Distribution of Pathogenic Bacteria and Its Influence on Expression of BCL-2 and BAX Protein after HSCT in the Patients with Hematological Malignancies / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the distribution of pathogenic bacteria in the patients with hematologic malignancies received hematopoietic stem cell transplantation (HSCT) and its influence on the expression of BCL-2 and BAX proteins.</p><p><b>METHODS</b>The clinical data of 64 patients with malignant lymphoma (ML) received auto-HSCT from January 2011 to December 2015 in our hospital were analyzed. On basis of post-treansplant infection, the patients were divided into infection group (36 cases) and non-infection group (28 cases). The distribution of pathogenic bacteria in 2 groups was identified, the T lymphocyte subsets of peripheral blood, expression level of apoptotic proteins and C-reaction protein (CRP) in 2 group were detected.</p><p><b>RESULTS</b>Thirty-six strains of pathogenic bacteria were isolated from 36 case of hematological malignancy after HSCT, including 24 strains of Gram-negative bacteria (66.67%) with predominamce of klebsiella pneumoniae (19.44%). The periperal blood CD4+ (t=2.637, P<0.01), CD4+/CD8+ ratio (t=8.223, P<0.01), BCL-2 protein (t=5.852, P<0.05), BCL-2/BAX ratio (t=14.56, P<0.01) in infection group were significantly lower than those in non-infection group, while CD8+ (t=2.285, P=<0.01), CRP (t=39.71, P<0.01), BAX level in infection group were higher than those in non-infection group. The pearson correcation analysis showed that the CD4+/CD8+ ratio in infection group positively correlated with BCL-2/BAX ratio (t=0.341, P<0.05), while serum CRP level in infection group negatively correlated with BCL-2/BAX ratio (t=-0.362, P<0.05).</p><p><b>CONCLUSION</b>The pathogenic bacteria infecting ML patients after HSCT were mainly Gram-negative bacteria. The post-transplant infection can promote the expression up-regulation of related inflammatory factors and apoptotic proteins. The pathogens may be involved in cell apoptisis that provides a new strategy to treat the hematologic malignancies.</p>

Comparison of curative efficacy after G-CSF-mobilized sibling HLA-matched peripheral blood hematopoietic stem cell transplantation versus that combined with BMT for patients with hematologic malignancies in a single center / 中国实验血液学杂志

This study was aimed to retrospectively analyze and compare the clinical curative efficacy of patients with hematologic malignancies after G-CSF-mobilized sibling HLA-matched (sm) peripheral blood hematopoietic stem cell transplantation (sm-allo-PBHSCT) and sm-allo-PBHSCT combined with bone marrow transplantation (BMT). 100 patients received sm-allo-HSCT in a single center from October 2001 to October to 2010, included 38 patients received sm-allo-PBHSCT and 62 patients received sm-allo-PBHSCT combined with BMT. The myeloablative or reduced intensity conditioning regimens were chosen according to the condition of patients. All patients received standard cyclosporine (CsA) and mycophenolate mofetil (MMF) as prophylaxis for GVHD. The results showed that the rapid hematopoietic reconstitution was observed in all patients. The median time of ANC ≥ 0.5 × 10(9)/L in both groups were 12 days, the median time of platelet count ≥ 20 × 10(9)/L was 15 days in sm-allo-PBHSCT group and 16 days in sm-allo-PBHSCT + BMT group. The incidence of acute GVHD, acute GVHD of III-IV grade and chronic GVHD in sm-allo-PBHSCT and sm-allo-PBHSCT + BMT groups were 37.1% and 34.2%, 7.89% and 8.06%, 36.11% and 41.38% respectively, there were no statistical differences. The relapse rates were similar in two groups (sm-allo-PBHSCT 13.16% vs sm-allo-PBHSCT + BMT 12.9%). The 3-year disease-free survivals in sm-allo-PBHSC and sm-allo-PBHSCT + BMT groups were 57.1 ± 8.7% and 61.3 ± 6.4% respectively (p = 0.852). The 2-year overall survival of high-risk patients was 41.4 ± 12.8% in sm-allo-PBHSCT group, while 60.9 ± 9.6% in sm-allo-PBHSCT + BMT group (p = 0.071). It is concluded that the rhG-CSF mobilized sibling matched allo-PBHSCT + BMT is superior to the rhG-CSF mobilized sibling matched allo-PBHSCT in increasing the overall survival of high-risk hematologic malignancies.

Detection of Th17/treg cell-associated cytokines in peripheral blood of patients with graft-versus-host disease and its clinical significance / 中国实验血液学杂志

To investigate the peripheral levels and clinical significance of Th17/Treg cell-associated cytokines in patients with acute graft versus host disease (aGVHD) or chronic GVHD (cGVHD), blood samples were collected from 39 hematopoietic stem-cell transplantation patients and 20 healthy donors. The patients included 10 patients with aGVHD, 13 patients with cGVHD and 16 patients without evidence of GVHD. Th17/Treg cell-associated cytokines such as IFNγ, IL-4, IL-6, IL-10, TGF-β(1), IL-17 and IL-23 were detected by ELISA. The results showed that the plasma levels of IFN-γ, IL-4, IL-6, IL-17 and IL-23 significantly increased in patients with aGVHD or cGVHD, compared with the patients without clinical signs of GVHD and the healthy donors (p < 0.05), while IL-10 and TGF-β(1) were obviously lower than that of them (p < 0.05). After aGVHD and cGVHD patients were treated effectively, the plasma levels of IL-6, IL-17 and IL-23 were significantly decreased, and IL-10, TGF-β(1) were significantly increased, while the levels of IFN-γ and IL-4 did not markedly change. The TGF-β(1) level were negatively correlated with IL-6 (r = -0.36, p < 0.05), IL-17 (r = -0.51, p < 0.05) and IL-23 (r = -0.44, p < 0.05) respectively, while there were positive correlations between IL-6 and IL-17 (r = 0.62, p < 0.05), IL-6 and IL-23 (r = 0.71, p < 0.05), IL-17 and IL-23 (r = 0.93, p < 0.05). It is concluded that Th17/Treg cell-associated cytokines may play an important role in the development of a/cGVHD, which helps to find novel targets for developing new strategies of GVHD treatment.

Influence of TLR2 and TLR4 agonists on migration of cord blood CD34(+) cells / 中国实验血液学杂志

This study was aimed to investigate the influence of TLR2 and TLR4 agonists on the migration and adhesion activity of umbilical cord blood (UCB) CD34(+) cells and to explore the underlying mechanism. The expression of TLR2 and TLR4 on UCB CD34(+) cells was detected with flow cytometry. The effect of TLR2 agonist (PAM3CSK4) and TLR2 agonist (LPS) on the migration and adhesion ability of UCB CD34(+) cells was evaluated with chemotaxis and adhesion assays. The results indicated that expression levels of TLR2 and TLR4 were (14.2 ± 3.8)%, (19.6 ± 4.1)% respectively. Compared with the control group, the migration activity of UCB CD34(+) cells toward SDF-1 decreased significantly in LPS group (p < 0.01). The adhesion activity was not altered significantly in LPS group. However, both the migration activity towards SDF-1 and the adhesion activity of UCB CD34(+) cells were not changed significantly in PAM3CSK4 group. Further study found that LPS did not affect the expression level of CXCR4 on CD34(+) cells, but could inhibit the spontaneous migration ability of CD34(+) cells. It is concluded that TLR4 activation can decrease the chemotaxis function of CD34(+) cells towards SDF-1, which may associate with the decreased spontaneous migration ability of CD34(+) cells.

Expression and abscission of activated receptors and their ligands on/from NK cells in peripheral blood of patients with acute leukemia / 中国实验血液学杂志

This study was aimed to explore the immune escaping mechanisms based on expression and abscission of human natural killer (NK) cell activating receptors NKG2D and their ligands MICA/B, ULBP-1, 2, 3 in patients with acute leukemia (AL). 30 de novo AL patients and 10 healthy persons (control) were enrolled in study. Flow cytometry was used to detect the expression levels of MICA/B, ULBP-1, 2, 3 on leukemic cells. ELISA was used to detect the levels of soluble MICA (sMICA), solube MICB (sMICB) and soluble ULBP-1, -2, -3 in the serum. The results showed that sMICA, sMICB and ULBP-1, -2, -3 were not expressed or expressed at very low levels on leukemia cells of the patients; the levels of free sMICA and sMICB in serum of AL patients were higher than that in serum of healthy persons, there was significant difference (p<0.01). But the levels of ULBP 1-3 in serum of AL patients did not show obvious statistical difference as compared with healthy persons (p>0.05). It is concluded that the negative or low expression of NKG2D ligands (MICA, MICB and ULBPs) on surface of acute leukemia cells may lead to the immune escape of leukemia cells, the abscission of MICA and MICB, and the deficiency of ULBP expression on leukemia cells may be one of immune escape mechanisms of leukemia cells.

Functional defect of partial homing receptor on human cord blood hematopoietic stem/progenitor cells / 中国实验血液学杂志

This study was aimed to investigate the function defect of partial homing receptor on cord blood hematopoietic stem cells (CBHSC) and explore efficacy and feasibility of intervention in vitro. The expression and activity of active groups in P, E-selectin ligands on CD34+ cells from cord blood, bone marrow and peripheral blood were detected by flow cytometry; meanwhile the expression of active groups in selectin ligands on CD34+ cells treated by fucosyl transferase in vitro was determined by flow cytometry. The results indicated that the expression levels of CD26 on the surface of stem/progenitor cells (CD34+) from cord blood, bone marrow and peripheral blood were (7.62+/-0.63)%, (6.35+/-0.89)% and (6.18+/-0.91)% (p>0.05) respectively. And the activities of CD26 of the three sources of stem cells were 67.15 U/1000 cells (1 U=1 pmol/min), 26.85 U/1000 cells and 20.95 U/1000 cells respectively, in which the activity of CD26 on surface of CD34+ from cord blood was significantly higher than that from other both sources (p<0.01). The expression levels of P-selectin ligand on the stem/progenitor cells three kinds were (83.46+/-6.33)%, (15.65+/-0.89)% and (80.17+/-6.85)%, and the expression levels of E-selectin ligand on stem/progenitor cells of three kinds were (25.31+/-1.03)%, (26.34+/-0.89)% and (29.79+/-1.78)% respectively. The expression of E-selectin ligand on the surface of cord blood stem/progenitor cell CD34+ increased from (25.31+/-1.03)% to (63.23+/-1.08)% after glycosylation engineering. It is concluded that there is no significant difference of the expression of CD26 between the three sources of stem/progenitor cells, but the activity of CD26 in cord blood was obviously higher than that in bone marrow and peripheral blood. The expression of P-selectin ligand on bone marrow stem/progenitor cell was lower than that on stem cells of cord blood and peripheral blood. Glycosylation engineering can promote and elevate the expression of E-selectin ligand on the surface of CD34+ cells from cord blood.

Unrelated cord blood transplantation in adult patients with hematologic malignancies / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyse the engraftment, transplant-related complications and survival after unrelated cord blood transplantation (UCBT) in patients with hematologic malignancies.</p><p><b>METHODS</b>Twenty eight consecutive adult patients with hematological malignancies were treated with UCBT and 20 of them were advanced-stage diseases. Double or multiple UCB grafts were used for 18 patients, while single UCB graft for 10 patients. Myeloablative conditioning regimens were given to 26 cases and nonmyeloablative regimens to 2 cases. All patients were given a combination of cyclosporine (CsA) and mycophenolate mofetil (MMF) for graft-versus-host disease (GVHD) prophylaxis.</p><p><b>RESULTS</b>Median time to neutrophil engraftment (≥ 0.5 × 10(9)/L) in 26 patients was 18 (14 - 37) days and platelet engraftment (≥ 20 × 10(9)/L) in 22 patients was 30 (25 - 49) days. Chimerism was weekly assessed by PCR analysis of short tandem repeat (STR) sequences in whole blood or bone marrow and 22 cases were confirmed of fully donor chimeric from 7 to 21 days after transplantation. Eighteen cases developed acute GVHD, greater than grade II in 1, and 6 of 22 patients who survived more than 100 days developed limited chronic GVHD. Eighteen cases were alive in hematologic remission at a median follow-up of 9.5 (2.5 - 72.0) months. The probability of event-free survival at 3 years was 56.7%. Two cases relapsed and 8 of 10 cases died of transplant related complications.</p><p><b>CONCLUSIONS</b>UCBT could be safely and effectively used for adult patients with hematologic malignancies. Use of double UCB units is a strategy extending the feasibility of UCBT.</p>

Reconstitution of NK cells and their receptors in patients with acute leukemia following unrelated cord blood stem cell transplantation / 中国实验血液学杂志

This study was to investigate the reconstitution of NK cells and their receptors after unrelated cord blood stem cell transplantation (UCBT) and its clinical importance. 11 cases of acute leukemia underwent UCBT were enrolled in this study. The reconstitution of NK cells and their surface receptors as well as the the recovery of T and B cells within 90 days after clinical engraftment following UCBT were measured and analysed by flow cytometry. The results indicated that the recovery of NK cells appears to be relatively early. CD3(-)56(+) NK cell count was (35.12 +/- 18.66)% of peripheral blood (PB) lymphocytes on the day of clinical engraftment and higher than that in normal. The peak of the NK cells reached to (37.8 +/- 17.52)% of lymphocyte at 30 days after clinical engraftment. NK count was (30.4 +/- 19.14)% at 60 days after clinical engraftment when the absolute NK cell count reached to the peak (up to 544 cells/microl) in PB. The activated receptor NKG2D was reconstituted fast and high expressed [(79.58 +/- 8.71)%] at the time of clinical engraftment with a tendency of gradual elevation, which reached to peak value (82.55 +/- 9.10)% at day 60. Another activated receptor NKp46 also reconstituted fast, and maintained at a high level even at 90 days after clinical engraftment. The expression of NKG2A was lower than that of the activated receptor of NK cells, which tendency lasted for at least 90 days after clinical engraftment. The reconstitution of T cells in PB after UCBT was relatively slow with lower expression rate. It is concluded that the reconstitution of NK cells in patients with acute leukemia is earlier following UCBT. The earlier recovery of activated receptor of NK cells, especially NKG2D, suggests that the activation of NK cells may play a role in graft versus leukemia (GVL) effect in the early period after UCBT.

Successful treatment of post-transplant lymphoproliferative disorder in allogeneic hemopoietic stem cell transplant recipient for severe aplastic anemia / 中华器官移植杂志

Objective To study early diagnosis and treatment of post transplant lymphoprolifer- ative disorder in allogeneic hemopoietic stem cell transplant recipients.Methods A 16 years old patient with severe aplastic anemia received HLA-mismatched sibling allogeneic hemopoietic stem cell trans- plant after conditioning with cyclophosphamide/antithymocyte globulin/methylprednisolone(CY/ ATG/MP)regimen.Results On the day 72 posttransplantation,he developed lymphoproliferative disorder.After withdrawal of CsA,he was treated with methylprednisolone,intravenous immune globulin and IFN alpha,and recovered completely from PTLD.Conclusions PTLD is a rare and fatal complication of both solid-organ and hemopoietic stem cell transplantation.Surveillance for PTLD by PCR for circulating EBV-DNA may be appropriate in high risk settings.Early diagnosis,immunosup- pression therapy reduction or even withdrawal in time is important.