Linear Relationship Between Lymphocyte Subpopulations Counts in Peripheral Blood and Buffy-Coat during Extracorporeal Photopheresis in Patients with Graft-Versus-Hot-Disease Using an Off-Line System

Abstract Introduction: Extracorporeal photopheresis (ECP) is a treatment modality that entails leukapheresis followed by mixing the buffy coat with 8-methoxypsoralen (8-MOP) and exposing it to UVA light. The buffy coat is then returned to the patient. ECP may be performed employing two different techniques: an on-line and off-line procedure. The off-line system includes two steps: a processing of buffy coat before reinfused to the patient. The treatment is thought to have an immunomodulatory effect and is most commonly used to treat cutaneous T cell lymphoma, acute and chronic graft-versus-host disease (GVHD), and heart and lung allograft rejection. The exact mechanism and optimal cells dose to be treated is unknown. At the present time, a standard protocol is used generally without considering peripheral blood (PB) leukocytes counts or lymphocyte subpopulations (LP) of the patient. With the off-line system, PB leukocytes counts and LP analysis may be useful to choose the amount and distribution of cells to be infused. The first objective of this study is to examine the quantitative correlation between PB LP and the buffy-coat in order to set individualized guidelines of treatment. Once this relationship is understood, the PB LP may serve as a surrogate marker for cell dose treated and help predicting the efficiency of ECP. The second objetive of this study is to examine the mean performance of the buffy coat LP categorized according to PB leukocytes counts (<1.5 x 109/L and >1.5x 109/L). Patients and methods: Twenty two consecutive patients with refractory GVHD were prospectively studied, from november 2009 to may 2014. Apheresis procedures were perfomed with COBE Spectra system (Terumo BCT®, Lakewood, CO, USA; version 7.0) by processing 1.5-2 times the patient blood volume. The product was transferred to a UVA-permeable bag (UVA, Macopharma, France), added 5 mL (0.1 mg) of 8-methoxypsoralen (8-MOP) aqueous solution (S.A.L.F.®, Cenate Sotto, Italy), exposed to UVA irradiation (Macogenic G2, Macopharma®), and then reinfused. Peripheral blood sample was drawn before ECP. Just before reinfused, buffy coat sample was drawn. Spearman correlation analysis was performed between the LP CD3+CD4+, CD3+CD8+, CD19+, NK of the preapheresis peripheral blood patient and the buffy coat infused analyzed by multiparameter flow cytometry 5 colors (FC500-Beckman Coulter®) in the total sample and in three groups according to preapheresis leukocytes counts (<2.5, 2.5-7.5, >7.5x 109/L). The mean performance was calculated: CD3+CD4+, CD3+CD8+, CD19+, NK+ LP count in buffy coat/ CD3+CD4+, CD3+CD8+, CD19+, NK+ PB /ml x treatment volume (ml) x 100. The mean performance of the buffy coat LP categorized according to preapheresis leukocytes counts (<1.5x 109/L and >1.5 109/L) were compared by Mann-Whitney test. Results: A total of 22 patients and 136 procedures were included in the final analysis. CD3+CD4+, CD3+CD8+, CD19+, NK LP in peripheral blood significantly correlated with those in buffy coat collected by COBE Spectra system, r= 0,74, 0.78, 0,92, 0,39 respectively (table 1). The LP mean doses and mean performance in buffy coat infused are specified in Table 1. The correlations was stronger in all LP with PB leukocytes counts <2.5 x 109/L (table2). We have not found any statistical correlation between the performance of the LP CD3+CD4+, CD3+CD8+, CD19+, NK according to PB leukocytes counts (<1.5x109/L and >1.5x109/L). Conclusions: The buffy coat contains great variability in lymphocyte subpopulations with predominant levels of CD3+CD8+. There is a robust linear relationship between all PB and buffy coat LP. The mean performance LP was around 40% and it was not related to very low PB leukocyte count (<1.5x109/L). The correlation was stronger with lower leukocytes counts in PB. If we could demonstrate a relationship between cell doses infused and clinical response, we could plan the necessary dose for each patient according to the PB leukocyte count and LP preapheresis. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures No relevant conflicts of interest to declare.

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Linear relationship between lymphocyte counts in peripheral blood and buffy coat collected during extracorporeal photopheresis

Transfusion ◽

10.1111/trf.12114 ◽

2013 ◽

Vol 53 (11) ◽

pp. 2635-2643 ◽

Cited By ~ 14

Author(s):

Chang Liu ◽

Kalpna Shah ◽

Marian Dynis ◽

Charles S. Eby ◽

Brenda J. Grossman

Keyword(s):

Linear Relationship ◽

Peripheral Blood ◽

Buffy Coat ◽

Extracorporeal Photopheresis

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Carfilzomib and Bortezomib Containing Regimens Yield High Rates Of MRD Negative Autologous Hematopoietic Progenitor Cell (HPC) Grafts In Multiple Myeloma (MM) and High Risk Smoldering Myeloma (SMM)

Blood ◽

10.1182/blood.v122.21.2030.2030 ◽

2013 ◽

Vol 122 (21) ◽

pp. 2030-2030

Author(s):

Nishant Tageja ◽

Neha Korde ◽

Constance Yuan ◽

Kristen Cole ◽

Jennifer Hsu ◽

...

Keyword(s):

Flow Cytometry ◽

Stem Cell ◽

High Risk ◽

Tumor Cell ◽

Peripheral Blood ◽

Conflicts Of Interest ◽

Hematopoietic Progenitor Cell ◽

Myeloma Cells ◽

Cd34 Cells ◽

The Mean

Abstract Background Regimens incorporating modern anti-myeloma drugs, such as carfilzomib (CFZ) and bortezomib (BOR), produce rapid, deep and durable responses in newly diagnosed myeloma patients but their effect on collection of autologous HPC is not well known, including minimal residual disease (MRD) testing of stem cell grafts. Employing older induction regimens (such as VAD), less sensitive flow cytometry techniques detected circulating myeloma cells in 38-46% of autologous HPC grafts (Stewart, et al. JCO. 2001 and Bourhis, et al. Haematologica. 2007). We hypothesized that the use of modern CRd combination therapy including Carfilzomib (CFZ)-Lenalidomide (LEN)-Dexamethasone (DEX) would significantly lower the rates of HPC product contamination. Methods Thirty-six patients, including 29 with MM and 7 with high-risk SMM, underwent HPC mobilization and collection following induction with CRd (n=30), LEN-BOR-DEX (RVd, n=4), Cyclophosphamide-BOR-DEX (CyBorD, n=1) and Cyclophosphamide-BOR-Prednisone (CyBorP, n=1). For HPC mobilization, all patients received 5 days of filgrastim at 10-16 mcg/kg/dose. A combination of the patient’s weight and a peripheral blood CD34 count after 4 doses was used to determine the likelihood of collecting > 4 x106 CD34+ cells/ kg in a single apheresis procedure after a fifth filgrastim dose, according to a previously published algorithm from our institution. Only subjects predicted to require > 1 apheresis by the algorithm received Plerixafor (PLX) at 240 mcg/kg/dose on the fifth day along with the fifth filgrastim dose. HPC collection occurred on day 6, 8 hours after the last mobilizing agent(s) administration. Product contamination with myeloma cells (i.e. MRD status) was evaluated using multi-parameter flow cytometry with a minimum of 3 x 106 events obtained (sensitivity detection rate 1 x 10-5) to examine expression of 9 antigens by the plasma cells. Results The median age at mobilization was 56.2 years (range 40-73) and 19 (53%) were male. At the time of HPC collection, 20 (55%) patients were in sCR/CR/nCR, 11 (30%) had VGPR with 4 PR (11%) and 1 SD (3%). The mean CD34+ cells in the peripheral blood were 33/uL on day 5 and 55/uL on day 6 for the whole cohort. Thirteen (36%) patients did not need PLX. Interestingly, the mean CD34+ count dropped by a mean of 2% from D5 to D6 in patients not receiving PLX while, as expected, it increased by 304% in those who did. The median number of CD34+ cells collected was 6.86 million/kg (range 2.6-12.5) for the whole cohort, (6.6 million/kg without PLX and 7.52 million with PLX p=0.46). Thirty-three of 36 patients (92%) achieved a collection of > 4 million cells /kg in a single apheresis procedure. The 30 patients treated with CRd had a median of 5 (range = 3-7) prior cycles containing LEN with a median of 12 days (range 1-34) between mobilization and last LEN dose. Only 2 of 36 (5%) products were found to have evidence of tumor cell contamination (i.e. MRD positive) using sensitive multiparameter flow cytometry, one patient in PR after 6 cycles of CRd and a second patient in CR after 5 cycles of RVd. Conclusions Modern anti-myeloma therapies, such as CRd and RVd, allow adequate HPC collection in a single apheresis procedure in most cases and improve the quality of the HPC product with greatly reduced tumor cell contamination compared to historical controls. Indeed, 34/36 (94%) patients treated with modern anti-myeloma therapy collected an MRD negative HPC product. Future prospective studies are needed to assess whether autologous stem cell transplants (ASCT) using tumor-free HPC products collected in the era of modern induction therapies have better outcomes. Disclosures: No relevant conflicts of interest to declare.

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HASIL HITUNG NORMOBLAS ANTARA SEDIAAN HAPUSAN DARAH TEPI PENDERITA AML DENGAN ALL

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v16i1.997 ◽

2018 ◽

Vol 16 (1) ◽

pp. 39

Author(s):

Hidayat . ◽

Nina Susana Dewi ◽

Nadjwa Zamalek Dalimoenthe

Keyword(s):

Peripheral Blood ◽

Leukocyte Count ◽

Lymphoblastic Leukemia ◽

Clinical Pathology ◽

Mean Value ◽

Blood Smears ◽

Immature Form ◽

The Subject ◽

The Mean ◽

Peripheral Blood Smears

Normoblast is an immature form of erythrocyte in erythropoietin system. Normally, normoblast can be found in peripheral blood healthy neonates. The existence of normoblast in peripheral blood might be the sign of pathologic conditions such as hemolytic anemia,acute blood loss, and ischemia and bone marrows abnormalities like malignancy or leukemia. In acute leukemia (Acute MyeloblasticLeukemia and Acute Lymphoblastic Leukemia), normoblast existence in peripheral blood may due to erythropoietin system suppression.The aim of this study is to compare normoblast count between AML and ALL, and also to find out the correlation between leukocyte andnormoblast count in AML and ALL. The subject of this study were patient diagnosed as AML (30) and ALL (30) in Hematology Divisionof Clinical Pathology Department at Dr.Hasan Sadikin Hospital Bandung in July 2006–August 2008. In this study we examined 30peripheral blood smears from AML and 30 peripheral blood smears from ALL. Leukocyte count result was derived from CBC performedwith Sysmex KX-21. The mean value of normoblast count from AML blood smear patients is 1930.60 (3.60/100 WBC) while ALL bloodsmear patients is 309.60 (0.43/100 WBC). Statistically this difference is significant (p < 0.001). There are strong correlation betweenleukocyte count and normoblast count within both group (r = 0.851, r = 0.948; p < 0.001).

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FA02.07: BARRIER STRUCTURE AND FUNCTION OF THE NEO-SQUAMOUS EPITHELIUM FOLLOWING SUCCESSFUL BARRETT’S ENDOTHERAPY IS IMPAIRED

Diseases of the Esophagus ◽

10.1093/dote/doy089.fa02.07 ◽

2018 ◽

Vol 31 (Supplement_1) ◽

pp. 5-5

Author(s):

Adharsh Ravindran ◽

Christopher Blevins ◽

Michele Johnson ◽

Ramona Lansing ◽

Kenneth Wang ◽

...

Keyword(s):

Intestinal Metaplasia ◽

Intercellular Space ◽

Structural Integrity ◽

Squamous Epithelium ◽

Conflicts Of Interest ◽

Surrogate Marker ◽

Functional Integrity ◽

Successful Ablation ◽

Endoscopic Ablation ◽

The Mean

Abstract Background The neo-squamous epithelium that replaces columnar epithelium after successful endoscopic ablation of Barrett's esophagus (BE) has not been fully characterized in regard to its structural and functional integrity. Intercellular space is a surrogate marker for epithelial structural integrity and mucosal impedance (MI) is a surrogate for transepithelial resistance and thus epithelial functional integrity. Impairment of the structural and functional integrity may influence recurrence following successful ablation. We aimed to measure the esophageal epithelial intercellular space diameter and MI in the neo-squamous and normal squamous epithelium of patients who underwent successful radiofrequency ablation for dysplastic BE. Methods Forty patients with complete remission of intestinal metaplasia (CRIM) (defined as two negative surveillance endoscopies for intestinal metaplasia) were prospectively recruited. Patients were excluded if they had endoscopic evidence of esophagitis. MI measurements were taken at 1 cm above the GE junction and 2 cm above the prior BE segment (targeting uninvolved squamous epithelium), using a novel through the scope MI probe. Endoscopic biopsies were also taken at these corresponding sites to assess intercellular space diameter using transmission electron microscopy (TEM) using standard techniques. Results 80% of patients were male with a mean age of 69 years. Mean follow since achieving CRIM was 30 months. 39 patients underwent RFA for dysplasia. The mean intercellular space diameter was higher in the neo-squamous epithelium (1.01 μm) when compared to the untreated native squamous epithelium (0.96 μm) [Difference of 0.056 μm, 95% CI of -0.036 to 0.148, P = 0.228]. The mean MI was significantly lower in the neosquamous epithelium (4001.3 Ω) when compared to the untreated native epithelium (5168.1 Ω) [Difference of 1166.7 Ω, 95% CI 418.8 to 1914.6, P = 0.003]. Conclusion Neo-squamous epithelium that replaces BE epithelium following successful endoscopic therapy for BE appears to be functionally and likely structurally deficient with regards to acid reflux barrier function, when compared to native squamous esophageal epithelium. This impairment may be associated with recurrence following successful ablation. Disclosure All authors have declared no conflicts of interest.

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Circulating Mesenchymal Stromal Cells As a New Prospective Cancer Marker,

Blood ◽

10.1182/blood.v118.21.3404.3404 ◽

2011 ◽

Vol 118 (21) ◽

pp. 3404-3404

Author(s):

Mikhail Kolonin ◽

Yan Zhang ◽

Paul J. Simmons ◽

Charles Bellows

Keyword(s):

Endothelial Cells ◽

Progenitor Cells ◽

Cancer Patients ◽

Cancer Progression ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Conflicts Of Interest ◽

Surrogate Marker ◽

Response To Therapy ◽

Peripheral Blood Mononuclear

Abstract Abstract 3404 Mobilization of progenitor cells is implicated in pathology and can be indicative of disease progression. Recently, we reported the influence of body mass index (BMI) on level of circulating progenitor cells (Bellows et al., Obesity 2011). Comparative analysis of peripheral blood mononuclear cells (PBMC) from 12 non-obese (BMI < 30) and 14 obese (BMI > 30) disease-free donors by flow cytometry revealed that obesity is associated with a 10-fold increased frequency of circulating mesenchymal stromal progenitor cells (MSC), which circulate at a very low level in healthy lean individuals. We showed that obesity is also associated with a 5-fold increased frequency of circulating progenitor cells (CPC), a population consisting of hematopoietic and endothelial precursors, while the frequencies of mature endothelial cells (EC) and CD34-bright leukocytes (CD34b LC) are unaffected by BMI. Here, we followed up on the assessment of circulating MSC as a potential surrogate pathology marker by analyzing the frequency of circulating CD34-positive progenitor and endothelial cells in a cohort of colorectal cancer patients. PBMC were collected from 45 obese and lean cancer patients and compared to control cancer-free donors. Flow cytometric enumeration of cells was performed based on established immunophenotypes: CD34brightCD31dimCD45dim (CPC), CD34dimCD31brightCD45- (EC), CD34brightCD31-CD45- (MSC) and CD34brightCD45bright CD34b (LC). Groups were compared using multivariate regression analysis. After adjusting for co-founders such as age and BMI, the mean frequencies of MSC and CD34bLC, but not of CPC and EC, were found to be significantly higher in the circulation of CRC patients compared to cancer-free donors. Interestingly, the frequency of circulating MSC, but not of the other cell populations, was also found to be significantly higher in the circulation of obese CRC patients compared to lean CRC patients and obese cancer-free controls. We conclude that markedly increased frequency of MSC in the peripheral blood may represent a new diagnostic CRC marker. BMI-dependent changes in circulating MSC, potentially mobilized from adipose tissue may reveal their trafficking to tumors, which could be one of the mechanistic links between obesity and cancer progression. Validation of MSC as a new surrogate marker of cancer could provide a tool for determining prognosis, predicting response to therapy, and detecting relapse following treatment. Disclosures: No relevant conflicts of interest to declare.

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Leukoreduction System Chambers as a Source of Viable Human Peripheral Blood Mononuclear Cells.

Blood ◽

10.1182/blood.v110.11.4912.4912 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4912-4912

Author(s):

Sinyoung Kim ◽

Han-Soo Kim ◽

Yangsoon Lee ◽

Jaewoo Song ◽

Hyun Ok Kim ◽

...

Keyword(s):

Cell Count ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Buffy Coat ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

The Mean

Abstract Many blood banks are using apheresis machines to collect blood components such as platelets (PLTs), RBCs, or plasma. Especially, leukoreduced plateletpheresis using apheresis instrument (Trima Accel, Gambro BCT, Lakewood, CO) provided subsidiary cell products retained in leukoreduction system (LRS) chamber that was originally discarded. The LRS chamber is a conical-shaped chamber that uses saturated, fluidized, particle bed filtration technology to remove WBCs from PLTs. In the current study, a total of 24 LRS chambers from different donors were investigated to determine it would be a valuable source of viable human peripheral blood mononuclear cells (MNCs). The proportions of CD3+, CD19+, CD16+/CD56+, CD14+, CD45+ cells, and absolute CD34+ cell count within the LRS chambers were determined by flow cytometry. Dendritic cells (DCs) were generated from the immunomagnetically purified CD14+ cells from LRS chamber and characterized by phenotypic surface marker and stimulatory capacity in an allogeneic mixed lymphocyte reaction. In the LRS chamber, the total number of WBC count was 1.1 × 109 ± 0.3 × 109 and the mean percentage of MNCs was 80.6 ± 13.1%. The mean proportion of T cells, B cells, NK cells, CD14+ monocytes among CD45+ cells was 54.3 ± 11.5%, 6.4 ± 3.1%, 14.6 ± 3.9%, 12.9 ± 7.5%, respectively. Total absolute CD34+ cell count in LRS chamber was 0.95 × 106 ± 0.65 × 106. Also, we could demonstrate CD14+ cells isolated from LRS chamber was capable of differentiating into functionally mature DCs in vitro. LRS chambers are a valuable and convenient source of viable human peripheral blood mononuclear cell population and could replace standard buffy coat preparations for research applications.

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Leukocyte Integrin CD18 Expression Mediates Transient Neutropenia Following G-CSF Administration.

Blood ◽

10.1182/blood.v114.22.3587.3587 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3587-3587

Author(s):

Laura Tuschong ◽

Catherine E. Dejesus ◽

Meredith Adams ◽

Aylin C. Bonifacino ◽

Dennis D. Hickstein ◽

...

Keyword(s):

Peripheral Blood ◽

Conflicts Of Interest ◽

Leukocyte Adhesion ◽

Surface Expression ◽

Peripheral Blood Leukocytes ◽

Blood Leukocytes ◽

Cell Counts ◽

Transient Loss ◽

The Mean ◽

White Blood Cell Counts

Abstract Abstract 3587 Poster Board III-524 The mechanism whereby neutrophils traffic from the circulation in response to G-CSF has remained unclear despite the observation of ourselves and others that there is a dramatic, yet transient, loss of circulating neutrophils shortly following the administration of G-CSF in humans, non-human primates, and mice (Gordon BC, et al. Exp Hematol. 35:872-8, 2007). To determine the role of the CD18 leukocyte integrin on neutrophils in the egress of neutrophils from the circulation, we used dogs with canine leukocyte adhesion deficiency (CLAD), a genetic disease in which a mutation in CD18 prevents CD18 surface expression. We selected CLAD dogs who had 5-10% CD18+ neutrophils following either matched littermate allogeneic transplant or autologous gene therapy for CLAD. Three CLAD dogs meeting these criteria were evaluated. Three carrier dogs served as controls. G-CSF was administered at 10μg/kg SQ to all six animals. Peripheral blood samples (EDTA) were taken immediately prior to G-CSF administration, and at 15, 30, 60, 120, 240 minutes, and 24 hours following G-CSF administration. Total white blood cell counts, neutrophil counts, and the number and percentage of CD18+ peripheral blood leukocytes were assessed. As anticipated, the control dogs had a 60% decrease in circulating neutrophils 30 minutes following G-CSF administration: the mean +/− standard of deviation (SD) absolute neutrophil baseline count decreased from 6806+/−1072/μL to 2727+/−767/μL. In five control animals the neutrophil nadir occurred at 30 minutes post-G-CSF, and in one control dogs it occurred 15 minutes following G-CSF administration. Experimental CLAD dogs had only a 35% decline in neutrophil numbers at 30 minutes, from a mean baseline of 6777+/− 672/μL to 4433+/−265/μL. In these dogs the neutrophil count returned to pre-G-CSF levels by 60 minutes post-G-CSF. By 24 hours after G-CSF, the neutrophil level was increased 3-fold from baseline. Immunophenotyping using an anti-CD18 and a canine specific anti-neutrophil PE conjugated antibody indicated that only the CD18+ neutrophils disappeared from the circulation following G-CSF administration. At baseline the transplanted CLAD dogs had a mean of 15.2+/−3.9% CD18+ peripheral blood leukocytes, of these 50.7+/−7.1% were CD18+ neutrophils. Thirty minutes following G-CSF administration the mean+/−SD percentage of CD18+ leukocytes declined to 13.7+/−3.7% with 33.4+/−5.8% being neutrophils. There was also a slight decline in CD14+CD18+ monocytes from 6.2 +/− 1.5% to 4.0 +/− 1.2%, which was not observed in the controls. There was no change in CD18- leukocyte numbers. The percentage of CD18+ neutrophils returned to baseline by 60 minutes and remained there at subsequent time points. These results demonstrate that the CD18 leukocyte integrin on circulating neutrophils mediates the transient neutropenia associated with G-CSF administration. Disclosures: No relevant conflicts of interest to declare.

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Multimodal Bcl-2 Sub-Populations in CLL.

Blood ◽

10.1182/blood.v114.22.4391.4391 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4391-4391

Author(s):

Heba A. Degheidy ◽

Shahinaz M. Gadalla ◽

Elizabeth Raveche ◽

MA Stetler-Stevenson ◽

Gerald E. Marti

Keyword(s):

B Cells ◽

Conflicts Of Interest ◽

Rank Order ◽

Surrogate Marker ◽

Homozygous Deletion ◽

Direct Result ◽

Fish Analysis ◽

Normal Donor ◽

Significant Difference ◽

The Mean

Abstract Abstract 4391 Introduction The Recently described NZB miRNA15a/16-1 mutation is syntenic to 13q14 deletion in CLL. The mir15a/16-1 complex is thought to regulate bcl-2 expression. Therefore the level of bcl-2 expression was examined more closely as it might be used to differentiate mono-allelic deletion from the diploid CLL clone. Patients and Methods Multicolor flow-cytometry using a panel of anti-bcl-2 FITC (clone 124, DAKO Cytomation, Glostrup, Denmark) and CD19, CD20, CD5, CD23, CD38, kappa, lambda, and CD45 was used. Bcl-2 expression was evaluated using the conventional bcl-2 index (Mean fluorescence intensity (MFI) B cell/ MFI T cells) in 25 untreated CLL patients, and 10 normal donor controls. A modified ratio based on normal remaining (NR) polyclonal B cells was also used (MFI clone/MFI NR) with a new gating strategy for bcl-2 expression analysis. Results The mean of the conventional bcl-2 index is significantly higher in CLL patients [1.52 ± 0.53(SD)] compared to the control [0.5 ± 0.19(SD)], p<0.001 Student‘s t test. When comparing the mean of conventional bcl-2 index (clone/T cell) versus the mean of the modified bcl-2 index (clone/NR) [1.78 ±0.61 (SD)] in only CLL patients there was no significant difference. On the other hand, the rank order analysis of the conventional and modified bcl-2 index showed an r=0.7, p< 0.001 indicating that both methods give similar results. In addition to the high correlation between the conventional and the modified ratio, the modified ratio was multimodal in distribution. Conclusions Bcl-2 expression varies largely among the lymphocyte sub-sets. The choice of the polyclonal B cells allows better segregation of multimodal bcl-2 expression. Since bcl-2 mRNA is a validated mir15a/16 target, increased bcl-2 expression maybe a direct result of and a surrogate marker for mir15a/16 levels and/or the 13q14 status. There are three possibilities: no deletion (diploid), heterozygous deletion, or homozygous deletion. This needs to be verified by fluorescence in situ hybridization (FISH) analysis of sorted CLL B cells based on bcl-2 expression. Disclosures: No relevant conflicts of interest to declare.

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Kajian Tentang Hematologi Penderita Plasmodium vivax di Laboratorium Inti Farma Jayapura-Papua

JURNAL BIOLOGI PAPUA ◽

10.31957/jbp.1363 ◽

2021 ◽

Vol 13 (1) ◽

pp. 36-43

Author(s):

Elieser Elieser ◽

Dais Iswanto

Keyword(s):

Plasmodium Vivax ◽

Peripheral Blood ◽

Vivax Malaria ◽

Blood Smear ◽

Leukocyte Count ◽

Peripheral Blood Smear ◽

Descriptive Study ◽

Hemoglobin Level ◽

Descriptive Statistics ◽

The Mean

Malaria due to Plasmodium vivax infection is a species that attacks humans more than other species. The purpose of this study was to determine the hematological description of Plasmodium vivax sufferers in the Inti Farma Jayapura Laboratory for the period January - June 2020. The descriptive study design used malaria examination using the peripheral blood smear method which was examined using a microscope at the Inti Farma Jayapura laboratory during the period January to June of the year. 2020. The results of the hematological examination were analyzed with descriptive statistics and crosstabulation using the SPSS version 25 program. The results revealed that the mean hemoglobin level of patients with vivax malaria was 12.12, the number of erythrocytes was 5.2750 million / miuL, the leukocyte count was 5,500 miuL, and the trombosiy was 303 thousand. / miuL. These findings provide important information for malaria management in general in Kota Jayapura.Key words: Laboratory, Plasmodium vivax, hematology, peripheral blood smear

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Linear Relationship Between Lymphocyte Counts in Peripheral Blood and Buffy Coat During Extracorporeal Photopheresis in Patients with Graft-Versus-Host Disease

Blood ◽

10.1182/blood.v120.21.4171.4171 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4171-4171

Author(s):

Chang Liu ◽

Kalpna Shah ◽

Marian Dynis ◽

Charles S. Eby ◽

Brenda J. Grossman

Keyword(s):

Graft Versus Host Disease ◽

Peripheral Blood ◽

Lymphocyte Count ◽

Collection Efficiency ◽

Buffy Coat ◽

Total Blood Volume ◽

Extracorporeal Photopheresis ◽

Total Blood ◽

Graft Versus Host ◽

Cell Counts

Abstract Abstract 4171 Background Extracorporeal photopheresis (ECP) is a treatment modality that entails leukapheresis followed by mixing the buffy coat with methoxsalen (8MOP) and exposing it to UVA light. The buffy coat is then returned to the patient. The treatment is thought to have an immunomodulatory effect and is most commonly used to treat cutaneous T cell lymphoma, acute and chronic graft-versus-host disease (GVHD), and heart and lung allograft rejection. The exact mechanism and optimal dose of cells to be treated is unknown. The primary goal of this study is to examine the quantitative relationship between cell counts in peripheral blood (PBL) and buffy coat collected during ECP. Once this relationship is understood, the PBL cell count may serve as a surrogate marker for cell dose treated and help predicting the efficacy of ECP. The secondary goal of the study is to compare the cell collection efficiency of the two instruments currently used for ECP in the United States, UVAR XTS and CELLEX (Therakos, Exton, PA). Methods Twenty six patients with GVHD were prospectively recruited for this study which was approved by the institutional review board. ECP was performed with either UVAR XTS (5 cycles with small bowl) or CELLEX (1500 mL of whole blood processed). PBL was drawn within 24 hours prior to the procedure for complete cell count with automated differentials (CBC). At the end of cell collection but prior to 8MOP treatment, 1 mL of buffy coat was drawn from well-mixed treatment bag for CBC. Spearman correlation analysis was performed between cell counts in PBL and buffy coat, including leukocyte counts (WBC), differential counts (neutrophil, lymphocyte and monocyte), and red cell counts (RBC). Collection efficiency was assessed in two ways: (1) the ratio of lymphocyte count in buffy coat to that in PBL (fold enrichment), and (2) the percentage of total lymphocytes collected [(lymphocyte count in buffy coat × treatment volume × 100)/(lymphocyte count in PBL × total blood volume estimated based on height and weight)]. These two variables were compared between UVAR XTS and CELLEX by Mann Whitney test. A double-sided p value was set at 0.05. Data visualization and statistical analysis were performed using GraphPad Prism 5 and IBM SPSS 19. Results A total of 22 patients were included in the final analysis after excluding 4 patients due to incorrect sampling of buffy coat. Seven patients were treated with UVAR XTS and 15 patients with CELLEX. The demographics (gender, race, age, weight, height, and estimated total blood volume), peripheral WBCs, Hemoglobin and hematocrits were not significantly different between the two groups. Lymphocyte counts in PBL significantly correlated with those in buffy coat collected by UVAR XTS (r=0.79, p<0.05) and CELLEX (r=0.92, p<0.0001). Linear relationships were observed with a slope of 1.7 (95% CI, 1.3–2.2) for UVAR XTS and a slope of 5.1 (95% CI, 4.7–5.4) for CELLEX (Figure 1A). Monocyte counts in PBL and buffy coat collected by UVAR XTS (r=0.82, p<0.05) and CELLEX (r=0.84, p<0.0001) also had robust correlations. There were no significant correlations for RBCs or neutrophil counts between PBL and buffy coat collected by either instrument. For the overall WBCs, there was only a weak but significant correlation for CELLEX (r=0.53, p<0.05). UVAR XTS enriched the lymphocyte counts by 2.2 ± 0.4 folds in buffy coat compared to PBL during ECP while CELLEX enriched the lymphocyte counts by 5.3 ± 0.4 folds (Mean ± SE, p<0.001, Figure 1B). UVAR XTS collected 9.9 ± 5.2% of total lymphocytes during each treatment while CELLEX collected 19.8 ± 8.6% of total lymphocytes (Mean ± SE, p<0.005, Figure 1C). Conclusion There is a robust linear relationship between PBL and buffy coat lymphocyte counts, which may allow PBL lymphocyte count to be used in future studies as a surrogate for number of cells treated. This relationship also ensures a practical way to correlate cell dose with outcome. CELLEX is significantly more efficient in collecting lymphocytes than UVAR XTS. Approximately twice as many lymphocytes are treated with CELLEX than with UVAR XTS during each procedure. Disclosures: No relevant conflicts of interest to declare.

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