Absolute Number Of Granulocytes Research Articles

Changes in the cellular composition of blood, phagocytic function of neutrophils in patients with acute coronary syndrome

The aim of this study was to determine changes in the cellular composition of the neutrophil blood, the phagocytic function in patients with acute coronary syndrome (ACS), who underwent coronary artery (CA) stenting. We examined 40 patients with acute coronary syndrome aged 40 to 65 years (mean age 52.58±6.82), who immediately after admission or within the next 72 hours, depending on the clinical situation, underwent coronary angiography and implanted stents with drug Xience Alpine finish. The control group consisted of 30 healthy men who had no contact with harmful production factors, and in whom no clinical, laboratory and instrumental deviations from the norm were detected during medical examinations. Complete blood count (25 parameters: leukocyte, erythrocyte, and platelet hematopoietic sprouts), quantitative and qualitative composition of hematopoietic sprouts was performed by a standardized method on a Medonic M20 hematological analyzer (Sweden). Evaluation of neutrophil phagocytic activity of latex particles with a diameter of 1.7 μm by neutrophils (phagocytosis activity, phagocytosis intensity, phagocytosis number), spontaneous and induced neutrophil HBT activity were recorded by a morphological method (light microscopy using Olimpus microscopes (Japan). Conventional methods were used to assess the functional activity of phagocytes. In patients with ACS, compared to the control group, leukocytosis was observed (10.34±0.764, p<0.001), a relative increase in the number of stab and segmented neutrophils in % (4.98±0.736, p<0.05 and 61.09±1.908, p<0.001, respectively), absolute number of granulocytes -7.30±0.711×10⁹/l (p<0.001), absolute increase in monocytes — 0.92±0.093 (p<0.05). Decreased percentage of lymphocytes -22.43±2.013 (p<0.001). The index of the neutrophil-to-lymphocyte ratio was 2.95 in the examined patients. rophils in % (38.55±3.230 — ACS, 50.6±3.44 — control, p<0.05), but higher NST induced activity in % (59.73±2.936 versus 39.4±1.51 , respectively, p<0.001) and the index of NBT-induced cell activity (0.90±0.065 vs. 0.63±0.031, respectively, p<0.001). These data indicate the severity of the process and characterize the exacerbation of inflammation in these patients against the background of a decrease in the immune activity of the phagocytic link.

DIAGNOSTIC UTILITY OF LEUKOCYTE PARAMETERS IN THE PATIENTS WITH ACUTE MYOCARDIAL INFARCTION

Background. Inflammation is one of the key players in acute myocardial infarction (AMI). One of the ways to evaluate it indirectly is to analyze leukocyte parameters of complete blood count (CBC), which is a routine and affordable method of diagnosis. Leukocyte counts can provide additional information about the course, as well as a potential prognosis for complications of AMI. We suggest that the dynamic changes of CBC during the treatment of the patients with AMI may be of value to assess the prognosis of the course of the disease, and therefore require further study. Objective. The aim of the study to evaluate the diagnostic and prognostic potential of leukocyte indexes of CBC, in particular the levels of leukocytes, lymphocytes, neutrophils and N/L, WBC/MPV, PLT/L ratios in the patients with AMI at the time of hospitalization and on the 7th day of hospital stay. Methods. The study involved 204 individuals: 152 patients with AMI (Group 1), 30 patients with stable coronary heart disease (Group 2) and 24 healthy volunteers (Group 3). Hemogram parameters and their ratios, in particular the levels of leukocytes, lymphocytes, neutrophils, ESR, as well as the ratios of N/L and PLT/L were studied. Results. Levels of leukocytes, neutrophils, lymphocytes, as well as N/L, WBC/MPV, MPV/L ratios were significantly higher in the patients with AMI compared to other groups. The best diagnostic value had such indicators as the total number of leukocytes (sensitivity 71.7%, specificity 69.7%, AUC 0.794), the absolute number of granulocytes (sensitivity 81.7%, specificity 77.4%, AUC 0.803), the N/L ratio (sensitivity 75.0%, specificity 71.7%, AUC 0.791) and the WBC/MPV ratio (sensitivity 76.7%, specificity 62.3%, AUC 0.760). The PLT/L ratio calculated on the 7th day of hospital stay correlated with the risk of in-hospital (r=0.369, p=0.002) and 6-month mortality (r=0.338, p=0.004) according to the GRACE score. Conclusions. Leukocytes, granulocytes, N/L and WBC/MPV ratios had a fairly high diagnostic value for the patients with AMI. Regarding the prognostic potential assessment, only the PLT/L ratio on the 7th day of hospitalization correlated with the risk of in-hospital and 6-month mortality. This proves the importance of assessing CBC parameters not only at the time of hospitalization, but also in the dynamics of AMI.

Immune Phenotype by Flow Cytometry of Paediatric Kidney Transplant Recipients and Healthy Adult Controls

Introduction The proportion of immune cell subsets in paediatric kidney transplant recipients could help monitor the best approach to allow tolerance to renal allografts. Aim To determine/establish the differences in the immunophenotype of pediatric kidney transplant recipients and healthy adult controls. Methods Absolute cell count (TruCount) and seven leukocyte-profiling panels containing 8-10 marker-antigens (46 of which were unique within the panels) consisting of Tregs, NKT, B, NK, DCs, and monocyte subsets were used to monitor the immune profiles of 9 paediatric kidney transplant and 8 adult control samples. Whole-blood samples, 100-300μl/panel, were stained and acquired on a BD-LSRFortessa and Flowjo was used for data analysis. Results Differences in subpopulations between pediatric and healthy adult control can be seen in Table 1. This showed a significant increase in absolute numbers of Granulocytes, CD14+Monocytes, double negative NKT cells and CD56+HICD16+ Intermediate NK cells. B cell panel showed a significant increase in naïve B cells, IgD+IgM+ B cells, and IgM+CD27- B cells. The naïve CD4+ (CD45RA+CCR7+/CD62+) and naïve CD8+ (CD45RA+CCR7+/CD62+) T cells, and naïve Tregs (CD4+CD25+hiCD127lowCD45RA+CCR7+/CD62+ and CD4+FOXP3+CD45RO-) which were higher than in adult controls. Moreover, naïve Foxp3 Tregs in pediatric transplanted patients had higher CD25 expression than naive Tregs in adult controls. Memory CD4+ T cells (CD45RA-CD4+) in pediatric patients has a similar HLA-DR expression as adult controls (Fig. 1).Further we found a significant decrease in the following cell populations in our pediatric kidney transplant patients compare to healthy adult controls: Non-classical monocytes, CD8+NKT, memory B cells (IgD-CD27+), IgD-IgM- B cells, CD27+CD38low Class-switched memory. A decreased proportion in CD27-CD28+, CD25+ of CD4+, CD57+CD27-CD28+ and CD25+ of CD8, CCR7-/CD62L-CD45RA- of CD4 and CD8, and CXCR3+CD45RO+ of CD4 and of CD8 (Fig.2), and CD127+CD45RO+ and CD25+CD45RO+ on CD4 T cells (effector Foxp3 Tregs) suggested less memory and effector memory T cells in our pediatric kidney transplant patients than controls (Fig.3). Conclusion Immune profiling of pediatric kidney transplant recipients demonstrated more naïve T cells, B cells and Tregs and less memory and effector memory T cells compared to healthy adult controls. Immunophenotyping by flow cytometry is a non-invasive technique requiring less than 1.5ml of whole blood and maybe a useful tool to monitor determine the differences in the immunophenotype in transplant recipients. Birgit Sawitzki. Mathias Streitz.

Invitro and invivo study on the effect of antifungal agents on hematopoietic cells in mice.

Liposomal amphotericin B, voriconazole, and caspofungin are currently used for systemic and severe fungal infections. Patients with malignant diseases are treated with granulocyte-colony stimulating factor (G-CSF) for the recovery of granulocytes after chemotherapy or hematopoietic cell (HC) transplantation. Since they have a high incidence of fungal infections, they inevitably receive antifungal drugs for treatment and prophylaxis. Despite their proven less toxicity for various cell types comparatively with amphotericin B and the decrease in the number of leukocytes that has been reported as a possible complication in clinical studies, the effect of liposomal amphotericin B, voriconazole, and caspofungin on HCs has not been clarified. The present study aimed to examine the invitro and invivo effect of these three modern antifungals on HCs. Colony-forming unit (CFU) assays of murine bone marrow cells were performed in methylcellulose medium with or without cytokines and in the presence or absence of various concentrations of liposomal amphotericin B, voriconazole, and caspofungin. In the invivo experiments, the absolute number of granulocytes was determined during leukocyte recovery in sublethally irradiated mice receiving each antifungal agent separately, with or without G-CSF. Invitro, all three antifungal drugs were nontoxic and, interestingly, they significantly increased the number of CFU-granulocyte-macrophage colonies in the presence of cytokines, at all concentrations tested. This was contrary to the concentration-dependent toxicity and the significant decrease caused by conventional amphotericin B. Invivo, the number of granulocytes was significantly higher with caspofungin plus G-CSF treatment, higher and to a lesser extent higher, but not statistically significantly, with voriconazole plus G-CSF and liposomal amphotericin B plus G-CSF treatments, respectively, as compared with G-CSF alone. These data indicate a potential synergistic effect of these antifungals with the cytokines, invitro and invivo, with subsequent positive effect on hematopoiesis.

Bone marrow and peripheral white blood cells number is affected by sleep deprivation in a murine experimental model

Sleep deficit and related disorders are becoming increasingly prevalent in modern life and an extensive literature has documented that acute or chronic sleep deprivation can lead to several physiological consequences. Here, we evaluated the effects of sleep deprivation on hematopoietic composition of either bone marrow or peripheral blood. Mice were subjected to paradoxical sleep deprivation (PSD) for 72 h by modified multiple platform method, with or without an additional sleep recovery (SR) period of 10 days. PSD decreased total cellularity of the bone marrow and peripheral blood concomitantly. Subsequent analysis of cell composition showed that absolute number of hematopoietic stem/progenitor cells and colony-forming units was decreased. Moreover, the absolute number of granulocytes and monocytes in bone marrow was reduced in PSD group. These alterations were paralleled by an accumulation of neutrophils and monocytes in peripheral blood. PSD also induced lymphopenia in the circulation. To the best of our knowledge, this is the first study that demonstrates the importance of sleep on the hematopoietic microenvironment and provides new insights into the relationship between sleep and the immune system.

Finger-Stick Complete Blood Counts

Background: A complete blood count (CBC) including an automated differential is currently performed on complex analyzers in either central or satellite laboratories. Previously, we reported that CBC measurements determined by Chempaq XBC point-of-care analyzer compare favorably with laboratory-based Beckman Coulter LH750 system. This study describes comparison of capillary finger-stick versus venous blood CBC analyses. Methods: Both finger-stick and venous blood samples obtained simultaneously from 310 patients from 5 different patient clinics were evaluated for white blood cell (WBC), hemoglobin (HGB), and differential cell counts. Finger-stick blood samples were collected and analyzed immediately by phlebotomy or nursing staff using a Chempaq XBC analyzer, whereas venous blood was sent to the main laboratory for the analyses on Beckman Coulter LH750 system. Results: There are no significant differences between the mean levels of CBC parameters analyzed between the finger-stick and venous blood: the mean and bias between finger-stick and venous blood for WBCs (7.33, 7.25, 0.08), HGB (13.47, 13.20; 0.27), absolute number of granulocytes (4.96, 4.83; 0.14), lymphocytes (1.70, 1.81; −0.10), and monocytes (0.62, 0.57; 0.05), respectively. There is also a good correlation between finger-stick and venous blood samples except for monocytes: the slope, intercept, and r value for WBCs (1.04, −0.24, 0.92), HGB (1.13, −1.48, 0.94), number of granulocytes (1.01, 0.08, 0.91), number of lymphocytes (0.87, 0.12, 0.86), and number of monocytes (1.01, 0.04, 0.63), respectively. Conclusions: Finger-stick CBC analyses offer a simple point-of-care in-office test. By following appropriate guidelines, like any flagged or abnormal results, being confirmed by the main laboratory, finger-stick blood testing for CBCs may be useful in remote sites, rural clinics, physician offices, and in developing countries.

Delayed recovery after autologous peripheral hematopoietic cell transplantation: potential effect of a high number of total nucleated cells in the graft

Some patients demonstrate delayed recoveries after autologous hematopoietic stem cell transplantation despite infusion of an adequate number of CD34+ cells/kg and clinically stable status. Factors considered being possible predictors of this outcome in this context were explored. A total of 246 patients were evaluated in terms of engraftment. Delayed recovery was defined by white blood cell recovery time exceeding mean+1 SEM. Clinical factors and graft characteristics were examined. Comparisons between patients with normal or delayed engraftment were made. Proinflammatory cytokines and proteolytic enzyme quantification and CXCR4+ and CD44+ cell enumeration were performed on peripheral hematopoietic stem cells (PHSC) product samples of patients with delayed engraftment and patients with usual engraftment time. Sixteen patients, who received at least 3 × 10(6) CD34+ cells/kg without known clinical factors likely to affect engraftment, demonstrated a delayed recovery time of over 20 days. Some graft variables were found to be significantly increased in these patients by univariate analysis. One variable was the total number of nucleated cells cryopreserved and infused. Among the nucleated cells, the absolute number of granulocytes before and after cryopreservation also differed significantly between the two groups. A multivariate analysis showed that the main predictive factor for delayed recovery was the number of nucleated cells in the graft (p=0.0044). The influence of contaminating cells might be related to the release of elastase, matrix metalloproteinase-9, interleukin (IL)-1β, and IL-6 involved in stem cell homing. Therefore, the numeration of total nucleated cells and granulocytes should be considered as a possible quality control variable of PHSCs submitted for cryopreservation.

Expression of Endothelial Selectin Ligands on Human Leukocytes Following Dive

The fact that impaired endothelial-dependent vasodilatation after scuba diving often occurs without visible changes in the endothelial layer implies its biochemical origin. Since Lewisx(CD15) and sialyl-Lewisx(CD15s) are granulocyte and monocyte carbohydrate antigens recognized as ligands by endothelial selectins, we assumed that they could be sensitive markers for impaired vasodilatation following diving. Using flow cytometry, we determined the CD15 and CD15s peripheral blood mononuclear cells of eight divers, 30 mins before and 50 mins after a single dive to 54 m for 20 mins bottom time. The number of gas bubbles in the right heart was monitored by ultrasound. Gas bubbles were seen in all eight divers, with the average number of bubbles/cm2 1.9+/-1.9. The proportion of CD15+monocytes increased 2-fold after the dive as well as the subpopulation of monocytes highly expressing CD15s. The absolute number of monocytes was slightly, but not significantly, increased after the dive, whereas the absolute number of granulocytes was markedly elevated (up to 61%). There were no significant correlations between bubble formation and CD15+monocyte expression (r=-0.56; P=0.17), as well as with monocytes highly expressing CD15s (r=0.43; P=0.29). This study suggests that biochemical changes induced by scuba diving primarily activate existing monocytes rather than increase the number of monocytes at a time of acute arterial endothelial dysfunction.

A role for leptin in sustaining lymphopoiesis and myelopoiesis

Although leptin is known for its regulation of food intake, it has many emerging roles in immune function. To better define the role of leptin in hematopoietic processes, a leptin-deficient obese mouse (ob/ob) and C57BL/6 lean wild-type controls were compared. Despite their large size and consumption of substantial amounts of nutrients, the ob/ob mice had only 60% as many nucleated cells in their marrow as controls. The greatest impact of leptin deficiency was on the B cell compartment that had 70% fewer cells, reducing the absolute number of pre-B and immature B cells to 21% and 12% of normal, respectively, and indicating a significant reduction in lymphopoiesis in ob/ob mice. Whereas the proportion of myeloids remained nearly normal in the obese mice, they also exhibited a reduction of 40% and 25%, respectively, in absolute numbers of granulocytes and monocytes. Seven days of provision of recombinant leptin promoted substantial lymphopoiesis, increasing the numbers of B cells in the marrow of the obese mice twofold, while doubling and tripling, respectively, the numbers of pre-B and immature B cells. Twelve days of supplementation brought these subpopulations to near-normal proportions. Leptin treatment also facilitated myelopoiesis such that the marrow of the obese mice contained normal numbers of monocytes and granulocytes after 7 days. Taken together, the data support an important role for leptin in sustaining lymphopoiesis and myelopoiesis.

The Effects of Echinacea Root Extract in Plasmacytoma-Bearing Mice: Enhancement of Non-Specific Immunity

ABSTRACT Alternative and complementary therapies, involving a host of herbal products are becoming increasingly popular. Information on such treatments is anecdotal for the most part, poorly understood, and lacks adequate preclinical and/or clinical empirical substantiation. Formal, controlled, laboratory studies on the effects of herbs on cells responsible for disease defense (granulocytes, natural killer cells, monocytes, lymphocytes) in pathogenic conditions in vivo, are, to date, almost completely lacking. The present study aimed to investigate the effect of long-term dietary administration of the herb, Echinacea, on these cell populations in the spleen and bone marrow of mice while bearing a plasmacytoma, known as a highly immunotoxic tumor. Mice in the present study, were given 4 pristane injections (days 0, 21, 42, and 100) to establish plasmacytoma, and the herb was provided daily in the chow from day 49 (above) until the day of sampling. At the time of euthanasia, the absolute numbers of granulocytes, natural killer (NK) cells, monocytes and lymphocytes, were recorded in both the spleen (a organ housing the mature, functional forms of theses cells), and the bone marrow (generation site of these cells). NK cells are responsible for host innate (nonspecific) immunity and represent the first line of defense against a wide variety of pathogens. The results revealed quantitative, cell lineage-specific changes in both organs in plasmacytoma-bearing mice consuming Echinacea-treated diets versus those consuming the control diet (untreated chow). Echinacea in the diet neutralized the inhibitory effects which the tumor's presence had on the absolute numbers of NK cells in both organs, returning them to control (normal) levels. This study provides the first evidence that NK cells, in vivo, in the presence of a chronic pathological condition, that is, an established tumor, react very positively to Echinacea, and may provide a means of immune support in the immunotoxic environment of chemotherapy administered chronically to tumor-afflicted humans.

Long-term effects of perinatal nutrition on T lymphocyte kinetics in young Gambian men

Background:Nutritional status is highly dependent on season in countries such as The Gambia. In a rural Gambian setting, individuals born during periods of seasonal nutritional deprivation (“hungry seasons”) are susceptible to mortality from infectious diseases in adult life.Objective:We investigated the hypothesis that impaired immunocompetence in those born in the hungry season results from an underlying defect in immunologic memory, similar to the immunosenescence of old age, which is likely to be reflected in the phenotype and kinetics of T lymphocytes in young adults.Design:T cell phenotype in terms of CD3, CD4, CD8, CD45RA, and CD45R0 expression and in vivo dynamics measured by stable isotope labeling of T cell subsets combined with gas chromatography–mass spectrometry and frequency of T cell receptor excision circles were measured in 25 young (18–24-y-old) Gambian men. Thirteen of these 25 men were exposed to perinatal malnutrition as defined by birth season and birth weight.Results:In persons born in the hungry season with low birth weight, no differences in the proportions of memory or naive T cells were found. Kinetic analysis showed higher proliferation rates in memory (CD45R0+) subsets of T cells than in naïve (CD45R0−) cells, which is consistent with previous studies, but no evidence was found for an effect of birth weight or season on T lymphocyte proliferation and disappearance rates. No significant correlations were found between in vivo T cell kinetics and frequency of T cell receptor excision circles. Only absolute numbers of granulocytes were elevated in those born in the nutritionally deprived season.Conclusion:In healthy young Gambian men, T lymphocyte homeostasis is extremely robust regardless of perinatal nutritional compromise.

Tumor Necrosis Factor-α Increases Circulating Osteoclast Precursor Numbers by Promoting Their Proliferation and Differentiation in the Bone Marrow through Up-regulation of c-Fms Expression

Osteoclasts are essential cells for bone erosion in inflammatory arthritis and are derived from cells in the myeloid lineage. Recently, we reported that tumor necrosis factor-alpha (TNFalpha) increases the blood osteoclast precursor (OCP) numbers in arthritic patients and animals, which are reduced by anti-TNF therapy, implying that circulating OCPs may have an important role in the pathogenesis of erosive arthritis. The aim of this study is to investigate the mechanism by which TNFalpha induces this increase in OCP frequency. We found that TNFalpha stimulated cell division and conversion of CD11b+/Gr-1-/lo/c-Fms- to CD11b+/Gr-1-/lo/c-Fms+ cells, which was not blocked by neutralizing macrophage colony-stimulating factor (M-CSF) antibody. Ex vivo analysis of monocytes demonstrated the following: (i) blood CD11b+/Gr-1-/lo but not CD11b-/Gr-1- cells give rise to osteoclasts when they were cultured with receptor activator NF-kappaB ligand and M-CSF; and (ii) TNF-transgenic mice have a significant increase in blood CD11b+/Gr-1-/lo cells and bone marrow proliferating CD11b+/Gr-1-/lo cells. Administration of TNFalpha to wild type mice induced bone marrow CD11b+/Gr-1-/lo cell proliferation, which was associated with an increase in CD11b+/Gr-1-/lo OCPs in the circulation. Thus, TNFalpha directly stimulates bone marrow OCP genesis by enhancing c-Fms expression. This results in progenitor cell proliferation and differentiation in response to M-CSF, leading to an enlargement of the marrow OCP pool. Increased marrow OCPs subsequently egress to the circulation, forming a basis for elevated OCP frequency. Therefore, the first step of TNF-induced osteoclastogenesis is at the level of OCP genesis in the bone marrow, which represents another layer of regulation to control erosive disease.

Radioprotective properties of nutraceutical Gonebazol: In vivo study

BACKGROUND: In vitro investigation of radioprotective properties of novel nutraceutical Gonebazol (Biofarm, Belgrade) has displayed its remarkable potential to reduce chromosomal aberrations and micronuclei induced by x rays. The goal of this study was to evaluate its protective properties in vivo. For this purpose, a group of medical staff performing invasive radiological diagnostics, that was identified to carry dicentrics in their lymphocytes, was selected for further monitoring. They consumed this nutraceutical for 3 weeks, 2 x 10 g per day (2 x 2 teaspoons dissolved in a glass of water). METHODS: Hematological parameters, chromosomal aberrations and micronucleus frequency were examined at the beginning of monitoring, on day 10 and 21 of the treatment. RESULTS: In the course of three weeks the incidence of chromosomal aberrations and micronuclei was significantly reduced (50%-80%), absolute number of granulocytes was lowered whereas the number of monocytes significantly was enhanced. This study has revealed that nutraceutic posses immunomodulatory properties seen as an improving monocyte-macrophage activity, which inversely correlates with incidence of lymphocyte micronuclei (r=0.75, p<0.05). CONCLUSION: Observed finding could be of particular importance in reducing cumulative effects of ionizing radiation in radiosensitive tissues and in preventing adverse health effects. No side effects were evidenced. Mechanisms of its immunomodulatory effects should be further examined.

Impaired maturation of myeloid progenitors in mice lacking novel Polycomb group protein MBT-1

Polycomb group (PcG) proteins participate in DNA-binding complexes with gene-repressing activity, many of which have been highlighted for their involvement in hematopoiesis. We have identified a putative PcG protein, termed MBT-1, that is associated with Rnf2, an in vivo interactor of PcG proteins. MBT-1 structurally resembles the H-L(3)MBT protein, whose deletion is predicted to be responsible for myeloid hematopoietic malignancies. The human MBT-1 gene is located on chromosome 6q23, a region frequently deleted in leukemia cells, and shows a transient expression spike in response to maturation-inducing stimuli in myeloid leukemia cells. MBT-1(-/-) myeloid progenitor cells exhibit a maturational deficiency but maintain normal proliferative activities. This results in the accumulation of immature myeloid progenitors and hence, a marked decrease of mature myeloid blood cells, causing the MBT-1(-/-) mice to die of anemia during a late embryonic stage. Together, we conclude that MBT-1 specifically regulates the maturational advancement of myeloid progenitor cells during transitions between two developmental stages. We also show that MBT-1 appears to influence myelopoiesis by transiently enhancing p57(KIP2) expression levels.

Phenotypical characterization of peripheral blood leucocytes in the newborn calf.

The present study was undertaken to establish reference values for the composition of blood leucocyte populations in neonatal calves by differential leucocyte counts and immunophenotyping. Neonatal calves 1 h post partum (p.p.) were found to have a very high absolute number of granulocytes while the number of peripheral blood mononuclear cells was lower than in calves aged 3-9 weeks. The relative numbers of T cell subpopulations were similar in newborn and older calves, but newborn calves had lower percentages of B cells and MHC class II positive cells. Within the first 4 h of life the relative numbers of CD2+, CD6+, and CD8+ T cells declined in colostrum-fed as well as in colostrum-deprived calves. In contrast, the percentage of MHC class II positive cells and monocytes increased from 1 h to 4 h p.p. particularly in colostrum-fed calves. Although there is some evidence for immaturity of lymphocytes in neonatal calves, the immune system of these animals seems to be fully present at birth.

Effects of isoniazid (INH) on the BCG-induced local immune response after intravesical BCG therapy for superficial bladder cancer.

Because recent investigations showed that the use of isoniazid (INH) severely impaired the local immune reaction to intravesical bacillus Calmette-Guérin (BCG) in the bladder of guinea pigs, in this study the effect of INH in man has been investigated. Patients were treated with BCG with or without oral INH. The concentration of free INH in most urine samples of patients treated with BCG/INH was much higher (mean 38.0 +/- 60.9 micrograms INH/ml) than the minimal inhibitory concentration (MIC; 0.1 microgram INH/ml), suggesting at least a bacteriostatic potential of the INH present. However, in vitro studies showed that these urinary concentrations of INH did not kill BCG organisms effectively, even at a concentration of 150 micrograms/ml for 24 h. After the fifth and sixth BCG instillations a significant increase in the concentration of cytokines (IL2, IL6, IL8 and TNFa), IgG and IgA antibodies to BCG and the number of leukocytes in urine was observed. The leukocytes mainly consisted of granulocytes, besides monocytes/macrophages and, in lower amounts, T- and B-lymphocytes and natural killer (NK) cells. The absolute number of granulocytes and the concentration of IgG antibodies after BCG instillation were significantly suppressed by INH, whereas INH appeared to have no effect on the urinary cytokine and IgA antibody concentrations or the total number and phenotype of the leukocytes present. In conclusion, the results of this study indicate that INH does not impair the local immunological stimulation after BCG instillation in man as severely as was observed in the guinea pig and it may be expected that INH does not impair the antitumor efficacy of BCG.

Natural killer cell activity, phagocytosis, and number of peripheral blood cells in breast cancer patients treated with tamoxifen

The number of leukocytes, proportion and absolute number of granulocytes, lymphocytes, CD4+ cells, CD8+ cells, CD16+ cells, B-lymphocytes, monocytes, natural killer cell (NK) activity, and granulocyte and monocyte phagocytic functions--ingestion and intracellular killing--were determined in a group of 27 patients with ductal invasive breast carcinoma, stage I-III, before and 7 months following postsurgical telecobalt radiotherapy, divided into two subgroups, one of them receiving tamoxifen (TMX group) and the other one not receiving any further therapy (control group). In control group, proportion of all lymphocytes and CD8+ cells as well as absolute number of all lymphocytes, CD4+, CD8+, CD16+ and B lymphocytes were decreased following TCT in comparison to their pre-TCT values, while in TMX group only absolute number of all lymphocytes remained decreased following TCT. Moreover, post-TCT proportions of all and CD8+ lymphocytes as well as absolute numbers of all and CD4+ and CD8+ lymphocytes in TMX patients were significantly increased in comparison to the same parameters in control post-TCT patients, although there was no difference between the two subgroups before TCT. At the other hand, granulocyte ingestion was decreased in post-TCT TMX patients compared to post-TCT values in control patients and NK cell activity showed a similar, although statistically not significant, tendency. It seems that TMX helps recovery of lymphocyte populations decreased by radiotherapy, probably by stimulation of cells carrying estrogen receptors, but its effects on phagocytic functions and probably NK cell activity seemed to be rather inhibitory than stimulatory.

Alpha interferon in chronic lymphocytic leukaemia

The role of alpha interferon in patients with chronic lymphocytic leukaemia (CLL) has yet to be well established. In studies carried out to date, a significantly higher response rate has been observed in previously untreated patients compared to those who have received prior chemotherapy. Patients with early-stage CLL also respond better than patients with advanced disease. Responses to alpha interferon are transient and complete responses are rare. It is not yet known whether alpha interferon can induce clonal remission, and response is usually measured in terms of the reduction in peripheral blood lymphocyte levels. In one study, a normalization of immunoglobulin levels was observed, and in another there was an increase in the absolute number of granulocytes. Further studies are needed to investigate the role of combined therapy with alpha interferon and cytotoxic agents or other cytokines, and to assess the ability of alpha interferon to prolong response duration after remission induction with chemotherapy. Toxicity is tolerable when alpha interferon is given in a low dose (e.g., 2 million units (MU)/m 2 three times a week) and low doses have been shown to be as effective as high doses in CLL patients.

Monocyte Function in Chickens with Hereditary Muscular Dystrophy

Monocyte functions of Storrs strain muscular dystrophic (MD) chickens and Storrs control strain were compared. After short-term in vitro culture of monocytes, cells taken from Storrs MD chickens had a significantly lower incidence of cells capable of phagocytizing sheep red blood cells than did cells cultured from the control strain. In contrast, no difference was found in the ability of MD vs. control strain blood mononuclear cells to undergo in vitro chemotaxis in response to a bacterial culture supernatant. The study also revealed a distinct peripheral blood leukocyte profile of young MD chickens: elevated percentages and absolute numbers of granulocytes. The total leukocyte count in MD chickens was not significantly different from that of normal chickens. These findings extend previous observations concerning altered reticuloendothelial system function in MD strain chickens.

Recombinant α2-Interferon in the Treatment of B Chronic Lymphocytic Leukemia in Early Stages

Ten previously untreated patients with early B cell chronic lymphocytic leukemia (B-CLL) (seven in Rai's stage 0, three in stage I) were given recombinant alpha 2-interferon (alpha 2IF) (2 X 10(6) U/m2 intramuscularly three times a week for a minimum of 14 weeks) to assess its effectiveness. All patients were evaluable for response to therapy and toxicity. No complete response was achieved. In all cases a definite, although transient reduction in the absolute number of peripheral blood lymphocytes was observed. In eight patients an increase in the absolute number of granulocytes was detected. None of the patients experienced severe hematologic toxicity. Fatigue, malaise, and fever were the more common side effects, but all patients were able to finish their treatment as planned. The results of this pilot study suggest that low doses of recombinant alpha 2-IF have some activity in early and previously untreated B-CLL and that further studies of IF effectiveness in B-CLL seem warranted.