Identification and cloning of a brain autoantigen in neuro-behavioral SLE

METHODS: The study reports the analytical and diagnostic performances of Cell Dyn 3500 (Abbott), an automated haematology analyser that provides the electrical impedance and the optical detection of total leukocyte and a five population leukocyte differential count. The evaluation of complete blood count and differential leukocyte count parameters was performed following the guidelines of the International Council for Standardization in Haematology and those of the National Committee for Clinical Laboratory Standards document H20-A. RESULTS: The CD 3500 demonstrated a good linearity, minimal carry-over, as well as acceptable levels of within and between-batch imprecision and stability. An excellent correlation between CD 3500 and the routine systems used in our laboratory (Technicon H*2 and Coulter STKS) was found for the major haematological indices, though agreement was not as good for MCHC. There was a good correlation between CD 3500 and manual reference differential method for neutrophils, lymphocytes, eosinophils and basophils, with the exception of monocytes. CONCLUSIONS: Regarding clinical sensitivity, the CD 3500 showed a high specificity to detect morphological abnormalities, but low sensitivity especially for the identification of immature granulocytes and nucleated red blood cells; if we overall evaluate any qualitative flags, we found an appreciable increase in sensitivity.     

Crucial suppressive role of renal kallikrein-kinin system in development of salt-sensitive hypertension

The performance of the SE-9000 automated haematology analyser in a laboratory receiving a high number of abnormal specimens from haematological oncology patients was assessed according to formal protocols for the evaluation of blood cell counters. Linearity over a useful working range, precision in clinically important ranges and negligible carry-over were demonstrated in this group of patient samples confirming the results of previous investigators. The comparability of instrument derived differential leucocyte counts from both normal and distributionally abnormal samples with those obtained by visual microscopy using the NCCLS H-20 A protocol was very good. The sensitivity of flags for the detection of immature granulocytes and myeloid blast cells was high and this can be attributed to the incorporation of a new measuring channel (Immature Myeloid Information or IMI channel). The number of unrecognized abnormalities was low and when compared with the poor sensitivity of the routine 100-cell visual differential leucocyte count, the analyser was judged suitable for monitoring patients with haematological malignancies. The performance of flags such as 'left shift' and 'atypical lymphocytes' can be improved by taking into consideration distributional abnormalities such as neutrophilia and lymphocytosis. The trigger level for these flags should be adapted to the clinical need particularly in cases of neutropenia following chemotherapy, and in lymphoproliferative disorders and infection.     

Comparison of tacrolimus absorption in transplant patients receiving continuous versus interrupted enteral nutritional feeding

1. The obese gene product leptin, secreted exclusively from adipocytes, was discovered to serve as a satiety factor and to play an important role in regulating body weight. In adults, the serum leptin level reportedly increases with the degree of obesity. Leptin receptors are expressed in various tissues, and recent in vitro studies suggest a role for leptin in haematopoiesis. 2. The present study was designed to clarify the relationship between serum leptin and body mass index, peripheral blood cell counts, serum cholesterol, high-density lipoprotein-cholesterol, insulin and cortisol levels in 299 Japanese male adolescents aged 15-16 years. 3. With simple linear correlation, log [serum leptin] showed a strong correlation with body mass index (r = 0.56), log [insulin] (r = 0.36) and leucocyte count (r = 0.22) (P < 0.001 for all). There were also correlations with systolic blood pressure, erythrocyte count, haematocrit and high-density lipoprotein-cholesterol (P < 0.01 for all). Even after adjustment for body mass index and log [insulin], log [leptin] correlated with leucocyte (P = 0.004) and erythrocyte (P = 0.057) counts. Stepwise multiple regression analyses revealed log [leptin] to correlate significantly with body mass index, log [insulin] and the leucocyte count (P < 0.005 for all, r2 = 0.399). 4. To our knowledge, this is the first clinical study to show the possible association of serum leptin level with blood cell counts, independent of body mass index and serum insulin. We conclude that these data further support a role for leptin in haematopoiesis.     

Pediatric experience with autologous peripheral blood progenitor cell transplantation: influence of CD34+ cell dose in engraftment kinetics

The aim of this study was to analyze factors affecting mobilization and engraftment in 40 children undergoing autologous peripheral blood progenitor cell transplantation for different malignancies: 19 patients with haematological malignancies and 21 patients with solid tumors. Patients received 4-5 days of rhG-CSF (12 micrograms/kg/day) subcutaneously. Apheresis was performed by continuous flow blood cell separation beginning on the fifth day of rhG-CSF. For patients weighing < or = 25 kg, the extracorporeal line was primed with irradiated red blood cells. After myeloablative conditioning regimens, patients were grafted with 7.21 +/- 7.8 x 10(6)/kg CD34+ cells. Days to achieve an absolute neutrophil count > 0.5 x 10(9)/1 and a platelet count > 20 x 10(9)/1 without platelet support were 9.50 +/- 1.2 (range 7-13) and 18.1 +/- 8.3 (range 9-37), respectively. The number of CD34+ cells infused was highly correlated with engraftment kinetics (P = 0.0001). The patient's body weight and the number of previous chemotherapy courses had a negative influence on CD34+ cells collected.     

Rhabdoid tumours of the central nervous system. Report of three cases with immunocytochemical and ultrastructural findings

Blood samples from malaria-infected patients and from in vitro culture were analyzed using the H*1 hematology analyzer. An attempt to find a hematologic parameter for detecting the malaria infection and to characterize the pathophysiological changes of red cells was made. The study included 18 malaria-infected patients (10 with Plasmodium falciparum and 8 with Plasmodium vivax) and 52 normal, healthy volunteers. Increased young large lymphocyte or large unstained cell count (LUC over 3%) in the peripheral blood of malaria-infected patients was evidence for malaria infection. Increased population dispersions of red cell volume (red cell distribution width: RDW) and intraerythrocytic hemoglobin concentration (hemoglobin distribution width: HDW) were demonstrated both in clinical samples and cultured blood. The increased RDW correlated with an increased percentage of macrocytes (r = 0.64, P = 0.004). Comparison of HDW and percentage of hypochromic red cells between the clinical specimen and the cultured blood supports the finding that changes in red cell hemoglobin concentration were mainly due to the response of the patient to malaria infection and partly due to the effect of malaria parasites on the red cells.     

Field evaluation of the NOVA Celltrak 12 hematology analyzer

An evaluation of the NOVA Celltrak 12 was performed over a 6-week period. Precision, linearity, carryover, and reproducibility of values compared favorably to manufacturers' claims. The correlation was performed using the Coulter S Plus VI as the reference instrument and yielded coefficients of correlation for measured parameters of greater than .98 with the exception of RDW at r = .84, MCV at r = 0.96, and MPV at r = .92. Three-part differential information and an expanded flagging system affords increased clinical information and trouble-shooting guides. The three-part differential information compared favorably with that of the Coulter S Plus VI with lymphocyte percentage r = .96, granulocyte percentage r = .94, and monocyte percentage r = .50. We conclude that this instrument performed well in a clinical setting.     

Effect of a constant magnetic field of ultra-high intensity on the morphologic composition of peripheral blood

An exposure of mice to a constant magnetic field (CMF) of 9.9, 25.4 and 39.4 kersted produced changes in the leucocyte and reticulocyte counts and caused no changes in the red blood cell count and hemoglobin. The changes noted during and after the exposure were of a cyclic pattern. During a 24-hour exposure of animals to a CMF of 39.4 kersted a short-term increase in the reticulocyte and leucocyte counts and a concomitant decrease of the neutrophil and lymphocyte counts were noted. Afterwards the reticulocyte count decreased and increased again by the end of the exposure. Following a 3-hour exposure to CMFs of the above intensities the changes persisted for 20-35 days. Normally the parameters increased during the 1-2 days of the exposure and decreased thereafter. The level of changes was not correlated with the CMF intensity. The increase in the reticulocyte count during and exposure to a CMF of 39.4 kersted was identical to that during an exposure to a CMF of I kersted.     

What''s new in syndactyly?

The collection of peripheral blood stem cells (PBSC) is a crucial step for successful PBSC transplantation. Routine hematological parameters utilized for predicting the optimal timing of collection include white blood cell (WBC) counts, high fluorescence ratio (HFR) of reticulocytes, and platelet counts. We compared these parameters with the CD34-positive rates in the peripheral blood. In regimen with high-dose chemotherapy where the WBC count at nadir was lower than 1,000/microliter, we found that the maximum mobilization of PBSC was observed on the day when the WBC count reached 10,000/microliter. This coincidence was within about one day (mean 0.44, standard deviation 0.53). However, the reliability of the WBC count as a marker of PBSC mobilization varied among different harvest regimens. In the regimen with regular-dose chemotherapy where the WBC count at nadir was above 1,000/microliter, we could not find such a tight coincidence between the WBC count and PBSC mobilization. These results suggested that, in some situations, the measurement of the CD34-positive rate is mandatory for an efficient PBSC collection. We also found that the number of CD34-positive cells in the peripheral blood correlated (x) well to the amount of the CD34-positive cells actually harvested (y) (y = 0.524x + 0.249, r = 0.787). Thus, rapid fluorescence activated cell sorter (FACS) analysis of peripheral CD34-positive rates seemed to be extremely useful to predict the yield of PBSC collection.     

A randomized trial of granulocyte colony-stimulating factor (Neupogen) starting day 1 vs day 7 post-autologous stem cell transplantation

The purpose of the study was to evaluate the effect of delayed granulocyte colony-stimulating factor (G-CSF) use on hematopoietic recovery post-autologous peripheral blood progenitor cell (PBPC) transplantation. Patients were randomized to begin G-CSF on day +1 or day +7 post transplantation. Thirty-seven patients with lymphoma or myeloma undergoing high-dose therapy and autologous PBPC rescue were randomized to daily subcutaneous G-CSF beginning on day +1 or day +7 post-transplant. Patients < or =70 kg received 300 microg/day and >70 kg 480 microg/day. All patients were reinfused with PBPCs with a CD34+ cell count >2.0 x 10(6)/kg. Baseline characteristics of age, sex and CD34+ cell count were similar between the two arms, the median CD34+ cell count being 5.87 x 10(6)/kg in the day +1 group and 7.70 x 10(6)/kg in the day +7 group (P=0.7). The median time to reach a neutrophil count of >0.5 x 10(9)/l was 9 days in the day +1 arm and 10 days in the day +7 arm, a difference which was not statistically significant (P=0.68). Similarly, there was no difference in median days to platelet recovery >20000 x 10(9)/l, which was 10 days in the day +1 arm and 11 days in the day +7 arm (P=0.83). There was also no significant difference in the median duration of febrile neutropenia (4 vs 6 days; P=0.7), intravenous antibiotic use (7 vs 8 days; P=0.54) or median number of red blood cell transfusions (4 vs 7 units; P=0.82) between the two arms. Median length of hospital stay was 11 days post-PBPC reinfusion in both groups. The median number of G-CSF injections used was 8 in the day +1 group and 3 in the day +7 group (P < 0.0001). There is no significant difference in time to neutrophil or platelet recovery when G-CSF is initiated on day +7 compared to day +1 post-autologous PBPC transplantation. There is also no difference in number of febrile neutropenic or antibiotic days, number of red blood cell transfusions or length of hospital stay. The number of doses of G-CSF used per transplant is significantly reduced with delayed initiation, resulting in a significant reduction in drug costs. For patients with an adequately mobilized PBPC graft, the initiation of G-CSF can be delayed until day +7 post-PBPC reinfusion.     

Cytogenetic findings in invasive breast carcinomas with prognostically favourable histology: a less complex karyotypic pattern?

Highly fluorescent reticulocyte (HFR) counts were evaluated in 13 consecutive patients affected by hematological malignancies and submitted to autologous selected CD34+ peripheral blood progenitor cell (PBPC) transplantation. Results were compared with a historical group of patients comparable for age, disease and conditioning regimen submitted to unfractionated PBPC transplantation. HFR counts of the CD34+ group declined to an undetectable level from day +4 to day +10 when they became detectable and reached 5% of total reticulocyte count by day +12. In the historical group, the nadir was identical but the recovery was faster (day +9). Total reticulocyte count > 1% was achieved at days +17 and +11, respectively. The absolute neutrophil count (ANC) recovery was identical in both groups, achieving a value > 0.5 x 10(9)/l at day +13 after reinfusion. Hence, in the historical group, HFR count gave advance notice of complete and stable hemopoietic engraftment while in the CD34+ group HFR and ANC count showed almost simultaneous recovery.     

Poly(ADP-ribose) polymerase gene expression status and genomic instability in human breast cancer

OBJECTIVE: To compare point-of-care results obtained from an on-site hemocytometer with values provided by an institutional laboratory instrument. DESIGN: A prospective laboratory evaluation. SETTING: The central laboratory and cardiac surgical intensive care unit of a university-affiliated tertiary care center. PATIENTS: Normal range comparison was performed using blood specimens routinely obtained from 48 hospitalized patients for complete blood count analysis. The second evaluation was performed on blood specimens routinely obtained (in the intensive care unit) after cardiac surgery involving extracorporeal circulation in a series of 187 consecutive patients. MEASUREMENTS AND MAIN RESULTS: Hemoglobin concentration, platelet count, mean corpuscular volume, mean platelet volume, and red and white blood cell counts were measured with both on-site (MD 16, Coulter Electronics, Hialeah, FL) and laboratory (STKS, Coulter Electronics) instruments. Hematocrit and red cell distribution width were calculated using measured variables. Blood specimens were obtained from two distinct patients series. To evaluate measurement values within the normal range, a series of 48 routinely obtained blood specimens for complete blood count analysis in our institutional laboratory were utilized for concurrent analysis with the on-site hemocytometer. To evaluate measurement values out of the normal range, a second comparison involved measurements performed on blood specimens obtained in the cardiac surgical intensive care unit for complete blood count analysis. Linear regression demonstrated good correlations between on-site and laboratory hemoglobin concentration (r2 = .97), hematocrit (r2 = .95), platelet count (r2 = .97), mean corpuscular volume (r2 = .91), red cell distribution width (r2 = .80), and red (r2 = .95) and white (r2 = .96) blood cell count results. A marginal correlation was observed between mean platelet volume values (r2 = .47). Bias analysis (mean +/- 2 SD) demonstrated similar measurements between on-site and laboratory hemoglobin concentration, hematocrit, platelet count, red blood cell count, white blood cell count, mean platelet volume, mean corpuscular volume, and red cell distribution width. CONCLUSIONS: On-site hemoglobin concentration, hematocrit, white blood cell count, red blood cell count, red cell distribution width, and platelet count values compare well with those results obtained from the laboratory. The MD 16 hemocytometer (Coulter Electronics) provides on-site hematologic results that can provide an accurate and rapid quantitative assessment of platelets, and red and white blood cells. Rapid access to information obtained from this type of system may be clinically useful, especially in critically ill patients.     

High-dose chemotherapy with autologous stem cell support in poor-risk germ cell tumors

We propose a simple and fast method of detecting apoptosis using an automated hematology analyzer. Detection is based on cellular optical light scatter properties and demonstration of the membrane fragility which characterizes cells undergoing the process of apoptosis. As part of it's routine leucocyte differential analysis, the Abbott Cell-Dyn 4000 collects multi-angle cellular light scatter data. In addition red fluorescence (FL3) emitted by cells following propidium iodide labeling is collected. This provides quantitation of both the erythroblast count and a leukocyte viability index (WVF). Fresh or cryopreserved peripheral blood cells from 17 B-chronic lymphocytic leukemia (B-CLL) patients were incubated in presence of theophylline, fludarabine or in medium alone. After 36-hrs of culture the percentage of apoptotic cells of the sample was determined from the parameters of the CD 4000 described above and thereafter this was compared with reference methods for estimation of apoptosis. The reference methods used were in situ detection of cell death on slides (TUNEL test) and also flow cytometry (Annexin V). Results showed an excellent correlation between the 3 techniques. This rapid, easy and reliable method of quantifying apoptosis may be very useful means of routinely predicting the response to chemotherapy.     

Tinnitus guideline. German Society of Otorhinolaryngology, Head and Neck Surgery

We reviewed a consecutive case series of 178 immunocompetent children aged 3-36 months without central venous lines who had blood cultures positive for Streptococcus pneumoniae by either of paired broth and quantitative culture methods. The incidence of accompanying focal infection was significantly greater in patients with > 10 colony-forming units (cfu)/mL than in patients with < or = 10 cfu/mL (30.4% vs 12.9% respectively, p = 0.04). No significant relationships existed between the magnitude of bacteremia and the age, gender, presenting temperature, interval until the blood culture turned positive, total peripheral blood white cell count, absolute neutrophil count, or absolute band count. Overall, the quantitative method detected 59/178 (33.1%) of the isolates, including five isolates (2.8%) that the broth method failed to detect.     

供者外周血干细胞动员采集198例临床分析

目的 研究外周血造血干细胞移植( PBSCT)供者外周血干细胞动员采集的方法及其效 果。方法 198名健康供者每天皮下注射重组人粒细胞集落刺激因子(rhG-CSF)(5～10) μg/kg进行外周血干细胞动员,第5天开始采集。采用血细胞分析仪行单个核细胞( MNC)计数,流式细胞术(FCM)行CD34+细胞计数。分析供者性别、身高、年龄、采集当天外周血白细胞(WBC)计数对动员采集效果的影响。结果所有供者均成功动员采集,采集当天的MNC计数平均为(4.19±1.96)×108/kg,CD34+细胞计数平均为(2.98±1.40) ×106/kg; MNC和CD34+细胞计数与供者性别、身高、年龄无关;采集当天外周血WBC计数与MNC、CD34+细胞计数呈正相关(r= 0.9201,P=0.0035;r=0.8420,P= 0.0149);采集当天外周血WBC计数≥20.0×109/L的供者比＜20.0×109/L的供者采集效果更显著(F=4.688,P= 0.0013;F= 4.622,P=0.0006)。结论rhG-CSF动员的健康供者采集当天外周血WBC计数是一项预测CD34+细胞采集数量的简单、可行的指标。     

Neonatal neutrophil activation is a function of labor length in preterm infants

To understand better the development of the neonatal immune system, we evaluated the role of labor length, gestational age, and mode of delivery on the expression of the neonatal neutrophil cell surface antigens CD11b, CD11c, CD15, CD33, and CD66b in premature newborns. Peripheral blood samples from 68 apparently healthy preterm infants were obtained within 12 h of birth and incubated with MAb to the CD antigens. Samples were lysed, fixed, and analyzed by flow cytometry. Multivariate analysis was used to study the simultaneous effect of the labor length and gestational age on the neonatal neutrophil cell surface antigen expression. A positive correlation was demonstrated between neutrophil antigen expression and labor length (p < 0.001-0.026) but not with the mode of delivery (p = 0.191-0.638). There was no significant correlation between expression of neutrophil antigens and gestational age at delivery (p = 0.057-0.866), except for CD15 (p = 0.010). Our results indicate labor length is a significant factor in neonatal neutrophil activation at birth. These findings are independent of gestational age in preterm newborns. Mode of delivery does not seem to influence neonatal neutrophil activation. The neutrophils of premature infants can be activated antenatally and/or during labor.     

Effect of specimen storage on absolute CD4 counts

We have evaluated the effect of specimen storage on absolute CD4 counts by a commercially available manual assay. This assay utilizes latex particles coated with CD4 monoclonal antibodies that are mixed with lymphocytes in whole blood. Thirty blood samples were analyzed on days 1, 2, 4, and 7 postcollection. Linear regression analysis and Pearson correlation coefficients were used to determine the relationship between the absolute CD4 count and the storage time after sample collection. There was a significant decrease in absolute CD4 counts from baseline over time, dropping 3.6% at day 2, 10.1% at day 4, and 18.8% at day 7. However, the standard error of the B coefficient was constant [SE (B) = 0.031] up to day 4, indicating that reliable estimates of the baseline CD4 counts could be made from the CD4 counts determined up to day 4 from the time of sample collection. In addition to being sample, rapid, and inexpensive, the manual assay is capable of giving a reliable absolute CD4 count after specimen storage of up to 4 days. The application of this assay in the limited facilities of developing countries' laboratories is attractive.     

Microvascular function and rheologic changes in hyperdynamic sepsis

OBJECTIVE: To investigate the rheologic changes and circulatory abnormalities at the microvascular level during severe sepsis. DESIGN: Prospective, controlled trial. SETTING: Medical and surgical intensive care units of a university-affiliated hospital. PATIENTS: Nine normal controls and eight adult patients with severe sepsis who met the study entrance criteria. INTERVENTIONS: Forearm blood flow was measured at rest and during reactive hyperemia by air plethysmography. Simultaneous hemodynamic measurements and blood samples for rheologic measurements were taken. MEASUREMENTS AND MAIN RESULTS: Red blood cell deformability index was determined using a simple filtration procedure. Leukocyte aggregation in whole human blood was detected by using a leukergy test. Expression of the neutrophil adhesion molecule CD11b/CD18 was measured using a monoclonal antibody and flow cytometry. All data were taken within 24 hrs of the patient meeting criteria for entrance into the study. Cardiac output, oxygen delivery, and oxygen consumption measurements were consistent with the hyperdynamic phase of severe sepsis. Forearm blood flow was significantly (p < .05) greater in septic patients (21 +/- 3 mL/min) than in controls (12 +/- 2 to 36 +/- 5 mL/min (p < .05), while in the septic patients, forearm blood flow during reactive hyperemia increased from 21 +/- 3 to 32 +/- 4 mL/min. The ratio of forearm blood flow during reactive hyperemia to forearm blood flow at rest was 3.2 +/- 0.1 in the controls and 1.6 +/- 0.1 in the septic patients (p < .01). The red blood cell deformability index in whole blood was significantly (p < .01) decreased in the septic patients compared with the control subjects (0.41 +/- 0.07 vs. 0.98 +/- 0.08 mL/min). This difference remained true when the hematocrit was adjusted to 45% (0.82 +/- 0.06 vs. 1.04 +/- 0.06 mL/min; p < .05). Increased expression of the neutrophil adhesion molecule CD11b/CD18 was observed in septic patients (349 +/- 46 logarithmic fluorescence units) as compared with control subjects (233 +/- 26 logarithmic fluorescence units; p < .05). Leukergy was also significantly (p < .05) increased in septic patients (17.7 +/- 3.8%) as compared with control subjects (8.9 +/- 1.6%). A significant correlation was observed between leukergy and the expression of the neutrophil adhesion molecule CD11b/CD18 in controls and septic patients (r2 = .62; p < .01). Leukergy was also inversely correlated with whole blood red blood cell deformability index (r2 = .28; p < .05). CONCLUSIONS: Reactive hyperemia in the forearm is significantly diminished in patients with sepsis, suggesting impaired microvascular blood flow. Rheologic changes, including impaired red blood cell deformability, increased leukocyte aggregation, and endothelial adherence, may contribute to this abnormality by compromising effective capillary cross-sectional area.     

Effects of trauma and sepsis on soluble L-selectin and cell surface expression of L-selectin and CD11b

OBJECTIVES: To examine (1) the effects of trauma on changes in neutrophil L-selectin and CD11b expression and on the levels of soluble L-selectin and (2) whether these alterations are different on leukocyte subpopulations in those patients who develop multiple organ dysfunction syndrome. MATERIALS AND METHODS: Twenty patients with Injury Severity Score (ISS) > or = 16 and 15 patients with ISS score < 16 were studied. Arterial blood were collected serially after injury. The staining of leukocyte surface adhesion molecules was performed with antibodies against L-selectin and CD11b. Positive cell count and mean fluorescence intensity were determined by flow cytometry. Soluble L-selectin was measured using enzyme-linked immunosorbent assay. RESULTS: In patients with ISS > or = 16, neutrophil L-selectin expression showed an immediate increase, reaching peak levels between 3 to 4 hours after injury (p < 0.05 vs. patients with ISS < 16), followed by a gradual decrease. Plasma levels of soluble L-selectin reached peak levels at 6 hours after injury. However, in patients with ISS < 16, minimal changes in L-selectin expression and soluble L-selectin were observed. Neutrophil CD11b expression showed an immediate increase for the first 3 hours followed by a gradual increase up to 24 hours after injury. In patients who developed multiple organ dysfunction syndrome, CD11b both on neutrophils and lymphocytes remained elevated for 120 hours. CONCLUSIONS: These findings suggest that acute neutrophil activation is an early event after trauma and may be implicated as "a vulnerable window" for leukocyte-mediated end organ injury.     

Do viral load and CD8 cell count at initiation of tritherapy influence the increase of CD4 T-cell count?

BACKGROUND: Tritherapies including protease inhibitors improve clinical status and usually increase CD4 T cell count. However, the dissociation between the marked decreases in viral load and the incomplete restoration of CD4 cell counts with a three-drug combination has been reported. We assessed this potential difference among our patients. METHODS: Patients were enrolled when a protease inhibitor was prescribed to them for the first time. Using a computerized medical record (ADDIS), we retrospectively assessed a potential relationship between the increase in CD4 T cells (deltaCD4) at M3, M6 and variables including sex, age, CDC staging, protease inhibitor, prior antiviral therapy, CD8 and viral load at baseline. We used Epi-Info 6.4 and BMDP software. RESULTS: Data were analyzed on 154 patients. The median CD4 T cell count was 157 at baseline, 215 at month 3 and 202 at month 6. The median viral load was 52000 copies at baseline, 530 at month 3 and 500 at month 6. In a univariate analysis, a significant relationship was found between deltaCD4 and CD8 at baseline. A statistically significant negative correlation appeared between the CD8 cell count at baseline and deltaCD4 at M6 (r=-0.28, Pearson). Moreover, we found that there also was a relationship between deltaCD4 and viral load at baseline. There was a correlation between deltaCD4 at M6 and the viral load at M0 (r=0.37, Pearson). In a multiple regression model, after CD8 count at baseline had been accounted for, we found a significant correlation between deltaCD4 and viral load at baseline (multiple r=0.33 at M3, and 0.40 at M6). CONCLUSIONS: Patients with a low viral load do not benefit from as great an increase in CD4 T cell count as others when they receive a tritherapy including protease inhibitors. These results suggest that another mechanism rather than direct viral pathogenicity leads to CD4 T cell destruction. This mechanism may not be efficiently stopped by antiviral therapy, especially protease inhibitors.     

Establishing an umbilical cord blood bank for unrelated allogenic stem cell transplantation

We report the establishment of a cord-blood bank in a routine hematological laboratory. Cord-blood collection was performed with placenta in utero by a trained team and immediately sent to the cord-blood bank. There, 6.8 ml cord-blood was used for analysis of nucleated cell counts, counts of CD34-positive cells, CFU's, complete HLA-typing, ABO and Rhesus blood groups, bacteriologic cultures and serology for HIV 1 and 2, HbsAg, HVC, CMV, syphilis and toxoplasmosis. The cord-blood collection was frozen and conserved at -192 degrees C. From each cord-blood vials of DNA, viable cells and plasma were cryopreserved. Between June 1997 and April 1998, 54 cord-bloods were collected. 40 of them were cryo-preserved, and 14 discarded because of low cell counts. The median volume was 109 ml with 1.4 x 10(9) nucleated cells. The in vitro capacity of proliferation of the cord-blood correlated well with the absolute counts of CD34-positive cells (r = 0.93), moderately with the relative counts of CD34 (r = 0.68) as well as the nucleated cells (r = 0.70), poorly with the volume (r = 0.44). Three of the 40 (7.5%) cord-blood products contained a bacterial contamination. This study shows that a cord-blood bank can be organised in a routine hematological laboratory, which is familiar with transplantation products. However, the procedure is time consuming, expensive and requires a highly qualified team and specialised technical equipment.