Total Mononuclear Cell Count Research Articles

Use of Sysmex Hematology Analyzer based Hematopoietic Progenitor Cell (XN-HPC) Count in Allogeneic Peripheral Blood Stem Cell Transplant Setting; Is It Substantially Reliable?

Allogeneic stem cell transplantation remains the only curative option for various benign and malignant hematological disorders. The procedure of allogeneic stem cell transplantation entails the administration of adequate doses of hematopoietic progenitor cells, aiming for complete and sustained hematopoietic reconstitution. Hematopoietic progenitors are obtained either through a bone marrow harvest or from GCSF-mobilized peripheral blood of the identified donor. Over the years various modalities have been utilized to establish the adequacy of harvested stem cell product e.g. Colony-forming unit– granulocyte monocyte (CFU-GM) assay, total mononuclear cell count (MNC), and CD34+ cell count

Effect of the umbilical cord blood acid‐base status and gas values on the yield of mononuclear cells and CD34+ cells

To investigate the influence of umbilical cord blood (CB) acid-base status and gas values on the yield of mononuclear cells and CD34⁺ cells, pH, pCO₂, pO₂, HCO₃⁻ and base excess were measured in arterial CB samples obtained from normal full-term deliveries. The relationship of these values with the yield of mononuclear cells and CD34⁺ cells detected in venous CB was analyzed. A total of 145 CB units were collected from full-term vaginal deliveries at a single hospital. Immediately after delivery, a segment of the umbilical cord was double clamped, and arterial CB was analyzed to determine the acid-base status and gases. Venous CB was collected in a sterile collection bag and processed for cell separation within 24 h of collection. The relationship between umbilical arterial acid-base status, each gas value, and the yield of mononuclear cells and CD34⁺ cells was analyzed. Statistically significant correlations were observed between the net weight of CB and the total mononuclear and CD34⁺ cell counts. In addition, there was a negative correlation between the mononuclear cell counts and pH, but a positive correlation between the mononuclear cell counts and pCO₂. However, no significant differences were observed between the primipara and multipara groups in terms of the net weight of CB, total mononuclear cell counts and total CD34⁺ cell counts. The findings of the present study show that the mononuclear cell counts are correlated with arterial CB pH and pCO₂, suggesting the involvement of fetal hypoxia on the yield of mononuclear cells.

SP6-28 Effect of umbilical cord blood acid-base status and gas values on the yield of mononuclear cells and cd34 cells

PurposeTo investigate the influence of umbilical cord blood (CB) acid-base status and gas values on the yield of mononuclear cells and CD34 cells, pH, pCO2, pO2, HCO3− and base excess...

Implications of CD34+ cell dose on clinical and haematological outcome of allo-SCT for acquired aplastic anaemia

The precise effects of CD34+ cell dose on the outcome of allogeneic transplantation for aplastic anaemia (AA) are not known. Previous studies have used the total mononuclear cell count to quantify stem cell dose. We evaluated the effects of CD34+ cell dose on the clinical and haematological end points of transplantation. The transplant variables and outcome parameters on 46 patients with acquired AA were assessed by comparing low vs high CD34+ cell doses. Infusion of less than 2 x 10(6)/kg of CD34+ cells was associated with an increased incidence of graft failures (P=0.03), higher incidence of bacterial infections (P=0.006) and a delay in the engraftment of neutrophils (P=0.046). The latter was found to be an effect of stem cell source (non-PBSC) rather than the CD34+ count. Other parameters, such as plt engraftment (P=0.63), red cell (P=0.94) and plt (P=0.31) transfusion independence, chimerism, acute and chronic GVHD (P=1.0) and OS (P=0.57), were not significantly influenced by the CD34+ cell dose. These findings are different to the published studies on the relevance of CD34+ cell dose in allogeneic transplantation for haematological cancers.

Stem cell harvesting protocol research in autologous transplantation setting: Large volume vs. conventional cytapheresis

The use of peripheral blood as a source of hematopoietic stem cells (SCs) is progressively increasing and has nearly supplanted bone marrow transplantation. Interpatient variability in the degree and kinetics of SC mobilization into peripheral blood is an expected event after conventional chemotherapy-based treatment, followed by sequential administration of recombinant granulocyte-colony-stimulating factor (rHu-CSF). In this study, specific factors associated with the application of two different SC-harvesting approaches, including the use of large volume leukapheresis (LVL) vs. repetitive conventional apheresis (RCA), were analyzed. The basic goal of the study was to evaluate the influence of apheresis protocol (collection timing, processed blood volume and cell yield) upon the clinical outcome of transplantation. Results obtained by LVL (76 pts) and RCA (20 pts--control group) were compared. The SC mobilizing regimen used was cyclophosphamide (4-7 g/m2) or polychemotherapy and rHuG-CSF 10-16 microg/kg of body mess (bm) per day. Cell harvesting was performed using Caridian-BCT Spectra (Gambro, USA). The volume of processed blood in LVL setting was > or = 3.5-fold of the patient's circulating blood quantity (ranged from 12.7 to 37.8 l). All patients tolerated well the use of intensive treatment, without any side or adverse effects. Our original controlled-rate cryopreservation was carried out with 10% dimethyl sulfoxide (DMSO) using Planer R203/200R or Planer 560-16 equipments (Planer Products Ltd, UK). Total nucleated cell (NC) and mononuclear cell (MNC) counts were examined by flow cytometry (Advia-2120 Bayer, Germany; Technicon H-3 System, USA). The CD34+ cell surface antigen was investigated by the EPICS XL-MCL device (Coulter, Germany). Performing LVL-apheresis, high-level MNC and CD34+ cell yields (7.6 +/- 4.6 x 10(8)/kg bm and 11.8 +/- 6.5 x 10(6)/kg bm, respectively) were obtained. As a result, rapid hematopoietic reconstitution ("graft-healing")--on the 9.4th and 12.4th day for granulocytes and platelets, respectively was achieved. Using repetitive conventional apheresis (2-3 procedures), the total MNC count was high (8.2 +/- 7.0 x 10(8)/kg bm), but the total CD34+ yield was lower 10.8 +/- 9.9 due to inferior CD34+ vs. MNC ratio. The results obtained suggest that well-timed LVL-apheresis increased SC-yield in cell harvest, resulting in faster bone marrow repopulation and hematological reconstitution, as well as better overall clinical outcome of transplantation. These results necessitate additional examinations of CD34+ subsets ratio in cell harvest.

Blood and spleen lymphocytes as targets for immunotoxic effects in the rat—a comparison

Traditionally, immunotoxicological studies in the rat have been performed by measuring the effect of chemical substances on spleen lymphocytes in vivo and in vitro. However, rat blood lymphocytes may be more relevant than spleen cells for comparison with human blood lymphocytes. Further, lymphocytes in blood may be a more sensitive indicator of immunotoxic effects than spleen lymphocytes. Finally, in longitudinal studies peripheral blood specimens can be collected repeatedly from the same animals, thereby reducing the number of animals sacrificed and, possibly, experimental variation. We compared blood and spleen lymphocyte parameters in rats treated with a single dose of the immunosuppressant cyclophosphamide (CY), monitoring effects on blood and spleen lymphocytes by immunophenotyping. We also performed repeated bleedings to demonstrate the feasibility of following the time course of induced changes in the same animals. Immunophenotyping as well as total mononuclear cell counts consistently showed as large or lager effects of CY in blood lymphocytes than in spleen cells. Further, the measured effects in blood lymphocytes became statistically significant at an earlier time point, compared to spleen cells. Repeated bleedings of the same animals illustrated that blood specimens drawn from a peripheral vein give sufficient numbers of cells to perform immunotoxicological tests.

Current antipsychotic dose correlates to mononuclear cell counts in the cerebrospinal fluid of psychotic patients

Elevated cerebrospinal fluid (CSF) angiotensin I-converting enzyme (ACE) levels have been evidenced in patients with schizophrenia who have been treated with antipsychotics. In order to explore a possible mononuclear cell origin of CSF ACE, the authors determined CSF ACE and CSF mononuclear cell counts from 25 acutely psychotic patients, who had been drug-free for at least 4 months but started on conventional antipsychotic medication within a few days before sampling. No correlations were found between CSF to serum ACE ratio and CSF mononuclear cell counts. However, CSF total mononuclear cell count, CSF lymphocyte count, and CSF mononuclear phagocyte count evidenced significant positive correlations with current dose of antipsychotic medication expressed as chlorpromazine equivalents. The authors conclude that no indication of a relationship between mononuclear cells and CSF ACE activity was found. Surprisingly, a relationship between chlorpromazine dose and CSF mononuclear cell counts was found, which may indicate drug-related changes in cell-mediated immunity. This finding needs replication and further corroboration in well-designed studies.

Recombinant Methionyl Human Stem Cell Factor and Filgrastim for Peripheral Blood Progenitor Cell Mobilization and Transplantation in Non-Hodgkin's Lymphoma Patients — Results of a Phase I/II Trial

AbstractTo examine the safety and efficacy of recombinant-methionyl human stem cell factor (r-metHuSCF), 38 patients with intermediate-grade or immunoblastic high-grade non-Hodgkin's lymphoma who were eligible for autologous transplantation were randomized to receive r-metHuSCF (5, 10, 15, or 20 μg/kg/d) plus Filgrastim (10 μg/kg/d) or Filgrastim (10 μg/kg/d) alone to mobilize peripheral blood progenitor cells. Subcutaneous administration of r-metHuSCF was well tolerated in conjunction with a multi-agent pre-medication regimen; local injection site reactions were the most commonly seen adverse event. The total mononuclear cell count, CD34+ cell content, granulocyte-macrophage colony-forming cells (GM-CFC), and burst-forming units-erythroid (BFU-E) per kilogram in the apheresis product was similar when all patients were analyzed by treatment cohort and mobilization regimen (Filgrastim or r-metHuSCF in combination with Filgrastim); however, when prior chemotherapy was taken into account in a supplementary analysis, clinically important differences were observed. Extensive prior therapy was defined as the amount of exposure to specific stem cell toxic chemotherapeutic agents that patients received. These agents include procarbazine, nitrogen mustard, melphalan, nitrosoureas (≥2 cycles of any of these drugs) or greater than 7.5 g of cytosine arabinoside. In these patients, there was an increased number of CD34+ cells (1.76 v 0.28 × 106/kg), GM-CFC (20.5 v 5.0 × 104/kg), and BFU-E (36.9 v 8.9 × 104/kg) in patients receiving r-metHuSCF and Filgrastim (N = 18) compared with Filgrastim alone (N = 5). These patients also had a decreased time to an untransfused platelet count of 20 × 109/L that was 10.5 days shorter in the patients who received r-metHuSCF and Filgrastim (12.5 v 23 days). These differences were not found to be statistically significant, possibly because of small size, but are clinically important.

Diagnostic features of tuberculous peritonitis in the absence and presence of chronic liver disease: A case control study

To determine diagnostic features of tuberculous peritonitis (TBP) in the absence and presence of chronic liver disease. Thirty-four patients with TBP (13 without [Group I] and 21 with chronic liver disease [Group II] and 26 controls with cirrhosis and uninfected ascites (Group III) were studied. The clinical features in Groups I and II were similar and all patients had elevated ascitic fluid total mononuclear cell count. In Groups I, II, and III, respectively, ascitic fluid protein was > 25 g/L in 100% (13/13), 70% (14/20), and 0% (0/26); serum-ascites albumin gradient (SAAG) was > 11 g/L in 0% (0/13), 52% (11/21), and 96% (25/26), (0% [0/13], 71% [15/21], and 96% [25/26] after correction for serum globulin); and ascitic fluid lactate dehydrogenase (LDH) level was > 90 U/L in 100% (12/12), 84% (16/19), and 0% (0/20), respectively. In Groups I and II combined, ascitic fluid acid-fast stain was negative in all but Mycobacterium tuberculosis culture was positive in 45% (10/22); peritoneal nodules occurred in 94% (31/33), granulomas in 93% (28/30), and positive peritoneal M tuberculosis culture in 63% (10/16). In patients with suspected TBP, ascitic fluid protein of > 25 g/L, SAAG of < 11 g/L and LDH of > 90 U/L have high sensitivity for the disease. With coexistent chronic liver disease, a lower protein level and higher SAAG are usually not helpful but LDH > 90 U/L is a useful parameter for screening. Diagnosis is best confirmed by laparoscopy with peritoneal biopsy and M tuberculosis culture.

Prediction of cytomegalovirus pneumonia after marrow transplantation from cellular characteristics and cytomegalovirus culture of bronchoalveolar lavage fluid.

Bronchoalveolar lavage (BAL) was performed between days 35 and 45 posttransplant in 37 marrow recipients without clinical or radiologic evidence of cytomegalovirus (CMV) pneumonia to determine if the presence of CMV or cellular characteristics was predictive of subsequent development of CMV pneumonia. Analysis of BAL fluid included culture and direct fluorescent antibody staining for CMV, total mononuclear cell count, lymphocyte count, and lymphocyte subsets. Patients were followed to day 110 after transplant for development of CMV pneumonia. Weekly cultures of blood, urine, and throat were also obtained. BAL was positive for CMV in 7 patients, of whom 3 developed CMV pneumonia. It was negative for CMV in 30 patients, of whom 7 developed CMV pneumonia. The positive and negative predictive values for detection of CMV in day 35-45 BAL were 43% and 78%, for CMV viremia 66% and 87%, and for detection of CMV in urine and throat 41% and 94%, respectively, for subsequent CMV pneumonia. In 9/10 patients who subsequently developed CMV pneumonia, excretion of CMV from BAL, blood, urine or throat preceded CMV pneumonia, and BAL was the earliest site of CMV excretion in all nine. Only one patient did not excrete before developing CMV pneumonia. In a stepwise logistic regression of risk factors for developing CMV pneumonia, only CMV viremia was significant (P = 0.002) although the total BAL mononuclear cell count approached significance (P = 0.053). The percentage of lymphocytes in BAL and CMV excretion in BAL, urine, and throat were not significant. The positive predictive value of BAL for subsequent CMV pneumonia was no greater than that of culture of urine and throat and less than that of viremia in our patients.

Prostaglandin and tumor necrosis factor levels in early wound inflammatory fluid: Effects of parenteral omega-3 and omega-6 fatty acid administration

Monokines are important mediators of wound healing. Specifically, the proportions of proinflammatory (tumor necrosis factor and PGE 2) and antiinflammatory (PGF 2α) monokines may modulate its early phases. Using a polyvinyl alcohol sponge model of rat wounding, the authors determined the temporal changes in the levels of monokines in wound inflammatory fluid, and examined whether dietary manipulation for 6 days with the precursors (ω6 fatty acids) and inhibitors (fish oil ω3 fatty acids) of the prostaglandin-2 series influenced monokine composition of wound fluid. For 3 days before the wounding, adult rats received isocaloric, isovolemic, and isonitrogenous total parenteral nutrition (TPN), in which lipids supplied either 35% (Intralipid [IL] or fish oil emulsion [FO]) or 8% (minimal essential fatty acid; EFA) of the total calories. Control rats received isocaloric enteral chow. The controls were studied at 24, 48, 72, and 96 hours, and the experimentals at 72 hours after wounding. Cell counts were performed, and cell-free fluid was analyzed for PGE 2, PGF 2α, and TNF. In control rats, the total WBC count was highest at 24 to 48 hours, and decreased significantly by 96 hours. The percentage of mononuclear cells progressively increased throughout the 96 hours, and the total mononuclear cell count peaked at 72 hours. The TNF and prostaglandin levels were highest at 24 hours; these decreased rapidly by 72 hours. At all time-points, the levels of PGE 2 remained higher than those of PGF 2α. Seventy-two hours postwounding, the different intravenous lipid preparations did not significantly after the WBC count, mononuclear cell count, or prostaglandin levels when compared with the 72-hour control, or with each other. However, TNF levels were significantly higher in the EFA low-fat group than in the high-fat IL or FO groups. This study confirms TNF's early role in wound healing, and provides previously undescribed evidence suggesting that PGE 2 and PGF 2α also help initiate the early inflammatory phase of wound healing. That all monokines peaked before the mononuclear cell peak suggests a functionally diverse, tightly regulated monocyte population differing in degree of activation over time. The lack of a temporal inversion of the PGE 2:PGF 2α ratio differs from previous studies that suggest that this ratio inverts as the inflammatory phase wanes. Interestingly, neither of the high-fat TPN preparations (IL or FO) resulted in significantly different monokine levels. However, the low-fat TPN preparation (EFA) did result in significantly increased wound TNF levels, which suggests that low quantities of exogenous fat may be more important than the type of fat for optimal wound healing.

Experience with autologous bone marrow harvesting and transfusion of marrow-derived red cells.

Marrow red cells (MRBCs) from 56 autologous bone marrow harvests were rescued after processing and transfused as the sole transfusion support after surgery. There was no correlation between volume of harvest and total mononuclear cell (MNC) count. The marrow collection induced a significant decrease in hematocrit values (mean, 6.1; range, -0.3-12.3; p less than 0.001) unrelated to the patient's diagnosis, age, or gender. Processing of the marrows resulted in a mean transfused MRBC mass of 258 mL (range, 101-494 mL), representing 78 percent (range, 43-100%) of the MRBC mass collected. The amount of MRBCs transfused correlated with total MNC count (p less than 0.01). All autologous MRBC transfusions were well tolerated. It can be concluded that autologous MRBC transfusions are safe and may eliminate the need for homologous blood transfusions after marrow harvest.

Biphasic adrenergic modulation of beta-adrenergic receptors in man. Agonist-induced early increment and late decrement in beta-adrenergic receptor number.

beta-Adrenergic receptors in mononuclear leukocyte preparations were assessed with (-)[(3)H]-dihydroalprenolol binding studies during the infusion of adrenergic agonists into normal human subjects. During the infusion of isoproterenol into seven subjects, mean (+/-SE) (-)[(3)H]dihydroalprenolol binding increased from 25+/-3 fmol/mg protein to 47+/-8 fmol/mg protein (P < 0.02) at 0.5 h and 40+/-3 fmol/mg protein (P < 0.01) at 1 h and decreased to 12+/-1 fmol/mg protein (P < 0.01) at 4-6 h. During the infusion of epinephrine into three subjects, mean (-)[(3)H]dihydroalprenolol binding increased from 32+/-3 to 63+/-3 fmol/mg protein (P < 0.01) at 0.5-1 h. By Scatchard plot analysis, these changes were attributable to changes in the number of available binding sites rather than changes in binding affinity. The observed changes in the number of (-)[(3)H]dihydroalprenolol binding sites were not paralleled by changes in total mononuclear cell counts or in T lymphocyte, B lymphocyte, and monocyte distributions. Thus, we conclude that adrenergic agonists modulate the number of available beta-adrenergic receptors on circulating mononuclear cells in a biphasic manner, with an early increment and a late decrement, in man. Further, the finding that the increase in pulse rate in response to a "pulse" infusion of isoproterenol was significantly greater after 0.5-1 h of agonist infusion suggests that the observed early agonist-induced increment in beta-adrenergic receptor number on circulating cells is paralleled by increments in extra-vascular beta-adrenergic receptor sensitivity.

Cellular Immunity in Infectious Mononucleosis

Abstract Infectious mononucleosis (IM) patients, Epstein-Barr virus (EBV)-seropositive and seronegative healthy donors, and patients with other viral infections were tested for lymphocyte blastogenesis (LB) with phytohemagglutinin and six EBV (virus concentrate, culture supernatant, and soluble [S] antigen) or control antigens. Fluorescent antibodies to EBV viral capsid antigen of IgG, IgM, IgA specificities, to nuclear antigen (EBNA), and heterophile antibodies were also assayed. These were correlated with clinical parameters (fever, pharyngitis, adenopathy, hepatitis, splenomegaly, atypical lymphocytes, and total mononuclear cell counts). EBV viral and S antigen-induced LB was significantly greater in seropositive donors. IM patients had antigen specific LB below that of seropositive donors initially and low responses for the acute phase of illness when clinical symptoms were present and antibody titers were maximal. Specific LB rose to a peak at 3.5 to 9 weeks when the patients had recovered, most laboratory findings had returned to normal, and antibodies had declined. At peak, specific LB in IM patients exceeded that of seropositive donors, but later declined. These results demonstrate specific cell-mediated immunity (CMI) to EBV, and indicate that this develops slowly in IM and contrasts with the evolution of the clinical events and humoral immunity. This correlation supports the hypothesis that CMI is the mechanism of terminating lymphoproliferation in IM.

Circulating deoxyribonucleic acid--synthesizing mononuclear leukocytes. I. Increased numbers of proliferating mononuclear leukocytes in inflammatory disease.

Abstract To quantitate deoxyribonucleic acid (DNA) synthesis by circulating human blood mononuclear leukocytes, buffy coat suspensions were incubated with tritiated thymidine in vitro, and incorporation was measured by liquid-scintillation counting. Elevated mononuclear leukocyte DNA synthesis was found in a wide spectrum of inflammatory diseases, including 2 patients with agammaglobulinemia and infection. Increased thymidine uptake was found especially in the acute phase of disease and tended to decrease with, clinical improvement of the patient. Uptake was lower in patients with chronic inflammatory diseases. No correlation was found between the mononuclear leukocyte thymidine uptake and the erythrocyte sedimentation rate or the total mononuclear cell count. Autoradiographic studies revealed that the tritium-labeled leukocytes had the appearance of atypical lymphocytes and did not phagocytize latex particles. Similar labeled mononuclear cells were found in the blood of a patient with Swiss-type agammaglobulinemia and pneumonia. It was suggested from the data obtained that proliferating cells may appear in the blood in response to inflammation. Some may represent lymphoid cells which participate in an immune response associated with the disease process involved, and some may be precursors of monocytes released in response to acute tissue injury.