and causes of platelet counts100 × 10(9)/L in state health facilities in Johannesburg, South Africa, as well as the quality of the subsequent laboratory work-up in this setting.Full blood counts (FBCs) performed on 7 randomly selected days at the National Health Laboratory Service laboratory at Chris Hani Baragwanath Academic Hospital were retrospectively reviewed. Samples with platelet counts100 × 109/L were identified, and pertinent information was extracted from the laboratory database.Of 4 456 FBCs included, 381 (8.6%) had a platelet count of100 × 10(9)/L. Thrombocytopenia prevalence rates were high in haematology/oncology wards (34.4%), intensive care units (20.5%) and medical wards (18.7%) and among neonatal inpatients (16.5%), and were lowest in outpatient clinics (1-2%). A cause was apparent in ~60% of patients, the commonest causes being chemotherapy and sepsis (each comprising20% of the recognised causes). Spurious thrombocytopenia, disseminated tuberculosis, aplastic anaemia, immune thrombocytopenia and malignant marrow infiltration each accounted for 5 - 10% of the causes, while malaria, thrombotic thrombocytopenic purpura, HIV effect and liver disease were each identified in5% of cases. HIV status was documented in ~70% of the patients, of whom ~50% tested positive. The quality of the laboratory work-up showed differences between specialties within the hospital setting, and was poorest in the primary healthcare clinic sector.Thrombocytopenia is common in hospitalised patients in the Johannesburg academic state sector. Differences in the quality of the laboratory work-up emphasise the need for a standardised approach to thrombocytopenia investigation and increased awareness among clinicians.