A Simple and Efficient Method of Separating Peripheral-Blood Leucocytes for in-Vitro Studies

Hulliger, Lotte; Blazkovec, A.A.

doi:10.1016/s0140-6736(67)91596-6

Cited by 58 publications

(18 citation statements)

References 2 publications

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“…preservative-free heparin (Weddel Pharmaceuticals Ltd, London). Excess erythrocytes wNere removed by layering these samples over methyl-cellulose/triosil (Hullinger and Blaztiovec, 1967) and allowing them to sediment at room temperature for 30-50 min. The leucocyte-rich upper layer was collected, the leucocytes washed once, resuspended in collecting medium and a nucleated-cell count performed.…”

Section: Methodsmentioning

confidence: 99%

Changes in granulopoiesis detected by in vitro colony formation in acute lymphatic leukaemia

Morris¹,

Butler²,

Muldrew³

et al. 1977

Br J Cancer

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Summary.-Patients with acute lymphatic leukaemia (ALL) could be divided into two groups at diagnosis-those whose peripheral blood and/or bone marrow exhibited in vitro colony formation and those in whom it did not, but this finding did not appear to correlate with any clinical or haematological parameter, or with prognosis. The colony-forming potential of patients with ALL in their first full remission, early relapse or second remission did not deviate significantly from previously established normal values, but the colony-forming potential of patients in early remission was very significantly reduced. No loss of colony-forming potential of normal marrow cells was noted when they were cultured with cells from patients with ALL.

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Section: Methodsmentioning

confidence: 99%

Changes in granulopoiesis detected by in vitro colony formation in acute lymphatic leukaemia

Morris¹,

Butler²,

Muldrew³

et al. 1977

Br J Cancer

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“…To wash them before incubation with iodinated antigen, suspensions were centrifuged through FCS gradients as described by Byrt and Ada (11). Lymphocytes were obtained from peripheral blood by sedimentation of heparinized samples on a methylcellulose-Urographin gradient as described by Hulliger and Blazhovec (12). They were washed three times before use.…”

Section: Methodsmentioning

confidence: 99%

“…After hatching all birds were housed in cages and maintained on water and chick feed ad libitum. Blood was collected by venepuncture at approximately 2 months of age and lymphocytes separated as described previously (12). Bursectomized chickens without detectable circulating immunoglobulin were selected as donors.…”

Section: Identification Of the Subpopulation Of Lymphocytes Binding Amentioning

confidence: 99%

A Receptor for Antibody on B Lymphocytes

Basten

Miller

Sprent

et al. 1972

The Journal of Experimental Medicine

397

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Evidence is presented for the existence on all B lymphocytes, but not on T lymphocytes, of a membrane-associated receptor for antibody. The receptor was detected by a radioautographic technique in which lymphoid cells were incubated with antibody followed by the corresponding radioiodinated antigen. The ease with which antibody eluted during washing indicated that the bond between antibody and cell was weak. The formation of an antibody-antigen complex on the cell surface, however, stabilized the bond and permitted accurate quantitation of cells with adherent antibody. The ability of several combinations of antibody and antigen to adhere to the cells demonstrated the nonspecificity of the phenomenon and emphasized the need for care in interpretation of antigen-binding studies particularly when immune cells are being used. The identity of antibody-binding lymphocytes was established by two different approaches. In the first, mouse lymphocyte populations greatly enriched for either T cells or B cells were examined. Their T cell content was assessed by means of well-established markers such as the θ C3H isoantigen. When this was compared with the number of antibody-binding cells, an inverse relationship was obtained in each instance; thus almost all thoracic duct cells from athymic mice labeled with an immune complex although none were θ positive. The striking reduction in antibody-binding cells observed in bursectomized chickens provided a second and independent line of evidence suggesting that B cells, not T cells, bind antibody. The ability of B cells from primed animals to bind antibody in vivo made it important to test whether this phenomenon was related to the carriage of immunological memory. No correlation was, however, found between membrane-bound antibody and memory. It was proposed that the existence of a receptor of this kind may provide a rational explanation for antibody-dependent killing of target cells and may prove of importance in antigen concentration particularly during the secondary response.

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“…Lipoxygenase was obtained from human leucocytes by a modification of the method of Hulliger & Blazkovec (1967) and assayed by the method of Ku et al (1983).…”

Section: Animalsmentioning

confidence: 99%

CGS 12970: a novel, long acting thromboxane synthetase inhibitor

Ambler

Butler

Ku³

et al. 1985

British J Pharmacology

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1 CGS 12970 is a potent selective inhibitor of human platelet thromboxane synthestase in vitro (IC50 = 12 nM). It is four orders of magnitude less potent as an inhibitor of sheep seminal vesicle cyclooxygenase, bovine aorta prostacyclin synthetase and human leucocyte 15-lipoxygenase. 2 The compound inhibited collagen-induced thromboxane B2 production by human platelets in vitro without an effect on the accompanying platelet aggregation induced by collagen, ADP, platelet activating factor, thrombin, arachidonic acid or the prostaglandin mimetic, U 46619. 3 Administration of CGS 12970 to rats inhibited collagen-induced thromboxane B2 production but had no effect on platelet aggregation ex vivo. It also had no effect on platelet aggregation induced by thrombin or on plasma clotting times. 4 A single oral dose of 1 or 3 mg kg-' to rabbits inhibited thromboxane B2 production in clotting blood ex vivo for 12 or 24 h respectively. 5 CGS 12970 inhibited thromboxane B2 production in vivo induced by intravenous administration of collagen to rats or calcium ionophore to guinea-pigs. In both cases there was a concomitant elevation of immunoreactive 6-keto-prostaglandin Flr. but no effect on the induced thrombocytopenia.6 As with other thromboxane synthetase inhibitors, CGS 12970 prolonged tail bleeding time in the rat. However, CGS 12970 was not as potent as other thromboxane synthetase inhibitors in this test. 7 In addition to these usual effects of thromboxane synthetase inhibitors, CGS 12970 inhibited the thrombocytopenia induced by the Forssman reaction or ADP administration. In these tests the effect of the compound was of short duration. 8 CGS 12970 had no effect on the thrombocytopenia associated with the Arthus reaction which distinguishes it from cyclo-oxygenase inhibitors. It also had no effect on thrombus formation on a cotton thread in an arteriovenous shunt in the rat.

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A Simple and Efficient Method of Separating Peripheral-Blood Leucocytes for in-Vitro Studies

Cited by 58 publications

References 2 publications

Changes in granulopoiesis detected by in vitro colony formation in acute lymphatic leukaemia

Changes in granulopoiesis detected by in vitro colony formation in acute lymphatic leukaemia

A Receptor for Antibody on B Lymphocytes

CGS 12970: a novel, long acting thromboxane synthetase inhibitor

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