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D F Stroncek

Publications and source records attributed to D F Stroncek.

At least 37 records · Page 2Linked to original sources
PubMed1999

Granulocyte storage and antigen stability.

BACKGROUND: Current methods for the detection of granulocyte antibodies require panels of freshly isolated cells. This makes these assays time-consuming, costly, and technically difficult. STUDY DESIGN AND METHODS: The immunofluorescence method of detecting the binding of antibodies to granulocytes was modified for use with a flow cytometer, and methods were tested to store granulocytes for use in that assay. Granulocytes were stored at 4 degrees C for 7 days under three conditions: 1 -percent formaldehyde-fixed cells were stored in Hanks' balanced salt solution (HBSS); untreated cells were stored in tissue culture medium (RPMI-1640); and cells were fixed and stored with a commercial white cell-storage solution (Cyto-Chex Reagent). Antigen stability was evaluated by using monoclonal antibodies (MoAbs) and alloantibodies. Serologic studies were done by an indirect immunofluorescence assay and assessed by flow cytometric analysis. RESULTS: On Day 2, only 2 to 7 percent of granulocytes stored in RPMI-1640 remained. On Day 7, 67 to 76 percent of granulocytes fixed in formaldehyde and stored in HBSS remained, and 47 to 87 percent of granulocytes stored in a white cell-storage solution remained. All antigens were detectable by the MoAbs and alloantisera on Day 7. However, nonspecific staining by the fluorescein isothiocyanate (FITC)-conjugated secondary antibody hindered interpretation of test results on Day 4. Non-specific staining occurred over time and was associated with increased cell permeability during storage. Two sources of nonspecific staining were identified. The first source was the FITC-conjugated secondary antibody; it was eliminated by switching to a phycoerythrin conjugate. The second source was factors in human serum; it was resolved by examining only viable, impermeable cells identified by using 7-aminoactinomycin-D. CONCLUSION: Granulocytes and their antigens can be preserved for at least 7 days, but evaluation of antibody reactions was possible for only 4 days as a result of non-specific staining due to enhanced membrane permeability of dying cells.

Animals↗
PubMed1999

Biotinylation modifies red cell antigens.

BACKGROUND: Chemical biotinylation of red cell membranes may be useful for several clinical applications, including red cell survival studies. STUDY DESIGN AND METHODS: To examine the possible effects of biotinylation on red cell antigens, standard hemagglutination assays were performed on matched sets of control and biotinylated red cells. The red cells were biotinylated at a final concentration of 2.0 pg of sulfo-N-hydroxysuccinimide-biotin per cell, and antigen-negative cells were directly compared to antigen-positive cells when possible. The hemagglutination assays were graded in a blinded fashion. Forty-one red cell antigens from 21 of the 23 established blood group systems were tested. RESULTS: Hemagglutination based upon antibody binding to A, A1, M, N, S, s, P1, D, C, E, c, e, C(w), Lu(b), K, k, Kp(b), Le(a), Le(b), Fy(a), Fy(b), Jk(a), Jk(b), Di(a), Wr(a), Wr(b), Yt(a), Xg(a), Sc1, Do(b), Co(a), Ch, H, Ge2, Cr(a), Kn(a), I, and P was not affected by biotinylation. Unexpectedly, the hemagglutination of Di(b+) and LW(a+) red cells was blocked after biotinylation. Conversely, MH04 monoclonal anti-A agglutinated red cells expressing B only after biotinylation. BIRMA-1 monoclonal anti-A and polyclonal anti-A from sera did not agglutinate the biotinylated B red cells. CONCLUSION: Biotinylation of human red cells specifically modified their antigenicity, as measured by standard hemagglutination assays.

Biotinylation↗
PubMed1999

Evaluation of the gel system for ABO grouping and D typing.

BACKGROUND: The gel agglutination assay has been approved by the Food and Drug Administration as an alternative to the tube assay for the detection of red cell antibodies. It has also been approved recently by the Food and Drug Administration for ABO blood grouping and D typing. STUDY DESIGN AND METHODS: Tube and gel agglutination assays were compared for ABO grouping and D typing of 100 donor and 100 patient specimens. ABO grouping of 14 specimens of known ABO groups and D typing of 10 specimens with weak D were also compared. When antigen typing or isohemagglutinin results differed, gel testing was repeated by the use of modified incubation times, reagent or specimen volumes, and red cell concentrations. RESULTS: ABO grouping and D typing in all patient and donor specimens concurred. B isohemagglutinins were not detected in seven group A specimens. Six of seven discrepancies were resolved when gel tests were incubated at room temperature with increased serum or plasma volume. Weak D was detected in all 10 specimens tested by both assays. When weak A and/or B were tested with monoclonal antibody reagents, the correct phenotypes were identified in 9 specimens by gel assay and in 10 by tube assay. Using human antisera, 6 specimens were correctly phenotyped by gel assay and 7 by tube assay. CONCLUSION: The gel assay performed as well as the tube assay in detection of A, B, and D, but the tube assay was slightly better at detecting B isohemagglutinins. The gel assay can be used in place of the tube assay for ABO blood grouping and D typing.

ABO Blood-Group System↗
PubMed1999

Retroviral transduction and expansion of peripheral blood lymphocytes for the treatment of mucopolysaccharidosis type II, Hunter's syndrome.

BACKGROUND: Gene therapy using autologous peripheral blood lymphocytes (PBLs) has been used to produce adenosine deaminase with which to treat patients with severe combined immunodeficiency. Patients with mucopolysaccharidosis type II (MPS II) lack iduronate-2-sulfatase (IDS), and serial PBL gene therapy may benefit these patients. STUDY DESIGN AND METHODS: The purpose of these studies was to develop a method to transduce PBLs from a patient with MPS II by using a retroviral vector, LS2N, containing the IDS gene. PBLs were collected by apheresis and cryopreserved in aliquots for the performance of multiple transductions and expansions. The PBLs were expanded in number and then transduced in a hollow-fiber bioreactor (HFBR). Additional culture allowed for further expansion. RESULTS: Fresh PBLs (6.2 x 10(7)) from a patient with MPS II were transduced with L2SN and expanded in an HFBR with an extracapillary space of 11 mL. After 10 days of culture, 4.1 x 10(9) cells were harvested. Cryopreserved MPS II PBLs could not be reliably expanded if they were placed in the HFBR immediately after being thawed; however, cells were successfully transduced and expanded in the HFBR if they were first cultured in a bag. To increase the cell yield, PBLs were expanded in a 60-mL HFBR after transduction and expansion in an 11-mL HFBR. In four separate experiments, 2 x 10(8) cryopreserved PBL were cultured for 3 days in a bag and transferred to an 11-mL HFBR, where they were transduced daily with L2SN for 3 days and then expanded for 4 additional days. Cells were then transferred into a 60-mL HFBR and expanded for an additional 7 days. In the four experiments, 5.5 x 10(9), 7.4 x 10(9), 1.12 x 10(9), and 19.4 x 1(9) cells were produced. The vector was detected in the harvested cells, but the proportion of cells transduced was less than 2.5 percent, the lowest standard used in the assay. In two of the experiments, cells harvested from the HFBR were used in a gene therapy clinical trial. CONCLUSION: Autologous cryopreserved PBLs can be transduced and expanded to produce >1 x 10(10) cells. This procedure is being used for a Phase I/II clinical trial of lymphocyte gene therapy.

Cells, Cultured↗
PubMed1999

Changes in serum osteocalcin and bone-specific alkaline phosphatase are associated with bone pain in donors receiving granulocyte-colony-stimulating factor for peripheral blood stem and progenitor cell collection.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) has been used to increase the number of CD34+ peripheral blood stem and progenitor cells collected by apheresis for use in autologous or allogeneic progenitor cell transplantation. The most frequent side effect of G-CSF treatment is bone pain, which occurs in over 80 percent of healthy progenitor cell donors. STUDY DESIGN AND METHODS: The possible mechanism of bone pain was investigated by measuring serum levels of osteocalcin (OC), bone-specific alkaline phosphatase (BAP), acid phosphatase (ACP), and tartrate-resistant acid phosphatase (TRAP) in seven healthy progenitor cell donors treated with human recombinant G-CSF administered subcutaneously for 5 consecutive days. RESULTS: All seven patients experienced bone pain during the treatment period. Serum levels of OC, BAP, ACP, and TRAP were measured in blood samples drawn on Days 0, 4, 5, 6, and 14. Levels of BAP were increased (p<0.05) over baseline on Days 4, 5, and 6, while those of OC decreased on Days 4, 5, and 6 (p<0.05). No significant changes occurred in ACP or TRAP levels. OC and BAP are considered markers of bone formation (osteoblast activity), and they correlate in many patients with metabolic bone disorders. The pattern of increased BAP and decreased OC has been reported in patients with osteolytic bone metastases. CONCLUSION: G-CSF treatment in healthy stem and progenitor cell donors may affect osteoblastic activity, and this activity may be associated with bone pain.

Alkaline Phosphatase↗
PubMed1998

Therapeutic apheresis for babesiosis.

Infection with the tick-borne protozoa Babesia is becoming more common. Babesiosis is usually successfully treated with antibiotics but, in some cases, apheresis may also be indicated. We report two patients with babesiosis and hemolysis treated by apheresis and antibiotics. One case had traditional indications for red blood cell (RBC) exchange, and a second patient was treated with RBC exchange, and plasmapheresis for hemolysis, probably secondary to Babesia parasitemia. Case 1 involved a 44-year-old man with chronic relapsing pancreatitis who had become infected with Babesia from a unit of RBCs transfused during surgery. At 5 weeks after surgery, fever and severe hemolysis developed, along with a hemoglobin of 69 g/L; 30% of his RBCs were found to be infected with Babesia. This patient had several postoperative complications; the babesiosis was treated with clindamycin, quinine, and three RBC exchanges. Parasitemia fell to less then 1% of RBCs, but the patient died of pancreatitis. Case 2 was a 47-year-old man with a renal transplant who had been receiving immunosuppressive therapy for 8 years. He had a history of tick bites, fever, and hemolytic anemia. Analysis of a peripheral blood smear detected Babesia. He was initially treated with antibiotic therapy and two RBC exchanges. Hemolysis improved transiently but worsening parasitemia developed later, as well as an IgG RBC autoantibody. He was then treated by plasmapheresis and RBC exchange. Although his condition improved, he had a third hemolytic episode, which was treated with plasmapheresis and RBC exchange before the parasitemia and autoimmune hemolytic anemia disappeared. In conclusion, immunosuppressed or severely ill people who become infected with Babesia may benefit from RBC exchange or plasmapheresis, or both.

Adult↗
PubMed1998

The expression of the NB1 antigen on myeloid precursors and neutrophils from children and umbilical cords.

The neutrophil-specific antigen NB1 is expressed by neutrophils from 97% of healthy adults. However, membrane expression of this molecule is unique in that it is found on only a subpopulation of neutrophils present in NB1-positive adults. We have investigated the ontogeny of NB1 antigen expression by haematopoietic progenitor cells to determine the stage and pattern of antigen expression during granulocytic cell differentiation. In addition, we examined whether the ontogeny and frequency of granulocytic cells expressing the NB1 antigen might vary in subjects according to age. A monoclonal antibody (MoAb) specific for NB1 (1B5) and flow cytometry was used to assess the frequency and characteristics of the NB1-positive cells found in umbilical cord blood (n = 11), children (n = 37), healthy adults (n = 46) and patients with chronic myelogenous leukaemia (n = 8). We also used flow cytometry to isolate NB1-positive and NB1-negative bone marrow and peripheral blood cells from various tissue sources. The separated subpopulations were then analysed by Wright stain and light microscopy. The size of the NB1-positive neutrophil subpopulation in 46 healthy adults (56 +/- 19%) was identical to that found for neutrophils from 36 children ranging in age from 8 months to 18 years (56 +/- 11%). In contrast, expression of the NB1 antigen by the neutrophils present in umbilical cord blood (91 +/- 3%, n = 11) was significantly greater than that in adults (P < 0.002) or children (P < 0.002). We also examined the size of the NB1-positive subpopulation among neutrophils from eight patients with chronic myelogenous leukaemia (CML). The NB1-positive subset in CML subjects (29.5 +/- 22.4%) was significantly less that in healthy adults (P < 0.02) or children (P < 0.02). Marrow cells from eight adults were similarly separated and analysed. We found that 69 +/- 17% of segmented and band forms of neutrophils, 70 +/- 2% of metamyelocytes and 61 +/- 23% of myelocytes were NB1-positive. In fetal bone marrow, 86 +/- 9% of the segmented and band forms, 82 +/- 10% of the metamyelocytes and 3 +/- 4% of myelocytes were NB1-positive. In conclusion, neutrophil-specific antigen NB1 is first expressed at the myelocyte stage of myeloid differentiation. In adult bone marrow, the percentages of myelocytes, metamyelocytes and segmented or band cells that expressed this antigen were similar and comparable in magnitude to the frequency of NB1-positive neutrophils found in the circulation. Although the size of the NB1-positive neutrophil subpopulation was the same in healthy adults and children, it was significantly increased in umbilical cord blood, and in fetal marrow cells.

Adolescent↗
PubMed1998

Expression of neutrophil antigens after 10 days of granulocyte-colony-stimulating factor.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) is becoming the standard agent for mobilizing granulocytes. Most granulocyte donors are given a single dose of G-CSF, but in some cases they are given G-CSF for several days, and multiple granulocyte concentrates are collected. The administration of a single dose of G-CSF induces several changes in the expression of neutrophil antigens, but the effects of multiple daily doses of G-CSF are not known. STUDY DESIGN AND METHODS: Seven healthy people received 5 microg per kg of G-CSF for 10 days. Their expression of several neutrophil antigens before, during, and after the administration of G-CSF was analyzed through the use of flow cytometry. RESULTS: The expression of L-selectin (CD62L), Fcgamma receptor (FcgammaR) III (FcgammaRIII, CD16), and the leukocyte function antigen (CD11a) decreased throughout the course of G-CSF administration, while the expression of FgammaR I (FcgammaRI, CD64) and lipopolysaccharide-binding protein receptor (CD14) increased. The expression of FcgammaR II (FcgammaRII, CD32) also increased, but not until the fourth day of G-CSF administration. The expression of amino peptidase N (CD13), C3bi receptor (CD11b), and the neutrophil beta2 integrin unit (CD18) did not change during the administration of G-CSF, but that of both CD13 and CD18 increased 3 days after the last dose. The expression of neutrophil-specific antigen NB1 initially increased, returned to pre-G-CSF levels after 4 days, and then increased again after 10 days of G-CSF administration. CONCLUSION: Changes in the expression of several neutrophil antigens occurred throughout a 10-day course of G-CSF Most of the changes occurred after one dose, but additional changes occurred later in the 10-day course and after its completion. These changes may affect the function of G-CSF-mobilized granulocytes.

Adult↗
PubMed1998

Comparison of tube and gel red blood cell agglutination techniques in detecting chimeras after major ABO-mismatched allogeneic hematopoietic stem cell transplantation.

We compared the ability of tube and gel red blood cell (RBC) agglutination techniques to follow erythroid engraftment in a patient who received a major ABO-mismatched peripheral blood stem cell transplant and bone marrow transplant. Tube and gel RBC agglutination techniques were used to detect mixed-field reactivity in cell mixtures containing A/O and c+/c- RBCs and the ability of these two technologies to detect RBC chimeras were compared. We detected c+ RBCs in c+/c- RBC populations microscopically at 1% by the tube RBC agglutination technique, but not until 10% by the gel technique. Group A RBCs in A/O RBC populations were detected at 10% by both techniques. In the patient studied, group A RBCs and c+ RBCs were detected on Days 20 and 14, respectively, with the tube RBC agglutination technique, but neither marker was detected until Day 26 with the gel technique. Tube and gel RBC agglutination techniques comparably identified ABO mixed fields. Although the tube RBC agglutination technique showed greater sensitivity than the gel technique in detecting the c antigen, the gel technique was easier to use and allowed more reliable interpretation of mixed fields by the technologist.

Journal Article↗
PubMed1997

Comparison of two blood cell separators in collecting peripheral blood stem cell components.

To ensure that a sufficient number of CD34+ cells are collected for an allogeneic blood progenitor cell transplant, the most effective blood cell separator should be used to collect peripheral blood stem cell (PBSC) components. We compared the effectiveness of two blood cell separators. We gave 29 healthy people 7.5 or 10 micrograms kg-1 of granulocyte colony stimulating factor (G-CSF) daily for 5 days and collected one PBSC component with either a Fenwal CS3000 (n = 15) or a Cobe Spectra (n = 14) blood cell separator. The volume of blood processed was the same for each machine (8.4 +/- 1.0 L; range = 4.9-9.4 L for the CS3000 and 8.9 +/- 1.0 L; range 6.7-10.9 L; P = 0.71). The components collected with the CS3000 contained more mononuclear cells (39.6 +/- 21.9 x 10(9) compared with 26.9 +/- 5.6 x 10(9), P = 0.02) and fewer neutrophils (1.38 +/- 1.88 x 10(9) compared with 5.53 +/- 8.71 x 10(9), P = 0.001). The total number of CD34+ cells collected with the two instruments was the same (470 +/- 353 x 10(6) for the CS3000 and 419 +/- 351 x 10(6) for the Spectra; P = 0.64) as was the number of CD34+ cells collected per litre of whole blood processed (55.9 +/- 42.0 x 10(6) L-1 compared with 45.9 +/- 37.9 x 10(6) L-1; P = 0.59). The mononuclear cell collection efficiency was greater for the CS3000 (82.4 +/- 54.9% compared with 53.3 +/- 14.1; P = 0.04) but the CD34+ cell collection efficiencies were the same (87.4 +/- 61.1% for the CS3000 compared with 56.3 +/- 23.5% for the Spectra; P = 0.07). In conclusion, both blood cell separators collected components which contained large numbers of CD34+ cells, but those collected with the CS3000 contained fewer neutrophils and the CS3000 was more efficient at collecting mononuclear cells.

Adult↗
PubMed1997

The kinetics of G-CSF mobilization of CD34+ cells in healthy people.

When healthy people are given granulocyte colony stimulating factor (G-CSF) for 10 days the number of CD34+ cells in the peripheral blood begins to increase on the fourth day, reaches a maximum on the sixth day and then decreases. In this study, we further define the time and variability of peak mobilization of CD34+ cells. Twenty-two healthy people were given G-CSF (7.5 or 10 micrograms kg-1 day-1) subcutaneously each morning for 5 days and peripheral blood CD34+ cell counts were analysed immediately prior to the fourth (day 4) and fifth (day 5) G-CSF injection and 24 h after the fifth injection (Day 6). White blood cell (WBC) and neutrophil counts were greatest on day 6 [WBC = 43.8 +/- 13.9 x 10(9) L-1 (mean +/- 1 SD) and neutrophils = 36.6 +/- 12.8 x 10(9) L-1]. In contrast the CD34+ cell counts on day 6 (107 +/- 104 x 10(6) L-1) were less than on day 5 (128 +/- 136 x 10(6) L-1) (P = 0.048) but still greater than on day 4 (60.7 +/- 40.2 x 10(6) L-1) (P < 0.0001). The CD34+ cell counts of 10 donors were measured 2, 4 and 6 h after the fifth injection to determine if the counts increased further between days 5 and 6. The number of CD34+ cells in the blood on day 5 2 h after the fifth injection (193 +/- 277 x 10(6) L-1) was greater than the number prior to the injection (158 +/- 190 x 10(6) L-1), 4 h post-injection (139 +/- 158 x 10(6) L-1) and 6 h post-injection (170 +/- 236 x 10(6) L-1), but the differences were not significant (P = 0.29, 0.25 and 0.45). The number of CD34+ cells in the blood of 12 people were measured before and after the fourth G-CSF dose. Prior to the day 4 injection the CD34+ count was 61 +/- 40 x 10(6) L-1. At 2, 4 and 6 h the counts were 60 +/- 40, 61 +/- 29 and 64 +/- 30 x 10(6) L-1, respectively, and the differences were not significant (P = 0.99, P = 0.98, and P = 0.73). In conclusion, when healthy volunteers are given daily G-CSF injections, the number of mobilized CD34+ cells was the greatest on day 5, slightly less on day 6 and the least on day 4. If only one PBSC component is needed, PBSCs can be collected on day 5 after only 4 days of G-CSF. If PBSC components are collected on both days 5 and 6, the fifth dose can be given either before or after the collection of the first PBSC component.

Adult↗
PubMed1997

Blood counts in healthy donors 1 year after the collection of granulocyte-colony-stimulating factor-mobilized progenitor cells and the results of a second mobilization and collection.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF)-mobilized blood cells are being used for allogeneic transplants, but the long-term effects of G-CSF on healthy individuals are not known. Furthermore, it is not certain how many CD34+ cells can be collected in a second mobilization and collection procedure. STUDY DESIGN AND METHODS: Nineteen people were given 2, 5, 7.5, or 10 micrograms of G-CSF per kg per day for 5 days, and blood progenitor cells were collected by apheresis on the sixth day; this was done on two occasions separated by at least 12 months. Blood counts obtained before and after each course of G-CSF and the quantity of cells collected were compared. RESULTS: There were no differences in white cell (WBC), platelet, red cell, and WBC differential counts measured before each course of G-CSF, and all the values were in the normal range. In a subset of 12 people who received 7.5 or 10 micrograms of G-CSF per kg per day for both courses, the numbers of neutrophils, mononuclear cells, and CD34+ cells in the blood after each course were similar (34.1 +/- 7.31 x 10(9)/L vs. 36.4 +/- 12.3 x 10(9)/L, p = 0.24; 6.59 +/- 2.28 x 10(9)/L vs. 5.63 +/- 2.11 x 10(9)/L, p = 0.24; and 92.0 +/- 55.6 x 10(5)/L vs. 119.2 +/- 104.6 x 10(6)/L; p = 0.48, respectively), as were the quantities of mononuclear cells (31.0 +/- 8.4 x 10(9) vs. 31.0 +/- 6.1 x 10(9); p = 0.64) and CD34+ cells (417 +/- 353 x 10(6) vs. 449 +/- 286 x 10(6); p = 0.53) collected in the two apheresis procedures. Furthermore, there was a positive correlation between the quantity of CD34+ cells collected from each of the 12 people per liter of whole blood processed in the two procedures (r2 = 0.86, p < 0.001). CONCLUSION: One year after the administration of G-CSF to healthy people, their blood counts were normal and unchanged from pretreatment counts. If healthy people donate blood progenitor cells after a second G-CSF course the quantity of CD34+ cells collected will be similar to that obtained in the first collection.

Adult↗
PubMed1997

Composition of peripheral blood progenitor cell components collected from healthy donors.

BACKGROUND: Peripheral blood progenitor cell (PBPC) components are being collected from healthy donors for allogeneic transplantation, but the quantity, quality, composition, and variability of PBPCs collected from healthy people given granulocyte-colony-stimulating factor (G-CSF) have not been evaluated. STUDY DESIGN AND METHODS: PBPC components were collected from 150 healthy people who were given G-CSF (5, 7.5, or 10 microg/kg/day) for 5 days. The components were evaluated for white cell (WBC), mononuclear cell, CD34+ cell, neutrophil, platelet, and red cell (RBC) composition. RESULTS: The quantities collected were: WBCs, 35.0 +/- 16.4 x 10(9) (range, 11.9-163.3 x 10(9)); mononuclear cells, 33.3 +/- 14.4 x 10(9) (range, 11.9-139.6 x 10(9)); CD34+ cells, 412 +/- 287 x 10(6) (range, 70-1658 x 10(6)); neutrophils, 1.71 +/- 3.59 x 10(9) (range, 0-27.6 x 10(9)); RBCs, 7.2 +/- 4.0 mL (range, 0-22.1 mL); and platelets, 480 +/- 110 x 10(9) (range, 250-920 x 10(9)). PBPC components collected from people given G-CSF at 7.5 or 10 microg per kg per day contained significantly more CD34+ cells (respectively, 428 +/- 300 x 10(6); range, 70-1658 x 10(6) and 452 +/- 294 x 10(6); range, 78-1380 x 10(6)) than those from people given G-CSF at 5 microg per kg per day (276 +/- 186 x 10(6); range, 91-767 x 10(6)) (p = 0.007 and p = 0.002). When 10 microg per kg per day of G-CSF was given, 50 percent of the components contained enough CD34+ cells for transplantation to a 75-kg recipient (375 x 10(6) CD34+ cells), but 10.6 percent of the components contained less than 150 x 10(6) CD34+ cells and thus would provide a transplantable dose only for a 30-kg patient. CONCLUSION: One PBPC component collected from a healthy donor given 7.5 or 10 microg per kg per day of G-CSF should contain 70 to 1660 x 10(6) CD34+ cells, with 0 to 22 mL of RBCs. Because of the great variability in the number of CD34+ cells collected, the quantity of CD34+ cells in each component should be measured after each procedure to ensure that sufficient quantities of cells are present for a successful transplant.

Antigens, CD34↗
PubMed1997

Lack of relation of granulocyte antibodies (antineutrophil antibodies) to neutropenia in children with human immunodeficiency virus infection.

BACKGROUND: Neutropenia in children and adults with HIV infection is frequently observed, perhaps as a result of impaired myelopoiesis, drug myelotoxicity, immune destruction or opportunistic infection. The presence of antineutrophil antibodies (granulocyte antibodies) has been associated with severe neutropenia in some reports but not in others, and such antibody assays can be confounded by the presence of immune complexes and HLA antibodies. METHODS: To determine both the prevalence of granulocyte antibodies in children with HIV infection and whether such antibodies were related to neutropenia, we screened the sera of 30 HIV-infected children by performing granulocyte immunofluorescence, granulocyte agglutination and lymphocytotoxic anti-HLA antibody assays. Reactivity was graded by a standard numeric score calculated per number of reactive cells. RESULTS: Of 26 evaluable sera, 16 (62%) had granulocyte antibodies, 6 (23%) had HLA antibodies and 4 (15%) had neither. There was no correlation between presence of granulocyte antibodies and degree of neutropenia. CONCLUSIONS: We conclude that granulocyte antibodies are highly prevalent in children with HIV infection but do not correlate with the degree of neutropenia. Antineutrophil antibody determination as currently performed does not appear to be useful in the evaluation of the HIV-infected neutropenic child.

Adolescent↗
PubMed1997

Neutrophil antibodies.

Several advances have been made in understanding the polymorphisms of the neutrophil Fc-gamma-receptor IIIb (Fc gamma RIIIb). In one recent study, 21 individuals whose neutrophils lack Fc gamma RIIIb were found to be missing the entire Fc gamma RIIIB and Fc gamma RIIC genes. Another polymorphism of Fc gamma RIIIb, SH, has been characterized. New methods to determine the genotype of Fc gamma RIIIB for NA1, NA2, and SH using leukocyte genomic DNA have been described. A new monoclonal antibody to neutrophil-specific antigen NB1 was produced. Advances have been made in understanding alloimmunization in granulocyte transfusion recipients and the treatment of autoimmune neutropenia with granulocyte colony-stimulating factor (G-CSF). Granulocyte transfusion recipients were found to be alloimmunized both to neutrophil-specific and HLA antigens, suggesting that the transfusion of these patients with granulocytes matched only for HLA antigens will not be effective. A case report suggests that the beneficial effects of G-CSF on patients with autoimmune neutropenia is due in part to G-CSF's action of increasing plasma levels of soluble Fc gamma RIIIb.

Autoimmune Diseases↗