Phase I/II trial of neutrophil transfusions from donors stimulated with G-CSF and dexamethasone for treatment of patients with infections in hematopoietic stem cell transplantation

We examined the feasibility of a community blood bank granulocyte transfusion program utilizing community donors stimulated with a single-dose regimen of subcutaneous granulocyte colony-stimulating factor (G-CSF) plus oral dexamethasone. The recipients of these transfusions were neutropenic stem cell transplantation patients with severe bacterial or fungal infection. Nineteen patients received 165 transfusions (mean 8.6 transfusions/patient, range 1-25). Community donors provided 94% of the transfusions; relatives accounted for only 6% of the transfusions. Sixty percent of the community donors initially contacted agreed to participate, and 98% of these individuals indicated willingness to participate again. Transfusion of 81.9 +/- 2.3 x 10(9) neutrophils (mean +/- SD) resulted in a mean 1-hour posttransfusion neutrophil increment of 2. 6 +/- 2.6 x 10(3)/microL and restored the peripheral neutrophil count to the normal range in 17 of the 19 patients. The buccal neutrophil response, a measure of the capacity of neutrophils to migrate to tissue sites in vivo, was restored to normal in most patients following the transfusion. Chills, fever, and arterial oxygen desaturation of >/= 3% occurred in 7% of the transfusions, but these changes were not sufficient to limit therapy. Infection resolved in 8 of 11 patients with invasive bacterial infections or candidemia. These studies indicate that transfusion of neutrophils from donors stimulated with G-CSF plus dexamethasone can restore a severely neutropenic patient's blood neutrophil supply and neutrophil inflammation response. Further studies are needed to evaluate the clinical efficacy of this therapy.     

Granulocyte transfusion

PURPOSE OF REVIEW: Granulocyte transfusions have been used for more than four decades. Several issues have complicated the analysis of previous studies, including the utilization of improved antimicrobials, the effects of recipient alloimmunization and variable cell dose. The use of granulocyte colony-stimulating factor for donor stimulation has revived interest in granulocyte transfusion. The aim of this review is to evaluate the most recent studies in granulocyte transfusion therapy and their clinical applicability. RECENT FINDINGS: Granulocyte colony-stimulating factor use has increased the granulocyte yield by approximately fourfold. Multiple recent studies have shown that granulocyte transfusions can be helpful in controlling severe infections progressing despite the use of appropriate antibiotics, with a response rate of 40-80% with variability in results depending on patient characteristics. This benefit is limited to a small patient population as the incidence of prolonged reversible neutropenia is relatively small. Severe side effects have been rare in those studies. SUMMARY: Granulocyte transfusions are beneficial in neutropenic patients with severe uncontrolled infection. The underlying disease process is the major determinant of outcome in these patients. Because granulocyte transfusions are not commonly used, centers are not currently able to provide transfusions in a timely fashion. Nonalloimmunized patients can receive cells from nonmatched ABO compatible donors, while alloimmunized patients should receive granulocytes from either HLA-matched donors or donors selected by leukoagglutination or lymphocytotoxicity crossmatching. Further studies are needed to clarify the optimal starting time and frequency of transfusions, and the best method for identifying donor-recipient compatibility.     

Therapeutic role of granulocyte transfusions

Granulocyte transfusions are used as adjuvant therapy for infection in neutropenic patients with underlying neoplastic disease, neutropenic infants, and patients with qualitative white blood cell disorders. In addition, prophylactic leukocyte transfusions have been administered to patients during remission induction for acute leukemia or after bone marrow transplantation. The role of granulocyte therapy will need constant reassessment as new antibiotics and other forms of treatment are developed. At present, granulocyte transfusions are indicated in the treatment of severely neutropenic patients with documented bacterial infection who are unlikely to recover hematopoietic function over the next week and are deteriorating despite 48-72 hr of optimal antibiotic therapy. Under these conditions, they improve the rate of survival from the infectious episode without clearly affecting the longer-term survival of the patient. Only a small minority of neutropenic patients will require granulocyte transfusions.     

Granulocyte transfusions in neutropenic patients

Patients with severe neutropenia are at increased risk for systemic infection with bacteria or fungi. This risk is in proportion to both the degree and duration of the neutropenic process. Although granulocyte transfusion as a means of augmenting host defenses would appear to be a logical therapeutic intervention in clinical contexts involving severe and prolonged neutropenia, several features of granulocyte physiology and collection complicate such considerations. These include the large numbers of granulocytes normally produced by healthy hosts, the short survival of the granulocyte in the circulation after transfusion, the relatively small number of granulocytes which can be collected using currently available pheresis techniques, problems associated with alloimmunization, and the possibility of transferring disease (CMV, toxoplasmosis, hepatitis) by means of these transfusions. In the mid-1970s, well-designed clinical studies strongly suggested that patients with documented Gram-negative sepsis or tissue infection that failed to respond to appropriate antibiotics were significantly benefited by granulocyte transfusions. With recent advances in potent, broad-spectrum antibiotic availability, some have questioned whether these observations remain valid. Several studies regarding the prophylactic use of granulocyte transfusions in patients undergoing allogeneic bone marrow transplantation and/or induction therapy for leukemia have failed to reveal therapeutic benefits and suggested the possibility of significant side effects. These studies are reviewed.     

Granulocyte transfusions: current status

Since granulocyte transfusions first became widely used in clinical medicine, there have been advances in the treatment of acute leukemia and improvement in prevention and management of infection in neutropenic patients. Improved understanding now exists concerning prognosis of infections in such patients, and advances have been made in procurement of granulocytes. Granulocyte transfusions should be given for specific indications, and used adjunctively to other established antiinfective therapy. Once initiated, transfusions should be given in adequate doses at daily intervals (at least) with ongoing evaluation and periodic reassessment of the whole antiinfective program. Serious complications of granulocyte transfusion therapy are relatively rare, but the physician should be prepared to manage them intelligently. Research continues in discerning exactly how granulocyte transfusion work, in preservation of granulocytes, and in delineation of immunologic phenomena affecting the efficiacy of such therapy. Granulocyte transfusions will continue to be important in the management of acute leukemia, and other reversible bone marrow failure states, and in marrow transplantation and autotransplantation.     

Granulocyte transfusion: revisited

Neutrophils are the immune system's main cellular defense against bacterial and fungal infections. Transfusion of granulocytes has been considered a therapeutic modality for severe bacterial and fungal infections in patients with prolonged neutropenia and with functional neutrophil disorders. Good theoretic and experimental evidence demonstrating granulocyte transfusion efficacy exists in preventing and treating severe infection. However, clinical evidence has been more difficult to interpret, with efficacy equivocal in many studies and further trials hindered by limitations in collecting adequate doses of leukocytes from healthy steroid-mobilized donors. The development and use of granulocyte colony-stimulating factor to stimulate normal donors has generated renewed interest in granulocyte transfusions. In clinical studies, granulocyte colony-stimulating factor has markedly enhanced the yield of leukocytes collected from normal donors, which may improve clinical outcomes in patients with severe infections and neutropenia who receive granulocyte transfusions. Preliminary clinical evidence, when correct granulocyte dose per patient body weight is optimized, suggests efficacy. However, well-designed randomized clinical trials are necessary to definitively establish granulocyte transfusions as a viable therapeutic modality in the treatment of severe bacterial and fungal infections in patients with functional neutrophil disorders or neutropenia.     

Granulocyte transfusion in the G-CSF era

Granulocyte transfusions have been used since the 1960s with varying degrees of clinical success in the treatment of infection in patients with neutropenia or inherited granulocyte disorders. A number of studies have indicated that efficacy may well be associated with the dose of granulocytes delivered. Collection of granulocytes using modern apheresis machines and corticosteroid administration yields approximately 20 to approximately 30 x 10(9) neutrophils, unlikely to be adequate for treating an established infection. The administration of G-CSF to healthy donors has resulted in average granulocyte yields up to 8 x 10(10) cells. Normal or near normal blood neutrophil counts are often attained when these concentrates are transfused to neutropenic recipients, and these levels are sustained for up to 24 h. G-CSF-primed granulocytes appear to be functionally normal by both in vitro and in vivo measurements. Adverse effects experienced by recipients are similar to those seen with traditional doses of granulocytes. G-CSF administration to donors is well tolerated. Controlled clinical trials are needed to determine the therapeutic efficacy of G-CSF-primed granulocyte transfusions.     

Granulocyte transfusion in the G-CSF era

Granulocyte transfusions have been used since the 1960s with varying degrees of clinical success in the treatment of infection in patients with neutropenia or inherited granulocyte disorders. A number of studies have indicated that efficacy may well be associated with the dose of granulocytes delivered. Collection of granulocytes using modern apheresis machines and corticosteroid administration yields approximately 20～30×10⁹ neutrophils, unlikely to be adequate for treating an established infection. The administration of G-CSF to healthy donors has resulted in average granulocyte yields up to 8×10¹⁰ cells. Normal or near normal blood neutrophil counts are often attained when these concentrates are transfused to neutropenic recipients, and these levels are sustained for up to 24 h. G-CSF-primed granulocytes appear to be functionally normal by both in vitro and in vivo measurements. Adverse effects experienced by recipients are similar to those seen with traditional doses of granulocytes. G-CSF administration to donors is well tolerated. Controlled clinical trials are needed to determine the therapeutic efficacy of G-CSF-primed granulocyte transfusions.     

A controlled study of the efficacy of granulocyte transfusions in patients with neutropenia

A randomized clinical trial to determine the efficacy of granulocyte transfusions in neutropenic patients with infection was conducted. Criteria for patient selection included a proved infection, a granulocyte count of less than 300/mm³, availability of a suitable donor and failure to respond to at least 72 hours of appropriate antibiotic therapy. Thirty patients were assigned at random to receive either granulocyte transfusions or to serve as a control group. Antibiotic therapy was continued in both groups. Responses were judged by the degree of diminution of infectious episodes and survival. The results showed that 11 of 13 control patients failed to respond during the period of observation, whereas 10 of 17 patients given transfusions responded. The results were statistically significantly different (p < 0.05). The median survival was 22.5 days in the group given transfusions (group 2) and 7.7 in the control group (group 1) (p < 0.01). The granulocyte transfusions were most effective in patients with hypocellular marrows who failed to recover during the period of observation. These results indicate that granulocyte transfusions are effective in the short-term control of infections in neutropenic patients.     

The white cells: use of granulocyte transfusions

Since serious infections are major complications in patients with fewer than 200 phagocytic cells per microliter or in patients with dysfunctional phagocytes, granulocyte transfusions have been used in an attempt to improve clinical outcome. After two decades of trial and clinical use, the role of granulocyte transfusions for therapy of serious infections has not been clearly established. The methods of harvest, storage, and transfusion of granulocytes are acceptable; however, the quantities that are obtained from donors restrict numbers of cells that may be transfused. Limited clinical response has diminished enthusiasm for the use of granulocyte transfusions as therapy, and their use as prophylaxis has been ineffective. Reported clinical data suggest that patients with persisting granulocytopenia with sepsis due to gram-negative bacteria and patients with chronic granulomatous disease with life-threatening infections unresponsive to aggressive antimicrobial therapy may benefit from granulocyte transfusions.     

Granulocyte transfusions in children with chronic granulomatous disease and invasive aspergillosis

The transfusion of granulocytes to restore host defenses in severely granulocytopenic patients or in patients with defective granulocyte functions has been studied for more than 60 years. However, inadequate dosage of cells and inconsistent efficacy has limited the usage of these transfusions. Recently, the use of mobilizing agents such as granulocyte colony stimulating factors and dexamethasone has renewed interest in these treatment modalities. The present study is conducted to determine an appropriate method of enriched granulocyte collection with Fresenius AS.TEC.204 cell separator (Fresenius, Bad Homburg, Germany) and to evaluate the preliminary clinical results of granulocyte transfusion therapy in patients with chronic granulomatous disease and invasive Aspergillosis in parallel with in vitro granulocyte function. Three patients who have been treated for chronic granulomatous disease and invasive Aspergillosis received a total of 20 granulocyte transfusions. To mobilize granulocytes, healthy donors were given 450 microg of granulocyte colony-stimulating factor (G-CSF) subcutaneously and 8 mg of dexamethasone orally approximately 12 h before collection. Five microg/kg/day of G-CSF was also subcutaneously administered prior to granulocyte transfusions. The first patient received 4; the second, 14 and the third, 2 transfusions. The granulocyte count given to these patients ranged between 0.4 and 3.0 x 10(9)/kg. Most transfusions were well tolerated. The nitroblue tetrazolium (NBT) tests that were done 16-24 h after the transfusion showed 14-46% dye reduction. Two of the three patients survived the infection. Granulocyte transfusions from G-CSF and dexamethasone stimulated donors could be a choice of treatment in chronic granulomatous disease patients, especially with disseminated invasive Aspergillosis.     

Gram-negative bacterial pneumonia in the immunocompromised host

Gram-negative bacillary pneumonia is common in all groups of iatrogenically immunosuppressed patients. Mortalities are directly proportional to the degree of neutropenia. Those at particular risk for gram-negative pneumonia are neutropenic patients, patients residing in the hospital setting for prolonged periods, and patients in postoperative periods (eg, organ transplant recipients). The most frequent pathogenesis for pneumonia appears to be airway colonization with gram-negative bacilli, followed by lowe respiratory tract infection. Thus, attention to infection control measures and surveillance culture data is important. Because sputum production is scant or absent, and blood cultures positive in only 30% to 40% of patients, it is often difficult to identify specific etiologic agents. If bacterial pneumonia is suspected in the immunocompromised host, empiric antibiotic coverage should include drugs active against all common aerobic gram-negative bacilli (including P aeruginosa), plus S aureus. Most advocate a beta-lactam plus aminoglycoside combination. Adjunctive treatment with granulocyte transfusions should be reserved for patients not responding to traditional regimens. Immune therapy or prophylaxis has not been fully evaluated for the immunocompromised patient population.     

Granulocyte transfusions: Current science and perspectives

Severe neutropenia renders patients susceptible to life-threatening bacterial and fungal infections. Despite improvements in supportive care and antimicrobial therapy, morbidity and mortality remains significant. Since the 1960s, granulocyte transfusions have been used to either treat or prevent serious infections in patients with neutropenia or neutrophil dysfunction. Despite significant optimizations in product collection, the practice of granulocyte transfusion therapy remains controversial. The use of granulocytes varies widely across institutions and countries in terms of indications, procurement, dose, infusion frequency, and duration of therapy. There are limited and conflicting data concerning its clinical effectiveness; current evidence from clinical trials does not support or refute efficacy. In this narrative review, we summarize the current evidence, discuss persistent concerns and consider future possibilities of the role of granulocyte transfusions.     

Long-term granulocyte transfusion in patients with malignant neoplasms.

Thirty-eight episodes of culture-documented antibiotic-resistant bacterial or fungal infection in patients with malignant neoplasms were treated with daily granulocyte transfusions until the infection improved or the patient died. Cumulative summation temperature plotting allowed easier interpretation of recipient fever response. Seventy-one percent of recipients had a favorable response to transfusion. There was no difference in mortality between patients treated with cells collected by filtration (FL) or intermittent flow centrifugation (IFCL) leukapheresis techniques. Transfusion reactions were more than twice as common with FL than IFCL collected cells. Seventy-four percent of recipients were alive 21 days after completion of transfusions; of the ten deaths, five could be classified as granulocyte transfusion failures. This study suggests that long-term granulocyte transfusion may be required in infected recipients when autologous granulocytes do not return after chemotherapy.     

Leukocyte transfusions: Thinking twice

There is convincing evidence that leukocyte transfusions have a definite, albeit restricted, role in managing profoundly neutropenic patients and certain rare patients with qualitative disorders of neutrophil function. Nonetheless, leukocyte transfusion techniques have yet to meet a number of important conceptual challenges of neutrophil replacement. Even with pharmacologic manipulation of leukocyte donors, current methods of leukapheresis cannot provide neutrophils in numbers that match the body's normal use and turnover of these cells. Neutrophils have also proved to be fragile cells in the blood bank, readily losing normal function with certain collection procedures and during short periods of storage. Moreover, transfused neutrophils that are unable to function and circulate normally in recipients can produce serious toxic reactions. In light of the marginal demonstrable benefits of leukocyte transfusions, potential toxicity is necessarily an important consideration in decisions to use this form of hematologic support. The fact that leukocyte recipients are usually extremely ill does not alter the physician's responsibility to avoid bringing harm to the patient without benefit.     

Effect of leukocyte compatibility on neutrophil increment after transfusion of granulocyte colony-stimulating factor-mobilized prophylactic granulocyte transfusions and on clinical outcomes after stem cell transplantation

The primary limitations of granulocyte transfusions include low component cell dose and leukocyte incompatibility. Component cell dose improved with granulocyte colony-stimulating factor (G-CSF) mobilization, and the transfusion of G-CSF-mobilized, human leukocyte antigen (HLA)-matched granulocyte components resulted in significant, sustained absolute neutrophil count (ANC) increments. However, the effect of leukocyte compatibility on outcomes with G-CSF-mobilized granulocyte transfusions is unclear. The objectives were to determine the effect of leukocyte compatibility on ANC increments and selected clinical outcomes after transfusion of prophylactic, G-CSF-mobilized granulocyte components into neutropenic recipients of autologous peripheral blood stem cell (PBSC) transplants. Beginning on transplant day 2, 23 evaluable recipients were scheduled to receive 4 alternate-day transfusions of granulocyte components apheresed from a single donor given G-CSF. G-CSF was also given to recipients after transplantation. Recipient ANC was determined before and sequentially after each granulocyte transfusion to determine the peak ANC increment. Leukocyte compatibility was determined at study entry only by a lymphocytotoxicity screening assay (s-LCA) against a panel of HLA-defined cells. Eight recipients had positive s-LCA. On days 2 and 4, the mean peak ANC increments after granulocyte transfusion were comparable between the cohorts with positive and negative s-LCA. However, the mean peak ANC increments on day 6 (246/microL vs 724/microL; P =.05) and day 8 (283/microL vs 1079/microL; P =.06) were lower in the cohort with positive s-LCA, in spite of the transfusion of comparable component cell doses. Adverse reactions occurred with only 5 of 87 (5.7%) granulocyte transfusions and were not associated with leukocyte compatibility test results. Platelet increments, determined 1 hour after granulocyte transfusion, were comparable between the cohorts. Although the 2 cohorts received PBSC components with similar CD34(+) cell doses, the cohort with a positive s-LCA had delayed neutrophil engraftment and a greater number of febrile days and required more days of intravenous antibiotics and platelet transfusions. Leukocyte incompatibility adversely affected ANC increments after the transfusion of G-CSF-mobilized granulocyte components and clinical outcomes after PBSC transplantation.     

The effects of daily recombinant human granulocyte colony-stimulating factor administration on normal granulocyte donors undergoing leukapheresis [see comments]

The effects of daily administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) to eight normal volunteers donating granulocytes for neutropenic relatives undergoing marrow transplantation were studied. Granulocyte donors consisted of seven marrow donors (5 syngeneic, 2 HLA identical) and one haploidentical son who had not donated marrow. All donors were administered daily rhG-CSF at a mean dose of 5 micrograms/kg/d (range 3.5 to 6.0) for a mean of 11.75 days (range 9 to 14 days), and granulocytes were collected a mean of 7.6 times (range 4 to 12). RhG-CSF was well tolerated and only minor side effects were observed. All donors became anemic from marrow donation and the removal of red blood cells during the collection procedures. Red blood cell transfusions were not given. All donors had a decrease in platelet counts and the magnitude of the decrement appeared to be greater than in historical donors. This was due in part to increased removal of platelets with the collection product, but a direct effect of rhG-CSF on platelet production cannot be excluded. The mean precollection granulocyte level was 29.6 x 10(9)/L (range 11.8 to 79.8), which was a 10-fold increase over baseline. The mean number of granulocytes collected was 41.6 x 10(9) (range 1.3 to 144.1), which was a six-fold increase over historical donors not receiving rhG-CSF. The mean granulocyte level 24 hours after transfusion into neutropenic recipients was 0.95 x 10(9)/L (median 0.57 and range .06 to 9.47). This study indicates that rhG-CSF is safe to administer to normal individuals, significantly improves the quantity of granulocytes collected, and results in significant circulating levels of granulocytes in neutropenic recipients. Further studies to evaluate rhG- CSF in normal granulocyte donors are warranted.     

Current status of granulocyte (neutrophil) transfusion therapy for infectious diseases

The transfusion of neutrophils, or granulocyte transfusion therapy, has long been considered as a logical approach to the treatment of severe bacterial and fungal infections in patients with prolonged neutropenia or intrinsic defects in neutrophil function. However, despite numerous clinical trials, the efficacy and safety of granulocyte transfusion therapy remain controversial. Efficacy has been compromised largely by the inability to transfuse sufficient quantities of functionally active neutrophils to patients. The recent use of recombinant granulocyte colony-stimulating factor (G-CSF) to mobilize neutrophils in donors before centrifugation leukapheresis has rekindled interest in the potential clinical applications of granulocyte transfusion therapy. This review focuses on the use of G-CSF for donor stimulation and summarizes the current status of granulocyte transfusion therapy for treatment of infectious diseases.     

Prevention of cytomegalovirus infection by cytomegalovirus immune globulin after marrow transplantation

In an effort to prevent cytomegalovirus infection among seronegative patients having marrow transplants, a globulin with high antibody levels against cytomegalovirus was given before and for 11 weeks after transplantation in a randomized trial. Among 36 patients who received no prophylactic granulocyte transfusions, globulin recipients had significantly fewer infections than controls (2 of 17 versus 8 of 19, p = 0.05 by Fisher's exact test and p = 0.03 by Mantel-Cox test). Conversely, infection rates were high and unchanged by globulin use among patients who received granulocytes from seropositive donors (7 of 8 recipients versus 6 of 7 controls). The lack of effect of the globulin among patients receiving transfusions of granulocytes from seropositive donors may suggest that the dose of antibody was insufficient or that antibody is ineffective against virus transmitted in granulocytes. We conclude that cytomegalovirus infection can be prevented by immunoprophylaxis in seronegative patients having marrow transplants who are not given granulocyte transfusions.     

Granulocyte transfusion therapy: update on potential clinical applications

The clinical usefulness of granulocyte transfusions for treatment or prevention of life-threatening bacterial and fungal infections remains controversial. Clinical benefit has long been limited by insufficient donor stimulation regimens and suboptimal leukapheresis techniques. Methodologic progress, in particular mobilization of neutrophils in healthy donors by administration of G-CSF, has significantly enhanced leukapheresis yields. A newly published study indicates that unrelated community donors can be effectively and safely used as an alternative to related family donors. Furthermore, several recent studies suggest that it may be possible to store granulocyte concentrates for 24 to 48 hours with adequate preservation of neutrophil function. This review summarizes the current role of granulocyte transfusion therapy in infectious diseases and highlights important recent advances.