Leukocyte-rich Plasma Research Articles

Mixed leukocyte culture in uraemia.

The activities of mixed leucocyte cultures (MLC) prepared from pairs of healthy subjects were estimated from the uptake of tritiated thymidine by the cultures. It was confirmed that MLC is a practical method for detecting identical twins amongst siblings. MLC has not often been employed for donor selection for renal transplantation, mainly because of the impaired MLC reactivity of leukocytes from uraemic patients. An attempt was made to separate lymphocytes from uraemic leukocyte-rich plasma, which always has a high proportion of neutrophils. A lymphocyterich fraction of uraemic leukocytes has MLC reactivity comparable to that of non-uraemic leukocytes.

Reactivity of Human Lymphocytes to Autologous and Homologous Serum

Abstract The mixed lymphocyte reaction initially described by Bain et al. (1) and Hirschhorn et al. (2) is currently being evaluated as a tool in many laboratories for histocompatibility testing. Johnson and Russell (3) have demonstrated that fetal calf serum, penicillin and streptomycin—substances routinely part of the culture media used in this reaction—may be stimulatory to lymphocytes. Such stimulation in the mixed lymphocyte reaction may mask fine differences which are essential in choosing a suitable donor for tissue transplantation. The present experiments were designed to investigate whether pooled human serum might also be a factor responsible for increased proliferative activity of human peripheral white blood cells. Heparinized blood (1000 units/60 ml of whole blood) drawn by venipuncture from normal human donors was allowed to sediment at room temperature for 2.5 hr. The leukocyte-rich plasma was aspirated and cell counts were carried out with cetrimide diluent (Fisher Scientific) in a model B Coulter Counter.

Chromosome damage induced by plasma of x-rayed patients: an indirect effect of x-ray.

Radiation can damage human chromosomes, both in vivo (1-5) and in vitro (6-8). This report describes more fully the results of an experiment devised to distinguish between the direct effects of X-rays on leukocyte chromosomes and a possible indirect effect mediated by way of blood plasma. Blood plasma from patients who had received large doses of X-ray enhanced the degree and frequency of chromosomal aberrations of normal nonirradiated lymphocytes in short-term cell culture (9). Materials and Methods. Six patients with a variety of tumors were chosen because they had received large doses of X-rays through several different ports (see Table I). X-ray integral doses varying from 2.88 to 25.1 × 106 gR were given with a 2 meV van de Graaf generator. Six other patients with similar tumors, studied prior to X-ray, served as controls for the first group. Three of the control patients with tumors were studied before and after X-ray. None of the patients received chemotherapeutic agents or had symptoms of viral infections during the studies. Blood for the studies was drawn from the irradiated patients 24−72 hr after irradiation. Subjects who had not received therapeutic X-ray furnished the normal lymphocytes and plasma. Chromosomes were studied by a modified method of Moorhead et al. (10). Plasma from the following sources was added to normal nonirradiated lymphocytes in short-term culture: (i) plasma from irradiated tumor patients “irradiated tumor plasma”, (ii) plasma from tumor patients before irradiation “nonirradiated tumor plasma”, (iii) homologous “normal plasma,” or (iv) autologous “normal plasma.” The cells for culture were obtained from leukocyte-rich plasma after sedimentation of heparinized blood. The leukocyte-containing plasma remaining after removal of erythrocytes, was centrifuged at 2400 rpm (IEC Universal model UV, head no. 240) for 20 min to obtain leukocyte-free plasma.

CULTURE AND SLIDE PREPARATION OF LEUKOCYTES FROM BLOOD OF MACACA.

Leukocyte-rich plasma is recovered from monkey blood by centrifugation. Portions of the plasma are planted in a medium consisting of 70% NCTC 109, 10% F 10, 15% fetal bovine serum, 5% human cord serum, and phytohemagglutinin (0.1 ml/5.0 ml of medium). Incubation is 48–72 hr, followed by colchicine 24 hr, hypotonic treatment with water 1–1 1/4 hr, and fixation in absolute methanol-glacial acetic acid (3:1). Permanent spreads are made by ejecting drops of suspension onto iced slides and flame drying. Staining is done in cresyl violet acetate. Because of some inconsistencies encountered in the spreading of monkey cells, alternate methods have been suggested for trial until one is found that works uniformly for the technician.

Electrodiagnosis in the neuromuscular disorders of childhood.

Previous work suggests that peripheral blood lymphocytes from patients with rheumatoid arthritis have altered reactivity. The purpose of this study has been to observe the behavior of cultures of lymphocytes from patients with rheumatoid arthritis when stimulated with autologous 19S rheumatoid factor and with rabbit serum against autologous IgG fraction. The peripheral blood lymphocytes (PBL) from 15 patients with classical rheumatoid arthritis were obtained by passage of the leukocyte-rich plasma through a polyamide fiber column. The purified lymphocytes were cultured with rheumatoid factor obtained from the same donor through Sephadex G-200 chromatography. In another set of experiments rheumatoid lymphocytes were stimulated with rabbit serum containing antibodies against the IgG fraction of the same patient obtained through DEAE chromatography. After five days the lymphocytes were examined for large cells plus cells in mitosis. A positive response was defined as an increase of 5% or more in the experimental tubes when compared with the cells cultured alone. Ten healthy normal subjects were used also as lymphocyte and serum donors. These lymphocytes were cultured alone and with rabbit serum, anti-autologous IgG. 80% of rheumatoid lymphocytes responded with an average of 37% of enlarging and dividing cells when stimulated by autologous RF. The same pannel of lymphocytes responded only with an average of 14% stimulation when cultured with rabbit serum anti-autologous IgG. The response to the 7S antigammaglobulin factor from rabbit in 7 normal PBL was similar to that of rheumatoid lymphocytes. There was some relation between activity of the RA and lymphocyte response to RF and rabbit antiserum. These results could explain the lymphoid infiltration and hyperplasia of patients with RA and, on the other hand, would support the idea that entrapment of RF by leukocytes ('RA cell') would be a protective mechanism.