Peripheral White Cell Count Research Articles

Administration of interleukin-6 stimulates multilineage hematopoiesis and accelerates recovery from radiation-induced hematopoietic depression

Hematopoietic depression and subsequent susceptibility to potentially lethal opportunistic infections are well-documented phenomena following radiotherapy. Methods to therapeutically mitigate radiation-induced myelosuppression could offer great clinical value. In vivo studies in our laboratory have demonstrated that interleukin-6 (IL-6) stimulates pluripotent hematopoietic stem cell (CFU-s), granulocyte-macrophage progenitor cell (GM-CFC), and erythroid progenitor cell (CFU-e) proliferation in normal mice. Based on these results, the ability of IL- 6 to stimulate hematopoietic regeneration following radiation-induced hematopoietic injury was also evaluated. C3H/HeN female mice were exposed to 6.5 Gy 60Co radiation and subcutaneously administered either saline or IL-6 (1,000 micrograms/kg) on days 1 through 3 or 1 through 6 postexposure. On days 7, 10, 14, 17, and 22, femoral and splenic CFU-s, GM-CFC, and CFU-e contents and peripheral blood white cell, red cell, and platelet counts were determined. Compared with saline treatment, both 3-day and 6-day IL-6 treatments accelerated hematopoietic recovery; 6-day treatment produced the greater effects. For example, compared with normal control values (N), femoral and splenic CFU-s numbers in IL-6-treated mice 17 days postirradiation were 27% N and 136% N versus 2% N and 10% N in saline-treated mice. At the same time, bone marrow and splenic GM-CFC values were 58% N and 473% N versus 6% N and 196% N in saline-treated mice; bone marrow and splenic CFU-e numbers were 91% N and 250% N versus 31% N and 130% N in saline-treated mice; and peripheral blood white cell, red cell, and platelet values were 210% N, 60% N, and 24% N versus 18% N, 39% N, and 7% N in saline- treated mice. These studies demonstrate that therapeutically administered IL-6 can effectively accelerate multilineage hematopoietic recovery following radiation-induced hematopoietic injury.

Administration of Interleukin-6 Stimulates Multilineage Hematopoiesis and Accelerates Recovery From Radiation-Induced Hematopoietic Depression

Hematopoietic depression and subsequent susceptibility to potentially lethal opportunistic infections are well-documented phenomena following radiotherapy. Methods to therapeutically mitigate radiation-induced myelosuppression could offer great clinical value. In vivo studies in our laboratory have demonstrated that interleukin-6 (IL-6) stimulates pluripotent hematopoietic stem cell (CFU-s), granulocyte-macrophage progenitor cell (GM-CFC), and erythroid progenitor cell (CFU-e) proliferation in normal mice. Based on these results, the ability of IL-6 to stimulate hematopoietic regeneration following radiation-induced hematopoietic injury was also evaluated. C3H/HeN female mice were exposed to 6.5 Gy 60Co radiation and subcutaneously administered either saline or IL-6 (1,000 micrograms/kg) on days 1 through 3 or 1 through 6 postexposure. On days 7, 10, 14, 17, and 22, femoral and splenic CFU-s, GM-CFC, and CFU-e contents and peripheral blood white cell, red cell, and platelet counts were determined. Compared with saline treatment, both 3-day and 6-day IL-6 treatments accelerated hematopoietic recovery; 6-day treatment produced the greater effects. For example, compared with normal control values (N), femoral and splenic CFU-s numbers in IL-6-treated mice 17 days postirradiation were 27% N and 136% N versus 2% N and 10% N in saline-treated mice. At the same time, bone marrow and splenic GM-CFC values were 58% N and 473% N versus 6% N and 196% N in saline-treated mice; bone marrow and splenic CFU-e numbers were 91% N and 250% N versus 31% N and 130% N in saline-treated mice; and peripheral blood white cell, red cell, and platelet values were 210% N, 60% N, and 24% N versus 18% N, 39% N, and 7% N in saline-treated mice. These studies demonstrate that therapeutically administered IL-6 can effectively accelerate multilineage hematopoietic recovery following radiation-induced hematopoietic injury.

In vivo therapy of non hodgkins lymphoma with a cytotoxic murine monoclonal antibody; a preliminary report

A murine IgM monoclonal antibody termed Wm-66 has been produced which binda to a novel uncluttered 65 Kitodalton leucocyte surface membrane antigen present on virtually all normal lymphocytes and monocytei. This antibody has been shown to react strongly with most types of chronic B cell malignancies (CLL and NHL) and to kill target cells in the presence of human complement. These properties suggested a potential role for WM-66 as a therapeutic agent in CLL and lymphoma. The antibody was prepared in mice as ascitet, separated by gel filtration and hydroxylapatite chromatography to 88% purity, and checked for immunoreactivity, sterility, and pyrogenicity. A 59 year old male with advanced refractory well-differentiated lymphocytic lymphoma in leukaemic phase was selected for treatment with WM-66 He had generalised lymphadenopathy and 19cms palpable splenomegaly Antibody was infused daily intravenously in saline over 1-2 hrs in increasing doses up to 30 mg with a total of 65 mg given in 5 days No significant toxic side effects were seen. The peripheral blood white cell count, initially 68 × 10 9 /L (90% lymphocytes), fell by 25-50% immediately after each infusion, although with some rebound by 24 hours, reaching a minimum of 34 × 10 9 /L at the completion of treatment Leucocytes showed coating with mouse immunoglobulin after the 20 mg and 30 mg doses. The spleen sise was recuded to 14 cms after 5 days. We conclude that WM-66 antibody produces cytoreduction when infused intravenously in a patient with NHL without evidence of toxicity Further studies with higher doses and different schedules are planned.

Fever and Petechiae in Children

To the Editor.— I read the article on "Fever and Petechiae in Children" by Baker et al1 with great interest. The results of Baker et al support the teaching I have received and my clinical impression that children having fever and petechiae most often do not have meningococcemia. Further, it appears that consideration of initial clinical features, peripheral white cell count, and examination of cerebrospinal fluid should allow a definitive assessment of whether invasive bacterial disease is present.

Neutrophil Kinetics in the Pulmonary Microcirculation: Effects of Pressure and Flow in the Dependent Lung

Increases in pulmonary arterial pressure or blood flow raise peripheral white cell count by releasing sequestered leukocytes from the lung. The effects of altered hemodynamics, however, on the leukocyte sequestration site and on the distribution of leukocyte transit times through the pulmonary microcirculation are unknown. We used in vivo fluorescence videomicroscopy to study the passage of individual, fluorescein-isothiocyanate-labeled neutrophils through the pulmonary microcirculation of anesthetized dogs. Pulmonary hemodynamics were altered over a wide range. Regardless of the hemodynamic conditions, the only place that any of the 2,919 observed neutrophils stopped was in the capillaries. The periods of immobility had a wide range, from less than 1 to greater than 1,200 s. Because the cells remained motionless once they stopped and then accelerated suddenly as they regained the free-flowing stream, the obstructions must have been discrete. About a quarter of the capillary pathways had one site of high resistance. Another quarter offered two or more obstructions. In the remaining half, the neutrophils passed rapidly and without pause from arteriole to venule. Increases in pressure and flow decreased the number of times that individual cells stopped. These changes altered the median transit time by shifting the distribution of transit times between the slowest and fastest groups. We conclude that most of the total pathlength of perfused capillaries offers little resistance even to neutrophils. There are, however, focal areas in individual capillaries that offer high resistance to neutrophil passage.

Normal T Lymphocyte Function in Patients with End-Stage Renal Disease Hemodialyzed with ‘High-Flux’ Polysulfone Membranes

T lymphocyte function was analyzed in patients hemodialyzed with 'high-flux' polysulfone membranes, which have been reported to improve the patients' overall clinical condition and well-being. For comparison purposes, patients treated by the use of 'low-flux' cuprophane membranes were also studied. Peripheral blood white cell counts, numbers of lymphocytes as well as the numbers of T cells and their CD4 and CD8 subsets were within normal range in both patient groups. The absolute number of B cells was slightly decreased in cuprophane-membrane- but not polysulfone-membrane-treated patients. The proliferative response of T lymphocytes after stimulation with optimal concentration of phytohemagglutinin (PHA) was normal in patients treated with 'high-flux' membrane dialysis but significantly reduced in those treated with cuprophane membranes. The generation of interleukin-2 (IL-2) receptor on T lymphocytes after PHA stimulation was normal in the polysulfone-membrane-treated group and slightly impaired in the cuprophane-membrane-dialyzed patients. Production of both IL-2 and interleukin-1, as well as the natural killer cell activity, in patients treated by 'high-flux' membrane dialysis were also comparable to controls. The levels of serum beta 2-microglobulin were significantly elevated in patients-maintained on 'high-flux' dialysis membranes but did not reach the levels seen in patients dialyzed by cuprophane membranes. The beta 2-microglobulin at levels seen in patients on cuprophane dialysis had no effects on activation and proliferation of control lymphocytes in vitro. These results suggest that impaired functional responses of T lymphocytes seen in end-stage disease patients on prolonged hemodialysis with cuprophane membranes are not seen in similar patients hemodialyzed with polysulfone membranes.

Typhoid fever in Hong Kong children.

Experience with typhoid fever in 111 children over a 5-year period was reviewed. There were 66 boys and 45 girls, ranging in age from 1 to 11.5 years. The symptoms of typhoid fever were quite non-specific. Fever was the most common presenting symptom (in 98.3%). Other common presenting features were diarrhoea (25.7%), constipation (22%), vomiting (21.1%), cough (25%), abdominal pain (27.5%), headache (9.2%), epistaxis, meningism and convulsions. Rose spots were detected in 20% of cases, occurring mainly during the first 2 weeks of illness. Significant Widal reactions were present in 84.7% of cases. Blood and stool cultures were positive in 57% and 44% of cases, respectively. Peripheral blood white cell counts were not found to be of great diagnostic value. Chloramphenicol remained the drug of choice in the treatment of typhoid fever. It was more effective than ampicillin or co-trimoxazole. Complications were uncommon, occurring in only two patients. There were two deaths; both were admitted late and in moribund state. Early diagnosis and treatment is vital in typhoid fever and, as the presenting features are non-specific, a high index of suspicion is required.

The potentiation of the antitumor activity but not toxicity of bleomycin by 3-aminobenzamide.

Our previous studies have demonstrated that 3-aminobenzamide (3AB), an inhibitor of adenosine diphosphate-ribosyl transferase (ADPRT) could enhance the cytotoxicity (in vitro) and antitumor activity (in vivo) of bleomycin (BLM) A5 and peplomycin (PEP) against S-180, hepatoma and Ehrlich ascites carcinoma (EAC). In this study, it was shown that the inhibition rates (INR's) of S-180 in two experiments were increased from 42.5 and 46.1% to 66.2 and 75.9% when BLM 2.5 mg/kg/day x 8 was combined with 3AB 385.4 mg/kg/day x 8, while the decrease of body weight could not be enhanced. BLM at a dose of 5 mg/kg/day x 8 gave INR's of 64.8 and 75%, similar to the combined group but decreased the body weight more significantly. However, the addition of 3AB 385.4 mg/kg/day to BLM did not increase the acute toxicity of BLM alone. There was no significant difference of change of the body weight and subacute toxicity between the BLM and BLM + 3AB group. There was no difference of peripheral blood white cell count and the pathomorphological and ultrastructural change, wet weight and hydroxyproline content (to reflect the collagen content) of the lung of the mice between BLM alone and BLM + 3AB group. Therefore, the study provided experimental evidences for the reasonable use of nontoxic ADPRT inhibitors in adjunct to the chemotherapy of BLM in cancers.

Immune competence in 90Sr-exposed, adult thymectomized and antilymphocyteglobulin-treated CBA mice. I. Allogenic skin graft reaction.

CBA mice subjected to either adult thymectomy, internal exposure to 90Sr or antilymphocyteglobulin treatment separately, or to combinations of the three were tested for cellular immune competence using their reaction to allogenic skin grafts. Peripheral blood white cell counts did not reveal any obvious correlation between the degree of mononuclear cell depletion and the ability to accept grafts, suggesting that the particular treatments depleted specific fractions of mononuclear cells, differing in their extent of involvement in the rejection process. No single treatment alone induced a significant prolongation in the time elapsed before graft rejection. Adult thymectomy followed by appropriate antilymphocyteglobulin treatment induced severe lymphocytopenia and a profound suppression of the cell-mediated immune system, as evidenced by the acceptance of allogenic skin grafts. When applied to 90Sr-preexposed mice the same treatment induced lifelong acceptance of grafts, indicating a similar, though weaker immunosuppressive impact of 90Sr. Hence it was possible to significantly enhance immunosuppression in 90Sr-exposed mice. This in vivo model should be useful when investigating the role of immunological responsiveness in radiation carcinogenesis.

Immune status in tuberculosis and response to treatment

Thirty hospitalized patients with newly diagnosed tuberculosis were studied prospectively with a range of in vitro and in vivo tests of immune function. Responses were compared with those of healthy controls matched for age, sex, ethnic group and diet. A series of metabolic and immunologic abnormalities was found, including evidence of undernutrition, anaemia, neutrophil leucocytosis, monocytosis, lymphopenia, hyperglobulinaemia and raised erythrocyte sedimentation rate. Some patients had accelerated, others diminished, cutaneous tuberculin hypersensitivity, and some had diminished mononuclear cell proliferative and lymphokine responses to tuberculin (purified protein derivative, PPD). The patients were not uniform in their responsiveness, but could be arranged within a spectrum which showed a relationship to crude bacillary excretion and response to treatment. 27% of patients were characterized by hypersensitivity, with normal in vitro cellular responses and skin tests to PPD, scanty bacillary excretion and rapid bacteriologic sputum conversion to negative cultures with treatment. In contrast, 30% of patients were relatively anergic with negative skin tests, reduced or absent in vitro cellular reactivity to PPD, moderate or heavy bacillary excretion and later (>4 weeks) bacteriologic sputum conversion. The remainder of the patients fell between these two groups. There were no correlations between cellular immunity on the one hand, and radiological extent of disease, levels of serum immunoglobulins, peripheral white cell counts or ESR on the other. In those patients followed throughout treatment, all the abnormalities with the exceptions of arm muscle circumference and serum albumin, reverted to the normal ranges established in the control group.

Prognosis of patients receiving intensive care for lifethreatening medical complications of haematological malignancy.

The mortality of patients admitted to intensive care units with haematological malignancy is high. A humane approach to the management of the critically ill as well as efficient use of limited resources requires careful selection of those patients who are most likely to benefit from intensive care. To delineate more accurately the factors influencing outcome in these patients the records of 60 consecutive admissions to the intensive care unit (37 male, 23 female) with haematological malignancy were reviewed retrospectively. Fifty patients were in acute respiratory failure, most commonly (34 patients) with a combination of pneumonia and septicaemic shock. The severity of the acute illness was assessed by the APACHE II (acute physiology and chronic health evaluation II) score and number of organ systems affected. Thirteen patients survived to leave hospital. The mortality of patients with haematological malignancy was consistently higher than predicted from a large validation study of APACHE II in a mixed population of critically ill patients. Moreover, no patient with an APACHE II score of greater than 26 survived. Mortality among the 22 patients with relapsed malignancy (21 deaths), was significantly higher than among the 35 patients at first presentation (26 deaths). On discharge from the intensive care unit all survivors had responded well to chemotherapy and had normal or raised peripheral white cell counts. They included seven patients who had recovered from leucopenia (white cell count less than 0.5 X 10(9)/l). In contrast, 36 of the 47 patients who died were leucopenic at the time of death. The overall mortality of critically ill patients with haematological malignancy is higher than equivalently ill patients without cancer. The dysfunction of an increasing number of organ systems, an APACHE II score of greater than 30, failure of the malignancy to respond to chemotherapy, and persistent leucopenia all point to a poor outcome.

Oral Idarubicin in Combination with Cytosine-Arabinoside in Previously Untreated Patients with Acute Non-lymphocytic Leukemia

Twenty-six previously untreated patients with acute non-lymphocytic leukemia (ANLL) were treated with oral idarubicin and cytosine-arabinoside (Ara-C). The median age of the patients was 44 years (range, 11-72). In 23 of the 26 patients a hypoplastic marrow, with a peripheral white cell count of less than 1,000/mm3 after treatment, was documented. Treatment was well tolerated with minimal symptoms of nausea and vomiting. Diarrhea was observed in three patients and stomatitis in nine patients. Alopecia was documented in only six patients. A complete remission (CR) was obtained in 12 patients (median duration 25 weeks). The median time to CR was 3.4 weeks (range, 1.4-5). Ten of the 26 patients were alive 6 months after the start of induction treatment, while a further four patients who were in the study for less than 6 months are alive and in remission at 5, 4, 3, and 3 months, respectively. Eight of 12 patients in whom bone marrow aplasia was documented achieved a CR; perhaps the drug dosages used in this study were suboptimal.

In vivo modulation of glutathione by buthionine sulfoximine: Effect on marrow response to melphalan

The effect of giving buthionine sulfoximine (BSO), 0.0265 g/mouse (6 mM), at 12 and 6 hr before treatment with melphalan-0.0 mg, 3 mg, 6 mg, and 9 mg/kg, was studied in C3H mice, and was compared with control groups that received normal saline 12 and 6 hr before identical melphalan treatment. BSO treatment resulted in depletion of GSH levels in bone marrow, liver, and muscle to 65, 13, and 41% of control levels, respectively. Hematological toxicity was assessed by measurement of CFU-S survival and peripheral white cell counts. CFU-S survival decreased with increasing doses of melphalan, but no difference was observed with BSO pre-treatment. Likewise, WBC counts following melphalan 9 mg/kg, were similar irrespective of BSO pre-treatment. These data suggest that the marrow toxicity seen with melphalan is not worsened by pre-treatment with BSO and that if tumors can be pre-sensitized with BSO, there may be a clinical role for melphalan/BSO drug combination.

Intensive therapy for life-threatening medical complications of haematological malignancy.

The medical records of 22 consecutive adult admissions to an intensive care unit (ICU) with life-threatening complications of haematological malignancy, or its treatment, are reviewed. Twenty patients (91%) were in acute respiratory failure, and 17 of the 22 patients required intermittent positive pressure ventilation (IPPV). The in-unit mortality was 55%, but only 4 patients (18%) survived to leave the hospital. Although the unit mortality appeared to be related to the acute physiology score (APS), this small series did not demonstrate a clear relationship between the APS and long-term survival (discharge from hospital). There were, however, significant differences in the number of organ systems involved between those who died on the ICU and those who returned to the ward, as well as between those who survived to leave hospital and those who died. No patient with more than three systems involved became a long-term survivor. All long-term survivors had either reasonable peripheral white cell counts throughout or their bone marrow showed significant recovery prior to discharge from the ICU. Unresponsive malignant disease and a failure to recover bone marrow function were major factors in those patients who died shortly after discharge from the ICU. Although long-term survival rates are low and are probably largely determined by the progress of the underlying malignancy, intensive care was life-saving in four patients, three of whom are alive several years after discharge.

Bronchoalveolar lavage in ponies with recurrent airway obstruction (heaves).

We performed bronchoalveolar lavage (BAL) in 2 groups of ponies. Principal ponies had a history of heaves, a disease characterized by recurrent airway obstruction and airway hyperreactivity when ponies are housed in a barn and fed hay; control ponies had no history of airway obstruction. Ponies were paired (principal and control), and BAL was performed after 2 months of being pastured when principal ponies were in clinical remission (Period A), after barn housing when principal ponies had acute airway obstruction (Period B), and after a 1- and a 2-wk recovery phase of pasture grazing (Periods C and D). At Periods A, C, and D, white blood cell counts and immunoglobulin/albumin ratios in peripheral blood and BAL fluid of principal and control animals did not differ. Peripheral blood white cell counts and immunoglobulin/albumin ratios were unaffected by barn exposure and return to pasture. However, at Period B, neutrophil numbers in the BAL fluid were increased in the principal but not in the control animals. The IgG/albumin ratio of the principal animals increased at Period C. We conclude that in ponies with a history of heaves, barn exposure results in increased neutrophils in BAL fluid. In this pony model of lung disease, pulmonary leukostasis follows a time course similar to that of airway obstruction and airway hyperreactivity.

The gastro-duodenal epithelium in peptic ulceration.

Mucosa-related bacteria, intra-epithelial lymphocytes and intra-epithelial polymorphonuclear leucocytes have been studied in 35 patients with duodenal ulceration, 27 patients with gastric ulceration and eight control subjects with normal gastro-duodenal mucosa. Mucosa-related bacteria were found in approximately 80 per cent of peptic ulcer patients and rarely in controls. The bacteria were most numerous at the sites of active chronic gastritis. There was a positive correlation between the number of bacteria and the number of intra-epithelial polymorphonuclear leucocytes. There was no correlation between the peripheral blood white cell count and the number of intra-epithelial polymorphonuclear leucocytes. The number of intra-epithelial lymphocytes was increased in peptic ulceration.

Physiologic consequences of status epilepticus.

Prolonged seizures produce central nervous system damage. Physiologic consequences of status epilepticus may exacerbate this damage or may mislead the physician into making inappropriate therapeutic decisions. Status results in an elevation of body temperature, an increase in the peripheral white cell count, and often a transient pleocytosis in the spinal fluid. A marked metabolic acidosis occurs routinely. Prominent elevations in plasma hormonal concentrations occur as well. Epinephrine levels are in the arrhythmogenic range and could play a role in sudden death. Transient but marked pressure responses occur in the systemic and pulmonary circulations. Pulmonary edema may result from these pressure transients.

Antitumor activity with nontoxic doses of protein A.

This report confirms our previous observation that IV inoculation of purified protein A causes regression of rat mammary adenocarcinomas. In treated tumors, we have obtained histological evidence of changes indicating tumor cell destruction. Protein A treatment does not cause reduction in the body weight or organ weights of rats; nor does it cause any decrease in activity of the enzymes of the microsomal mixed function oxidase system in the liver. Protein A stimulates peripheral white cell counts in normal rats, but not in tumor-bearing rats. We found that protein A infusion reduced (P less than 0.0005) the level of circulating plasma immune complex concentration. A homing study with 125I-labeled protein A indicated that liver, spleen, and kidney tissues are the major sites of protein A accumulation. Therefore, protein A seemed to exert its antitumor effects without causing any generalized toxicity to the system. It is postulated that the action of protein A may be related to its ability to cause a drastic reduction in circulating plasma immune complex concentration, thus potentiating the immune reactivity of the host observed earlier.

Haematology of subclinical fatty liver in dairy cows

The relationship between liver fat content and haematology was investigated in 369 cows from eight herds sampled in the second week after calving. High levels of fat in the liver were associated with a depression in total white cell count and in neutrophils, eosinophils and lymphocytes. There was no correlation between liver fat content and the percentage of E rosetting lymphocytes, packed cell volume or haemoglobin concentration. The changes in peripheral white cell counts may be related to the increased incidence of post parturient disease in cows with fatty liver.

Nitrous oxide reduces growth of experimental rat leukemia

The ability of nitrous oxide to inhibit the in vivo growth of hematological neoplasms was investigated in a rat model for acute myeloid leukemia (BNML). Nitrous oxide, administered in a concentration of 67% with 33% oxygen, resulted in a reduction of spleen and liver weights of approx. 30%, as compared with leukemic rats kept in ambient air. Peripheral white cell counts were also considerably lower in the treated rats. Plasma levels of vitamin B 12 were found to be elevated in untreated leukemia, but fell to about normal levels after nitrous oxide exposure. On the contrary, folic acid levels were low in untreated leukemic rats, and significantly higher in animals exposed to nitrous oxide. The observed effects of nitrous oxide appeared to be dose-dependent. The deoxyuridine suppression test performed with leukemic cells became abnormal after nitrous oxide inhalation, in accordance with the effect on normal bone marrow. These results indicate that the interference of nitrous oxide with vitamin B 12-related metabolism, which leads to impairment of de novo thymidine synthesis, has the potency to reduce leukemic proliferation in vivo.