Through thorough understanding of the pathophysiology of graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect, especial the effects of immune cells and cytokines, the article is designed to find out multiple ways to reduce GVHD and reserve or enhance GVL effect. Many kinds of cytokines are involved in the pathophysiologic mechanisms of GVHD and GVL effect. Different cytokines have different effects on GVHD and GVL effect. Immune cells play important roles in regulating GVHD and GVL effect. Studies shows that some cytokines and specific immune cells are able to reduce GVHD, as well as reserve or enhance GVL effect at the same time, which make the separation of GVL from GVHD can be achieved. On the whole, through thorough understanding of the mechanism of immune cells and cytokines in GVHD and GVL effect, we can find out multiple ways to reduce GVHD, as well as reserve or enhance GVL effect.

BACKGROUND: Hemorrhagic cystitis remains a common complication of hematopoietlc stem cell transplantation.Granulocyte-macrophage colony stimulating factor (GM-CSF) affects proliferation and differentiation of hematopoietic stem/progenitor cells, adjusts functions of monocytes, granulocytes, lymphocytes and endothelial cells.OBJECTIVE: To investigate the protective effects of GM-CSF bladder irrigation in hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.DESIGN: Case analysis.PARTICIPANTS: A total of 15 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation at the Zhongshan Hospital of Sun Yat-sen University from January 2004 to August 2006 (routine treatment group). A total of 16 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation from September 2006 to December 2008 (GM-CSF group).METHODS: In the routine treatment group, patients received mesna, hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis. In the GM-CSF group, GM-CSF was infused into the bladder in addition to mesna,hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis 24 hours before cyclophosphamide treatment. Catheter was extracted 3 days following cyclophosphamide withdraw. Following washing with saline, the bladder was emptied. 10 mL of saline and 5 mL of lidocaine were added into 300 μg of GM-CSF. The mixture was infused into the bladder for 60-120 minutes.MAIN OUTCOME MEASURES: The following parameters were measured: occurrence of hemorrhagic cystitis and its correlation to graft versus host disease, as well as the occurrence of cytomegalovirus infection and urinary system infection.RESULTS: Compared with routine treatment group, the occurrence rate of hemorrhagic cystitis was significantly decreased in the GM-CSF group (x2=4.39, P < 0.05), mean duration of hemorrhagic cystitis and duration of hospitalization were significantly shortened (t=3.97, P < 0.05; t=3.13, P < 0.05), and the occurrence rate of over grade HI hemorrhagic cystitis was significantly reduced (x2=5.04, P < 0.05). Cystitis degree was associated with degree and duration of graft-versus-host disease (r = 0.76).Compared with the routine treatment group, cytomegalovirus infection rate was slightly decreased in the GM-CSF group (x2=0.28, P> 0.05), and occurrence rate of over grade Ⅲ hemorrhagic cystitis was higher in patients with cytomegalovirus infection.Compared with the routine treatment group, the occurrence rate of urinary system infection was slightly reduced in the GM-CSF group (x2=0.28, P > 0.05).CONCLUSION: GM-CSF bladder irrigation is well tolerated and often effective, and should be considered as a preparative regimen of hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.

Objective To explore the relationship between immune functions of patients with acute leukemia (AL) and invasive fungat infection (IFI). Methods T lymphocyte subpopulations and natural killer (NK) cells in 61 AL patients complicated with IFI at first visit, AL remission, the time of IFI onset and 4 weeks after antifungal treatment were measured by flow cytometry. Meanwhile,levels of IgG, IgM and IgA were detected by immunoturbidimetry. The statistical analysis was done using ANOVA, t test and chi-square test. Results CD3+ , CD3+ CD4+ , CD8+ CD28+ T lymphocyte as well as CD4+/CD8+ ratio at the time of IFI onset in AL patients were all lower than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 25.6,26.6,13. 1,167.9; all P＜0.05), while CD8+ CD28- T lymphocyte were higher than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 220.2,P＜0.01). CD3+ , CD3+ CD4+ and CD4 + /CD8+ ratio of patients who responded effectively to antifungal treatment were all higher than those of non-responders (t=3.75,8. 61,3.17; all P＜0.05). The serum levels of IgG, Igm and IgA at first visit, ALremission, the time of IFI onset and 4 weeks after treatment were similar (F=0.78,0.72,0.81; all P ＞0.05). The effect rate of antifungal therapy in AL remission group was higher than that in nonremission group (87% vs 53%,x2 = 7.62, P＜0.05). Conclusions The cellular immune functions are impaired severely in AL patients complicated with IFI, while the levels of IgG, IgM and IgA are similar during IFI. Therefore, the efficacy of antifungal therapy may partly depend on the recovery of cellular immune functions and remission of AL.

BACKGROUND:Donor selection for high-risk acute leukemia is still controversial.OBJECTIVE:To compare the therapeutic efficacy of the unrelated donor peripheral blood and matched sibling allogeneic hematopoietic stem cell transplantation for high-risk acute leukemia.METHODS:Total 65 patients with high-risk acute leukemia treated during January 2008 to January 2016 were included,in which 30 patients chose the unrelated donor peripheral blood stem cell transplantation (UD), and other 35 chose the matched sibling allogeneic hematopoietic stem cell transplantation (MS) according to the wishes of patients and their own situation. After treatment, the chi-square test, Kaplan-Meier survival analysis method, and other methods were used to compare the implanted and hematopoietic reconstitution, the occurrence of graft-versus-host disease, relapse mortality and long-term survival between the two groups.RESULTS AND CONCLUSION:The implantation rate, platelet hematopoietic reconstitution time, the incidence of acute and chronic graft-versus-host disease, and its type exhibited no significant differences between the two groups (P > 0.05).The relapse rate, total death rate, and transplant-related mortality rates were 10.0%, 50.0%, and 40.0% in the UD group and 20.0%, 48.6%, and 25.7% in the MS groups, respectively, and the intergroup difference was insignificant (P > 0.05).The expected 2-year cumulative disease-free free survival and overall survival rates were (49.4±9.2)% and (52.6±9.2)% in the UD group and (53.9±8.5)% and (53.9±8.5)% in the MS group, respectively, and the intergroup difference was also insignificant (P > 0.05). Our experimental findings show that unrelated donor peripheral blood stem cell transplantation can be used as an effective alternative in the absence of sibling donors.

Objective: To investigate the synergistic lethal effect and mechanism of arsenic trioxide (ATO) and aclacinomycin (ACM) on human acute myeloid leukemia cell line KG-1a. Methods: Colony-forming assay was used to detect the proliferation of KG-1a cells treated with different concentration of ATO and ACM. Compusyn software was used to analyze the synergistic effect of ATO and ACM. Flow cytometry and Wright's staining were used to analyze the apoptotic rate of KG-1a cells induced by combined treatment of ATO and ACM. Western blot was used to determine the expression of proteins associated with apoptosis. Results: The cytotoxicity of arsenic trioxide or aclacinomycin alone was in a dose-dependent manner. Flow cytometry analysis showed that the apoptotic rate of KG-1a cells treated with both 0.4 μmol/L ATO and 10 nmol/L ACM was (34.5±3.1)%, significantly higher than (7.6±1.1)% of 0.4 μmol/L ATO treatment or (18.7±2.3) % of 10 nmol/L ACM treatment alone (P<0.05). The apoptotic rate of KG-1a cells treated with both 1.5 μmol/L ATO and 37.5 nmol/L ACM was (52.5±4.7)%, significantly higher than (19.1±3.2)% of 1.5 μmol/L ATO treatment or (27.7±2.2)% of 37.5 nmol/L ACM treatment alone (P<0.05). The apoptotic rate of KG-1a cells treated with both 3.0 μmol/L ATO and 75 nmol/L ACM was (61.3±4.5)%, significantly higher than (29.5±2.5)% of 3.0 μmol/L ATO treatment or (28.6±3.4) % of 75 nmol/L ACM treatment alone (P<0.05). In addition, the result of Wright's staining showed that combined treatment of ATO and ACM induced a more apparent phenotype of apoptosis when compared with single agent treatment. Compusyn software analysis showed that the combination index (CI) value of combined treatment group was less than 1, which indicated the synergistic effect of these two agents. Conclusions Combined treatment of ATO and ACM shows a synergistic lethal effect on human acute myeloid leukemia cell line KG-1a via activating the apoptotic pathway, which inhibits cell growth and induces apoptosis.

OBJECTIVETo study the effect of glycolytic inhibitor 3-Bromopyruvate (3-BrPA) on the proliferation and apoptosis of mouse spleen lymphocytes and explore its mechanism.

METHODSAn one-way mixed lymphocyte culture (MLC) system was established, including BALB/c mouse spleen cells (H-2d) as stimulator and C57BL/6 mouse spleen cells (H-2b) as responder. With treatment of 3-BrPA at different concentrations (0-200 μmol/L), lymphocyte proliferation capacity was detected by the CCK-8 method, the expression of CD3, CD4, and CD8 by flow cytometry, and the concentrations of cytokine interleukin (IL)-4 and interferon (IFN)-γ in the supernatant by ELISA.

RESULTSAt a middle or high dose (over 20 μmol/L), 3-BrPA displayed a dose-dependent inhibitory effect on lymphocyte proliferation in the MLC system. The 50% inhibitory concentration (IC50) were 48.6, 41.2, and 41.9 μmol/L after 24, 36, and 48 h culture, respectively. With treatment of 50 μmol/L 3-BrPA, the IFN-γ level [(164.25 ± 20.14) ng/L] was significantly lower, compared with control [(277.61 ± 18.46) ng/L]. The IL-4 level [(31.06 ± 6.06) ng/L] was significantly higher, compared with control [(28.64 ± 3.97) ng/L]. Consequently, the IFN-γ/IL-4 ratio decreased significantly.

CONCLUSIONThese results indicate that 3-BrPA had a significant inhibitory effect on the proliferation of mouse spleen lymphocytes cultured in MLC system, accompanied with the Th2-biased secretion of cytokines.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lymphocyte Culture Test, Mixed ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Pyruvates ; pharmacology ; Spleen ; cytology ; metabolism