Marginal Pool Research Articles

The toxicologic evaluation of marcellomycin--an antineoplastic anthracycline antibiotic.

Dose-related toxicologic effects of marcellomycin, an antineoplastic anthracycline antibiotic, were observed in single-dose studies in mice iv (43.05-67.65 mg/m2) and dogs iv (41.0-90.2 mg/m2), and in multiple-dose studies in dogs iv (9.8-29.6 mg/m2 2X/week for 6 weeks) and rats sc (9 weekly doses at 26.2-72.2 mg/m2). Toxicity was primarily manifested by suppression of myeloid tissue, especially the erythrocytic and thrombocytic series, and lymphoid tissues. Initially a neutrophilic leukocytosis was observed in dogs and rats, which was considered possibly to be due to mobilization of the marginal and bone marrow neutrophil pools. In dogs, this was followed by a marked, dose-related neutropenia; and, in rats that died, there was marked depletion of bone marrow cells. Other toxicities observed included enteropathy, severe subcutaneous serofibrinous inflammatory edema and necrosis at injection sites, prostate and seminal vesicle atrophy, uterine hypoplasia, testicular and pancreatic degeneration, thyroid follicular proliferation and hemorrhage in various organs. In general, these toxicities were reversible in surviving animals during recovery periods. Significant cardiotoxicity was not demonstrated.

Effect of pulmonary blood flow on the exchange between the circulating and marginating pool of polymorphonuclear leukocytes in dog lungs.

The effect of pulmonary blood flow on the exchange between the circulating was marginating pool of polymorphonuclear leukocytes (PMN) was examined in three sets of experiments. In the first we used the double indicator dilution technique with labeled PMN and erythrocytes (RBC) to calculate the percent extraction and percent recovery of PMN at different levels of cardiac output (CO). In the second group of experiments we took advantage of the wide range of blood flow in the lung to determine the effect of regional blood flow on regional PMN retention, and in the third set we measured total leukocyte (WBC) and PMN counts in simultaneous samples from the pulmonary artery and aorta over a wide range of cardiac output. The studies showed that 80-90% of the labeled PMN were removed in a single pass through the lung and that regional retention of labeled PMN and A-V differences for unlabeled PMN increased with decreasing blood flow. The data for regional retention of labeled PMN and the A-V differences observed for unlabeled cells both fit the equation Y = A + Be-cx (where A + B = 100), which showed that PMN accumulate in the lung as blood flow is reduced. We conclude that a dynamic equilibrium exists between the circulating and marginating pools of leukocytes in the lung and that blood flow primarily effects the reentry of cells into the circulating pool so that the marginating pool of PMN within the lung accumulates cells when blood flow is reduced below 7 ml/min per g.

Effect of physical effort on the white blood cells in benign familial leukopenia.

The components of the peripheral white blood cells (WBC) were determined before and after 10 min of submaximal ergometric work in two groups of subjects: 34 healthy Yemenite Jews with benign familial leukopenia (BFL) and 34 healthy Yemenite Jews without BFL. The mean +/- SEM of the increase in the peripheral WBC following the effort in the control group was 2,606 +/- 1,272 cells/mm3, due to the relative rise in the neutrophils, eosinophils, monocytes and lymphocytes. The identical ergometric work, performed by BFL subjects, brought about significantly lower increase (700 +/- 865 cells/mm3) in the peripheral WBC, the majority of which were lymphocytes. It seems that subjects with this form of BFL lack granulocytes in the marginal pool.

Effect of Hypertransfusion on Bone Marrow Regeneration in Sublethally Irradiated Mice. I. Enhanced Granulopoietic Recovery

AbstractHypertransfusion can enhance recovery from neutropenia in certain clinical and experimental situations. We have studied the pattern of myeloid recovery in mice hypertransfused after receiving 350 rads whole body irradiation. Both hypertransfused and control groups showed the degenerative phase, abortive rise, and regenerative phase that has been described following sublethal irradiation. The blood granulocyte counts in the hypertransfused group returned to normal more rapidly and were maintained at a significantly higher level during the regenerative phase. This difference is not the result of a shift in granulocytes from the marrow granulocyte reserve or marginal granulocyte pool to the circulating pool, but is associated with significantly enhanced bone marrow granulopoiesis. While the total bone marrow cellularity of the hypertransfused mice is less than that of the control mice, the hypertransfused group contains more CFU-GM and myeloid cells during the regenerative phase. The enhanced granulopoiesis is not due to increased colony-stimulating activity (CSA) levels in the hypertransfused mice, as the CSA levels were significantly lower in this group compared to the controls prior to and during the initial phase of granulopoietic recovery. This study suggests that hypertransfusion increases the rate of recovery of myelopoiesis by increasing the number of precursors available for myeloid differentiation from an earlier stem cell compartment.

Effect of hypertransfusion on bone marrow regeneration in sublethally irradiated mice. I. enhanced granulopoietic recovery

Hypertransfusion can enhance recovery from neutropenia in certain clinical and experimental situations. We have studied the pattern of myeloid recovery in mice hypertransfused after receiving 350 rads whole body irradiation. Both hypertransfused and control groups showed the degenerative phase, abortive rise, and regenerative phase that has been described following sublethal irradiation. The blood granulocyte counts in the hypertransfused group returned to normal more rapidly and were maintained at a significantly higher level during the regenerative phase. This difference is not the result of a shift in granulocytes from the marrow granulocyte reserve or marginal granulocyte pool to the circulating pool, but is associated with significantly enhanced bone marrow granulopoiesis. While the total bone marrow cellularity of the hypertransfused mice is less than that of the control mice, the hypertransfused group contains more CFU-GM and myeloid cells during the regenerative phase. The enhanced granulopoiesis is not due to increased colony-stimulating activity (CSA) levels in the hypertransfused mice, as the CSA levels were significantly lower in this group compared to the controls prior to and during the initial phase of granulopoietic recovery. This study suggests that hypertransfusion increases the rate of recovery of myelopoiesis by increasing the number of precursors available for myeloid differentiation from an earlier stem cell compartment.

Inhibition of granulocyte adherence by lithium: possible relationship to lithium-induced leukocytosis.

Many patients receiving lithium carbonate therapy for psychiatric disorders develop increased neutrophil counts in their peripheral blood (O’Connell, 1970; Shopsin et al., 1971). The mechanism for this neutrophilia is thought to involve the stimulation of increased neutrophil production by the marrow. Addition of lithium carbonate in therapeutic concentrations to soft agar cultures of bone marrow promotes granulocyte colony growth (Tisman et al., 1973) and this promotion has been shown to result from the production of increased colony-stimulating activity (Harker et al., 1977). A recent study of granulocyte kinetics in patients treated with lithium has shown that they develop an increased total blood granulocyte pool, an increased marginal pool, and accelerated granulocyte production (Rothstein et al., 1978). Thus, the mechanism for lithium-induced neutrophilia appears to be the stimulation of increased marrow production, with a resultant increase in total intravascular neutrophil pool.KeywordsLithium ConcentrationNylon FiberPlasma FactorSerum Lithium ConcentrationIncrease Neutrophil CountThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Disparity between circulating and marginated neutrophils: evidence from studies on the granulocyte alkaline phosphatase, a marker of cell maturity.

Since circulating and marginated human granulocytes are in rapid kinetic equilibrium, cells of these two compartments have been considered to be a homogeneous population. Our studies on the relationship between neutrophil maturity and the granulocyte alkaline phosphatase (GAP) activity cast doubt on this assumption. After iv administration of hydrocortisone, 12 male volunteers showed an augmentation in circulating granulocytes of 5730 cells/mm3, accompanied by an increase of band neutrophils from 18% to 33% (p less than 0.001). During this influx phase, the GAP activity decreased by 73% when measured cytochemically and by 28% when assayed biochemically (p less than 0.001 and less than 0.01, respectively). When granulocytes were demarginated by epinephrine, the mean count increased by 38%, accompanied by a rise in the portion of segmented forms from 74% to 79% (p less than 0.005) and by an increase of the cytochemical GAP activity by 24% (p less than 0.01). Exact complementary results were obtained during an excessive transient margination, the hemodialysis neutropenia: bands increased from 24% to 54% (p less than 0.02), while the cytochemical GAP dropped by 40% (p less than 0.005). Thus, our analysis of three situations with an acute transient shift of granulocytes indicates that functionally or chronologially "older" cells have a higher GAP activity, and that the transfer of granulocyte from the circulating to the marginal pool is selective.

Neutropenia, Inflammation, and the Kinetics of Transfused Neutrophils in Rabbits

A rabbit model was used to study the effects of neutropenia and inflammation on the intravascular distribution, survival, and tissue accumulation of transfused neutrophils. Donor blood labeled with [(3)H]thymidine was infused into normal or neutropenic (vinblastine treated) animals. Inflammation was created by subcutaneous implantation of polyvinyl sponges, some with added endotoxin. Initial circulating neutrophil pool recovery, survival, and inflammatory site accumulation of labeled neutrophils were measured. Neutropenia was associated with a relative increase in the marginal pool size, manifested by a diminished initial circulating pool (CNP) recovery of transfused cells. The CNP recovery was directly proportional to recipient neutrophil count. Neutropenia had no effect on the intravascular survival of transfused cells and was accompanied by only a modest decrease in the inflammatory site recovery of the transfused neutrophils (10.4+/-5.4 vs. 14.4+/-4.0% in normals). Inflammation in the form of subcutaneous polyvinyl sponges was accompanied by an increase in margination with initial CNP recoveries of 24.3+/-4.7 and 27.6+/-8.8% at zero and 4 h after implantation respectively (normal, 38.2+/-9.9%). Transit through the CNP was hastened by inflammation with a t((1/2)) of 2.02+/-0.72 h (normal, 3.2+/-1.0 h). Addition of endotoxin to the sponges further perturbed cell kinetics. CNP recoveries were considerably lower and half-lifes were initially shorter and subsequently uninterpretable in studies done after endotoxin sponge insertion. Inflammatory site accumulation was markedly diminished to 7.4+/-1.9% of injected neutrophil label in the endotoxin sponge animals, suggesting that many of the transfused cells were functionally unavailable rather than marginated. These studies demonstrate that neutropenia and inflammation with or without endotoxin markedly alter the kinetics of transfused neutrophils and that CNP recovery of transfused cells is not necessarily predictive of their inflammatory site accumulation.

FUNCTIONAL CELL COMPARTMENTS IN A RAT MODEL FOR HUMAN ACUTE MYELOCYTIC LEUKAEMIA

Functional cell compartments were studied in a rat model for human acute myelocytic leukaemia (AML). This was done by tracing the distribution of injected 51Chromium-labelled leukaemic cells in the body. It was concluded that two functional compartments can be distinguished in acute leukaemia, i.e., a rapidly exchangeable pool of cells (including the circulating blood pool, the marginal noncirculating blood pool and the rapidly exchangeable tissue pool; RETP) and a slowly exchangeable tissue pool (SETP). The sizes of these various compartments were roughly quantified at various stages of the disease by calculations based on the principle of isotope dilution and organ weight measurements. As the leukaemia progresses, the size of the SETP increased significantly relative to the size of the RETP. Simultaneously, the exchange rates of leukaemic cells between the organs and the blood decrease. The blood transit time of leukaemic cells was also significantly prolonged, as is the case in human AML.

Correlation of splenomegaly and abnormal neutrophil pooling (margination).

Abstract Splenomegaly causes abnormal pooling of platelets and RBCs, but it has not been clear whether or not the same is true for neutrophils. We have used in vitro labeling of whole blood with DF 32 P to investigate the recovery of labeled neutrophils in circulation immediately after reinfusion into 19 patients with normal-sized to "moderately enlarged" spleens. Twelve patients were neutropenic, and 18 later had a splenectomy. Eleven of these 18 patients were studied by the same technique after splenectomy. Labeled neutrophils which were not recovered were interpreted as being in the "marginal pool." Correction factors for the increased blood volume in splenomegaly were applied. Significant correlations were found between spleen weight or spleen size by external palpation and the marginal pool size. These correlations were evident whether or not blood volume corrections were made. Five patients with moderately enlarged spleens, the largest in the study, had an average of 80% of intravascular neutrophils in the marginal pool as opposed to a normal average of 58%. Following splenectomy, the marginal pool proportion decreased to an average of 45%. Thus the abnormally large marginal pool in splenomegaly can be a significant pathophysiological cause of neutropenia. This cause can be identified by physical examination.

Effect of acute alcohol intoxication on granulocyte mobilization and kinetics

Granulocyte mobilization into skin abrasions in human volunteers was significantly inhibited by acute alcohol intoxication (45,-800 cells in 8 hr versus 353,000 in normal controls). Alcohol applied locally did not inhibit granulocyte delivery, and protection of the abrasion against heat loss did not reduce the inhibited delivery in intoxicated volunteers. Intoxication inhibited granulocyte adherence and local mobilization in parallel. Alcohol administration to rabbits shifted granulocytes from marginal to circulating pool in a manner similar to epinephrine. Mobilization of bone marrow granulocytes by glucocorticoid or endotoxin administration was not inhibited by intoxication, nor did it prevent the endotoxin-induced shift of granulocytes from circulating to marginal pool.

Effect of Acute Alcohol Intoxication on Granulocyte Mobilization and Kinetics

Granulocyte mobilization into skin abrasions in human volunteers was significantly inhibited by acute alcohol intoxication (45,-800 cells in 8 hr versus 353,000 in normal controls). Alcohol applied locally did not inhibit granulocyte delivery, and protection of the abrasion against heat loss did not reduce the inhibited delivery in intoxicated volunteers. Intoxication inhibited granulocyte adherence and local mobilization in parallel. Alcohol administration to rabbits shifted granulocytes from marginal to circulating pool in a manner similar to epinephrine. Mobilization of bone marrow granulocytes by glucocorticoid or endotoxin administration was not inhibited by intoxication, nor did it prevent the endotoxin-induced shift of granulocytes from circulating to marginal pool.

THE HABITATS AND HABITAT PREFERENCES OFCLARIAS GARIEPINUS(PISCES: CLARIIDAE) IN A CLEAR COASTAL LAKE (LAKE SIBAYA, SOUTH AFRICA)

SUMMARY The habitat preferences of the sharptooth catfish, Clarias gariepinus, in a clear coastal lake, Lake Sibaya, are described. Larval and small juvenile catfish inhabit flotsam and the rootstocks of plants in very shallow marginal areas. Larger juveniles inhabit densely-vegetated marginal pools, and may venture into open areas at night. Adult catfish (over 350 mm TL) occupy offshore areas, where their density is highest in terrace and sheltered bay habitats, and least in slope and profundal habitats. Adult catfish undergo a diel migration into shallow water at night and into deeper water during the day which is more marked at low lake levels. The methods used to capture catfish, and the main habitats in Lake Sibaya, are also described.

Leukocyte mobilization by epinephrine and hydrocortisone in patients with chronic renal failure.

The mobilization of WBC from the bone marrow, as judged from hydrocortisone-induced leucocytosis, is markedly impaired in dialyzed and nondialyzed uremic patients. The release of WBC from the marginal pool by epinephrine was found to be normal.

Neutrophil leukocytes and inflammation of the bovine mammary gland

Observations on the production, distribution, structure, and function of neutrophils is briefly reviewed and related to the role of neutrophils in inflammation of the bovine mammary gland. The mature neutrophil is an end stage cell possessing an armament of two distinct types of granules containing a variety of enzymatic and non-enzymatic constituents. Neutrophils are produced in the bone marrow by an orderly process of mitosis and maturation. The developing neutrophils form a morphologic continuum with functional compartmentalization into mitotic, maturative and reserve pools. Within the blood vasculature, neutrophils are either freely circulating or remain marginated along vessel walls in capillary beds, particularly in the lungs, liver and spleen. However, neutrophils so distributed are in a dynamic equilibrium, and after a brief sojourn in blood, they migrate into body cavities and tissues constituting a physiologic barrier to microbial invasion. Neutrophils from the marrow reserve and marginal pools rapidly and preferentially mobilize into areas of acute inflammation in response to chemotactic factors generated locally. There, they phagocytize, kill, and digest bacteria in a complex but sophisticated manner utilizing energy and granular constituents. During this process neutrophils also release certain agents which by themselves are inflammatory. Studies in developing calves have shown that (a), there is a sizable marrow reserve of neutrophils, (b) newly formed neutrophils appear in blood in about 7 days, (c) the circulating and marginal pools of neutrophils are about equal in size, and (d) neutrophils disappear from blood randomly with a half-time of about 9 hours. Such information on mature cows is not available. Experimental studies on mastitis in normal and neutropenic cows have demonstrated the dual role of neutrophils also in this species viz. , in protection against mammary infections and as incitors of the inflammatory reaction. Neutrophils rapidly mobilize into the mammary parenchyma and subsequently into the milk in glands experiencing acute inflammatory reaction and continue to emigrate for a variable period after disappearance of clinical signs of mastitis. While much has become known about neutrophil chemotaxis, phagocytosis, and bacterial killing in other species, little is known about bovine neutrophils in this regard and, particularly, in relation to mastitis. The bovine mammary gland is an excellent model to study inflammation and the cow has a remarkable ability to adapt to a heavy daily loss of neutrophils in milk.

Intravascular fate of granulocytes administered by granujlocyte transfusions.

Granulocytes were harvested from hematologically normal individuals using continuous flow centrifugation (CFC) or filtration leukapheresis (FL). The isolated granulocytes were labeled in vitro by 3H-diisopropyl fluorophosphate (3H-DFP) and autotransfused. Their intravascular fate was analyzed by autoradiography. Immediately after autotransfusion the majority of granulocytes administered, collected in the marginal granulocyte pool. Margination was particularly prominent in granulocytes isolated by FL. The distribution of transfused granulocytes between the circulating and the marginal granulocyte pool showed wide and irregular fluctuations in time. Margination of transfused granulocytes was counterbalanced, and its fluctuation between the two intravascular pools was stabilized by prednisone treatment. The transit of transfused granulocytes from blood to tissue seemed to be governed by a random process, the half-disappearance time being either normal or prolonged. Compatible granulocytes administered to hematologically normal recipients circulated for at least 20 h.

Neutrophil leukocyte reserves in lymphoreticular malignancies.

The marginal neutrophil leukocyte pool and the bone marrow neutrophil reserves were assessed in 43 patients with lymphoreticular malignancies, using epinephrine and steroid stimulation. Thirty-three patients were studied before staging, following splenectomy, before and after nodal irradiation and during remission induction chemotherapy. Ten patients in partial remission were studied after 12 month or more of chemotherapy. A neutrophil count of 1,500-2,500 cells/mm3 is not a sufficient guide for midifying the chemotherapy. The response to steroid stimulation may be a more dependable guide to evaluate the patient's potential risk for infection.

Studies of the neutropenia in kala-azar: results in two patients

Two patients with kala-azar were studied, one with DF 32P (diisopropylfluorophosphate) and one with 51Cr (chromate), in an attempt to elucidate the mechanisms producing neutropenia in this disease. The granulocyte half-life was found to be reduced in both patients, with pooling and probable destruction occurring in the spleen and, to a lesser extent, in the liver. Bone marrow neutrophil reserve, estimated by the response to intravenous hydrocortisone hemisuccinate, was found to be markedly reduced in both patients. An enlarged marginal granulocyte pool in one patient indicated that the neutropenia may also be due to altered intravascular granulocyte distribution.

Metabolic and functional studies of monocytes in congenital neutropenia.

Summary. Children with the severe form of congenital neutropenia usually die from infection by 2 yr of age. Metabolic and functional studies of monocytes from three patients with this disease were undertaken to understand further the role of monocytes in the host's defences against bacterial infection. The bactericidal capacity of monocytes compared favourably to the bactericidal capacity of control neutrophils (PMN). Metabolic studies of monocytes and control leucocytes showed similar stimulation of oxygen consumption, hexose monophosphate pathway activity arid 125iodide fixation during phagocytosis. Further, myeloperoxidase (MPO) activity of patients’monocytes was comparable to the activity of control leucocytes. Using the Rebuck skin window, very few of the patients’mononuclcar cells were seen to migrate during the initial 2–4 hr of observation. During this time, the PMN migration of controls was maximal. Following the relatively acellular period observed in patients, a brisk mononuclear cell infiltration occurred apparently independent of the presence of neutrophils. Epinephrine injection into patients stimulated a marked increase of blood monocytes, suggesting the existence of a marginal monocyte pool. An injection of hydrocortisone caused an unexplained 3–8‐fold decrease in circulating monocytes. The delay of phagocytic cell migration to an inflammatory site, and the apparent sensitivity of monocytes to hydrocortisone may account in part for the increased susceptibility to infection in patients with severe congenital neutropenia.

Neutrophil and Monocyte Kinetics in a Case of Cyclic Neutropenia

Abstract The kinetics of neutrophilic granulocytes and monocytes were examined simultaneously in a patient suffering from cyclic neutropenia. During the remission phase of the disease the observed kinetic data for both cell types did not deviate significantly from the normal range. At a stage just before the minimal blood granulocyte count was reached, the total blood granulocyte pool was diminished and the granulocyte half disappearance time shortened to 2.3 hr. Moreover, the intravascular granulocyte distribution changed in favor of the marginal granulocyte pool. During the neutropenic phase, the intravascular monocyte pools were Increased and monocyte half-disappearance time was prolonged to 14 hr; the monocyte turnover rate was 2.4-fold higher than during remission.