First report on leucocytes cell population data as an applicable test for clinical stratification in people living with HIV

Background: Leukocyte cell population data (CPD) are parameters currently being studied as biomarkers in sepsis and other pathologies. CPD was procured with the BC 6800 Plus Mindray® hematology analyzer that also provides information on the internal cellular complexity, nucleic acid content, and size of leukocytes. This study aimed to assess the efficacy of CPD parameters as biomarkers in the diagnosis of sepsis by comparing these with the standard sepsis biomarker, procalcitonin (PCT). Methods: In total 107 patients with suspected sepsis were included in the study, and serum procalcitonin levels were measured and WBC cell population data (CPD) were analyzed in a hemogram to confirm the diagnostic accuracy of these biomarkers. ROC curves were plotted for all parameters and patients were categorized based on their serum procalcitonin levels. IBM SPSS Stadistics version 24 was used for statistical analysis of the data. Results: All parameters except the CPD NEU-X (neutrophil complexity), NEU-Z (neutrophil size), LYM-Z (lymphocyte size) and MON-Z (monocyte size) showed statistically significant results (p<0.05). ROC curve analysis showed that the CPD parameters MON-X and NEU-Y had area under the ROC curve (AUC) > 0.7, reflecting the better diagnostic performance in sepsis. A lower AUC of 0.650 was obtained for PCT; 87.8% of septic patients had serum PCT levels between ≥5 ng/mL and 51.2% ≥ 2 ng/mL. Conclusion: This study suggests that some of the new CPD parameters (MON-X and NEU-Y) have the potential to be a useful diagnostic marker for sepsis.

BackgroundMRSA bacteremia requires early anti-MRSA therapy, as beta-lactams are ineffective; delays for pathogen identification and susceptibility results can worsen outcomes. Rapid biomarkers are thus essential. Cell population data (CPD), captured via flow cytometry by automated hematology analyzers, appears earlier than conventional laboratory changes but is not reported to clinicians. CPD is readily available at no extra cost and shows promise for early exacerbation detection, though its utility remains underexplored.Figure 1:Study Flowchart for Classification of MRSA Bacteremia Patients Based on Initial Antimicrobial Therapy to Evaluate the Utility of Cell Population Data (CPD)Abbreviations: Abx, antibiotic; CBC, complete blood countTable 1:Comparison of Patient Characteristics Between the Appropriate Therapy Group (AT Group; Anti-MRSA Agents Initiated) and the Inappropriate Therapy Group (IT Group; Non-Anti-MRSA Antibiotics)1. Statistical analyses were performed using Stata/BE 18 (StataCorp., College Station, TX, USA).Continuous variables were analyzed using the t-test, and binary or categorical variables usingFisher’s exact test. A two-sided p-value of <0.05 was considered statistically significant. Variablesshowing statistical significance are indicated with an asterisk.2. Battle SE, et al. Prediction of mortality in Staphylococcus aureus bloodstream infection using quickPitt bacteremia score. J Infect. 2022 Feb;84(2):131–5.MethodsWe retrospectively analyzed MRSA bacteremia cases at Saitama Medical Center, a tertiary hospital in Japan (April 2018–March 2021). Patients were grouped into appropriate therapy (AT; anti-MRSA agents) or inappropriate therapy (IT; non-anti-MRSA agents) (Fig. 1). Hematologic parameters, including CPD from XN-9000 analyzers (Sysmex Corp., Japan), were collected before and after antibiotic initiation; post-/pre-treatment ratios were calculated to minimize interpatient variability. Univariate analyses assessed patient characteristics and laboratory value ratios between groups. Logistic regression adjusting for significant covariates evaluated therapy appropriateness. A multivariable model yielded a composite score, and its diagnostic performance was evaluated.Table 2:Comparison of Patient Characteristics Between the Appropriate Therapy Group (AT Group; Anti-MRSA Agents Initiated) and the Inappropriate Therapy Group (IT Group; Non-Anti-MRSA Antibiotics)1. Hematologic parameters, including cell population data (CPD) from XN-9000 analyzers (Sysmex Corp., Kobe, Japan), were collected before and after antibiotic initiation, and post-/pre-treatment ratios were calculated to reduce interpatient variability.2. Statistical analyses were performed using Stata/BE 18 (StataCorp., College Station, TX, USA).Continuous variables were analyzed using the t-test. A two-sided p-value of <0.05 was considered statistically significant. Variables showing statistical significance are indicated with an asterisk.3. The prefixes NE-, LY-, and MO- indicate that the corresponding CPD are derived from neutrophils, lymphocytes, and monocytes, respectively.Figure 2:Diagnostic Performance of a Composite Prediction Score for Identifying Appropriate Versus Inappropriate Therapy in MRSA Bacteremia 1The composite prediction score was derived from the following multivariable logistic regressionmodel:The composite prediction score = β0 + β1 × lymphocyte (%) + β2 × NE-SFL + β3 × NE-FSC+ β4 × interval1 + β5 × interval2β0 = intercept valueβ1 = coefficient for lymphocyte (%)β2 = coefficient for NE-SFLβ3 = coefficient for NE-FSC,β4 = coefficient for restricted cubic spline term (blood sampling interval1)β5 = coefficient for restricted cubic spline term (blood sampling interval2)To screen for the optimal cutoff of the composite prediction score, Liu’s method was applied. The final diagnostic performance of the composite prediction score was then derived from the receiver operating characteristic curve based on the Liu’s method-derived cutoff.2. Abbreviations: AUC, Area Under the Receiver Operating Characteristic Curve; PPV, PositivePredictive Value; NPV, Negative Predictive Value.ResultsFifty-two patients (AT 32, IT 20) were analyzed. Univariate analysis identified four significant factors: blood sampling interval and changes in lymphocyte (%), NE-SFL, and NE-FSC (both CPD parameters) (Table 1, 2). No strong multicollinearity was observed among these three markers (r = −0.11 to +0.35). A composite model from multivariable logistic regression adjusted for the interval and including lymphocyte (%), NE-SFL, and NE-FSC showed excellent discrimination (AUC 0.93, 95 % CI: 0.86–1.0), with sensitivity 0.90, specificity 0.88, PPV 0.82, and NPV 0.93 (Fig. 2).ConclusionNeutrophil-related CPD parameters quickly reflected treatment appropriateness in MRSA bacteremia. A composite score derived from lymphocyte (%), NE-SFL (nucleic acid content), and NE-FSC (cell size) may enable early recognition of ineffective therapy. As CPD is routinely collected without added cost or procedures, it offers a practical biomarker to prompt timely anti-MRSA adjustments.DisclosuresAll Authors: No reported disclosures

The aim of the study was to evaluate the predictive value of cell population data (CPD) parameters in comparison with procalcitonin (PCT) and C-reactive protein (CRP) for an early diagnosis of sepsis in intensive care unit (ICU). The effect of renal function on CPD, PCT and CRP, in septic and non-septic patients was also investigated. This is a retrospective, observational and single-center study, performed with data collected from patients consecutively admitted to the ICU of the Edoardo Bassini Hospital in Milan. Patients were divided in septic and non-septic according to Sepsis-III criteria. The control group was formed by critically ill patients without sepsis. Patients with sepsis were further divided in patients with sepsis and patients with septic shock. A significant difference between septic and non-septic patients was found for neutrophils complexity (NE-SSC), neutrophils fluorescence intensity (NE-SFL), width of dispersion of neutrophils fluorescence (NE-WY), monocytes complexity (MO-X), monocytes fluorescence intensity (MO-Y), PCT and CRP parameters. PCT, neutrophils sixe (NE-FSC), NE-WY, width of dispersion of neutrophils size (NE-WZ) and MO-X discriminated sepsis and septic-shock patients. CPD parameters were not influenced by renal function. CPD, PCT and CRP had a heterogeneous diagnostic performance efficiency in the prediction of sepsis. Overall, NE-SSC, NE-SFL, width of dispersion of neutrophils complexity (NE-WX), MO-X, MO-Y, PCT and CRP displayed the best diagnostic performance for sepsis. This study suggested that some CPD parameters (i.e.,NE-SFL and MO-X) might provide useful information for diagnosis and management of sepsis.

Background The cell population data (CPD) parameters reported by XN analyzers (Sysmex Corporation, Kobe, Japan) reflect the size and internal structure of leukocytes. We explored whether CPD values could contribute to recognize those patients with fever at risk to develop sepsis. A profile of sepsis was developed combining CPD parameters and other markers. Methods We recruited 295 patients at the onset of fever, with infection confirmed by positive cultures. We studied the diagnostic performance of the CPD parameters in the differential diagnosis of sepsis vs. non-systemic bacterial infection using receiver operating characteristic (ROC) curve analysis. Additionally, the K-means unsupervised clustering method was applied. Once the clusters had been defined, the relationship between them and the CPD parameter values was assessed with the non-parametric Wilcoxon test. Lastly, the relationship between the clusters obtained and the categorical variables was examined with the χ2-test (or Fisher's exact test). Results ROC analysis demonstrated that NE-FSL, NE-WY, NE-WZ and MO-WZ had areas under the curve (AUCs) >0.700 for predicting sepsis. Using the K-means clustering algorithm, 80 patients (66.67%) were assigned to Cluster 1 and the others to Cluster 2. Out of 80 of patients in Cluster 1, 45 (56.25%) presented a PCT value ≥2 ng/mL, whereas almost 80% of Cluster 2 patients had a PCT <2 ng/mL. Cluster 1 was characterized by high NE-SFL, NE-WY, MO-X, MO-WX and MO-Z values (p<0.05). Conclusions CPD related to monocyte complexity and neutrophil activation were found to be significant, with high values suggesting sepsis.

Quality Control of Structural Parameters (CELL POPULATION DATA) of Sysmex XN Series

Hitherto, there has been no comprehensive study on the usefulness of cell population data (CPD) parameters as a screening tool in the discrimination of non-neoplastic and neoplastic haematological disorders. Hence, we aimed to develop an algorithm derived from CPD parameters to enable robust screening of neoplastic from non-neoplastic samples and subsequently to aid in differentiating various neoplastic haematological disorders. In this study, the CPD parameters from 245 subtypes of leukaemia and lymphoma were compared against 1103 non-neoplastic cases, and those CPD parameters that were vigorous discriminants were selected for algorithm development. We devised a novel algorithm: [(SD-V-NE*MN-UMALS-LY*SD-AL2-MO)/MN-C-NE] to distinguish neoplastic from non-neoplastic cases. Following that, the single parameter MN-AL2-NE was used as a discriminant to rule out reactive cases from neoplastic cases. We then assessed CPD parameters that were useful in delineating leukaemia subtypes as follows: AML (SD-MALS-NE and SD-UMALS-NE), APL (MN-V-NE and SD-V-MO), ALL (MN-MALS-NE and MN-LMALS-NE) and CLL (SD-C-MO). Prospective studies were carried out to validate the algorithm and single parameter, MN-AL2-NE. We propose these CPD parameter-based discriminant strategies to be adopted as an initial screening and flagging system in the preliminary evaluation of leukocyte morphology.

Timely diagnosis of neonatal sepsis is crucial but remains challenging. Cell Population Data (CPD) provide high-resolution phenotyping of leukocytes, offering potential for sepsis detection. We aimed to establish neonatal CPD reference intervals and explore their capacity to detect sepsis and necrotizing enterocolitis (NEC). CPD from neutrophils, monocytes, and lymphocytes were analyzed in hospitalized newborns. Reference intervals (5-95th percentiles), at birth and during the first 28 days, were derived from newborns without conditions potentially impacting CPD (reference group). The performance of CPD in detecting blood culture-proven sepsis/NEC was evaluated against complete blood count (CBC) and C-reactive protein (CRP). Reference intervals from 905 neonates showed that mean CPD values followed distinct trajectories for each parameter, while distribution width generally decreased with increasing gestational and postnatal age. CPD in 39 sepsis/NEC cases, obtained on the day of clinical suspicion or from the closest CBC, differed from those in the reference group, particularly neutrophil fluorescence intensity (NE-SFL) (56.2 vs. 41.1 arbitrary units, P < 0.001). NE-SFL had superior accuracy compared to other CPD, CBC, and CRP, with 90% sensitivity and 76% specificity. This study establishes neonatal CPD reference intervals and identifies NE-SFL as a potential sepsis biomarker. This study establishes reference intervals for neonatal leukocyte Cell Population Data (CPD), providing a valuable resource for interpreting these preclinical parameters in newborns. Our findings highlight the impact of gestational and postnatal age on neutrophil, monocyte, and lymphocyte morphology, contributing to a better understanding of neonatal immune development. In an exploratory analysis, CPD parameters, particularly NE-SFL, had superior diagnostic accuracy for sepsis and necrotizing enterocolitis compared to traditional biomarkers. As CPD are automatically generated with CBC, they offer a cost-effective, real-time, and objective tool with potential for improving neonatal sepsis detection.

This study evaluated the clinical significance of cell population data (CPD) parameters obtained on Sysmex XN-9000 in septic patients admitted to intensive care unit (ICU) and stratified according to liver function. The study population consisted in 84 patients, 44 of whom did not develop sepsis (NS), whereas the remaining 40 developed sepsis (SE) (n=24) or septic shock (SS) (n=16). Two hundred ostensibly healthy blood donors [healthy subjects (HS)], undergoing routine blood testing before a regular blood donation, were studied. Except for neutrophils and lymphocytes cell size (NE-FCS and LY-Z), all other CPD values were significantly different in ICU patients compared to HS. Neutrophils and monocytes fluorescence intensity (NE-SFL and MO-X) values were significantly higher in SS compared to sepsis and not develop sepsis patients. The value of many parameters was also different according to liver function. Overall, MO-X and neutrophils fluorescence intensity (NE-SFL) exhibited the best performance for diagnosing sepsis in all patients (AUC, 0.75 and 0.72), as well as in those with (AUC, 0.95 and 0.89) or without (AUC, 0.72 for both) liver impairment. These parameters were also significantly correlated with Sequential Organ Failure Assessment (SOFA) score. This study suggested that some novel CPD parameters (namely NE-SFL and MO-X) may provide useful information for diagnosis and management of sepsis.

Myelodysplastic syndromes (MDS) are characterized by morphologic dysplasia and cytopenia and have a propensity for acute leukemic transformation. However, dysplasia is diagnosed by morphology, thus having cell population data (CPD) that can differentiate cytopenic patients with MDS from other conditions may facilitate accurate diagnosis. We assessed the utility of complete blood count (CBC) parameters and CPD derived from an Abbott Alinity hq analyzer to discriminate MDS-related cytopenia. The patient cohort (n=345) included 64 samples from patients with MDS, 162 from patients with other cytopenia, and 119 from healthy controls. The hematological parameters and research use-only parameters of the Abbott Alinity hq analyzer were compared between the cytopenic groups. The effectiveness of the individual standard and research CBC parameters to differentiate MDS from other forms of cytopenia was assessed through a receiver operating characteristics (ROC) analysis. The percentage of MAC (Macrocytic RBCs) and hemoglobin distribution width (HDW) were higher in the MDS group than in the other cytopenia group and showed the greatest difference between both groups, with an area under the curve (AUC) of 0.766 (0.678-0.855) and 0.786 (0.702-0.870), respectively. The platelet distribution width was higher in the MDS group than in the other cytopenia group, with an AUC of 0.697 (0.623-0.770). WBC CPD extracted from histograms, especially Atyp-PMN-loc and Neu-ALL-M, showed high AUCs of 0.815 (0.750-0.879) and 0.778 (0.711-0.845), respectively. Our findings demonstrate the clinical utility of CPD and hematology parameters of the Abbott Alinity hq analyzer in the differential diagnosis of MDS.

BackgroundWorldwide tuberculosis (TB) takes more lives than any other infectious diseases. WHO estimates around 68,000 incident TB cases in Nepal. However, in 2018 only around 27,232 new TB cases were reported in the national system, resulting around 40,768 incident TB cases missing every year in Nepal. National Tuberculosis Control Center carried out this study in anti-retroviral therapy (ART) sites to estimate the prevalence of TB and identify the associated risk factors for TB among the people living with Human Immunodeficiency Virus (PLHIVs) in Nepal.MethodsIt was a cross-sectional institution-based study conducted between March and August 2018. Six ART sites with high caseloads of PLHIVs were selected. PLHIVs who were equal or above 18 years of age and were in ART program at the selected study sites were considered eligible for the study. Diagnosis of tuberculosis among PLHIVs who agreed to participate in the study was carried out as per the National Tuberculosis Management Guideline of National Tuberculosis Program of Nepal.ResultsAmong 403 PLHIVs, tuberculosis was diagnosed in 40 (9.9%) individuals. Median age of the participants was 36 (30–43) years. Prevalence of TB was significantly higher among male PLHIVs than female PLHIVs (13.6% Vs 5.8%; P = 0.02) and Dalit ethnic group compared to Brahmin/Chettri (22.0%Vs5.9%, P = 0.01). The risk of developing TB was found significant among those with HIV stage progressed to WHO stage 3 and 4 (OR = 4.85, P<0.001) and with the family history of TB (OR = 4.50, P = 0.002).ConclusionsPrevalence of TB among PLHIVs in Nepal was found 9.9%. Risk of developing TB was higher among PLHIVs who were male, Dalit, with HIV stage progressed to WHO stage 3 and 4 and with family history of TB. Hence, targeted interventions are needed to prevent the risk of developing TB among PLHIVs. Similarly, integrated, and comprehensive TB and HIV diagnosis and treatment services are needed for the management of TB/HIV co-infection in Nepal.

The purpose of this study was to preliminarily evaluate the immunogenicity and immune persistence of inactivated SARS-CoV-2 vaccines in PLWH in the real world. We collected blood samples from 132 PLWH aged 18–59 years who were vaccinated with two doses of BBIBP-CorV vaccine (Sinopharm) or CoronaVac vaccine (SinoVac) at 28 ± 7 days and 180 ± 20 days the after second dose, to detect the level of Spike receptor binding domain-protein specific IgG (S-RBD-IgG) by using chemiluminescence. We found that the BBIBP-CorV vaccine or the CoronaVac vaccine induced lower S-RBD-IgG antibody seropositivity rates and levels in PLWH than in healthy controls (HCs). The BBIBP-CorV vaccine or the CoronaVac vaccine induced lower humoral immune responses in PLWH, having lower CD4+T cell counts (<350 cells/μL) compared to PLWH, and having higher CD4+T cell counts (≥350 cells/μL) after a second dose of vaccination. The BBIBP-CorV vaccine or the CoronaVac vaccine induced lower S-RBD-IgG antibody levels in PLWH, having CD4+T cell counts ≥350 cells/μL compared to HCs. No negative effects were observed in terms of the CD4+T cell counts and HIV RNA viral load (VL) of PLWH after vaccination. Ninety-nine PLWH and eighty-three HCs completed a second blood collection for testing; we found a statistically significant decrease in the humoral immune response both in PLWH and HCs from 28 days to 180 days after a second dose of BBIBP-CorV vaccine or CoronaVac vaccine. The S-RBD-IgG antibody induced by the BBIBP-CorV vaccine or the CoronaVac vaccine declined faster in the PLWH population than in the healthy population, and two doses of the BBIBP-CorV vaccine or the CoronaVac vaccine may not be enough to provide PLWH with persistent immunity against SARS-CoV-2. It is necessary for PLWH to be prioritized for a third dose over the healthy population, but the immunogenicity of the third dose of the homologous or heterologous vaccine requires further study.

This editorial comments on the article by Miller et al.

Absolute Lymphocyte Count Expansion Kinetics Can be Utilized to Assess Response to CAR T Cell Products

Fibroblast Growth Factor 21 Is Elevated in HIV and Associated With Interleukin-6.

Objectives: Innovative Cell Population Data (CPD) have been used as early biomarkers for diagnosing sepsis in adults. We assessed the usefulness of CPD in pediatric patients with sepsis/septic shock, in terms of early recognition and outcome prediction. We revised 54 patients (0–15 y) admitted to our Pediatric Intensive Care Unit (PICU) for sepsis/septic shock during a 4-year period. Twenty-eight patients were excluded, 26 septic patients were enrolled (G1). Forty children admitted for elective surgery served as controls (G2). Data on five selected CPD parameters, namely neutrophils fluorescence intensity (NE-SFL), monocytes cells complexity (MO-X), monocytes fluorescence intensity (MO-Y), monocytes complexity and width of dispersion of events measured (MO-WX), and monocytes cells size and width dispersion (MO-WZ), were obtained at time of PICU admission (t0) by a hematological analyzer (Sysmex XN 9000®). As the primary outcome we evaluated the relevance of CPD for diagnosing sepsis/septic shock on PICU admission. Furthermore, we investigated if CPD at t0 were correlated with C-reactive protein (CRP), patient survival, or complicated sepsis course.Results: On PICU admission (t0), NE-SFL, MO-WX, and MO-Y were higher in sepsis/septic shock patients compared to controls. NE-SFL values were correlated with CRP values in G1 patients (r = 0.83). None of the five CPD parameters was correlated with survival or complicated sepsis course.Conclusion: We found higher values of NE-SFL, MO-WX, and MO-Y in children with sepsis/septic shock upon PICU admission. These parameters may be a promising adjunct for early sepsis diagnosis in pediatric populations. Larger, prospective studies are needed to confirm our preliminary observations.