1 Comment


Williams Hematology



History of Granulocyte Transfusion
Preclinical Studies of Dogs with Granulocyte Transfusion
Clinical Experience with Granulocyte Transfusion

Granulocyte Concentrates from CML Donors

Controlled Trials of Granulocyte Concentrates from Healthy Donors

Mobilization of Donor Granulocytes and Collection

Indications for Granulocyte Transfusions

Prophylactic Transfusion

Storage of Granulocyte Concentrates
Apheresis-Derived Peripheral Blood Stem Cells

History of Peripheral Blood Stem Cell Transplantation

Stem Cell Mobilization Strategies

Stem Cell Collection for Allogeneic Transplantation

Dose and Enumeration of Stem Cells

Timing of Stem Cell Collections
Mononuclear Cells for Adoptive Immunotherapy

Allogeneic Adoptive Immunotherapy

Autologous Adoptive Immunotherapy
Chapter References

The availability of blood cell components including red cells, platelets, plasma, and plasma proteins has facilitated the treatment of hematologic diseases. Preparation of an effective granulocyte product has been elusive. The advent of recombinant granulocyte-mobilizing cytokines, which recruit large numbers of granulocytes into the blood, the use of hemapheresis instruments to cull the granulocytes from up to 15 liters of blood, and the use of starches to increase the efficiency of the separation of red cells from granulocytes have combined to make granulocyte transfusion of patients with severe reversible neutropenia and serious infections possible.
Stem cell transplantation from blood products has been made possible by using chemotherapeutic agents, cytokines, or combinations of the two to mobilize stem cells from marrow into blood, allowing recovery of sufficient numbers of stem cells by hemapheresis of several blood volumes. In so doing, sufficient CD34 antigen-positive cells can be recovered to engraft an allogeneic or autogeneic recipient.
Blood mononuclear cells of several types can be harvested by hemapheresis and used for immunotherapy. T lymphocytes can be used in their native state or after lymphokine activation. CD34 antigen-positive cells can be harvested and induced to transform into antigen-presenting (dendritic) cells by in vitro treatment with several cytokines. The use of mononuclear cells for immunotherapy of lymphoma, leukemia, or myeloma is becoming more frequent, and these approaches are dependent on hemapheresis to obtain sufficient quantities of cells to have therapeutic effects.

Acronyms and abbreviations that appear in this chapter include: BFU-E, burst-forming unit–erythroid; CD, cluster of differentiation; CFU-GEMM, colony forming unit–granulocyte-erythocyte-monocyte-megakaryocyte; CFU-GM, colony forming unit–granulocyte-macrophage; CML, chronic myelogenous leukemia; G-CSF, granulocyte colony stimulating factor; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL, interleukin; LAK, lymphokine-activated killer (cell); TIL, tumor-infiltrating lymphocyte.

Granulocyte transfusion for the treatment of neutropenia was first explored in 1934 but met with little success.1 It was not until 1953 that the first promising experiments with leukocytes were reported.2 In these experiments, leukocytes transfused into irradiated dogs migrated to sites of infection and were capable of normal phagocytic activity.2
Subsequently, interest in granulocyte transfusions has been cyclical. In the 1960s there was a strong sentiment for the use of granulocyte transfusions. This enthusiasm was fueled by the ability to collect large quantities of granulocytes from patients with CML and the early success with the infusion of these cells to patients with severe granulocytopenia and life-threatening infection.4 However, due to the unavailability of donors and concerns with transfusing malignant cells this practice was abandoned. In the 1970s, following the introduction of blood cell separators capable of harvesting large quantities of granulocytes from healthy donors, there was an overly optimistic view that granulocytes would have substantial clinical impact. However, in the 1980s disillusionment had set in following limited success with healthy-donor granulocyte therapy, and use of this therapy decreased. In the mid-1990s the introduction of recombinant cytokines capable of mobilizing large numbers of healthy-donor granulocytes led to renewed interest in granulocyte therapy.
Studies in neutropenic dogs with gram-negative sepsis,3,4,5,6 and 7 pneumonia,8,9 or candidemia10,11 and 12 found increased survival with or without antibiotic therapy. A threshold dose of 2 × 108 granulocytes/kg was protective against a lethal infection with Pseudomonas, while there was uniform mortality when doses less than 1.5 × 108 granulocytes/kg were administered. Mean 1 h posttransfusion increments of greater than 200/µl or greater than 500/µl were seen with infusions of at least 2 × 108 and 3 × 108 granulocytes/kg, respectively. Such studies suggested that a clinical benefit from granulocyte transfusions in neutropenic septic patients would be strongly dose-dependent.
Cell collections from donors with CML yielded as many as 1 × 1011 phagocytic cells,17,18 and transfusion of these concentrates increased white cell counts and led to improvement in septic neutropenic recipients.19,20 and 21 Leukocyte transfusions from CML donors also caused a more sustained elevation of the white cell count, perhaps because of their content of granulocytic progenitors capable of cell division.17 Patients with CML are rarely available and are not considered to be acceptable blood donors.
There have been seven controlled clinical trials of granulocyte transfusion therapy for the treatment of bacterial sepsis in adults.22,23,24,25,26,27 and 28 These trials were considered successful,23,25,26 partially successful,22,27 or unsuccessful.24,28 A meta-analysis of these studies concluded that the dose of granulocytes transfused (greater than 1 × 1010 granulocytes transfused per day on at least four successive days) and the survival rate of controls (efficacious when the survival of the controls was under 40 percent) were primarily responsible for the disagreements across the reports.29 When the analysis was limited to the five randomized controlled studies,23,24,25,26,27 and 28 granulocyte therapy conferred a significant survival benefit in the following settings: (1) low survival rate of controls (RR = 8.0, 95% CI, 1.5–42), (2) adequate dose of granulocytes (RR = 4.0, 95% CI, 1.2–12), (3) timely marrow recovery; and (4) pretransfusion assessment of compatibility of granulocytes (RR = 8.0, 95% CI, 1.5–42).
Bacterial infection in the neonatal period may be life-threatening because of inadequate granulocyte function or the infant’s inability to mount or sustain neutrophilia because of inadequate reserves of neutrophilic precursors. In septic neonates there have been five studies of the effects of granulocyte therapy,30,31,32,33 and 34 four of which were randomized.31,32,33 and 34 A meta-analysis of these studies found that the only significant predictor of success was a dose of cells greater than 0.5 × 109/kg (RR 18, 95% CI, 1.3–252).29 Granulocyte therapy was not efficacious when buffy coats (as contrasted with granulocyte concentrates) which contained less than 0.5 × 109/kg granulocytes were transfused (RR = 0.74, 95% CI, 0.20–2.7). The effect of a low survival rate of controls did not reach statistical significance; however, the relative risk was large (RR = 13, 95% CI, 0.39–462). The effect of leukocyte compatibility was not assessed in the studies of neonates.
Less than five percent of the total body neutrophil pool is in the intravascular space.36,37 These cells have a half-life of only 4 to 10 h in the intravascular compartment.38,39 The collection of large numbers of granulocytes requires the processing of 5 to 14 liters of blood per donation, which can be accomplished within 2 to 3 h. Apheresis procedures are performed with a special centrifuge.39,40 and 41 Donor blood is pumped through the centrifuge and separated, based on density, into three layers. The leukocyte-containing layer is harvested to form the granulocyte concentrate, and the red cells and plasma are returned to the donor. Of historical note is that an alternative, noncentrifugal method of granulocyte collection that exploited the ability of granulocytes to adhere to nylon in the presence of divalent cations was once employed.42 Although filtration leukapheresis was simple and yielded large quantities of granulocytes,27,39,40,42,43,44,45,46,47,48,49,50 and 51 the technique has been abandoned because of the need to heparinize the donor and because of reactions such as severe abdominal pain,52 priapism,53 complement activation in the donor,54,55,56,57 and 58 frequent shaking chills in the recipient,59 and poor survival, in vivo recovery, and chemotactic capability of the collected neutrophils.18,28,60,61,62,63 and 64
Granulocyte collections taken from healthy donors by centrifugal leukapheresis do not routinely yield adequate quantities of phagocytic cells (0.1–0.5 × 1010 granulocytes per liter of donor blood).29,40 This is due to two factors: (1) the difficulty of separating granulocytes from lighter red cells and lymphocytes because of the similarity in their density, and (2) the low number of granulocytes in the circulation. The addition of a rouleaux-inducing sedimenting agent to the blood before it enters the centrifuge enhances the separation of red cells from white cells and increases yields as much as twofold. The most common rouleaux-inducing agents employed are hydroxyethyl starches.17,19,20 and 21 Hydroxyethyl starches are branched-chain polyglucose molecules, or glucans, similar in structure to glycogen but having variable degrees of hydroxyl substitutions on the glucose residues (carbons 2, 3, and 6). Such substances are also used as a synthetic colloid volume expander. Virtually all centrifugal cell separators require the use of cell-sedimenting agents to achieve an adequate granulocyte yield. In the United States two hydroxyethyl starches, hetastarch (Hespan), sometimes referred to as high molecular weight hydroxyethyl starch or HMW-HES, and pentastarch (Pentaspan), sometimes referred to as low molecular weight hydroxyethyl starch, or LMW-HES, are approved and commercially available for human use. Pentastarch was thought to be a superior starch erythrocyte-sedimenting agent due to its lower molecular weight (average pentastarch 260 kDa, versus hetastarch 450 kDa) and consequently more rapid and complete clearance.65 A paired controlled prospective study involving 36 donors from whom collections were alternately made with both starch sedimenting agents found that in 33 of 36 (92%, P < .001) pairs of donors, hetastarch was significantly more efficient in granulocyte collection. Overall, hetastarch provided 1.6-fold greater collections than did pentastarch.66 Donor reactions are not common but can range from mild rashes, rhinorrhea, or paresthesias to anaphylaxis, circulatory failure, pulmonary edema, and others.
Donor granulocyte counts also can be increased prior to donation by the administration of drugs that either mobilize granulocytes from the marrow or cause a shift of granulocytes from the marginal pool to the circulating blood pool.67,68 At present, dexamethasone or another glucocorticoid is administered either orally or by intravenous infusion to raise the donor granulocyte count prior to leukapheresis.20,39,40,68,69 and 70 Various dose schedules have been used, but a typical regimen is 8 mg dexamethasone (or 20–30 mg prednisone) given orally 12 h prior to collection. Leukapheresis yields have increased by 50 to 100 percent after glucocorticoid administration, and their use is routine in most leukapheresis facilities. The effects of hydroxyethyl starch and glucocorticoid administration are additive, and neither has significant adverse effects on the function of the granulocytes.60,68,71,72
Despite the availability of glucocorticoids and erythrocyte sedimenting agents, a dose of greater than 1 × 1010 granulocytes frequently is not achieved with one apheresis collection. This level may be particularly difficult to achieve for the first collection when there has not been time to medicate the donor with glucocorticoids prior to the apheresis collection.
The introduction of cytokines such as G-CSF (Neupogen) for the mobilization of hematopoietic cells has allowed the collection of much larger numbers of granulocytes. Multiple studies employing G-CSF at a dose of 5 to 10 µg/kg per day (300–600 µg given subcutaneously) administered 12 h prior to collection will result in a blood white count in the range of 20–50 × 109/liter range with a three- to sixfold increase in the number of granulocytes collected (4–8 × 1010 granulocytes).73,74,75 and 76 Combining glucocorticoids and cytokines results in even greater efficacy, with mean collections of 0.8–1.1 × 1011 granulocytes.76,77 and 78 Such doses approach the amount of granulocytes that a normal marrow produces in a day (1.5 × 109/kg).79,80
Use of G-CSF is not without problems. Donors complain of bone pain, headaches, fatigue, night sweats, nausea and vomiting, sleep disturbances, and redness or swelling at the injection site.77 Comparison of multiple mobilization schemes suggests that a dose of both G-CSF and glucocorticoids given once on the day prior to collection minimizes side effects and results in a fourfold increase in granulocytes collected over the use of glucocorticoids alone.76,77 and 78 A single dose of 300 µg of G-CSF results in nearly the same elevation in neutrophil count as a 600-µg dose 78. For repeated donations, administration of G-CSF on alternate days maintains the neutrophil count, with decreased side effects and lower cost than a daily regimen.75 In a paired three-arm study, the use of dexamethasone alone resulted in 25 percent of donors reporting insomnia or flushing. With G-CSF, 65 percent had bone pain, headache, insomnia, fatigue, nocturia, or diaphoresis; this increased to 85 percent with combination of dexamethasone and G-CSF.78 There are no known long-term sequelae after the transient administration of G-CSF to healthy donors.
The granulocytes collected following cytokine mobilization are not fully mature, but studies have not identified any significant dysfunction granulocyte dysfunction. Granulocytes appear activated (primed for a respiratory burst), have normal phagocytosis and staphylococcal killing capability in vitro, and have a prolonged half-life in the circulation.73,78,79,80 and 81 There is evidence of degranulation and decreased postinfusion recovery (31% versus 65%), but G-CSF-stimulated granulocytes migrate into skin-window chambers and into sites of inflammation and infection.78 After G-CSF priming of the donor, there are alterations of surface antigen expression with increased expression of adhesion molecules (CD11b, CD14, CD18) and Fc receptors (CD32, CD64).78 The clinical significance of these changes, if any, is not known.
The indications for granulocyte transfusion are not well defined. In general, the patient should have (1) an absolute granulocyte count less than 500/µl (0.5 × 109/liter), (2) fever, (3) an identified responsible microorganism, and (4) no decrease in fever after 48 h of antibiotic treatment. Also, the prognosis without granulocyte therapy in the specific setting should be poor and the prognosis of the underlying disorder should be favorable. When possible, attempts should be made to match for leukocyte antigens and transfuse cytomegalovirus-negative, irradiated cells.
The risk of infection in granulocytopenic patients is related to the severity and duration of the cytopenia.82 The risk of infection increases dramatically when the granulocyte count is below 500/µl (0.5 × 109/liter). An equally important consideration in determining the need for granulocyte transfusion is the status of the patient’s marrow. If early marrow recovery is expected, granulocyte concentrates are probably superfluous. Conversely, if the neutropenia is expected to last for more than several days the patient may benefit from granulocyte transfusions when serious infection is present.
Granulocyte transfusions also have been used successfully in the treatment of infected patients with chronic granulomatous disease. In these patients granulocyte transfusion is indicated if appropriate antibiotic therapy used alone is unsuccessful.83,84,85 and 86
The usefulness of giving prophylactic granulocyte transfusions to neutropenic patients without evidence of infection has been evaluated in several studies.86,87,88,89,90,91 and 92 In a large randomized study, neutropenic patients with leukemia who received prophylactic granulocyte transfusions did not have an overall reduced incidence of infections compared with controls, although the proportion of transfused patients with bacterial septicemia was reduced. Also, prophylactic granulocyte transfusions did not improve the rates of remission, survival, or time to marrow recovery.90 Most other studies agree with these findings. No randomized studies have been done with the G-CSF plus glucocorticoid stimulated granulocyte collections.
Two randomized studies have evaluated the use of prophylactic granulocyte transfusions in patients receiving marrow transplants. One study found fewer infections in transfused patients than in controls.88 In contrast, the other study89 found a decrease in bacterial septicemia in transfused patients but did not observe a reduction in overall infection; survival was not improved. Furthermore, 13 of the 18 patients receiving transfusions developed cytomegalovirus infections, compared with only 6 of 17 controls. Thus, viral infection is a significant additional hazard in severely immunosuppressed patients.
One major drawback to prophylactic granulocyte transfusion is the increased likelihood of transfusion reactions, alloimmunization, lymphocytotoxic antibodies, and refractoriness to future transfusions. Refractoriness to platelet transfusion occurs frequently in patients who are given granulocyte transfusions.87 Although there is some reduction in the incidence of infection, the occurrence of alloimmunization and the absence of clear benefit are major deterrents to the use of prophylactic granulocyte transfusions.
G-CSF mobilized, HLA-matched, ABO compatible granulocytes provided to allogeneic bone marrow transplant recipients resulted in significant and sustained increments in the neutrophil and the platelet count.75 The peak of the neutrophil increment was observed 4 to 12 h following infusion and the rise in the neutrophil count persisted for 25 to 37 h. Nevertheless, three of the ten recipients developed culture-proven bacterial infections during the period of granulocyte infusions.
The general hazards of blood transfusion also apply to granulocyte transfusions. Hemolytic reactions may occur because there are red cells in granulocyte concentrates. ABO-compatible concentrates should be given. Recovery of granulocytes in the recipient 1 h after transfusion is adversely affected by ABO incompatibility.22 The presence of erythrocyte antibodies in the recipient other than AB blood groups should also be ascertained prior to transfusion, but such erythrocyte incompatibilities are not an absolute contraindication to transfusion if the patient’s infection is severe. If necessary, most of the erythrocytes in the concentrate can be removed by differential centrifugation prior to transfusion.
Febrile transfusion reactions occur frequently with granulocyte transfusions.48,59 About 10 percent of recipients have chills and fever after transfusion of cells collected by a centrifugation method, as a result of antileukocytic antibodies in the recipient or pyrogen released from damaged white cells. Glucocorticoids, antihistamines, and meperidine can be given either prophylactically or therapeutically to reduce the frequency or severity of these reactions.
Acute pulmonary insufficiency has been reported with granulocyte transfusion.93 A number of different mechanisms have been proposed. In neutropenic patients with pneumonia it has been postulated that the rapid migration of transfused granulocytes into the infected lung may induce such a reaction.93 Leukoagglutinins in the recipient may result in leukocyte aggregates which embolize to the lung.94 Complement activation and the generation of C5a, which results in granulocyte aggregation or adhesion to endothelial cells, also have been postulated.95 Severe pulmonary reactions with the concomitant use of granulocyte transfusion and amphotericin B have been reported by some observers96 but not by others.97,98 As a precaution, however, amphotericin B and leukocyte transfusion should be administered at different times.
Severely immunosuppressed patients are at risk for other serious complications of granulocyte transfusion. Graft-versus-host disease, sometimes fatal, has occurred in patients receiving intensive chemotherapy and perhaps in neonates.99,100,101,102,103 and 104 In such cases, the concentrate should be irradiated with 2500 rad prior to administration to prevent this occurrence. Severely immunosuppressed patients, especially those undergoing marrow transplantation, also may develop a cytomegalovirus infection, particularly pneumonia, that may be fatal.89 Individuals who do not have serologic evidence of prior cytomegalovirus infection should be selected as donors for immunosuppressed patients.
It is best to transfuse granulocytes as soon as possible after collection.48,105 Granulocyte concentrates system should not be stored for more than 24 h.
Granulocyte chemotaxis is the first function altered during storage.72,106,107,108 and 109 Changes in phagocytic function, bactericidal capacity, and biochemical functions such as fructose-6-phosphate (hexose monophosphate) shunt activity and oxygen consumption are generally less sensitive indicators of cell damage during storage.
Cell viability and bactericidal capacity seem to be well maintained at either room temperature (22°C) or during refrigerator storage (5°C) for at least 24 h,106,107,108,109,110 and 111 but chemotaxis, chemotaxis-related functions, adherence to endothelial cells, and cell ATP levels are better preserved at 20–24°C.107,108,111,112,113,114,115,116 and 117 Storage of granulocytes is associated with a tendency for hyperadherence and spontaneous aggregation that is more pronounced at 5°C.116 Because of better preservation of function, current practice is to store granulocytes at room temperature when storage is required.
In general, granulocyte concentrates are better preserved in plasma than in other suspension fluids,111 but it is possible that improved storage can be achieved in other optimized solutions.118 Additional glucose may be required when the cell leukocyte count is high119 or when there are significant numbers of contaminating platelets and red cells that compete with the granulocytes for glucose.120 The type of plastic used in the storage bag is another important storage variable.121 Gentle agitation was found to be beneficial in one study122 but not in another.111 It is unclear whether irradiation of granulocyte concentrates prior to transfusion to prevent graft-versus-host disease is associated with alteration in cell function.123,124 The subject of granulocyte preservation has been reviewed.125
Cryopreservation of granulocytes would be an ideal method of storage because it would permit the “banking” of large quantities of cells that would be available when needed. Dimethyl sulfoxide is used as a cryoprotective agent, but there are no satisfactory methods for freezing granulocytes. Post-thaw recovery of viable cells, as measured by phagocytosis, rarely exceeds 25 percent.126,127 and 128
Granulocyte transfusion therapy has been full of promise for several decades. Now, with the ability to reliably collect G-CSF plus steroid-mobilized granulocytes from healthy donors with total doses exceeding 1 × 1011 granulocytes, large randomized trials are needed to define the efficacy, toxicity, and cost-effectiveness of granulocyte transfusions both for prophylaxis in transiently neutropenic patients and for neutropenic patients with progressive bacterial or fungal infections.129
Within the past 10 years the role of blood stem cells has evolved from research conducted in a few centers to a widely utilized therapeutic modality. Since the initial hypothesis in 1909 that hematopoietic stem cells circulate130 and subsequently, the demonstration of hematopoietic reconstitution following myeloablative irradiation and parabiosis in rodents by Brecher and Cronkite in 1951,131 the potential applications of blood stem cells in humans have emerged. Documentation of cell engraftment in a patient receiving granulocytes derived from a donor with chronic myelogenous leukemia in 1963132 paved the way for the first successfully performed autologous blood stem cell transplant in a patient with chronic myelogenous leukemia in 1979.133 In 1986, successful hematopoietic reconstitution by autologous blood stem cell transplantation was described in a patient with Burkitt lymphoma following marrow-ablative chemotherapy.134 Initially, autologous stem cells collected by leukapheresis were used to shorten the period of pancytopenia following myeloablative chemotherapy in patients undergoing bone marrow transplantation.135,136 Early successes with blood stem cell transplantation involved steady-state harvesting of the circulating hematopoietic stem cells by apheresis without specific mobilization strategies. Engraftment times were comparable to bone marrow transplantation.134,137,138 Unfortunately the number of collections, time, and cost of collection and transplantation of blood stem cells were not yet optimal for the evolving technology to emerge as an alternative to marrow transplantation.139 The development of mobilization techniques to enhance the circulating number of stem cells, optimizing of the timing of apheresis harvests, and progress on characterizing stem cells by immunophenotyping have enabled this transition to occur. Currently, autologous transplantation using blood stem cells is performed more frequently than autologous bone marrow transplants.139,140 and 141
Apheresis-derived blood stem cells offer several benefits over marrow transplants.142 Apheresis collection is feasible on an outpatient basis, avoids general anesthesia, and enables the processing of multiple blood volumes per collection. Blood stem cell collection is suitable for patients whose marrow is infiltrated by disease or fibrosis or who have a history of prior pelvic radiation. Blood stem cells result in a shortened cytopenic period following myeloablative treatment, fewer infectious complications, a lower incidence of tumor cell contamination, decreased transfusion requirements, and a shortened period of hospitalization.135,143,144 Disadvantages include a potentially longer period to obtain an adequate harvest; a greater volume to infuse; maintenance of central venous access, which predisposes to risks for infection and thrombosis145,146; and, in the allogeneic setting, the possibly greater risk of graft-versus-host disease due to the presence of mature lymphocytes in the stem cell suspension.
Technical advances have facilitated the application of peripheral blood stem cell transplantation. These include improved automated blood separators capable of efficient collection of mononuclear cells and the use of cryopreservation and sterile bag systems to pool components from multiple collections.147 Innovations in mobilization techniques, quantification of appropriate stem cell doses to ensure timely engraftment, and optimization of the timing and efficiency of apheresis collections have contributed to the growing use of this stem cell collection strategy.
Collection of adequate numbers of stem cells from blood requires large volumes to be processed (2–4 blood volumes). Retrieval of this small population of cells is challenging, since very primitive hematopoietic cells comprise only 0.01 to 0.1 percent of circulating leukocytes and 1 to 10 percent of cells obtained from bone marrow.148 Mobilization refers to those techniques which increase the proportion of circulating primitive cells, especially stem cells. Although the exact mechanism(s) by which mobilization occurs is unclear, glucocorticoids, endotoxin, stress, exercise, and dextran have been shown to cause alterations in the concentration of peripheral progenitor cells.149 Mobilization strategies have significantly improved the efficiency of peripheral blood stem cell collection by apheresis and have decreased the number of procedures necessary to achieve target stem cell collection amounts. As a consequence, transplantation using blood stem cells has become an attractive alternative to marrow transplantation.
Myelosuppressive (as opposed to myeloablative) chemotherapy is associated with a recovery phase with increased concentrations in circulating hematopoietic progenitor cells.150,151 A variety of chemotherapeutic agents/regimens can elicit mobilization, which is enhanced depending on the dose and timing of the preceding chemotherapy.152 The effectiveness of mobilization can be assessed by in vitro clonogenic progenitor cell assays of blood. These assays have also been instrumental for quantifying the harvested stem cells after apheresis. Morphologically distinct CFU-GM, BFU-E, and CFU-GEMM colony-forming units can be quantified by colonial growth on cytokine-supplemented semisolid media after incubating 7 to 20 days. However, due to the time required for these assays they primarily provide retrospective information. Flow cytometric “real time” assessment of stem cell content using CD34 antigen has greatly enhanced the clinical utility of stem cell enumeration (see “Dose and Enumeration of Stem Cells”).
Use of high-dose cyclophosphamide to mobilize progenitor cells has been effective in patients with lymphoma, myeloma, and solid tumors.134,153,154 A 14-fold increase in the mean CFU-GM in blood of patients with lymphoma, myeloma, and solid tumors occurred two days after recovering leukocyte counts exceeded 1 × 109/liter.154 Comparison of hematologic recovery times between autologous chemotherapy mobilized stem cell transplants and marrow autotransplants revealed significantly shorter time to attain a neutrophil count of 0.5 × 109/liter (mean of 11 days versus 22 days) and a platelet count of 50 × 109/liter (mean of 14 days versus 32 days).144 In addition, the number of hospital days and blood component transfusions were reduced in the group that received blood stem cells.
When mobilization regimens incorporate cytokines such as G-CSF or GM-CSF in addition to chemotherapy, progenitor cell concentrations increase dramatically over steady-state blood levels.155 A strategy combining chemotherapy and hematopoietic growth factors offers the most effective means for mobilization of progenitor cells. This strategy was first described in the treatment of patients with sarcoma.156 GM-CSF when combined with myelosuppressive chemotherapy produced a 63-fold elevation in circulating progenitors measured as CFU-GM by in vitro culture. GM-CSF priming alone produced a 13-fold increase in CFU-GM, and chemotherapy alone produced only a twofold rise in CFU-GM. The synergistic effect of chemotherapy plus cytokine administration becomes readily apparent when reviewing published experience. In general mobilization with chemotherapy alone affords a 20- to 50-fold increase in circulating CFU-GM. Cytokines or hematopoietic growth factors alone afford a comparable mobilization: G-CSF typically results in a 20- to 50-fold increase in CFU-GM, while GM-CSF results in a modest 10-fold increase in CFU-GM. When cytokines are combined with chemotherapy, mobilization of CFU-GM increases by approximately 70-fold (range 25 to 250 fold).153,157,158,159,160,161,162,163 and 164 A median time to a platelet count of greater than 20 × 109/liter occurred in 10 days in a chemotherapy plus G-CSF mobilized stem cell transplant group, as compared to 17 days in an autologous marrow transplant group.165 The preferred approach is to use harvested blood stem cells as the source of hematopoietic reconstitution.140,162 However, effective mobilization of autologous stem cells in patients may be unsuccessful, depending on the extent of previous chemotherapy/radiation or involvement of the marrow in disease.166
Blood stem cell transplantation in the allogeneic setting has evolved at a slower pace due to concerns of graft-versus-host disease that might arise from donor lymphocytes. Fortunately, experience with sibling HLA-matched donors, syngeneic donors, and a second autologous marrow transplantation revealed minimal graft-versus-host disease and few adverse effects on the donor.167,168 and 169 Allogeneic blood stem cell transplantation results in rapid, multi-lineage engraftment following successful mobilization of donor stem cells with G-CSF. Higher doses of G-CSF enhance the progenitor cell harvest beyond that obtained by marrow harvest and have been associated with more rapid engraftment of platelets.170 Initial enthusiasm was offset by reports of chronic graft-versus-host disease, which occurred despite the low numbers of CD3+ cells present in the blood stem-cell suspension following CD34+ cell selection.171,172 Diminishing the graft-versus-host effect while retaining the graft-versus-leukemia effect remains a challenge.173
Reports of significant donor reactions have been infrequent, although cytokine mobilization has been associated with mild bone pain, headache, body aches, fatigue, nausea and vomiting, local reactions at the injection site, insomnia, night sweats, dyspnea, transient changes in serum chemistries, and thrombocytopenia.73,74,143,174,175,176,177,178 and 179 Risks associated with apheresis include citrate toxicity, hypotension, fatigue, and if antecubital access is insufficient, the risks of central venous access. Large-volume leukapheresis collections are typically successful in harvesting sufficient progenitor cells in one or two collections. While clinically significant thrombocytopenia has not been described following administration of G-CSF, stem cell suspension products contain a substantial number of platelets. A drop from preapheresis platelet counts of approximately 28 percent per daily collection has been described, with recovery after about 10 days.179 A predictable 2–4% decrease of the platelet count per liter of whole blood processed occurs in donors.180 Close monitoring of pre-, post- and intraprocedure donor platelet counts is advisable. Strategies to minimize this problem may include shorter collections, longer recovery periods between collections, or the use of thrombopoietin. Salvaging of platelets may provide the most immediate practical solution.181
The long-term effects, and specifically the risk of subsequent hematologic consequences, remain a theoretical concern when exposing healthy donors to hematopoietic growth factors. Several hundred allogeneic blood stem cell donors followed for 2 to 5 years postdonation have not shown an increased risk of hematologic malignancy.139 A prospective study of 19 donors demonstrated that one year after the administration of G-CSF their blood counts were normal and unchanged from predonation counts. Furthermore, following a second course of G-CSF the progenitor content of the peripheral blood stem-cell product was comparable to the first.182
The CFU-GM count correlates inversely with the time to detectable engraftment, the length of hospitalization, and the amount of supportive care needed.136,144,183 The correlation between the CFU-GM and CD34 counts reported by several independent investigators,184,185,186 and 187 along with the advent of flow-cytometric quantitation of CD34 cells, has facilitated more rapid and reproducible measurement of putative stem cells in “real time.”
The CD34 antigen is a glycoprotein expressed on the surface of lymphohematopoietic self-renewing stem cells, lineage-committed progenitors, some endothelial cells, embryonic fibroblasts, and marrow stromal cells. Although its functional role has yet to be definitively established, it has been suggested that the CD34 antigen assists in the adhesion and routing of primitive hematopoietic cells during hematopoiesis and differentiation.139 Approximately 1.5 percent of the bone marrow mononuclear cells express CD34. Within this population reside the CFU-GM, BFU-E, and CFU-GEMM capable of short-term hematopoietic reconstitution,176,188,189 as well as the pluripotential self-renewing stem cell, the exact phenotype of which remains controversial. The pluripotential stem cell presumably resides within the CD34+, thy-1dim, CD38–, HLA-DR–, and lineage-specific negative subpopulation (see Chapter 14).162
The blood contains approximately 1 to 10 percent of the CD34+ population in the marrow in the steady state. Mobilization techniques, as discussed above, expand this circulating compartment and enable efficient collection by leukapheresis. The adequacy of a blood stem-cell collection is gauged by the number of CD34+ cells per kilogram of recipient body weight. The minimal threshold of CD34+ cells necessary for neutrophil and platelet recovery after autologous transplantation has ranged from 2 to 5 × 106 per kilogram.166,190,191,195 Higher stem-cell doses have been associated with accelerated platelet engraftment.170 In one study of breast cancer patients, 14 days after transplant, the probability of the platelet count being greater than 20 × 109/liter was 95, 85, 65, or 50 percent with a total infused doses of 10 × 106, 5 × 106, 2 × 106, or 1 × 106 CD34+ cells/kg, respectively. Some transplant physicians have made a distinction between an optimal cell dose for autologous versus allogeneic stem-cell grafts and recommend that the latter contain at least threefold more progenitor cells, although the necessity for this has not been proved.139
Increasing the total dose of CD34+ cells from apheresis-derived collections theoretically should result in a greater likelihood of enduring hematopoietic engraftment. The higher incidence of tumor cell contamination with increased numbers of apheresis collections for autologous stem-cell harvests of breast cancer patients gives pause to the notion that more is better.193 A shortened disease-free survival was observed among breast cancer patients undergoing autologous marrow transplantation who had evidence of micrometastatic disease in the marrow harvested, as measured by immunocytochemical methods.194
A variety of methods for enumeration of CD34+ cells by flow cytometry have been published.148,194,195,196,197,198,199,200,201 and 202 However, comparison of CD34 measurements from different centers is difficult due to differences in methods among laboratories. A multi-institutional study of 21 marrow and blood samples showed a twenty-fold range in results.203 These discrepant observations confirmed results from several smaller studies196,198,204 and emphasize the need for standardization. One approach has been suggested by the International Society for Hematotherapy and Graft Engineering (ISHAGE).205 Counting techniques that incorporate a standard bead into the analysis or volumetric capillary cytometry hold promise for increasing accuracy and reproducibility.206,207 and 208
At present, stem cell collections using continuous flow apheresis equipment depends on processing 1.5 to 4 blood volumes over several hours. Repeated daily collections over 2 to 5 days are used to reach an adequate threshold of stem cells. The adequacy of the collection is dependent on the mode of mobilization, the timing of the collection with respect to the recovering leukocytes, the volume of whole blood processed, and the amount of patient pretreatment.162
Recruitment of noncirculating hematopoietic progenitors during a single large-volume leukapheresis collection (2 to 3 times the donor blood volume) can occur, including a 2.5-fold increase in the CFU-GM when more than 15 liters of whole blood was processed.186,209,210 and 211 However, such results are dependent on the patient’s ability to tolerate prolonged leukapheresis, adequate mobilization, and timing.212
Healthy allogeneic donors treated with G-CSF have a predictable peak in circulating stem cells occurring 5 to 6 days after initiating daily subcutaneous doses of G-CSF of 5 to 10 µg/kg per day.134,143,176,214,216
Timing collections in the autologous setting has been more challenging. Early studies suggested that collections begin when the leukocyte count exceeded 1.0 × 109/L.139,214 While circulating progenitor cells and the total mononuclear count begin to rise concurrently, premature cessation of cytokine administration in these studies likely led to a decrease in circulating progenitors.139 Subsequently, cytokine administration has been continued longer throughout serial collection procedures. Peak levels of circulating progenitors have been reported 2 days after the WBC count exceeds 2 × 109/L,213 12 days after the WBC exceeds 10 × 109/L,167 or when the rebounding WBC reaches 5 × 109/L.157
Unfortunately, the blood white cell and mononuclear cell counts do not correlate with the number of hematopoietic progenitor cells in the blood.217,218 Quantitation of circulating CD34+ cells the day of leukapheresis is a more accurate predictor of the optimal time of stem cell harvest.217,218,219,220,221,222 and 223 A minimum threshold greater than 10 to 30 circulating CD34+ cells per microliter will afford a satisfactory stem cell harvest, although serial collections may still be necessary to attain the appropriate CD34+ dose for transplantation.219,220 and 221 When the circulating progenitors approach or exceed 100 CD34+ cells per microliter, a single leukapheresis may provide all of the CD34+ cells necessary.
The relationship of circulating CD34+ cells with the CD34+ cell yield allows for the optimal timing of stem cell harvests. Thus, fewer collections and a lower volume of pooled product to infuse following myeloablation can be achieved. By combining microvolume fluorometry and an automated analysis, accurate CD34+ cell counts can be made in 30 min. This assay has the potential to expedite optimal timing of stem cell harvests.207,208
Finally, awareness of the phenotype of the patient’s malignant cells is important, since lymphoblastic lymphomas and other hematologic malignancies can occasionally express the CD34 antigen and yield an erroneously high stem cell dose.
Allogeneic blood cell collection and reinfusion may benefit marrow transplant patients with recurrent leukemia. This form of adoptive immunotherapy utilizes the inherent immune reactivity of apheresis-derived allogeneic leukocytes from the original marrow donor to either prevent recurrence or induce remission. With strategies that manipulate the timing and the dose of donor T lymphocytes infused following transplantation, this therapy attempts to exploit the beneficial graft-versus-leukemia effect while minimizing the risks of graft-versus-host disease. Donor lymphocytes have been used to induce cytogenetic remission in patients with CML who relapsed after transplantation.173,224 Although the role of interferon-a in such therapy has been questioned, the antileukemia effect of reinfused donor lymphocytes in CML patients has been confirmed, yielding a composite clinical response rate of 77 percent.225 Unfortunately, transfusion of allogeneic mononuclear cells from the original donor has not been capable of inducing a durable remission with other hematologic malignancies.224,226 Graft-versus-host disease remains a major complication frequently accompanying this therapy. Of the CML patients who achieved remission, graft-versus-host disease developed in 73 percent (8/11) of patients receiving a total T-cell dose greater than 5 × 107/kg but only in 12.5 percent (1/8) of patients who received a total T-cell dose of 1 × 107/kg.173 Remission in CML can result from donor leukocytes containing as few as 1 × 107 T cells/kg.
Another application of mononuclear cell collection using apheresis cell separators has been in the preparation of autologous mononuclear cells with antitumor activity. Examples include the production of LAK cells,227,228 TIL,229 and activated dendritic cells (antigen-presenting cells). Each of these applications must be considered experimental at present.
The production of LAK cells may require pretreatment of the patient with IL-2, leukapheresis for the collection of mononuclear cells, ex vivo manipulation of the collected cells to activate and/or purify them, and reinfusion into the patient. In a study of 539 leukapheresis collections, a mean of 3.03 × 1010 lymphocytes were collected in a four-hour procedure.230 High yields of lymphocytes in these procedures result in part from recruitment of these cells into the circulation by prior IL-2 treatment. The culturing, harvesting, and washing of these lymphocyte preparations prior to infusion, rather elaborate processes, have been described.231,232 and 233
Due to the limited success observed with both LAK and TIL cell therapy for metastatic cancer, attention has turned to the collection, purification, activation, and reinfusion of dendritic cells as a means for the production of tumor-specific cytotoxic T cells. Experimental evidence suggests that the presence of tumor inhibits dendritic cell maturation and activation both locally and systemically.234 However, isolated dendritic cells (from marrow or blood) can be concentrated or cultured from CD34+ progenitor cells. These fully functional isolated dendritic cells are subjected to cytokines (such as GM-CSF, IL-4, and TNF-a) and tumor-associated antigen stimulation and have been used as a pulse therapy.233,234,235,236,237,238,239,240 and 241 The identification of antigens such as CD83 and CMRF-44 on dendritic cells may allow the optimization of cytokine mobilization and of the timing of collection of blood dendritic cells, and possibly lead to positive selection techniques similar to those currently used for CD34+ cells.242,243 Preliminary clinical trials with dendritic cells in low-grade B cell lymphoma, prostate cancer, and melanoma have been associated with apparent clinical responses in some patients.239,240 and 241 These early reports, while promising, require further clinical investigation to assess the clinical efficacy.

Strumia MM: Effect of leukocyte cream injection in treatment of neutropenias. Am J Med Sci 187:527, 1934.

Brecher G, Wilbur KM, Cronkite EP: Transfusion of separated leukocytes into irradiated dogs with aplastic marrows. Proc Soc Exp Biol Med 84:54, 1953.

Applebaum FR, Bolwes CA, Makuch RW, Deisseroth AB: Granulocyte transfusion therapy of experimental Pseudomonas septicemia: study of cell dose and collection technique. Blood 52:323, 1978.

Debelak KM, Epstein RB, Andersen RB: Granulocyte transfusion in leukopenic dogs: in vivo and in vitro function of granulocytes obtained by continuous-flow filtration leukopheresis. Blood 43:757, 1974.

Epstein RB, Waxman FJ, Bennett BT, Andersen BR: Pseudomonas septicemia in neutropenic dogs. I. Treatment with granulocyte transfusions. Transfusion 14:51, 1974.

Epstein RB, Clift RA, Thomas ED: The effect of leukocyte transfusions on experimental bacteremia in the dog. Blood 34:782, 1969.

Westrick MAMA, Debelak-Fehir KM, Epstein RB: The effect of prior whole blood transfusion on subsequent granulocyte support in leukopenic dogs. Transfusion 17:611, 1977.

Dale DC, Reynolds HY, Pennington JE, et al: Granulocyte transfusion therapy of experimental Pseudomonas pneumonia. J Clin Invest 54:664, 1974.

Dale DC, Reynolds HY, Pennington JE, et al: Experimental Pseudomonas pneumonia in leukopenic dogs: comparison of therapy with antibiotics and granulocyte transfusions. Blood 47:869, 1976.

Chow HS, Sarpel SC, Epstein RB: Pathophysiology of Candida albicans meningitis in normal, neutropenic, and granulocyte transfused dogs. Blood 55:546, 1980.

Chow HS, Sarpel SC, Epstein RB: Experimental candidiasis in neutropenic dogs: Tissue burden of infection and granulocyte transfusion effects. Blood 59:328, 1982.

Epstein RB, Chow HS: An analysis of quantitative relationships of granulocyte transfusion therapy in canines. Transfusion 21:360, 1981.

Buckner D, Graw RG, Eisel RJ, et al: Leukapheresis by continuous flow centrifugation (CFC) in patients with chronic myelocytic leukemia (CML). Blood 33:353, 1969.

Freireich EJ, Levin RH, Whang J, et al: The functions and fate of transfused leukocytes from donors with chronic myelocytic leukemia in leukopenic recipients. Ann N Y Acad Sci 113:1081, 1964.

Eyre HJ, Goldstein IM, Perry S, et al: Leukocyte transfusions: Function of transfused granulocytes from donors with chronic myelocytic leukemia. Blood 36:432, 1970.

Schiffer CA, Aisner J, Dutcher JP, et al: Sustained posttransfusion granulocyte count increments following transfusion of leukocytes obtained from donors with chronic myelogenous leukemia. Am J Hematol 15:65, 1983.

McCredie KB, Freireich EJ, Hester JP, et al: Increased granulocyte collection with the blood cell separator and the addition of etiocholanolone and hydroxyethyl starch. Transfusion 14:357, 1974.

Schiffer CA, Aisner J, Dutcher JP, Wiernik PH: Sustained post-transfusion granulocyte count increments following transfusion of leukocytes obtained from donors with chronic myelogenous leukemia. Am J Hematol 15:65, 1983.

Mishler JM, Hadlock DE, Fortuny IE, et al: Increased efficiency of leukocyte separation by the addition of hydroxyethyl starch to the continuous flow centrifuge. Blood 44:571, 1974.

Mishler JM, Higby DJ, Rhomberg W: Hydroxyethyl starch and dexamethasone as an adjunct to leukocyte separation with the IBM blood cell separator. Transfusion 14:352, 1974.

Mishler JM: Hydroxyethyl starch as an experimental adjunct to leukocyte separation by centrifugal means: Review of safety and efficacy. Transfusion 15:449, 1975.

Graw RG, Herzig G, Perry S, et al: Normal granulocyte transfusion therapy: Treatment of septicemia due to gram-negative bacteria. N Engl J Med 287:367, 1972.

Higby FJ, Yates JW, Henderson ES, et al: Filtration leukapheresis for granulocytic transfusion therapy. N Engl J Med 292:761, 1975.

Fortuny IE, Bloomfield CD, Hadlock DC, et al: Granulocyte transfusion. A controlled study in patients with acute non-lymphocytic leukemia. Transfusion 15:548, 1975.

Vogler WR, Wintron EF: A controlled study of the efficacy of granulocyte transfusions in patients with neutropenia. Am J Med 63:548, 1997.

Herzig RH, Herzig GP, Graw RG, et al: Successful granulocyte transfusion therapy for gram-negative septicemia: A prospectively randomized controlled study. N Engl J Med 296:701, 1977.

Alavi JB, Root RK, Djerassi I, et al: A randomized clinical trial of granulocyte transfusions for infection in acute leukemia. N Engl J Med 296:706, 1977.

Winston DJ, Ho WG, Gale RP: Therapeutic granulocyte transfusions for documented infections: A controlled trial in ninety-five infectious granulocytopenic patients with gram-negative bacteremia. Am J Med 68:643, 1980.

Vamvakas EC, Pineda A: Meta-analysis of clinical studies of the efficacy of granulocyte transfusions in the treatment of bacterial sepsis. J Clin Apheresis 11:1, 1996.

Laurenti F, Ferro R, Isacchi G, et al: Polymorphonuclear leukocyte transfusion for the treatment of sepsis in the newborn infant. J Pediatr 98:118, 1981.

Christensen RD, Rothstein G, Anstall HB, Bybee B. Granulocyte transfusions in neonates with bacterial infection, neutropenia, and depletion of mature marrow neutrophils. Pediatrics 70:1 1982.

Cairo MS, Worcester C, Rucker R, et al: Role of circulating complement and polymorphonuclear leukocyte transfusion in treatment and outcome in critically ill neonates with sepsis. J Pediatr 110:935, 1989.

Wheeler JG, Chauvenet AR, Johnson CA, Block SM, Dillard R, Abramson JS: Buffy coat transfusions in neonates with sepsis and neutrophil storage pool depletion. Pediatrics 79:422, 1987.

Baley JE, Stork EK, Warkentin PI, et al: Buffy coat transfusions in neutropenic neonates with presumed sepsis: A prospective, randomized trial. Pediatrics 80:712, 1987.

Donahue DM, Grabrio BW, Finch CA: Quantitative measurements of hematopoietic cells of the marrow. J Clin Invest 37:1564, 1958.

Donahue DM, Reiff RH, Henson ML, et al: Quantitative measurements of the erythrocytic and granulocytic cells of the marrow and blood. J Clin Invest 37:1571, 1958.

Athens JW, Haab OP, Mauer AM, et al: Leukokinetic studies. IV. The total blood circulating and marginal granulocyte pools and the granulocyte turnover rate in normal subjects. J Clin Invest 40:989, 1961.

Cartwright GE, Athens JW, Wintrobe MM: The kinetics of granulopoiesis in normal men. Blood 24:780, 1969.

MacPherson JL, Nusbacher J, Bennett JM: The acquisition of granulocytes by leukapheresis: A comparison of continuous flow centrifugation and filtration leukapheresis in normal and corticosteroid-stimulated donors. Transfusion 16:221, 1976.

Nusbacher J, McCullough J, Huestis DW: Granulocyte collection and processing, in The Granulocyte: Function and Clinical Utilization, edited by TJ Greenwalt, GA Jamieson, p 175. Liss, New York, 1977.

Kalmin ND, Grindon AJ: Pheresis with the IBM 2997. Transfusion 21:325, 1981.

Djerassi I, Kim JS, Suvansri U, et al: Continuous-flow filtration leukapheresis. Transfusion 12:75, 1972.

Herzig GP, Root RK, Graw RG: Granulocyte collection by continuous-flow filtration leukapheresis. Blood 39:554, 1972.

Higby DJ, Henderson ES, Burnett D, et al: Filtration leukapheresis: Effects of donor stimulation with dexamethasone. Blood 50:953, 1977.

Rubins JM, MacPherson JL, Nusbacher J, et al: Granulocyte kinetics in donors undergoing filtration leukapheresis. Transfusion 16:56, 1976.

Debalak KM, Epstein RB, Anderson BR: Granulocyte transfusions in leukopenic dogs: In vivo and in vitro function of granulocytes obtained by continuous-flow filtration leukapheresis. Blood 43:757, 1974.

Harris M, Djerassi I, Schwartz E: Polymorphonuclear leukocytes prepared by continuous flow filtration leukapheresis: Viability and function. Blood 44:707, 1974.

Higby DH, Burnette D: Granulocyte transfusions: Current status. Blood 55:2, 1980.

Epstein RB, Waxman FJ, Bennet BT, et al: Pseudomonas septicemia in neutropenic dogs. I. Treatment with granulocyte transfusions. Transfusion 14:51, 1974.

Higby DJ, Yates J, Henderson ES, et al: Filtration leukapheresis for granulocyte transfusion therapy. N Engl J Med 292:761, 1975.

Schiffer CA, Buchholz DH, Aisner J, et al: Clinical experience with transfusion of granulocytes obtained by continuous flow filtration leukapheresis. Am J Med 58:373, 1975.

Wiltbank TB, Nusbacher J, Higby DJ, et al: Abdominal pain in donors during filtration leukapheresis. Transfusion 17:159, 1977.

Dahlke MB, Shah SL, Sherwood WC, et al: Priapism during filtration leukapheresis. Transfusion 19:482, 1979.

Schiffer CA, Aisner AJ, Wiernik PH: Transient neutropenia induced by transfusion of blood exposed to nylon fiber filters. Blood 45:141, 1975.

Hammerschmidt DE, Craddock PR, McCullough J, et al: Complement activation and pulmonary leukostasis during nylon fiber filtration leukapheresis. Blood 51:721, 1978.

Nusbacher J, Rosenfeld SI, MacPherson JL, et al: Nylon fiber leukapheresis: Associated complement changes and granulocytopenia. Blood 51:359, 1978.

Craddock PR, Hammerschmidt DE, White JG, et al: Complement (C5a)-induced granulocyte aggregation in vitro: A possible mechanism of complement-mediated leukostasis and leukopenia. J Clin Invest 60:261, 1977.

Nusbacher J, Rosenfeld SI, Leddy JP, et al: The leukokinetic changes and complement activation associated with filtration leukapheresis. Exp Hematol 7(suppl 4):24, 1979.

French JE, Solomon JM, Fratantoni JC: Survey on the current use of leukapheresis and the collection of granulocyte concentrates. Transfusion 22:220, 1982.

McCullough J, Weiblin BJ, Deinard AR, et al: In vitro function and post-transfusion survival of granulocytes collected by continuous-flow centrifugation and by filtration leukapheresis. Blood 48:315, 1976.

Wright DG, Kauffman JC, Chusid MJ, et al: Functional abnormalities of human neutrophils collected by continuous flow filtration leukapheresis. Blood 46:901, 1975.

Applebaum FR, Norton L, Graw RG: Migration of transfused granulocytes in leukopenic dogs. Blood 49:483, 1977.

Price TH, Dale DC: Neutrophil transfusion: Effect of storage and of collection method on neutrophil blood kinetics. Blood 51:789, 1978.

Higby DJ: Granulocyte transfusions: Where now? N Engl J Med 305:636, 1981.

Strauss RG, Hester JP, Vogler WR, et al: A multicenter trial to document the efficacy and safety of a rapidly excreted analog of hydroxyethyl starch for leukapheresis with a note on steroid stimulation of granulocyte donors. Transfusion 26:258, 1986.

Lee JH, Leitman SF, Klein HG: A controlled study of the efficacy of hetastarch and pentastarch in granulocyte collections by centrifugal leukapheresis. Blood 86:4662, 1995.

Athens JW, Heab OP, Raab SO, et al: The mechanism of steroid granulocytosis. J Clin Invest 41:1342, 1962.

Mischler JM: The effects of corticosteroids on mobilization and function of neutrophils. Exp Hematol 5(suppl):15, 1977.

Higby DJ, Mishler JM, Rhomberg W, et al: The effect of a single or double dose of dexamethasone on granulocyte collection with a continuous flow centrifuge. Vox Sang 28:243, 1975.

Winton EF, Vogler WR: Development of a practical oral dexamethasone premedication schedule leading to improved granulocyte yields with the continuous-flow centrifugal blood cell separator. Blood 52:249, 1978.

Glasser L, Huestis DW, Jones JF: Functional capabilities of steroid-recruited neutrophils harvested for clinical transfusion. N Engl J Med 297:1037, 1977.

Steigbigel RT, Baum J, MacPherson JL, et al: Granulocyte bactericidal capacity and chemotaxis as affected by continuous-flow centrifugation and filtration leukapheresis, steroid administration, and storage. Blood 52:197, 1978.

Caspar CB, Segar RA, Burger J, Gmur J: Effective stimulation of donors for granulocyte transfusions with recombinant methionyl granulocyte colony-stimulating factor. Blood 81:2866, 1993.

Bensinger WI, Price TH, Dale DC: The effects of daily recombinant human granulocyte colony-stimulating factor administration on normal granulocyte donors undergoing leukapheresis. Blood 81:1883, 1993.

Adkins D, Spitzer G, Johnston M, et al: Transfusions of granulocyte-colony-stimulating factor-mobilized granulocyte components to allogeneic transplant recipients: analysis of kinetics and factors determining posttransfusion neutrophil and platelet counts. Transfusion 37:737, 1997

Jendiroba DB, Lichtiger B, Anaissie E, et al: Evaluation and comparison of three mobilization methods for the collection of granulocytes. Transfusion 38:722, 1998.

Leitman SF, Oblitas JM: Optimization of granulocytapheresis mobilization regimens using granulocyte colony stimulating factor (G-CSF) and dexamethasone (DEXA). Transfusion 37(suppl):67S, 1997.

Dale DC, Liles WC, Llewellyn C, Rodger E, Price TH: Neutrophil transfusions: kinetics and functions of neutrophils mobilized with granulocyte-colony-stimulating factor and dexamethasone. Transfusion 38:713, 1998.

Demitri GD, Griffin JD: Granulocyte colony stimulating factor and its receptor. Blood 78:2791, 1991.

Dancey JT, Deukelbeiss KA, Harker LA, Finch CA: Neutrophil kinetics in man. J Clin Invest 58:705, 1976.

Brach MA, DeVos S, Gruss HJ, Herrmann F: Prolongation of survival of human polymorphonuclear neutrophils by granulocyte-macrophage colony-stimulating factor is caused by inhibition of programmed cell death. Blood 80:1920 1992.

Bodey GP, Buckley M, Sathe YS, et al: Quantitative relationships between circulating leukocytes and infection in patients with acute leukemia. Ann Intern Med 64:328, 1966.

Raubitschek AA, Levin AS, Stites DP, et al: Normal granulocyte infusion therapy for aspergillosis in chronic granulomatous disease. Pediatrics 51:230, 1973.

Yomtovian R, Abramson J, Quie PG, McCullough J: Granulocyte transfusion therapy in chronic granulomatous disease: Report of a patient and review of the literature. Transfusion 21:739, 1981.

Elliot GR, Clay ME, Mills EL, et al: Granulocyte transfusion kinetics measured by chemiluminescence, nitroblue tetrazolium reduction, and recovery of indium-111-labeled granulocytes. Transfusion 26:23, 1986.

Ford JM, Cullen MH: Prophylactic granulocyte transfusions. Exp Hematol 5(suppl 1):65, 1977.

Schiffer CA, Aisner J, Daily PA, et al: Alloimmunization following prophylactic granulocyte transfusion. Blood 54:766, 1979.

Clift RA, Sanders JE, Thomas ED, et al: Granulocyte transfusions for the prevention of infection in patients receiving bone-marrow transplants. N Engl J Med 298:1052, 1978.

Winston DJ, Ho WG, Young LS, et al: Prophylactic granulocyte transfusions during human bone marrow transplantation. Am J Med 68:893, 1980.

Strauss RG, Connett JE, Gale RP, et al: A controlled trial of prophylactic granulocyte transfusions during initial induction chemotherapy for acute myelogenous leukemia. N Engl J Med 305:597, 1981.

Ford JM, Cullen MH, Roberts MM, et al: Prophylactic granulocyte transfusions: Results of a randomized controlled trial in patients with acute myelogenous leukemia. Transfusion 22:311, 1982.

Sutton DMC, Shumak KH, Baker MA: Prophylactic granulocyte transfusions in acute leukemia. Plasma Ther 3:45, 1983.

Higby DJ, Freeman AI, Henderson ES, et al: Granulocyte transfusions in children using filter-collected cells. Cancer 38:1407, 1976.

Ward HN: Pulmonary infiltrates associated with leukoagglutinin transfusion reaction. Ann Intern Med 73:689, 1970.

Jacob HS: Granulocyte-complement interaction. Arch Intern Med 138:461, 1978.

Wright DG, Robichaud KJ, Pizzo PA, et al: Lethal pulmonary reactions associated with the combined use of amphotericin B and leukocyte transfusions. N Engl J Med 304:1185, 1981.

Forman SJ, Robinson GV, Wolfe JL, et al: Pulmonary reactions associated with amphotericin B and leukocyte transfusions. N Engl J Med 305:584, 1981.

DeGregorio MW, Lee WMF, Ries CA: Pulmonary reactions associated with amphotericin B and leukocyte transfusions. N Engl J Med 305:584, 1981.

Ford JM, Lucey JJ, Cullen MH, et al: Fatal graft-versus-host disease following transfusion of granulocytes from normal donors. Lancet 2:1167, 1976.

Rosen RS, Huestis DW, Corrigan JJ: Acute leukemia and granulocyte transfusion: Fatal graft-versus-host reaction following transfusion of cells obtained from normal donors. J Pediatr 93:268, 1978.

Cohen D, Weinstein H, Mihm M, et al: Nonfatal graft-versus-host disease occurring after transfusion with leukocytes and platelets obtained from normal donors. Blood 53:1053, 1979.

Graw RG, Buchner CD, Whang-Peng J, et al: Complication of bone-marrow transplantation: Graft-versus-host disease resulting from chronic-myelogenous-leukaemia leucocyte transfusions. Lancet 2:338, 1970.

Weiden PL, Zuckerman N, Hansen JA, et al: Fatal graft-versus-host disease in a patient with lymphoblastic leukemia following normal granulocytic transfusions. Blood 57:328, 1981.

Cairo MS: Granulocyte transfusions in neonates with presumed sepsis. Pediatrics 80:738, 1987.

McCullough J: Liquid preservation of granulocytes. Transfusion 20:129, 1980.

Lane TA: Storage of granulocyte concentrates (GC): Bacterial killing and chemotaxis. Transfusion 18:650, 1978.

McCullough J, Weiblin BJ, Quie PG: Chemotactic activity of human granulocytes preserved in various anticoagulants. J Lab Clin Med 84:902, 1974.

McCullough J, Weiblin BJ: Relationship of granulocyte ATP to chemotactic response during storage. Transfusion 19:764, 1979.

Lane TA: Granulocyte concentrate preservation: 6°C versus room temperature. Transfusion 18:394, 1978.

Lane TA: Storage of granulocyte concentrates (GC): Bacterial killing and chemotaxis. Transfusion 18:650, 1978.

McCullough J, Weiblin BJ, Peterson PK, et al: Effects of temperature on granulocyte preservation. Blood 52:301, 1978.

McCullough J: Liquid preservation of granulocytes for transfusion. Prog Clin Biol Res 13:185, 1977.

Lane TA, Lamkin GE: Hydrogen ion maintenance improves the chemotaxis of stored granulocytes. Transfusion 24:231, 1984.

Glasser L, Lane TA, McCullough J, Price TH: Neutrophil concentrates: Functional considerations, storage and quality control. J Clin Apheresis 1:179, 1983.

Lane TA, Windle BE: Granulocyte concentrate function during preservation: Effect of temperature. Blood 54:216, 1979.

Lane TA, Lamkin GE: Adherence of fresh and stored granulocytes to endothelial cells. Effect of storage temperature. Transfusion 28:237, 1988.

Lane TA, Lamkin GE: Stimulus-response coupling in fresh and stored granulocytes. Transfusion 28:243, 1988.

Babior BM, Berkman E: Granulocyte storage. Lancet 1:50, 1990.

Glasser L, Fiederlein RL, Huestis DW: Granulocyte concentrates: Glucose concentrations and glucose utilization during storage at 22°C. Transfusion 25:68, 1985.

Glasser L, Fiederlein RL: The effect of platelets and red cells on granulocytes stored at 22°C. Transfusion 24:310, 1984.

Contreras TJ, Jemionek JF, French JE, et al: Effects of plastic polymer surfaces on the liquid preservation of human granulocytes. Transfusion 18:650, 1978.

Miyamoto M, Sasakawa S: Studies on granulocyte preservation. III. Effect of agitation on granulocyte concentrates. Transfusion 27:165, 1987.

Wolber RA, Duque RE, Robinson JP, Oberman HA: Oxidative product formation in irradiated neutrophils: A cytometric analysis. Transfusion 27:167, 1987.

Buescher ES, Gallin JI: Effects of storage and radiation on human neutrophil function in vitro. Inflammation 11:401, 1987.

Lane TA: Granulocyte storage. Transfus Med Rev 4:23, 1990.

Meryman HT, Howard J: Cryopreservation of granulocytes, in The Granulocyte: Function and Clinical Utilization, edited by TJ Greenwalt, GA Jamieson, p 193, Liss, New York, 1977.

Hill RS, MacKinder CA: Freeze preservation of human granulocytes. Lancet 1:878, 1980.

Lionetti FJ, Hunte SM, Gore JM, et al: Cryopreservation of human granulocytes. Cryobiology 12:181, 1975.

Strauss RG: Neutrophil (granulocyte) transfusions in the new millennium. Transfusion 38:710, 1998.

Maximow A: Der Lymphozyt als gemeinsame Stamzelle der verschiedenen Blutelemente in der embryonalen Entwicklung und im postfetalen Leben der Säugetiere. Folia Haematol 8, 125, 1909.

Brecher G, Cronkite EP: Post-irradiation parabiosis and survival in rats. Proc Soc Exp Biol Med 77:292, 1951.

Levin RH, Whang J, Tjio JH, et al: Persistent mitosis of transfused homologous leukocytes in children receiving antileukemic therapy. Science 142:1305, 1963.

Goldman JM, Catovsky D, Hows J, et al: Cryopreserved peripheral blood cells functioning as autografts in patients with chronic granulocyte leukemia in transformation. Br J Med 13:148, 1979.

Korbling M, Durkin B, Ho AD, et al: Autologous transplantation of blood derived hematopoietic stem cells after myeloablative therapy in a patient with Burkittí lymphoma. Blood 67:529, 1986.

Chao NJ, Schriber JR, Grimes K, et al. Granulocyte colony stimulating factor “mobilized” peripheral blood progenitor cells accelerate granulocyte and platelet recovery after high dose chemotherapy. Blood 81:2031, 1993.

Sheridan WP, Begley CG, Juttner CA, et al: Effect of peripheral-blood progenitor cells mobilized by filgrastim (G-CSF) on platelet recovery after high-dose chemotherapy. Lancet 339:640, 1992.

Kessinger A, Armitage JO, Landmark JD, et al: Autologous peripheral hematopoietic stem cell transplantation restores hematopoietic function following marrow ablative therapy. Blood 71:723, 1988.

Reiffers J, Bernard P, David B, et al: Successful autologous transplantation with peripheral blood hematopoietic cells in a patient with acute leukemia. Exp Hematol 14:312, 1986.

Leitman SF, Read SJ. Hematopoietic progenitor cells. Semin Hematol 33:341, 1996.

Bandarenko N, Owen HG, Mair DC, et al: Apheresis: new opportunities, in Clinics in Laboratory Medicine, edited by HF Polesky, EH Perry, SJ Ilstrup, pp 907–914. Philadelphia, WB Saunders, 1996.

McCullough J: Quality assurance and good manufacturing practices for processing hematopoietic progenitor cells. J Hematother 4:493, 1995.

Korbling M, Przepiorka D, Huh YO, et al: Allogeneic blood stem cell transplantation for refractory leukemia and lymphoma: potential advantage of blood over marrow allografts. Blood 85(6):1659, 1995.

Bensinger W, Singer J, Appelbaum F, et al: Autologous transplantation with peripheral blood mononuclear cells collected after administration of recombinant granulocyte stimulating factor. Blood 81:3158, 1993.

To LB, Roberts MM, Haylock DN, et al: Comparison of haematologic recovery times and supportive care requirements of autologous recovery phase peripheral blood stem cell transplants, autologous bone marrow transplants, and allogeneic bone marrow transplants. Bone Marrow Transplant 9:277, 1992.

Goldberg SL, Mangan KF, Klumpp TR, et al: Complications of peripheral blood stem cell harvesting: review of 554 PBSC leukaphereses. J Hematother 4:85, 1995.

Stephens LC, Haire WD, Schmit-Pokorny K, et al: Granulocyte macrophage colony stimulating factor: high incidence of apheresis catheter thrombosis during peripheral stem cell collection. Bone Marrow Transplant 11:51, 1993.

Brecher ME, Lasky LC, Sacher RA, Issit LA (eds): Hematopoietic Progenitor Cells: Processing, Standards and Practice. American Association of Blood Banks, Bethesda, MD, 1995.

Bender JG, Unverzagt KL, Walker DE, et al: Identification and comparison of CD34 positive cells and their subpopulations from normal peripheral blood and bone marrow using multicolor flow cytometry. Blood 77:2591, 1991.

Lasky LC: Peripheral blood stem cell collection and use, in Cellular and Humoral Immunotherapy and Apheresis, edited by RA Sacher, DB Brubaker, DO Kasprisin, LJ McCarthy, pp 73–85. American Association of Blood Banks, Arlington, VA 1991.

Richman CM, Weiner RS, Yankee RA: Increasing circulating stem cells following chemotherapy in man. Blood 47(6):1031, 1976.

To LB, Haylock DN, Kimber RJ, Juttner CA: High levels of circulating haematopoietic stem cells in the very early remission of acute non-lymphoblastic leukaemia and their collection and cryopreservation. Br J Haematol 58:399, 1984.

To LB, Roberts MM, Haylock DN, et al: The optimization of collection of peripheral blood stem cells for autotransplantation in acute myeloid leukemia. Bone Marrow Transplant 4:41, 1989.

Kosatec D, Shepherd KM, Sage RE, et al: Factors affecting blood stem cell collections following high-dose cyclophosphamide mobilization in lymphoma, myeloma, and solid tumors. Int J Cell Cloning (suppl 1):35, 1992.

To LB, Shepherd KM, Haylock DN, et al: Single high doses of cyclophosphamide enable the collection of high numbers of haemopoietic stem cells from the peripheral blood. Exp Hematol 18:442, 1990.

Gianni AM, Siena S, Bregni M, et al: Very rapid and complete hematopoietic reconstitution following myeloablative treatments: the role of circulating stem cells harvested after high-dose cyclophosphamide and GM-CSF, in Bone Marrow Transplantation, 4th ed, edited by KA Kicke, G Spitzer, S Jagannath, MJ Evinger-Hodges, pp 723–731. University of Texas Press, Austin, 1989.

Socinski MA, Elias A, Schnipper L, et al: Granulocyte-macrophage colony stimulating factor expands the circulating haematopoietic progenitor cell compartment in man. Lancet 1:1194, 1988.

Pettengell R, Testa NG, Swindell R, et al: Transplantation potential of hematopoietic cells released into the circulation during routine chemotherapy for non-Hodgkins lymphoma. Blood 82:2239, 1993.

Elias AD, Ayash L, Anderson KC, et al: Mobilization of peripheral blood progenitor cells by chemotherapy and granulocyte-macrophage colony stimulating factor for hematologic support after high-dose intensification for breast cancer. Blood 79:3036, 1992.

Gillespie TW, Hillyer CD: Peripheral blood progenitor cells for marrow reconstitution: mobilization and collection strategies. Transfusion 36:611, 1996.

Haas R, Hohaus S, Egerer G, et al: Recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) subsequent to chemotherapy improves collection of blood stem cells for autografting in patients not eligible for bone marrow harvest. Bone Marrow Transplant 9(6):459, 1992.

Lane TA: Mobilization of hematopoietic progenitor cells, in Hematopoietic Progenitor Cells: Processing, Standards and Practice, edited by ME Brecher, LC Lasky, RA Sacher, LA Issit, pp 59–108. American Association of Blood Banks, Bethesda, MD, 1995.

Lee JH, Klein HG: Collection and use of circulating hematopoietic progenitor cells in Hematology/Oncology Clinics of North America: Transfusion Medicine II, edited by PD Mintz, pp 1–22. WB Saunders, Philadelphia, 1995.

Siena S, Bregni M, Brando B, et al: Circulation of CD34+ hematopoietic stem cells in the peripheral blood of high-dose cyclophosphamide-treated patients; enhancement by intravenous recombinant human granulocyte-macrophage stimulating factor. Blood 74:1905, 1989.

Sutherland HJ, Eaves CJ, Lansdorp PM, et al: Kinetics of committed and primitive blood progenitor mobilization after chemotherapy and growth factor treatment and their use in autotransplants. Blood 83:3808, 1994.

Beyer J, Schwella N, Zingsem J, et al: Hematopoietic rescue after high-dose chemotherapy using autologous peripheral-blood progenitor cells or bone marrow: A randomized comparison. J Clin Oncol 13:1328, 1995.

Deeg HJ: Bone marrow and hematopoietic stem cell transplantation: Sorting the chaff from the grain, in Bone Marrow and Stem Cell Processing: A Manual of Current Techniques, edited by EM Areman, HJ Deeg, RA Sacher, pp 17–29. FA Davis, Philadelphia, 1992.

Dreger P, Suttorp M, Haferlach T, et al: Allogeneic granulocyte colony stimulating factor mobilized peripheral blood progenitor cells for treatment of engraftment failure after bone marrow transplantation. Blood 81:1404, 1993.

Russell NH, Hunter A, Rogers S, et al: Peripheral blood stem cells as an alternative of marrow for allogeneic transplantation [Letter]. Lancet 341:1482, 1993.

Weaver CH, Buckner CD, Longin K, et al: Syngeneic transplantation with peripheral blood mononuclear cells collected after the administration of recombinant human granulocyte colony-stimulating factor. Blood 82:1981, 1993.

Glaspy JA, Shpall EJ, LeMaistre CF, et al: Peripheral blood progenitor cell mobilization using stem cell factor in combination with Filgrastim in breast cancer patients. Blood 90:2939, 1997.

Link H, Arseniev L, Bahre O, et al: Transplantation of allogeneic CD34+ blood cells. Blood 87:4903, 1996.

Majolino I, Corradini P, Scime R, et al: Allogeneic transplantation of unmanipulated peripheral blood stem cells in patients with multiple myeloma. Bone Marrow Transplant 22(5):449, 1998.

MacKinnon S, Papadapoulos EB, Carabasi MH, et al: Adoptive immunotherapy evaluating doses of donor leukocytes for relapse of chronic myeloid leukemia after bone marrow transplantation: separation of graft-versus-leukemia responses from graft-versus-host disease. Blood 86(4):1261, 1995.

Majolino I, Buscemi F, Scime R, et al: Treatment of normal donors with rhG-CSF 16 micrograms/kg for mobilization of peripheral blood stem cells and their apheretic collections for allogeneic transplantation. Haematologica 80:219, 1995.

Bishop MR, Tarantolo SR, Jackson JD, et al: Allogeneic-blood stem-cell collection following mobilization with low-dose granulocyte colony-stimulating factor. J Clin Oncol 15:1601, 1997.

Schmitz N, Dreger P, Suttorp M, et al: Primary transplantation of allogeneic peripheral blood progenitor cells mobilized by filgrastim (granulocyte colony-stimulating factor). Blood 85(6):1666, 1995.

Stroncek D, Clay M, Petzoldt ML, et al: Treatment of normal individuals with granulocyte-colony-stimulating factor: donor experiences and the effects on peripheral blood CD34+ cell counts and on the collection of peripheral blood stem cells. Transfusion 36:601, 1996.

Anderlini P, Pzrepiorka D, Seong D, et al: Clinical toxicity and laboratory effects of mobilization and blood stem cell apheresis from normal donors, and analysis of charges for the procedure. Transfusion 36:590, 1996.

Stroncek D, McCullough J: Policies and procedures for the establishment of an allogeneic blood stem cell programme. Transfus Med 7:77, 1997.

Bandarenko N, Brecher M, Owen H, et al: Thrombocytopenia in allogeneic peripheral blood stem cell collections [Letter]. Transfusion 36:668, 1996.

Anderlini P, Korbling M, Dale D, et al: Allogeneic blood cell transplantation: Considerations for donors [Editorial]. Blood 90:903, 1997.

Stroncek D, Clay M, Herr G, et al: 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. Transfusion 37:304, 1997.

Reiffers J, Bernard P, Marit G, et al: Collection of blood-derived hematopoietic stem cells and applications for autologous transplantation. Bone Marrow Transplant 1(suppl 1):371, 1986.

Gorin NC, Lopez M, Laporte JP, et al: Preparation and successful engraftment of purified CD34+ bone marrow progenitor cells in patients with non-Hodgkin’s lymphoma. Blood 85(6):1647, 1995.

Hohaus S, Goldschmidt H, Ehrhardt R, Haas R: Successful autografting following myeloablative conditioning therapy with blood stem cells mobilized by chemotherapy plas rhG-CSF. Exp Hematol 21(4):508, 1993.

Jones HM, Jones SA, Watts MJ, et al: Development of a simplified single-apheresis approach for peripheral-blood progenitor cell transplantation in previously treated patients with lymphoma. J Clin Oncol 12(8):1693, 1994.

Urashima M, Uchiyama H, Hoshi Y, et al: Prediction of engraftment after peripheral blood stem cell transplantation by CD34+ cells in grafts. Acta Paediatr Jpn 35(4):325, 1993.

Carow CE, Hangoc G, Broxmeyer HE: Human multipotential progenitor cells (CFU-GEMM) have extensive replating capacity for secondary (CFU-GEMM): an effect enhanced by cord blood plasma. Blood 81:942, 1993.

Korbling M, Huh YO, Durett A, et al: Peripheralization and yield of donor derived primitive hematopoietic progenitor cells (CD34+ Thy-1dim) and lymphoid subsets, and possible predictors of engraftment and graft-versus-host disease. Blood 86:2842, 1995.

Bender JG, To LB, Williams S, Schwartzberg LS: Defining a therapeutic dose of peripheral blood stem cells. J Hematother 1:329, 1992.

Bensinger WI, Longin K, Appelbaum F, et al: Peripheral blood stem cells (PBSCs) collected after recombinant granulocyte colony stimulating factor (rhG-CSF): an analysis of factors correlating with the tempo to engraftment after transplantation. Br J Haematol 87(4):825, 1994.

Van der Wall E, Richel DJ, Holtkamp MJ, et al: Bone marrow reconstitution after high-dose chemotherapy and autologous peripheral progenitor cell transplantation: effect on graft size. Ann Oncol 5:795, 1994.

Kahn DG, Prilutskaya M, Cooper B, et al: The relationship between the incidence of tumor contamination and number of pheresis for stage IV breast cancer (abstr 2514). Blood 90(suppl 10):565a, 1997.

Moreb J, Cooper B, Holland K, et al: The prognostic value of immunocytochemical (ICC) analysis on bone marrow (BM) taken from patients with stage II/III breast cancer undergoing autologous transplant therapy (abstr 1703). Blood 90(suppl 10):383a, 1997.

Chen CH, Lin W, Shye S, et al: Automated enumeration of CD34+ cells in peripheral blood and bone marrow. J Hematother 3:3, 1994.

Johnsen HE: Report from a Nordic workshop on CD34+ cell analysis: technical recommendations for progenitor cell enumeration in leukapheresis from multiple myeloma patients. J Hematother 4:21, 1995.

Owens MA, Loken MR: Flow Cytometry Principles for Clinical Laboratory Practice: Quality Assurance for Quantitative Immunotyping, pp 111–127. Wiley-Liss, New York, 1995.

Sovalat H, Wunder E, Zimmerman R, Serke S: Multicentric determination of CD34+ cells, in Hematopoietic Stem Cells: The Mulhouse Manual, edited by E Wunder, H Sovalat, PR Henon, S Serke, p 61. AlphaMed Press, Dayton, OH, 1994.

Sutherland RE, Keating A, Nayar R, et al: Sensitive detection and enumeration of CD34+ cells in peripheral and cord blood by flow cytometry. Exp Hematol 22:1003, 1994.

Trischmann TM, Schepers KG, Civin CJ: Measurement of CD34+ cells in bone marrow by flow cytometry. J Hematother 2:305, 1993.

Wunder E, Sovolat H, Fritsch G, et al: Report on the European Workshop on Peripheral Blood Stem Cell Determination and Standardization—Mulhouse, France. J Hematother 1:131, 1992.

Siena S, Bregni M, Brando B, et al: Flow cytometry for clinical estimation of circulating hematopoietic progenitors for autologous transplantation in cancer patients. Blood 77:400, 1991.

Brecher ME, Sims L, Schmitz J, Shea T, Bentley SA: North American multicenter study on flow cytometric enumeration of CD34+ hematopoietic stem cells. J Hematother 5:227, 1996.

Chang A, McLachlan J, Ma DDF: Towards standardization of CD34+ cell determination: Australian Perspective (part I) [Abstract]. J Hematother 4:240, 1995.

Sutherland RE, Anderson L, Keeney M, et al: The ISHAGE guidelines for CD34+ cell determination by flow cytometry. J Hematother 5:213, 1996.

Dietz LJ, Dubrow RS, Manian BS, et al: Volumetric capillary cytometry: a new method for absolute cell enumeration. Cytometry 23:177, 1996.

Sims LC, Brecher ME, Gertis K, et al: Enumeration of CD34 positive stem cells: Evaluation and comparison of three methods. J Hematother 6:213, 1997.

Read EJ, Kunitake ST, Carter CS, et al: Enumeration of CD34+ hematopoietic progenitor cells in peripheral blood and leukapheresis products by microvolume fluorometry: a comparison with flow cytometry. J Hematother 6:291, 1997.

Hillyer CD, Lackey DA 3rd, Hart KK, et al: CD34+ progenitors and colony forming units–granulocyte macrophage are recruited during large-volume leukapheresis and concentrated by counterflow centrifugal elutriation. Transfusion 33(4):316, 1993.

Malachowski ME, Comenzo RL, Hillyer CD, et al: Large-volume leukapheresis for peripheral blood stem cell collection in patients with hematologic malignancies. Transfusion 32(8):732, 1992.

Pettengell R, Morgenstern GR, Woll PJ, et al: Peripheral blood progenitor cell transplantation in lymphoma and leukemia using a single apheresis. Blood 82(12):3770, 1993.

Bentley SA, Brecher ME, Powell E, et al: Long-term engraftment failure after marrow ablation and autologous hematopoietic reconstitution: differences between peripheral blood stem cell and bone marrow recipients. Bone Marrow Transplant 19:557, 1997.

Matsunaga T, Sakamaki S, Kohgo Y, et al: Recombinant human granulocyte colony-stimulating factor can mobilize sufficient amounts of peripheral blood stem cells in healthy volunteers for allogeneic transplantation. Bone Marrow Transplant 11(2):103, 1993.

Arseniev L, Hertenstei B, Link H, et al: Stem cell mobilization in normal donors [Letter]. J Hematother 7:5, 1998.

Gianni AM, Siena S, Bregni M, et al: Granulocyte-macrophage colony-stimulating factor to harvest haemopoietic stem cells for transplantation. Lancet 2:580, 1989.

Ho AD, Gluck S, Germond C, et al: Optimal timing for collections of blood progenitor cells following induction chemotherapy and granulocyte-macrophage colony-stimulating factor for autologous transplantation in advanced breast cancer. Leukemia 7:1738, 1993.

Bandarenko N, Sims L, Brecher M: Circulating CD34+ cell counts are predictive of CD34+ peripheral blood progenitor cell yields [Letter], Transfusion 37:1218, 1997.

Areman EM, Meehan KR, Sacher RA: Preapheresis levels of peripheral blood CD34+ cells correlate with CD34+ peripheral blood progenitor cells in autologous patients [Letter]. Transfusion 37:1217, 1997.

Elliot BC, Samson DM, Armitage S, et al: When to harvest peripheral blood stem cells after mobilization therapy: prediction of CD34+ cell yield by preceding day CD34+ concentration in peripheral blood. J Clin Oncol 14:970, 1996.

Mohle R, Murea S, Pforsich M, et al: Estimation of the progenitor cell yield in a leukapheresis product by previous measurement of CD34+ cells in the peripheral blood. Vox Sang 71:90, 1996.

Schots R, Van Riet I, Damiaens S, et al: The absolute number of circulating CD34+ cells predicts the number of hematopoietic stem cells that can be collected by apheresis. Bone Marrow Transplant 17:509, 1996.

Teshima T, Sunami K, Bessho A, et al: Circulating immature cell counts on the harvest day predict the yields of CD34+ cells collected after G-CSF plus chemotherapy-induced mobilization of peripheral blood stem cell. Blood 89:4660, 1997.

Benjamin RJ, Linsley L, Fountain D, et al: Preapheresis peripheral blood CD34+ mononuclear cell counts as predictors of progenitor cell yield. Transfusion 37:79, 1997.

Kolb HJ, Schattenberg A, Goldman JM, et al: Graft-versus-leukemia effect of donor lymphocytes marrow grafted patients. Blood 86(5):2041, 1995.

Lee JH, Klein HG: Mononuclear adoptive immunotherapy, in Hematology/Oncology Clinics of North America: Transfusion Medicine II, edited by PD Mintz, pp 1–22. WB Saunders, Philadelphia, 1995.

Collins RH, Piniero LA, Nemunaitis JJ, et al: Transfusion of donor buffy coat cells in the treatment of persistent or recurrent malignancy after allogeneic bone marrow transplantation. Transfusion 35: 1995.

Rosenberg SA, Lotze MT, Muul LM, et al: Observation on the systemic administration of autologous lymphokine-activated killer cells and recombinant interleukin-2 to patients with metastatic cancer. N Engl J Med 313:1485, 1985.

Rosenberg SA, Lotze MT, Muul LM, et al: A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone. N Engl J Med 316:889, 1986.

Klein HG, Leitman SF: Adoptive immunotherapy in the treatment of malignant disease. Transfusion 29:170, 1989.

Topalian SL, Muul LM, Solomon D, Rosenberg SA: Expansion of human tumor infiltrating lymphocytes for use in immunotherapy trials. J Immunol Methods 102:127, 1987.

Muul LM, Director EP, Hyatt C, Rosenberg SA: Large scale production of human lymphokine activated killer cells for use in adoptive immunotherapy. J Immunol Methods 88:265, 1986.

Carter CS, Leitman SF, Cullis H, et al: Use of a continuous-flow cell separator in density gradient isolation of lymphocytes. Transfusion 27:362, 1987.

Muul LM, Nason-Burchenal K, Carter CS, et al: Development of an automated closed system for generation of human lymphokine-activated killer (LAK) cells for use in adoptive immunotherapy. J Immunol Methods 101:171, 1987.

Troy AJ, Hart DNJ: Dendritic cells and cancer: Progress toward a new cellular therapy. J Hematother 6:523, 1997.

Young JW, Szabolics P, Moore MAS: Identification of dendritic cell colony-forming units among normal human CD34+ bone marrow progenitors that are expanded by c-kit ligand and yield pure dendritic cell colonies in the presence of granulocyte/macrophage colony-stimulating factor and tumor necrosis factor-a. J Exp Med 182:1120, 1995.

Szabolics P, Moore MA, Young JW: Expansion of immunostimulatory dendritic cells among the myeloid progeny of human CD34+ bone marrow precursors cultured with c-kit ligand, granulocyte/macrophage colony-stimulating factor and tumor necrosis factor a. J Immunol 154:5841, 1995.

Bernhard H, Disis ML, Heimfeld S, et al: Generation of immunostimulatory dendritic cells from human CD34+ hematopoietic progenitor cells in the bone marrow and peripheral blood. Cancer Res 55:1099, 1995.

Mackensen A, Herbst B, Kohlter G, et al: Delineation of the dendritic cell lineage by generating large numbers of Birbeck granule-positive Langerhans cells from human peripheral blood progenitor cells in vitro. Blood 86:2699, 1995.

Hsu FJ, Benike C, Fagonei F, et al: Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med 2:52, 1996.

Nestle FO, Alijagic S, Gilliet M, et al: Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4:328, 1998.

Tjoa BA, Simmons SJ, Bowes VA, et al: Evaluation of phase I/II clinical trials in prostate cancer with dendritic cells and PSMA peptides. Prostate 36:39, 1998.

Zhou LJ, Tedder TF: Human blood dendritic cells selectively express CD83, a member of the immunoglobulin superfamily. J Immunol 154:3821, 1995.

Hock BD, Starling GC, Daniel PB., et al: Characterization of CMRF-44, a novel monoclonal antibody to activation antigen expressed by the allostimulatory cells within peripheral blood, including dendritic cells. Immunology 83:573, 1974.
Copyright © 2001 McGraw-Hill
Ernest Beutler, Marshall A. Lichtman, Barry S. Coller, Thomas J. Kipps, and Uri Seligsohn
Williams Hematology



  1. […] Singapore HotelsMEDICALHotel At Singapore Planning With A Spending Budget, Midrange, Or LuxuryCHAPTER 141 COLLECTION AND TRANSFUSION OF LEUKOCYTES, DENDRITIC CELLS, AND STEM CELLS div.socialicons{float:left;display:block;margin-right: 10px;}div.socialicons p{margin-bottom: […]

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s

%d bloggers like this: