Buffy Coat Preparations Research Articles

Platelet to Leukocyte Adherence Phenomena Associated With Thrombocytopenia

Two patients developed thrombocytopenia associated with an in vitro platelet to leukocyte adherence phenomenon (PLAP). In one patient, whole serum, intact plasma, and the IgG fraction of the serum could induce the PLAP of normal platelets in normal whole blood or buffy coat preparations. Passage of serum over a Sepharose column coupled with antihuman IgG resulted in the loss of all the PLAP activity. Activity could not be recovered from the whole serum or the IgG fraction of serum of the second patient; however, the PLAP was transferred to her newborn infant. In both patients, an unusual pulmonary illness accompanied the PLAP and thrombocytopenia.

Carcinocythemia (carcinoma cell leukemia): An acute leukemia-like picture due to metastatic carcinoma cells

Observation of a unique population of cells on a Wright-stained blood smear of a patient with metastatic breast carcinoma prompted a study to determine their origin. The primary carcinoma contained a marker, the presence of “signet celte.” These were demonstrated in direct peripheral smears and buffy coat preparation of peripheral blood and confirmed histochemically by showing positive periodic acid-Schiff, alpha-napthol and beta-glucuronidase reactions. “Carcinocythemia” is suggested as a name for this unusual process observed over a six month period. Studies of the patient's immunocompetence, of circulating cell surface immunoglobulins and karyotype analysis were made. Postmortem examination revealed retroperitoneal fibrosis, splenic atrophy and extensive metastatic carcinoma but no evidence of leukemia. The cells will be contrasted to those seen in a second patient who appeared to have acute myelocytic leukemia complicating extensive cancer involving the bone marrow. The observations suggest that a leukemia-like blood picture due to circulating cancer cells may occur during the course of metastatic breast carcinoma.

The significance of abnormal circulating cells in patients with Hodgkin's disease.

Buffy coat preparations of peripheral blood from two patients with Stage IVB Hodgkin's disease, 33 patients with Hodgkin's disease undergoing complete staging including laparotomy, 12 normal controls and eight patients with viral upper respiratory infections were examined for the presence of abnormal cells. Primitive histiocytic cells were seen in the two patients with IVB disease and in two patients with IIIB disease all of whom proved to be refractory to chemotherapy. Reed-Sternberg cells were seen in one IVB patient. Large cells with basophilic, agranular cytoplasm and moderately convoluted nuclei often with a perinuclear halo and nucleoli were detected in 30 of 35 patients with Hodgkin's disease, five of eight patients with viral infections but in none of 12 normal controls. Other abnormal large cells with round nuclei, occasional prominent nucleoli, grey to lightly eosinophilic cytoplasm and diffuse granulation were seen almost exclusively in Hodgkin's disease. There was no correlation between histology, stage, splenic involvement, skin test positivity and response to therapy with the presence or frequency of either of these cell types. This observation suggests that these cells are more likely to be 'reactive' rather than malignant. It is unusual for abnormal histiocytic cells to circulate in Hodgkin's disease and their presence may be a poor prognostic sign. The presence of other types of large atypical cells is not indicative of either haematogenous spread of Hodgkin's disease or of poor prognosis after treatment with radiotherapy alone.

Morphological changes of basophils in immunological reactions--effect of sodium cromoglycate.

The morphological changes in blood basophils, from patients with bronchial asthma, were studied in vitro during immunological reactions, with particular reference to interaction with anti-IgE. Anti-human IgE results in an absolute reduction in basophil numbers in buffy coat preparations as well as basophil cytoplasmic degranulation. An increase in the number of pear-shaped (A-form) basophils is also observed. Sodium cromoglycate impairs all these anti-IgE induced basophil changes, providing further insight into the mechanism of action of this compound in bronchial asthma.

The mechanism of the entry of dye into neutrophils in the nitroblue tetrazolium (NBT) test.

1. Washed buffy-coat preparations of human blood suspended in different media were exposed to the Nitroblue Tetrazolium (NBT) and the mechanism of entry of the dye into neutrophil polymorphonuclear leucocytes was investigated. 2. Stimulation of neutrophils with endotoxin and the presence of heparin and/or fibrinogen in the suspending medium were necessary for dye reduction. 3. NBT complexed with and precipitated heparin and/or fibrinogen from solution. The percentage of cells reducing the dye and the degree of precipitation of fibrinogen and heparin depended on the concentration of dye; the critical concentration of NBT necessary for each effect was the same. 4. Electron microscopy of stimulated neutrophils revealed the presence of amorphous material, which was probably complexed NBT, outside the cell, in the process of endocytosis and within membrane-bound vacuoles within the cytoplasm. 5. Neither complement nor immunoglobulins were obligatory for dye reduction in this system. 6. It is suggested that NBT only enters neutrophils in quantities visible by light microscopy, after stimulation which produces phagocytosis of a macromolecular complex of the dye and heparin and/or fibrinogen.

Nucleoside phosphorylase activity in blood vessels and formed elements of the blood of the dog.

The distribution of nucleoside phosphorylase activity was investigated in dog coronary blood vessels and in buffy coat preparations of circulating blood cells of the dog using a histochemical method. The enzyme activity was found in the cytoplasmic sap of endothelial cells, adventitial fibroblasts and fibrocytes, and in the axonal cytoplasm of the nerves. Smooth muscle and macrophages revealed no enzyme activity. Of the circulating blood cells, neutrophils, basophils, monocytes and platelets showed a uniform enzyme reaction, whereas eosinophils and erythrocytes apparently lacked activity. A part of the lymphocyte population reacted strongly while some lymphocytes were completely devoid of activity.

A direct micromethod for the preparation of blood platelets and thrombocytes for electron microscopy

A micromethod for the preparation of small volumes of whole blood (down to .05ml) for ultrastructural studies is described. It uses standard haematological equipment with minimal centrifugation, and by employing immediate and brief pre-fixation in glutaraldehyde, avoids the use of anticoagulants, thus ensuring preservation of undistorted morphology and ultrastructure in all cell types. A large number of small compact buffy coat preparations can be produced rapidly and consistently, in which stratification is maintained during subsequent processing; this allows examination of the stratification at either the light or electron microscope level. The method makes haematological studies in small animals possible. Platelets and thrombocytes from a wide range of mammalian and avian species, having large or small blood volumes, have been examined.

Circulating iron-containing macrophages in hemochromatosis.

Iron-containing macrophages, present in various tissues in patients with iron-storage diseases, have been demonstrated to pass from the intestinal villus into the intestinal lumen, acting to transport iron as well as to store it. It was proposed that these cells may transport iron from one organ to another and thus might be present in the circulating blood. Therefore, they were sought in buffy-coat preparations of venous blood in normal subjects and in patients with hemochromatosis and other iron-storage conditions such as transfusion siderosis. Circulating hemosiderin-containing cells were demonstrated in eight of the nine untreated patients with hemochromatosis and in four of 20 with other iron-storage disorders. None were found in normal subjects. The discovery of such cells in the circulating blood suggests an iron-storage disease although their absence does not exclude such a possibility. The macrophages in hemochromatosis appear different from those in transfusion siderosis, containing smaller granules of iron.

Demonstration of L.E. cells in pericardial fluid; report of a case.

The value of the so-called lupus erythematosus (L. E.) inclusion phenomenon of bone marrow and blood as an aid in the diagnosis of acute disseminated lupus erythematosus has been amply demonstrated.1It is the purpose of this paper to report a case of pericardial effusion due to acute disseminated lupus erythematosus in which the diagnosis was first intimated by the finding of L. E. cells in the Wright-stained buffy coat smears of the patient's oxalated pericardial fluid. Subsequently, large numbers of L. E. cells were demonstrated in buffy coat preparations made from an oxalated marrow specimen from the same patient. Response to cortisone therapy was striking and led to rapid resorption of the pericardial effusion. REPORT OF A CASE A 38-year-old white woman was well until May, 1950, at which time pain developed in the right thigh, with subsequent pain and swelling in the right leg and foot. In