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At least 19 recordsLinked to original sources
PubMed1997

A three-step procedure for the purification of human basophils from buffy coat blood.

OBJECTIVE AND DESIGN: We report a method for basophil purification from buffy coats, which avoids positive selection of the cells and gives rise to good purity, yield and functional integrity of the cells. SUBJECTS: Buffy coat blood (concentrated leukocyte fraction derived from 450 ml venipuncture donations) obtained from healthy blood donors (n = 51). METHODS: Basophils were enriched by a three-step process starting with Ficoll density centrifugation (1.6 +/- 0.1% basophil purity) followed by counter current centrifugal elutriation (17.7 +/- 1.4% basophil purity). The final stage involved negative selection using Dynal immunomagnetic beads directed against CD2, CD14, CD16 and CD19 positive cell contaminants. Functional integrity of which was assessed by comparing the anti-IgE or calcium ionophore A23187 induced histamine release from basophils obtained from each enrichment step. Furthermore, basophil morphology was investigated using light and electron microscopy. RESULTS: The final mean basophil purity of 67.3 +/- 1.4% with a yield of 3.5 +/- 0.5 x 10(6) basophils and a recovery of 21.8 +/- 2.4% was achieved. Net histamine release from basophils stimulated with optimal concentrations of anti-human IgE was 39.1 +/- 6.5% after Ficoll centrifugation, 41.6 +/- 7.7% following elutriation and 35.7 +/- 6.8% from the final purified fraction. Additionally, basophils enriched with our method showed intact morphology by electron microscopy and were functionally active to non-immunological stimulation. CONCLUSIONS: These results compare favourably with previous studies, which have often required the use of positive selection via the Fc epsilon RI receptor, which may result in cell degranulation, or cell sorting, which cannot be applied to large cell numbers. Our method provides a reproducible technique for basophil enrichment when large numbers of functionally intact basophils are required.

Antigen-Antibody Reactions↗
PubMed1996

Transfusion of donor peripheral blood buffy coat cells as effective treatment for relapsed acute leukemia after transplantation of allogeneic bone marrow or peripheral blood stem cells from the same donor.

BACKGROUND: The transfusion of same-donor peripheral blood buffy coat (PBBC) cells to chronic myelocytic leukemia patients in relapse after bone marrow transplantation has been increasingly used as an effective antileukemic therapy. A graft-versus-leukemia effect mediated by immunocompetent donor T cells underlies its success. In acute leukemia, however, the effect of this adoptive cellular immunotherapy has not been established, and the results are generally poor. STUDY DESIGN AND METHODS: Five patients, three with acute lymphoblastic leukemia and two with acute myelocytic leukemia, who relapsed within 6 months after allogeneic marrow transplantation were enrolled in a nonrandomized pilot study to receive donor PBBC cell transfusions either before or after undergoing cytoreductive chemotherapy. ABO genotyping and polymerase chain reaction amplification of human tetrameric short tandem repeats DNA typing were used to test for marrow chimerism. RESULTS: Two acute lymphoblastic leukemia patients-both of whom underwent chemotherapy before PBBC cell transfusions, experienced a complete remission, and developed acute and then chronic, extensive graft-versus-host disease-have been leukemia-free for 9 and 7 months, respectively. Repeated molecular studies of their marrow as early as 2 weeks to 8 months after treatment confirmed that the marrow was of donor origin. The other three patients, who chose not to undergo chemotherapy before PBBC cell transfusions, failed to achieve remission and died 14, 16, and 30 days, respectively, after leukemia relapse. CONCLUSION: Adoptive cellular immunotherapy may be effective for acute lymphoblastic leukemia patients in relapse after bone marrow transplantation if chemotherapy is administered before PBBC cell transfusions are initiated.

Acute Disease↗
PubMed1994

Blood buffy coat from Alzheimer's disease patients and their relatives does not transmit spongiform encephalopathy to hamsters.

There was a report of spongiform encephalopathy transmitted to Syrian hamsters by intracerebral inoculation with the blood buffy coat of patients with Alzheimer's disease (AD) and their unaffected first-degree relatives. We attempted to verify that report, taking measures to reduce the risk of contaminating samples with agents causing spongiform encephalopathies. We obtained blood from 50 subjects, including six patients with familial AD, 21 unaffected first-degree relatives (siblings and offspring) of patients with familial AD, and 20 control subjects. We inoculated the buffy coats intracerebrally into Syrian LVG hamsters, observed them for signs of neurologic disease, examined their brains for neuropathologic changes at time of death, and performed serial (blind) passages by inoculating suspensions of all recovered brains into fresh LVG hamsters. We discerned no clinical illness or histopathologic changes resembling experimental spongiform encephalopathy in any hamster inoculated with human buffy coat nor in blind-passage hamsters, nor were the life spans of those hamsters shortened. We conclude that AD is not caused by an agent that transmits spongiform encephalopathy to hamsters.

Alzheimer Disease↗
PubMed1983

The leukocyte composition of peripheral blood buffy coat.

To assist in the interpretation of buffy coat smears from leukopenic patients, we determined the composition of the buffy coat of the blood of normal volunteers. Leukocytes were concentrated by centrifugation in microhematocrit tubes. Stained smears of these cells showed minor differences in the proportions of neutrophils, lymphocytes, and monocytes when compared to unconcentrated peripheral blood. Granulocytes of greater immaturity than the metamyelocyte were not encountered, and their presence should suggest the possibility of marrow dysfunction. Occasional young monocytoid cells and "stimulated" lymphocytes were seen.

Adult↗
PubMed1991

Molecular diagnosis of alcelaphine herpesvirus (malignant catarrhal fever) infections by nested amplification of viral DNA in bovine blood buffy coat specimens.

A fragment of alcelaphine herpesvirus-1 (AHV-1; malignant catarrhal fever) DNA was subcloned into pUC 18 and sequenced. The subclone hybridized strongly to AHV-1 DNA, weakly to alcelaphine herpesvirus-2 (AHV-2) DNA, and not at all to DNA from bovine herpesvirus-1 (BHV-1; infectious bovine rhinotracheitis [IBR] virus), bovine herpesvirus-2 (BHV-2; bovine herpes mamillitis [BHM] virus), and bovine herpesvirus-4 (BHV-4; isolate DN599). A 2-stage (nested) polymerase chain reaction (PCR) diagnostic test was devised based on a portion of the subcloned AHV-1 DNA sequence. First and second stage amplified AHV-1 DNA targets were 487 and 172 base pairs (bp) in length, respectively. Unique Pvu II and Stu I restriction endonuclease cleavage sites confirmed the identity of amplified AHV-1 DNA. Five AHV-1 and 2 AHV-2 isolates were identically and specifically PCR positive. BHV-1, BHV-2, and BHV-4 viruses were negative by the same procedure. As little as 0.01 TCID50 AHV-1 was detected using the nested amplification procedure. Simple methods of buffy coat isolation from bovine blood were employed to prepare specimens for PCR. An AHV-1-infected calf was PCR positive from 3 to 77 days postinoculation (PI), with rising seroconversion first noted 14 days PI. The AHV-1 DNA sequence was 62% homologous to a portion of the Epstein-Barr virus genome. The nested PCR procedure may improve the viral diagnosis of clinical and subclinical alcelaphine herpesvirus infections.

Animals↗
PubMed1999

Detection of Pneumococcus from whole blood, buffy coat and serum samples by PCR during bacteremia in mice.

Whole blood, purified leukocyte fraction and serum were investigated as specimens for the detection of pneumococcal bacteremia by polymerase chain reaction (PCR) in mice. The PCR findings were compared to the blood culture results. Samples were taken from animals 3 and 12 h after intraperitoneal bacterial challenge. The pneumococcal culture was positive in 27% and 77% of blood samples at 3 and 12 h after challenge, respectively. All whole blood samples were positive by PCR at both time points. Of the buffy coat samples, two of the three pools were PCR-positive at 3 h and all pools at 12 h after bacterial challenge. In the serum sample group, only 40% of the sera were PCR-positive at 3 h, while at 12 h 90% of the samples were PCR-positive. According to these results, whole blood seems to be the best specimen for the detection of pneumococcal DNA by PCR in bacteremic mice.

Animals↗
PubMedDate not supplied

A simple technique for preparation of bone marrow or peripheral blood buffy coat cells for electron microscopy.

A simple and rapid method of processing ethylene-diamine-tetra-acetic acid anticoagulated peripheral blood or aspirated bone marrow for electron microscopy is described. The resultant buffy coat pellet is easily processed into epoxy resin. Semithin sections (1 mu) stained with 1% toluidine blue reveal the various stratified cell layers allowing convenient selection for ultramicrotomy and ultrastructural evaluation.

Acute Disease↗
PubMed1984

Comparison of apheresis and other methods for separation and purification of hemopoietic stem cells: initial experience with a blood buffy coat model for the use of autologous bone marrow transplantation.

With an increasing number of bone marrow transplantations (BMT) being contemplated in leukemia and cancer patients, it is prudent for blood banks to develop a suitable program within their resources for harvesting, purifying and freezing bone marrow stem cells. In order to do this, initially a prototype has been developed involving buffy coat model (BC) using normal donor blood. Centrifugation, sedimentation and machine apheresis methods were separately evaluated leading to a combined and sequential handling procedure. Blood was passed through a cell separator resulting collection of BC with 90% reduction of the volume showing 80% recovery of total leucocytes and 87% yield of mononuclear cells. Following centrifugation the cells with DMSO were frozen in a controlled freezing system and stored in liquid nitrogen. After thawing 94% cells were recovered with 93% viability. The initial experience gained in the model system could be incorporated in autologous BMT program in patients but requires modifications for improved results; the latter will be described separately.

Blood Cells↗
PubMed1979

Autologous bone-marrow and peripheral blood buffy coat cell infusion in the treatment of chronic myeloid and acute leukemia.

Autologous marrow infusion has been attempted in three patients with chronic myeloid leukemia (two in blast crisis, one with severe myelofibrosis and pancytopenia) and one patient with acute lymphatic leukemia. One patient with blast crisis of CML expired prior to marrow infusion. One patient with myelofibrotic phase of CML is alive seven months post marrow infusion. The other two patients expired 6 and 16 days post marrow infusion. Bone-marrow repopulation is feasible in the face of severe myelofibrosis.

Adult↗
PubMed1991

[Isolation of thrombocyte concentrates from the leuko-thrombocytic buffy coats of donor blood and their preservation].

A new method has been developed for platelet concentrate (PC) preparation from the whole blood buffy coats. This method makes possible obtaining leukocyte-poor PC as compared to that prepared from platelet rich plasma. Functional characteristics of PC stored in Soviet plastic bags at room temperature (22 degrees C) under continuous agitation at circular rotator were studied during 7 days. Satisfactory maintenance of functional activity was noted during 5 days of PC storage. The method of PC preparation from the whole blood buffy coats can be also used for manual plateletpheresis of a single donor.

Blood Donors↗
PubMed1983

A comparison of peripheral blood and buffy coat smear examination for the prediction of bone marrow relapse of acute lymphoblastic leukaemia in childhood.

In an attempt to see if buffy coat smear examination might be an alternative to bone marrow aspiration for predicting relapse, 98 consecutive bone marrow aspirates from 96 children with acute lymphoblastic leukaemia were examined blind with buffy coat and peripheral blood from the same patients. The 28 bone marrow aspirates from children no longer on treatment were all normal, and routine aspirates would appear unjustified in these patients. Eight of the remaining marrows showed relapse, but only three were not predicted from the peripheral blood and buffy coat. In no case was buffy coat superior to peripheral blood in the detection of bone marrow relapse. Routine bone marrow aspirates are an inefficient way of diagnosing relapse in acute lymphoblastic leukaemia in childhood, despite their precision, and a prospective study is needed to determine their value.

Bone Marrow↗
PubMed2006

[Long-term survival rate of patients transfused with leucocyte-depleted and buffy-coat-poor blood during colorectal surgery--secondary publication. An eight-year post-trial follow-up study].

This study investigated the long-term survival rate of 589 patients enrolled in a trial in 1992-1995 who underwent colorectal surgery. The patients were randomised to receive leucocyte-depleted or buffy-coat-poor blood when transfusion was indicated. Significantly more of the non-transfused patients (59%) were alive seven years later compared to patients transfused with leucocyte-depleted blood (41%) and to patients transfused with buffy-coat-poor blood (45%).

Adult↗
PubMed1996

Randomised comparison of leucocyte-depleted versus buffy-coat-poor blood transfusion and complications after colorectal surgery.

BACKGROUND: Allogeneic blood transfusion is associated with an increased frequency of postoperative infection. We studied whether such events can be avoided by the intraoperative and postoperative use of leucocyte-depleted blood. METHODS: 589 consecutive patients scheduled for elective colorectal surgery were randomised to receive buffy-coat poor (n = 299) or filtered leucocyte-depleted red-cells (n = 290) when transfusion was indicated. 260 patients actually received blood transfusion. Three patients were excluded from analysis. FINDINGS: The 142 patients randomised to and transfused with buffy-coat-poor blood had a significantly higher frequency of wound infections and intra-abdominal abscesses than the 155 patients who were allocated to this group but who were not transfused. (12 vs 1%, p < 0.0001) and (5 vs 0%, p = 0.005), respectively. Those receiving buffy-coat-poor blood also had a significantly higher frequency of wound infections and intra-abdominal abscesses than the 118 randomised to and receiving leucocyte-depleted blood (12 vs 0%, p < 0.0001) and (5 vs 0%, p = 0.017), respectively. Postoperative pneumonia occurred at a significantly higher rate in patients receiving buffy-coat-poor blood: 23 versus 3% in non-transfused patients (p < 0.0001), and 23 versus 3% in patients transfused with leucocyte-depleted blood (p < 0.001). The mortality rate between the groups was not statistically different. INTERPRETATION: Our data suggests that the association between allogeneic blood transfusion and postoperative infection is limited to allogeneic blood products that are not adequately depleted of immunosuppressive leucocytes. This undesirable effect can be reduced by leucocyte depletion with high-efficiency filters.

Aged↗
PubMed1994

Randomised controlled trial comparing transfusion of leucocyte-depleted or buffy-coat-depleted blood in surgery for colorectal cancer.

In retrospective studies, perioperative blood transfusions were associated with poor prognosis after surgery for cancer and were a major independent risk factor for postoperative bacterial infection. Leucocyte-depleted, in contrast to buffy-coat-depleted, blood has no immunosuppressive effects in transplantation and so might lack detrimental effects on cancer prognosis and postoperative infections. We studied this hypothesis in a controlled trial by randomly allocating patients to receive either leucocyte-depleted red cells or packed cells without buffy coat when blood was needed. Between 1987 and 1990, 871 eligible patients with colorectal cancer, including 697 patients operated upon with curative intent, were randomised in the 16 participating hospitals. Neither the eligible group nor the curative group showed significant differences between the two trial transfusions in survival, disease-free survival, cancer recurrence rates, or overall infection rates after an average follow-up of 36 months. Patients who had a curative resection and who received blood of any sort had a lower 3-year survival than non-transfused patients (69% vs 81%, p = 0.001) and a higher infection rate (39% vs 24%, p < 0.001). Colorectal cancer recurrence rates, however, were not influenced by blood transfusion (30% vs 26%, p = 0.22). These combined observations confirm the association between blood transfusion and poor patient survival but indicate that the relation is not due to promotion of cancer.

Adenocarcinoma↗
PubMed2001

Major histocompatibility complex class II polymorphisms in Macaca mulatta: factors influencing comprehensive genotyping of Macaca mulatta (Mamu)-DQA1 alleles by PCR-RFLP in archival samples.

In the last 5 years, HLA class II genotyping methods have been adapted for genotyping of class II loci in rhesus macaques. Since previously published typing protocols were used on samples that were collected and stored under ideal conditions, it was of interest to determine if these methods were adequate for genotyping a large collection of archival samples from which DNA had been isolated and stored under various conditions. Established macaque DQA1 typing protocols were modified to optimize the typing procedure and enhance the ability to successfully genotype DNA from samples that were of poor quality and/or quantity. Long-term storage of whole-blood buffy coats or stored DNA extracted from whole-blood buffy coats did not affect typing success; however, amplification and typing of DNA extracted from archival samples of plasma were difficult and resulted in a low success rate. This suggests that amplification and DQA1-genotyping of archival samples is possible with a modified protocol, but is influenced by the age and source of the sample, and to a lesser extent, the method used to extract DNA from sample substrates.

Animals↗