CD4-positive Cell Count Research Articles

Improving cellular therapy operations through pre-harvest measurement of peripheral CD34-positive cell counts in allogeneic stem cell harvest.

Previously, our institution measured peripheral blood CD34 cell counts both pre- and post-peripheral blood stem cell harvest (PBSCH), with both samples analyzed simultaneously post-PBSCH. Since 2021, we have measured pre-CD34 cell counts during PBSCH, adjusting the processed blood volume based on these results. We retrospectively evaluated how this change impacted cellular therapy. Related healthy donors were included and divided into 1-day and 2-day harvest cohorts. Donors with CD34 cell counts measured post- and during PBSCH were categorized into the previous and current sub-cohorts, respectively. Regarding the 1-day cohort (n = 212), the current sub-cohort had a significantly shorter average harvest duration (151 [standard deviation, SD = 45.1] vs. 180 [SD = 27.8] minutes, respectively) and higher average infusion rates (87.6% [SD = 21.1] vs. 78.1% [SD = 25.7], respectively) than the previous sub-cohort. Adjusting the processed blood volume based on pre-PBSCH CD34 cell counts measured during the harvest may reduce donor burden and enhance workflow efficiency.

Circ VRK1/microRNA-17/PTEN axis modulates the angiogenesis of human brain microvascular endothelial cells to affect injury induced by oxygen-glucose deprivation/reperfusion

BackgroundCircular RNAs (circRNAs) can act as microRNA (miRNA) sponges, thus regulating gene expression. The role of circRNAs in the process of oxygen-glucose deprivation/reoxygenation (OGD/R) is unclear. Here, we explored the mechanism underlying Circ VRK1 in human brain microvascular endothelial cells (HBMVECs) injury induced by OGD/R.MethodsThe OGD/R cell model was established in HBMVECs. The microarray was applied to detect differentially expressed circRNAs, followed by subcellular fractionation assay. Colony formation assay, flow cytometry, ELISA, tube formation, Transwell and western blot assays were performed for loss-of-function assay. HE staining, TTC staining, immunohistochemistry and western blot were performed in an established mouse model. The relationships between Circ VRK1 and miR-17, and between miR-17 and PTEN were detected by bioinformatics and dual-luciferase assays. Rescue experiments were conducted in vitro and in vivo, and PI3K/AKT activity was detected by Western Blot.ResultsCirc VRK1, predominantly present in the cytoplasm of cells, was upregulated in the HBMVECs exposed to OGD/R. Circ VRK1 downregulation decreased proliferation, migration, tube formation, inflammatory factors and oxidative stress, while increased apoptosis in HBMVECs. Moreover, Circ VRK1 silencing reduced neurological damage, cerebral infarct size, CD34-positive cell counts and VEGF expression in mice. Circ VRK1 mediated PTEN expression and the PI3K/AKT pathway by targeting miR-17. Deletion of miR-17 inhibited the effects of Circ VRK1 siRNA, and silencing of PTEN suppressed the effects of miR-17 inhibitor.ConclusionCirc VRK1 was upregulated during OGD/R. Circ VRK1 downregulation regulates PTEN expression by targeting miR-17, thereby promoting PI3K/AKT pathway activity to alleviate OGD/R injury.

Mechanism underlying vascular remodeling in relation to circulating CD34-positive cells among older Japanese men

Development of structural atherosclerosis, an established cardiovascular risk factor, requires hematopoietic stem cells known as CD34-positive cells. However, an inverse association between circulating CD34-positive cell count and cardiovascular disease has been reported. These studies evoke a contradiction: characteristics associated with a low risk of developing structural atherosclerosis are also associated with a high risk of cardiovascular disease. To clarify the mechanisms underlying vascular remodeling, we conducted several epidemiological studies of Japanese men aged 60 to 69 years who participated in annual health check-ups. The present study summarizes those epidemiological studies and adds some discussion. From the perspective of endothelial repair activity, there are significant differences between functional versus structural atherosclerosis. Aggressive endothelial repair increases both functional and structural atherosclerosis. Deficient endothelial repair related to a shortage of CD34-positive cells due to consumption furthers functional atherosclerosis but not structural atherosclerosis. Therefore, the lack of structural atherosclerosis does not always reflect a favorable condition for the endothelium. Although further investigation is necessary, the present study suggests that higher endothelial repair activity that leads to structural atherosclerosis might have a beneficial effect on vascular health among older men.

Association between circulating CD34-positive cell count and height loss among older men

Height loss starting in middle age is reportedly significantly associated with death due to cardiovascular disease. Impaired blood flow is the main pathology in cardiovascular disease. Hematopoietic stem cells such as CD34-positive cells play an important role in maintaining the microcirculation and preventing impaired blood flow by activating endothelial repair and angiogenesis. Therefore, circulating CD34-positive cell count could be associated with height loss. To clarify the association between circulating CD34-positive cell count and height loss, we conducted a follow-up study of 363 Japanese men aged 60–69 years over 2 years. Height loss was defined as being in the highest quartile of height decrease per year. Independent of known cardiovascular risk factors, circulating CD34-positive cell count was significantly inversely associated with height loss. The fully adjusted odds ratio (OR) and 95% confidence interval (CI) of height loss for circulating CD34-positive cell count (logarithmic values) was 0.49 (0.32, 0.74). This study suggests that a lower capacity to maintain the microcirculation due to a fewer CD34-positive cells might affect height loss.

Correlation between recurrence of anorectal condyloma acuminatum and human papilloma virus subtype.

Anorectal condyloma is the representative venereal disease caused by human papilloma virus (HPV), which has more 180 subtypes. Although there are various known risk factors for recurrence, few studies have investigated the influence of HPV subtypes. We aimed to investigate the correlation between the recurrence of anorectal condyloma and HPV subtypes. We analyzed the clinical and histopathological information of 143 patients who underwent surgery for anorectal condyloma at the National Medical Center between March 2016 and September 2020. PCR analyses were performed to confirm the HPV subtype in 24 patients. Recurrence was confirmed in 63 patients through outpatient follow-up over a median of 31.7months (range: 0-56.2) after surgery. Recurrence was significantly associated with anorectal condyloma severity (p < 0.001), but there were no differences between the primary and recurrent groups, CD4-positive cell counts, or human immunodeficiency virus loads. The high-risk HPV subtype was associated with a high recurrence rate. Furthermore, the overall recurrence rate in anorectal condyloma patients was 44.1%. These results suggest that recurrence of anorectal condyloma has a significant association with high-risk HPV subtypes. Therefore, it is necessary to check for recurrences during follow-up after surgery.

Maternal and Neonatal Variables Affecting CD34+ Cell Count in the Umbilical Cord Blood

INTRODUCTION: Ease of collection, ready availability and lower graft-versus-host disease compared to peripheral blood stem cell favored umbilical cord blood stem cell transplantation. AIM: To assess the maternal and neonatal predictor affecting total nucleated and CD34+ cell count in cord blood collections. METHOD: A total of 200 Cord blood units were collected under aseptic conditions. Volume reduction was made by 6% hydroxyethyl starch followed by upright centrifugation of cord blood units to obtain a cellular pellet. Differential cell counts were done through hematology analyzer, viability testing by trypan blue exclusion test and percentage CD34, and CD45 estimation by flow cytometry. Two samples were HBsAg positive and not included in the study. RESULT: Total CD34 positive cell counts were more in cord blood units collected from younger age mother. Higher birth weight of newborns yielded a larger volume of cord blood unit and higher absolute TNC and CD34+ cell counts. The placental weight was positively correlated with the volume of cord blood collected, birth weight of new born, and total nucleated cell count but no correlation was observed with absolute CD34+ cell counts. The gravida status, gestational age and method of delivery were not significant with total nucleated cell (TNC), mononuclear cell (MNC), and CD34+ counts. CONCLUSION: Maternal age, birth weight, placental weight, and volume of CBUs were the most important predictor of getting increased nucleated and CD34+ cells in cord blood. Moreover, CBUs collected from pre-term deliveries were shown to have an approximately equal absolute number of CD34+ cells.

The Efficacy of Plerixafor Plus Recombinant Human Granulocyte Colony-Stimulating Factor for Autologous Stem-Cell Mobilization and Transplantation for Patients with Lymphoma and Plasma Cell Diseases

Objective: Plerixafor is a small-molecule inhibitor of CXCR4. This study evaluates the efficacy of plerixafor combined with recombinant human granulocytic colony stimulating factor(G-CSF), in mobilizing hematopoietic stem cells for autologous stem-cell transplantation (ASCT) in patients with lymphoma and plasma cell diseases.Methods: A retrospective analysis was performed on 41 patients with Plerixafor combined with G-CSF admitted to the Department of Hematology, Nanfang Hospital, Southern Medical University,China from July 2019 to February 2021. Mobilization protocol: all patients were given G-CSF 10μg/kg d1-4 and Plerixafor 20mg (all patients weighed less than 83kg) 11 hours before collection on day 5 (night of day 4).Results: The median age of 41 patients was 52 years (25-66 years), the median body weight was 57kg (41-80kg), and the male to female ratio was 1.16:1(22/19). The patients of plerixafor used in combination with G-CSF include Multiple Myeloma 75.6%(31/41), Diffuse Large B Cell Lymphoma was 9.8%(4/41), Peripheral T Cell Lymphoma (4.9%, 2/41), Angioimmunoblastic T Cell Lymphoma (2.4% ,1/41), Systemic light chain (AL) amyloidosis (2.4% ,1/41), POEMS syndrome (2.4%,1/41), and plasma cell leukemia (2.4%, 1/41). Above the patients got partial remission（PR） before mobilization. 17 patients got complete remission（CR）, 4 patients got very good PR(VGPR) and ten got PR. There were 5 lymphoma patients with CR and 2 with PR. The majority of patients, 25, received first-line chemotherapy. 11 patients received second-line chemotherapy, and five received third-line myeloma therapy or above. The median cycle of chemotherapy was 4 (range 4-41). Twenty-two patients with multiple myeloma received treatment with lenalidomide regimens. 26 of all (63.4%) had optimum mobilization (CD34 positive cell count ≥5 × 10 6 cells/kg), and 36 (87.8%) had minimum mobilization (CD34 positive cell count ≥2 × 10 6 cells/kg). The minimum and optimum mobilization rate of first-line chemotherapy regimen was 92% and 72% respectively. The rate of the minimum mobilization of the second line chemotherapy was 100%, the optimum mobilization rate was 63.6%. The minimum and the optimum mobilization rate were 60% and 20% for the third line chemotherapy and above. The minimum mobilization rate was 94.7% and the optimum mobilization rate was 57.9% in patients receiving lenalidomide containing chemotherapy less than the 4 cycles. The minimum and optimum mobilization rate were 66.7% and 66.7% for patients with more than 4 cycles of lenalidomide chemotherapy. Thirty-four patients received autologous hematopoietic stem cell transplantation. Conditioning regimens was TBI (total-body irradiation) + CY (CTX)+ VP-16 or BUCY+ Me-CCNU protocol in lymphoma patients, and Melphala 200mg/m 2 in Multiple myeloma patients. A reduction of melphalan dose(140mg/m 2) in patients with severe renal failure, here defined as iohexol clerarance <30 ml/min, is suggested. Neutrophil engraftment was defined as neutrophil count ≥ 0.5 × 10 9/L for 3 days or ≥ 1.0 × 10 9/L for 1 day. Platelet engraftment was defined as platelet count ≥ 20 × 10 9/L without a transfusion from the preceding 7 days. All patients who received transplantation had successful neutrophil engraftment and 94.1% had successful platelet engraftment. The median time to engraftment was 10 days(9-13 days) for neutrophil and 11 days (9-48 days) for platelet. Using all available laboratory measurements and clinical information to determine graft failures, there were two platelet graft failures in at 1 month follow-up visit. However they had successful platelet engraftment after treatment with Eltrombopag or Avatrombopag(43 days, 48 days).Conclusions: The mobilized cells after G-CSF plus Plerixafor contained high numbers of hematopoietic stem cells, and engrafted very efficiently. G-CSF plus Plerixafor may be preferred for ASCT in patients with lymphoma and plasma cell diseases. DisclosuresNo relevant conflicts of interest to declare.

Measurement of hematopoietic progenitor cells using XN2000 hematology analyzer.

Stem cell enumeration by the hematopoietic progenitor cells (HPC) mode is a novel method available from Sysmex XN2000 hematology analyzer. A small amount of blood (190μL) is required, and the results are available in a few minutes without manual gating or presample treatment. The present study compares stem cell measurements using XN2000 analyzer HPC mode and FC500 flow cytometry analyzer using peripheral blood (PB) specimens and apheresis products. In this prospective study, CD34-positive cell counts were enumerated using an FC500 flow cytometry analyzer and compared with XN2000 Sysmex analyzer (XN-HPC mode) in the same samples. Results were compared using Bland-Altman plots. A total of 103 samples were used. In the PB samples, the median HPC count and CD34-positive cells were 83.5×106 /L and 78.0×106 /L, respectively. The mean Bland-Altman difference was 4.5×106 /L (Limits: -51.7 to 60.7×106 /L), with a Pearson's correlation of 0.79. In the apheresis products, the median HPC count and CD34-positive cells were 1468×106 /L (IQR: 1049 - 1960×106 /L) and 1327×106 /L (IQR: 910 - 2001×106 /L), respectively. The mean Bland-Altman difference was 179.0×106 /L (Limits: -2022.2 - 2380.2×106 /L), with a Pearson's correlation of 0.58. The XN-HPC mode has an excellent correlation and minimal disagreement for stem cell enumeration in PB compared with flow cytometry and could replace it. There is high disagreement in apheresis products, and therefore, the XN-HPC mode cannot be recommended.

PD-L1 protein expression and copy number gains in HIV-positive locally advanced cervical cancer.

Background:The programmed death-1/programmed death-ligand-1 (PD-1/PD-L1) axis may represent a target for cervical cancer; however, it is poorly understood in human immunodeficiency virus (HIV)-infected patients.Methods:We evaluated HIV-positive (n = 42) and HIV-negative (n = 110) women with locally advanced cervical cancer regarding their PD-L1 expression, determined by combined positive score (CPS) ⩾ 1 and tumor proportion score (TPS) ⩾ 25%, and PD-L1 copy number alterations, assessed by fluorescence in situ hybridization.Results:Regardless of HIV status, 84.9% and 44.8% of cases were PD-L1-positive according to CPS ⩾ 1 and TPS ⩾ 25%. Per CPS ⩾ 1, PD-L1 positive rate was similar between HIV-positive and HIV-negative women, whereas a significant difference was seen per TPS ⩾ 25%. Tumor size and parametrial invasion were correlated with PD-L1 positivity in HIV-negative women, whereas anti-retroviral therapy (ART) was correlated with TPS < 25%. Low CD4-positive cell counts were associated with CPS < 1 in HIV-positive women. No significant difference was observed in PD-L1 copy number status between HIV-positive and HIV-negative women. PD-L1 amplification and polysomy were independently associated with TPS ⩾ 25%, whereas the presence of parametrial invasion was independently associated with CPS ⩾ 1. Cancer stage and PD-L1 amplification were identified as independent predictors of recurrence-free survival [hazard ratio (HR) = 2.40 (1.32–4.36) and HR = 5.33 (1.94–14.61)] and cancer-specific survival [HR = 13.62 (5.1–36.38) and HR = 3.53 (1.43–8.69)]. PD-L1 polysomy was an independent predictor of locoregional recurrence-free survival [HR = 3.27 (1.27–8.41)]. HIV status and PD-L1 expression (CPS ⩾ 1 or TPS ⩾ 25%) were not associated with poor patient outcomes.Conclusion:PD-L1 amplification and polysomy are the strongest drivers of PD-L1 expression in cervical cancer, and could represent prognostic biomarkers for anti-PD-1/PD-L1 therapy. Cervical cancer biology may be modulated by HIV infection, CD4-positive cells, and HIV treatments.

Antibody-Associated Autoimmune Disease (AAD) in Children with Cancer in Immune Recovery Phase Following Cessation of Chemotherapy

(Introduction) Outcome of pediatric cancer has improved dramatically by intensive chemotherapy, surgical treatment, radiotherapy, and stem cell transplantation (SCT). In children with cancer, immune system is damaged during treatment. Autoimmune diseases such as autoimmune hemolytic anemia (AIHA) during immune reconstitution were observed as complication after intensive chemotherapy in our institute. Although most of them recovered with supportive treatment, immunosuppressive agents including steroid were needed in some patients. Thus, we defined this complication as antibody-associated autoimmune disease (AAD), and we examined AAD in pediatric patients following cessation of treatment. (Patients and Methods) We reviewed medical records of patients who received intensive chemotherapy in Department of Pediatrics, Hokkaido University Hospital. Between January 2010 and December 2018, 130 patients with hematological malignancies, 127 patients with malignant solid tumor, 120 patients with brain tumor were treated in our institute. All of these patients were included in this study. The diagnoses of AAD were made by clinical criteria, combined with laboratory evidence of a definitive antibody in the serum of patients such as positive Coombs test, antineutrophil antibodies, antiplatelet antibodies, and thyroid-stimulating antibodies. In this study, apparent clinical GVHD and AAD associated with SCT were excluded. (Results) Among the 377 patients, 10 patients (2.7%) developed AAD. AAD was observed as ITP in 5 patients, AIHA in 2 patients, Hashimoto's diseases, multiple sclerosis, and neutropenia with antineutrophil antibody in one patient each. AAD developed a median of 4.0 months (range 2-27months) after cessation of treatment. At onset of AAD, the mean CD4 positive cell counts was 397 /ul and the mean serum IgG was 789 mg/dl. Prednisolone was administered in 3 patients (AIHA and ITP) to treat AAD. The clinical symptoms of AAD regressed with supportive therapy only in the other patients. In 3 out of 5 patients who developed AAD with CD4 positive cell count over 500 /ul, autoimmune antibody in serum of the patients was detected continuously. On the other hand, in 5 patients who developed AAD with CD4 positive cell count below 500 /ul, both clinical symptoms and laboratory examinations regressed, synchronizing with the immune recovery. The mean duration of CD4 positive cell count &lt;500/ul was significantly longer in patients with AAD (n=10) compared with that in patients without AAD (n=45) (p=0.013, 11.0±7.2 months vs 5.6±5.3 months), and the mean duration of serum IgG &lt;500mg/dl was significantly longer in patients with AAD, compared to patients without AAD (p=0.002, 11.0±6.3 months vs 4.3±4.4 months). (Discussion and Conclusion) AAD was observed in immune recovery phase following cessation of chemotherapy, which mimicked immune reconstitution syndrome (IRS) after antiretroviral therapy in patients with HIV infection. It was reported that IRS was also observed in HIV-negative patients who were administered alemtuzumab, and in those who received stem cell transplantation. It was interesting that patients who had smaller number of CD4-positive cells at diagnosis of AAD had a faster recovery of the diseases in our study. We also speculate that AAD mimic autoimmune cytopenia such as ITP, AIHA, and autoimmune neutropenia in infancy because the diseases are caused by autoantibodies, occurring in immune-naive age, and they naturally recover in most cases although some patients need immunosuppressive treatment. Further studies are needed to clarify the pathogenesis of AAD in children. Disclosures No relevant conflicts of interest to declare.

Impact of Combining Frailty Assessment Systems to Select Elderly Myeloma Patients Eligible for Autologous Peripheral Blood Stem Cell Transplantation

Introduction The International Myeloma Working Group (IMWG)-frailty index (IMWG-FI) and revised myeloma comorbidity index (R-MCI) are multiple myeloma (MM)-specific frailty scoring systems, which the European Myeloma Network advocates to assess patient frailty, and to select goals and treatments including autologous stem cell therapy (ASCT). In Japan, patients &lt;65 years old are recommended to receive ASCT under the guidelines of the Japanese Society of Hematology, The Japan Society for Hematopoietic Cell Transplantation, and Japanese Society of Myeloma. However, some Japanese institutes perform ASCTs on MM patients ≥65 years old, although the eligibility criteria and the ASCT methods are variable, or depending on the attending physicians' experience and preference, and doctor-patient relationships. In our institute, 17 MM patients ≥65 years old received ASCT and their frailty at the time of ASCT was retrospectively analyzed. Methods We performed 51 ASCTs on 43 MM patients between April 2017 and June 2019. Seventeen patients were elderly, and 34 were younger. The conditioning regimens were melphalan 200 mg/m2 for younger and 140 mg/m2 for the elderly patients. Both groups received the same supportive therapy. Frailty was compared using the IMWG-FI, R-MCI, and Hematopoietic Cell Transplantation-Comorbidity Index (HCT-CI), and the toxicity and efficacy of ASCT were retrospectively analyzed. Results The patients' ages at the time of ASCT were statistically different between groups (67.8 vs 57.8 years old, p&lt;0.001). The number of fit and intermediate-fit patients by IMWG-FI were 14 and 3 in the elderly group, and 28 and 6 in the younger group, respectively (p=1.0). Fit and intermediate-fit patients by R-MCI were 13 and 4 in the elderly group, and 27 and 7 in the younger group, respectively (p=1.0) (Table 1). No patients were classified as frail in either group by the IMWG-FI and R-MCI. The median scores (ranges) of HCT-CI were 1 (0-5) in the elderly group, and 1.5 (0-5) in the younger group (p=0.93). The number of CD34 positive cells harvested from peripheral blood was lower in the elderly than in the younger group (3.8 vs 5.4×106/kg, p=0.0027). For the elderly and younger groups, the infused CD34 positive cell count (2.4×106 vs 2.4×106/kg, p=0.87), days for neutrophil (11 vs 11 days, p=0.87) and platelet engraftments (15 vs 15 days, p=0.7), and the hospitalization periods (26 vs 27 days, p=0.27) were not statistically different. Both groups had no transplantation-related deaths, similar rates of the non-hematologic adverse events (grade ≥3), an improvement in response (≥very good partial response (VGPR); 49% vs 74%, p=0.003) before vs after ASCT. Post-ASCT ≥VGPR rates were similar between the elderly and younger groups (73% vs 75%, p=1.0). Discussion We safely performed ASCT for elderly MM patients. The frailty status of the younger and elderly groups were the same, as evaluated by the IMWG-FI, R-MCI, and HCT-CI. Our eligibility criteria for MM patients for ASCT was by biological and not chronological age. Thus, the prospective assessment of elderly MM patients by these tools may enable us to evaluate the risks of elderly ASCT, through which elderly ASCT-eligible cases can be selected. Eleven of the 17 elderly patients were termed fit by the IMWG-FI and R-MCI, and scored 0-3 by HCT-CI, 6 of whom were 0-1. Five patients were intermediate-fit by either IMWG-FI or R-MCI, with 0-3 by HCT-CI. Only 1 patient was intermediate-Fit by both IMWG-FI and R-CMI, and was 5 by HCT-CI (Table 1). Based on our data, we propose a stratification system that can determine ASCT eligibility in elderly MM patients. The elderly patients regarded fit by IMWG-FI and R-MCI with 0-1 by HCT-CI could be stratified as Low Risk for elderly ASCT. In addition, the frail patients by IMWG-FI or R-MCI or &gt;3 by HCT-CI could be stratified as High Risk ineligible for elderly ASCT. Other patients could be Intermediate-Risk for elderly ASCT. Further studies are needed to determine the adequate melphalan dosage such as 200 mg/m2 for 65-69 year-old patients, and 140 mg/m2 for 70- Conclusion We retrospectively analyzed the elderly patients receiving ASCT using two MM-specific frailty indices and HCT-CI. We propose a new stratification system to select elderly patients eligible for ASCT. ASCT issues for the elderly MM patients should be examined prospectively. Disclosures No relevant conflicts of interest to declare.

A Comparison of the Effect of Granulocyte-Colony Stimulating Factor (G-CSF) in Chemotherapy Versus Plerixafor for Hematopoietic Stem Sell Transplantation (HSCT) and Mobilization

INTRODUCTION: Peripheral Hematopoietic Stem Cells have been widely used as a source for Autologous Hematopoietic Stem Cell Transplantation (HSCT). The use of G-CSF associated with chemotherapy increases the number of circulating Hematopoietic Progenitor Cells; however, there is no harmony about the best medical strategy for the use of G-CSF associated with the mobilization conduct. The use of plerixafor for poorly mobilized patients has provided a safe and effective option for optimizing apheresis procedures despite the high cost. Thus, the evaluation of efficacy becomes essential for the incorporation of plerixafor into the HSCT routine. OBJECTIVE: To evaluate the mobilization of autologous HSCT in chemotherapy associated with G-CSF and cyclophosphamide (QT) versus plerixafor. METHODS: Retrospective evaluation of 66 patients mobilized for autologous HSCT with G-CSF associated with chemotherapy versus plerixafor (plerixafor was used in the first mobilization), also to evaluate the number of apheresis procedures, CD34-positive cells, cell viability and for the bone marrow engraftment date. Statistical analyses were performed with GraphPad Prism 5.0 (GraphPad Software, Inc., San Diego, CA), using the Mann-Whitney test for nonparametric data (p &lt;0.05). RESULTS: A mobilization protocol was performed in 83 patients (60% male), with a median age of 55 years (17 to 68 years). Multiple myeloma (55.7%), Hodgkin's lymphoma (14.7%), Non-Hodgkin's lymphoma (16.39%), primary amyloidosis (4.9%) were the most common hematologic malignancies. There was no statistical difference between G-CSF and QT group versus plerixafor groups regarding age, sex, type of disease and previous QT. The plerixafor group had better CD34 positive cell counts compared to G-CSF and QT (median 6.2 versus 3.8 x106, p&lt;0.004), shorter sessions of apheresis procedure (median 2 versus 4, p &lt; 0.0002) and a better cell viability rate (98 versus 95%, p &lt;0.0034). The bone marrow engraftment rate was 10 days for plerixafor group versus 15.05 days for G-CSF and QT group, p &lt;0.0055 (Figure 1).CONCLUSION: Our results showed the mobilization group using plerixafor had superior data and statistical significance regarding the number of apheresis procedures, cell viability, number of CD34 positive stem cells and earlier bone marrow engraftment rates when compared to the G-CSF + QT. These findings suggest and encourage the use of plerixafor in the first mobilization, as well as it should be incorporated into the standard routine. In addition, the number of apheresis procedures will be reduced, cell viability and CD34-positive cells rate will be increased. As a result, there will be a better medullary recovery and the reducing of both hospital costs and infectious complication rates. Disclosures No relevant conflicts of interest to declare.

Probable progressive multifocal leukoencephalopathy-immune reconstitution inflammatory syndrome with immunosuppressant dose reduction following lung transplantation: a case report and literature review

BackgroundProgressive multifocal leukoencephalopathy (PML) is a rapidly developing demyelinating disease in the cerebral white matter and is often caused by JC polyomavirus (JCV). PML after lung transplantation is rare and has a poor prognosis, with no established therapies. Reducing the patient’s immunosuppressant doses, thereby restoring immunity, could be used to treat PML. However, some patients develop immune reconstitution inflammatory syndrome (IRIS) with this treatment, an immune-induced inflammatory response to JCV that results in serious neuronal damage. We herein report a case of a 60-year-old female who suffered from PML 5 years after lung transplantation, had worsened brain lesions thought to be related to PML-IRIS at the time of immunosuppressant reduction, and missed treatment opportunities.Case presentationA 60-year-old female developed PML 5 years after lung transplantation. Fluid-attenuated inversion recovery and diffusion-weighted brain magnetic resonance imaging (MRI) revealed multiple high-signal lesions, mainly in the cerebral white matter. Polymerase chain reaction found 0.32 million copies/mL of JCV in the cerebrospinal fluid. Thus, she was given a diagnosis of PML. Mycophenolate mofetil and tacrolimus dosages were reduced, and CD4-positive cell counts and the blood concentration of each immunosuppressant were monitored. Mefloquine was also orally administered at a daily dose of 275 mg for 3 days and was then administered at a dose of 275 mg per week. Although the patient’s CD4-positive cell counts increased and her immune system recovered, her symptoms and brain MRI findings worsened. We suspected PML progression or a transition to PML-IRIS. Steroid pulse therapy to suppress the inflammatory lesions was not possible but was retrospectively indicated. The patient rapidly began to exhibit akinetic mutism and died 4 months after the onset of neurologic symptoms.ConclusionsWhen neurologic symptoms and abnormal brain MRI findings are noted during immune recovery, it is often difficult to distinguish between progressed PML and PML-IRIS. However, the pathogenesis of brain lesions usually involves inflammation and immune-reactive mechanisms for JCV. Steroid pulse therapy, which can reduce inflammation, should thus be administered in organ transplantation cases with differential diagnoses including PML-IRIS.

Assessment of haematopoietic progenitor cell counting with the Sysmex® XN-1000 to guide timing of apheresis of peripheral blood stem cells.

Successful peripheral blood stem cell (PBSC) collection depends on optimal timing of apheresis, as usually determined by flow cytometry CD34-positive (+) cell count in peripheral blood (PB). Since this method is costly and labour-intensive, we evaluated the use of the Hematopoietic Progenitor Cell count programme on a Sysmex® XN haematologic analyser (XN-HPC) as a rapid and inexpensive alternative for predicting CD34+ cell count in PB samples. Haematopoietic progenitor cell and CD34+ cell counts were compared using 273 PB samples collected from 78 healthy donors and 72 patients who underwent PBSC transplantation. We assessed the effectiveness of the XN-HPC in safely predicting pre-harvest CD34+ counts. The most efficient cut-off values of XNHPC were identified. We also evaluated the imprecision (coefficient of variation, CV) and functional sensitivity. Imprecision of the XN-HPC count was <6.3% on daily measurement of three levels of quality control material. Functional sensitivity was 8.9×106/L. A cut-off value of ≥62×106/L XN-HPC for multiple myeloma (MM) patients and ≥30×106/L for all other subjects had both 100% specificity and 100% positive predictive value for identifying samples with CD34+ cells ≥20×106/L. An XN-HPC threshold of <13×106/L identified preharvest CD34+ cell count <10×106/L with 100% sensitivity and 100% negative predictive value. The XN-HPC is a fast, easy and inexpensive test that can safely improve apheresis workflow thus possibly replacing other more expensive CD34 counts currently performed and promoting optimal timing of PBSC collection.

Cardio-ankle vascular index and circulating CD34-positive cell levels as indicators of endothelial repair activity in older Japanese men.

The cardio-ankle vascular index (CAVI) reflects functional arterial stiffness, which is related to endothelial dysfunction. CD34-positive cells carry out an important function in endothelial repair. However, there have been no reports assessing the association between CAVI and the number of circulating CD34-positive cells. We carried out a cross-sectional study of 249 Japanese men, aged 60-69 years, who underwent annual health checkups between 2013 and 2015. As individuals with high levels of circulating CD34-positive cells might indicate the influence of consumptive reduction of circulating CD34-positive cells as a result of aggressive endothelial repair, participants were stratified by circulating CD34-positive cell levels, using the median value in this population (0.95 cells/μL) as the cut-off. For participants with low circulating CD34-positive cell levels, logarithmic values of circulating CD34-positive cells were inversely associated with CAVI (multivariable standardized parameter estimate [β] = -0.22, P = 0.014), but not for participants with high levels (β = -0.04, P = 0.638). In addition, even when no significant associations between CAVI and carotid intima-media thickness were detected for participants with low circulating CD34-positive cell levels (β = -0.02, P = 0.865), significant positive associations were identified for participants with high levels (β = 0.22, P = 0.028). As circulating CD34-positive cell count might indicate endothelial repair activity, the present results show that CAVI is affected by insufficient endothelial repair in individuals with low circulating CD34-positive cell counts. Our results also show that a positive association between CAVI and carotid intima-media thickness exists only in individuals with aggressive endothelial repair, which indicates the presence of organic arterial disease, such as atherosclerosis. Geriatr Gerontol Int 2019; 19: 557-562.

Intestinal helminth infections in HIV-infected patients in Savannakhet after establishment of an HIV registration network in Lao People\u2019s Democratic Republic

BackgroundIn Lao People’s Democratic Republic (PDR), which borders China, Vietnam, Cambodia, Thailand, and Myanmar, the number of HIV-infected patients has increased in recent years. HIV-infected patients diagnosed in Lao PDR are enrolled in a registration network and receive antiretroviral therapy (ART) covered by governmental financial support. Based on the registration network, we investigated intestinal helminth infections and coinfection with HTLV-1 in HIV-infected patients treated with an early intervention using ART in Lao PDR.MethodsThis cross-sectional study of all 252 HIV-infected patients at Savannakhet Provincial Hospital, located in the southern part of Lao PDR, was conducted between February and March 2018. Socioepidemiological information and clinical information were collected from a registration network database and by questionnaire administered to participants. Microscopic examination of intestinal helminth infections in stool samples and particle agglutination for anti-HTLV-1 antibody in plasma were performed.ResultsThe median age of all 252 participants was 39 years old (range, 18–59). Based on the registration network database, there were 156 (61.9%) HIV-infected patients with a CD4-positive cell count ≥ 200 cells/μL and 146 (57.9%) with an HIV viral load < 250 copies/mL. Among 212 stool samples, 75 (35.4%) were found to contain one or more intestinal helminth species, including Opisthorchis viverrini (16.5%), Strongyloides stercoralis (10.8%), hookworm (10.4%), and Taenia saginata (3.3%). This rate of intestinal helminth infections was lower than that of a previous report conducted before the establishment of the registration network for HIV-infected patients in Lao PDR. There was no significant association between intestinal helminth infections and a lower CD4-positive T cell count or higher HIV viral load. HIV-infected patients with anti-HTLV-1 antibody positivity were not found in this cohort.ConclusionThe registration network and an early intervention using ART may provide good medical care and improve the clinical course of HIV-infected patients in Lao PDR. However, the incidence of intestinal helminth infections remains high at 35.4%. The development of a specific medical care system for helminth infection for HIV-infected patients is necessary.

Association between chronic kidney disease and carotid intima-media thickness in relation to circulating CD34-positive cell count among community-dwelling elderly Japanese men

Background and aimsEndothelial injury is well-known as a process that can lead to chronic kidney disease (CKD) and atherosclerosis. Hematopoietic activity is known to be associated inversely with CKD and positively with atherosclerosis. Since bone-derived progenitor cells (CD34-positive cells) contribute to endothelial repair (including the progression of atherosclerosis), understanding the association between CKD and carotid intima-media thickness (CIMT), in relation to circulating CD34-positive cell count, may be an efficient means of clarifying the mechanisms underlying endothelial activity. MethodsWe conducted a cross-sectional study of 570 elderly Japanese men aged 60–69 years, who underwent a general health check-up. Participants were stratified as per a median circulating CD34-positive cell count (1.01 cells/μL). ResultsIndependent of the known cardiovascular risk factors, CIMT was found to be positively associated with CKD in the participants with high circulating CD34-positive cell counts but not in participants with low counts. Odds ratios were 1.40 (1.04, 1.89) for participants with high and 1.01 (0.72, 1.43) for participants with low circulating CD34-positive cell counts after adjustment for known cardiovascular risk factors at 95% confidence intervals for CKD with one standard deviation increment of CIMT. ConclusionsA positive association between CIMT and CKD was observed among participants with high circulating CD34-positive cell counts but not among participants with low counts. Endothelial repair activity might determine the association between CKD and CIMT.

Radiographic Manifestations of Tuberculosis in HIV Positive Patients: Correlation with CD4 Positive Tcell Count

Background: Observations on Tuberculosis HIV co-infection in addition to epidemiologic molecular studies have recently provided strong evidence for the state of immune system as the major determinant of the TB imaging spectrum. However, the presence of any correlation between radiographic findings and the degree of suppression in HIV positive patients still remains controversial. The present study aimed to investigate the TB radiographic manifestation in HIV positive patients and its relationship to the CD4 positive cell count. Methods: Chest radiography of 15 HIV positive patients with a definite diagnosis of pulmonary Tuberculosis in Masih Daneshvari hospital, between 2013 - 2014, were retrospectively reviewed. Radiographic findings and severity were categorized as typical (upper lobe infiltration/cavity) and atypical (middle/lower lobe opacity, adenopathy, pleural effusion and normal X-ray). Demographics and CD4 positive cell count were also recorded. Data analysis was performed using SPSS version 23 (frequency and mean for descriptive quantitative variables and Logistic regression analysis for correlation, P < 0.05). Results: Of a total 15 patients (86.7% men and 13.3% women), 78.6% had CD4 positive counts below 350 (mean SD; 229.15 199.45). The most common radiographic findings in descending order of frequency were adenopathy (53.3%), pleural effusion (26.7%) and cavitation (6.7%) with an overall atypical presentation of 93.3%. This study failed to reveal any statistically significant correlation between CD4 positive cell count and radiographic manifestation as well as severity. Conclusions: In CD4 positive cell count below 500, the dominant radiographic pattern of Tuberculosis is atypical presentation. At this level of immunity, CD 4 positive T cell dysfunction may play a deterministic role in TB radiographic manifestation.

Platelets and circulating CD34-positive cells as an indicator of the activity of the vicious cycle between hypertension and endothelial dysfunction in elderly Japanese men

Background and aimsThe bi-directional association between hypertension and endothelial dysfunction (vicious cycle) is crucial for the development of cardiovascular disease. Since platelets and circulating CD34-positive cells have been reported to contribute to vascular repair (endothelial repair and the development of atherosclerosis), these two factors could act as an indicator for the activity of this vicious cycle. MethodsWe conducted a cross-sectional study of 580 Japanese men aged 60–69 years who underwent an annual health check-up. Since aggressive endothelial repair might cause consumptive reduction of circulating CD34-positive cells, subjects were stratified by circulating CD34-positive count (<1.01 cells/μL or ≥ 1.01 cells/μL). ResultsPlatelet count is positively associated with hypertension among participants with a low CD34-positive cell count (multivariable-OR (odds ratio) for hypertension of 1 standard deviation (SD) increments in platelets (5.24 × 104/μL) = 1.47 (1.12, 1.91)), but not among participants with a high CD34-positive cell count (multivariable-OR = 0.91 (0.71, 1.18)). We also found that platelet count is positively associated with carotid intima-media thickness related to endothelial dysfunction among hypertensive participants with a low CD34-positive cell count (standardized parameter estimate (β) = 0.26, p = 0.008), but not with a high CD34-positive cell count (β = 0.11, p = 0.234). ConclusionsOur results indicate that the number of platelets and circulating CD34-positive cells acts as an indicator of the activity of the vicious cycle that exists between hypertension and endothelial dysfunction in elderly Japanese men.

Single nucleotide polymorphisms of the DGKB and VCAM1 genes are associated with granulocyte colony stimulating factor-mediated peripheral blood stem cell mobilization

We previously reported the association between LDL cholesterol level (LDL-C) and granulocyte colony stimulating factor (G-CSF)-mobilized peripheral blood (PB) hematopoietic stem cells (HSC). In this study, we investigated the association between gene single nucleotide polymorphisms (SNPs) involved in hematopoiesis and lipid level and PBHSC mobilization. In 46 patients who underwent peripheral blood stem cell harvest (PBSCH), we measured CD34-positive cells in PB and PBSCH, and the patients were classified into good, intermediate, or poor mobilizer groups based on the CD34-positive cell counts. And SNPs of the OR4C12, ENO1, RERE, DGKB, DSC3, VCAM1, CD44, and FADS1 genes were investigated. The frequency of the TT type of the DGKB gene was higher in the poor mobilizer group compared to other groups (p<0.05), whereas that of the CC type of the VCAM1 gene was high in the good mobilizer group (p<0.05). Association with the efficiency of HSC mobilization to PB were found in the SNPs of the DGKB gene involved in cell transport and SDF-1-induced migration ability and of the VCAM1 gene which is essential for HSC homing, suggesting that SNPs involved in cell migration ability might be partly involved in HSC mobilization to PB.