Background: Neutropenic fever is the most important complication of chemotherapy and hematopoietic stem cell transplantation. The granulocyte transfusion (GT) has been used in neutropenic patients with severe bacterial infection. However, the data regarding the efficacy of the GT and the results are limited in children. The purpose of this study was to evaluate the efficacy of the GT in pediatric patients with neutropenic fever and clinical response to documented gram-negative sepsis. Methods: 11 patients received totally 70 courses of GT, were retrospectively identified in Department of Pediatrics, Pusan National University Children’s Hospital, from July 2014 to June 2017. Eight of 11 patients in nine episode had confirmed gram-negative sepsis. 15 patients of the control group were received a broad-spectrum antibiotics therapy alone. The granulocyte were collected by automated apheresis and approved donors applied 300 ug of G-CSF 12 hours before apheresis. Results: For 11 pediatric patients with neutropenic fever, the GT was performed at an average of 1-13 courses per episode, a total of 70 courses in 12 episodes. 10 of the patients were boys and one was girl, mean age was 11.34±1.02 years. Eight of the patients had acute leukemia, two patients had solid tumors and one had myelodysplastic syndrome (MDS). In 15 patients of the control group, 12 patients had acute leukemia, other three patients had aplastic anemia, lymphoma and MDS. Mean duration of neutropenic fever was 22.83±4.24 days. Infection were classified primary bacterial infection (83.3%, n=10) included gram-negative sepsis (75.0%, n=9) and clinically suspected infection (16.7%, n= 2). All 15 patients of the control group diagnosed gram-negative sepsis. The mean granulocyte yield was 2.22±0.14 x 1010/unit, the mean increment in absolute neutrophil count (ANC) was 209.23±94.46 x 103/uL and correlated with the granulocyte yield infused ( P=0.001). The WBC count and ANC after GT were significantly higher than the baseline values ( P0.001, P=0.004). 10 of 15 controls survived, and eight of nine received GT for revealed gram-negative sepsis survived ( P=0.037). Conclusion: The GT is safe and effective in pediatric hemato-oncology patients with neutropenic fever. Additionally, the GT combined with antibiotics improved survival from gram-negative sepsis in pediatric patients with neutropenia.