Early T-cell precursor acute lymphoblastic leukemia: report of 1 case and review of literature / 白血病·淋巴瘤

Objective:To improve the understanding of the diagnosis and treatment of early T-cell precursor acute lymphoblastic leukemia (ETP-ALL).Methods:The clinical data of a patient with ETP-ALL who was misdiagnosed as peripheral T-cell lymphoma-not otherwise specified (PTCL-NOS) admitted to the Second Hospital of Lanzhou University in October 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient who presented "inguinal lymphadenopathy" as the first symptom underwent lymph node biopsy and pathological examination at local hospital, and he was diagnosed as PTCL-NOS according to the consultation of another 2 hospitals. After 2 courses of chemotherapy (CHOPE regimen, GLD regimen, unknown specific medication and dosage), the therapeutic efficacy was poor. For further diagnosis and treatment, this patient came to Lanzhou University Second Hospital. Flow cytometry found blast cells in the bone marrow, and then other related examinations were completed, he was finally diagnosed as ETP-ALL. The chemotherapy regimens of Hyper-CVAD and EA were alternatively used, progressive disease (PD) occurred after 3 courses of treatment, and chidamide was added in the 4th and 5th courses of treatment, the disease still progressed, and the patient died after follow-up. The disease course of the patient was about 12 months.Conclusions:ETP-ALL has unique immunophenotypic characteristics. ETP-ALL patients have a low remission rate after conventional induction therapy, high recurrence rate and poor prognosis. Currently, there is no effective standard treatment regimen, and allogeneic hematopoietic stem cell transplantation or timely addition of new drugs may improve the prognosis.

The Influence of CONUT Score on the Prognosis of Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the influence of controlling nutritional status (CONUT) score on the prognosis of newly diagnosed patients with multiple myeloma (MM).@*METHODS@#The clinical data 119 patients with MM who were diagnosed according to the international myeloma diagnostic criteria in Lanzhou University Second Hospital from April 2010 to October 2018 were collected and retrospectively analyzed. The relationship between clinical indexes, including age, sex, MM type, absolute lymphocyte count (ALC), absolute neutrophil count (ANC), absolute monocyte count (AMC), hemoglobin (Hb), platelet (PLT), β@*RESULTS@#Compared with high-scoring group, low-scoring group had a better OS ［median OS was 43.3 months and 127.67 months, respectively, 95% confidence interval (CI): 57.065-78.345, P=0.038］. At the same time, the low-scoring group also had higher level of ALC, ANC, AMC, Hb, PLT, ALB, and CHO but lower of GLO. Multivariate survival analysis showed that age (HR=1.027, 95%CI: 1.000-1.054, P=0.048), AMC (HR=11.284, 95%CI: 22.968-42.897, P<0.001), CONUT score (HR=1.198, 95%CI: 1.036-1.385, P=0.015), M protein (non-IgG/IgG type) type (HR=0.503, 95%CI: 0.259-0.977, P=0.043) were independent factors affecting the prognosis of MM patients.@*CONCLUSION@#The CONUT score as an immune-nutrition score is a convenient and easy-to-obtain index to effectively predict the prognosis of MM patients.

Clinical Significance of sFLC/MP Level and Light Chain Escape in Patients with Multiple Myeloma Relapse / 中国实验血液学杂志

OBJECTIVE@#To explore the effects of serum free light chain (sFLC) and monoclonal protein (MP) on the efficacy and prognosis of patients with multiple myeloma relapse, and investigate the clinical value of light chain escape (LCE).@*METHODS@#The relationship between sFLC/MP levels and clinical features and outcomes in 71 patients with multiple myeloma after relapse were retrospective analyzed. The patients were divided into MO group, MLC group and LCE group according to different levels of sFLC/MP after relapse. Then the clinical indicators, efficacy, survival after relapse (SAR) and overall survival (OS) of the patients in each group were compared. Meanwhile a paired sample t test was used to analyze the relevant indicators of the patients before and after relapse in LCE group.@*RESULTS@#There were significant differences in ISS stage, the levels of Hb, PLT, ALB, SFLC/MP and the proportion of myeloplasma cells afte relapse (P<0.05). The initial treatment effect of the patients in MO group was better than those in the other groups, and the LCE group was the worst (P<0.05). Comparison of relevant indicators between the patients before and after relapse in LCE group showed that the levels of MP, Hb and PLT decreased significantly, while sFLC, LDH and Cr increased significantly (P<0.05). Multivariate analysis showed that MO was the independent risk factor affecting SAR, while MO and LCE were the independent risk factors affecting OS (P<0.05) of the patients.The average SAR of the patients in MO, MLC and LCE group was 41, 28.6 and 23.5 months (P=0.002), and the average OS was 79.6, 57.9 and 41 months (P<0.001), respectively. The patients in MO group showed longer SAR and OS, while the LCE group was the shortest.@*CONCLUSION@#After relapse, patients only with elevated MP levels have a better curative effect and prognosis, while only with elevated sFLC levels have poor curative effects and prognosis, which suggesting that sFLC/MP levels can be used as a good indicator for predicting the prognosis of multiple myeloma patients.The appearance of LCE indicates disease progression, poor prognosis and early relapse.

Value of Red Blood Cell Distribution Width and Fibrinogen Level for Evaluation of the Therapeutic Efficacy and Prognosis in Patients with Diffuse Large B-Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the value of red blood cell distribution width (RDW) and fibrinogen (Fib) level for the evaluation of therapeutic efficacy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL).@*METHODS@#The relationship between RDW/Fib at initial diagnosis and efficacy and the clinical outcome was retro-spectively analyzed based on the study of 105 patients with DLBCL. The patients were divided into two groups: low RDW group (≤15%) and high RDW group (>15%), low Fib group (Fib≤4 g/L) and high Fib group (Fib>4 g/L) according to the normal values of RDW and Fib. Therapeutic efficacy, overall survival (OS) time and progression free survival (PFS) time were compared between two groups. The correlation between each factors and efficacy, prognosis was analyzed by univariate and multivariate regression.@*RESULTS@#The therapeutic efficacy (P<0.001), OS time(P=0.004), and PFS time(P=0.007) were poorer in the high RDW group as compared with the low RDW group. The efficacy (P=0.015) and PFS time(P=0.04) were poorer in the high Fib group as compared with the low Fib group. Multivariate analysis showed that high RDW was the independent risk factor for efficacy of DLBCL patients (OR=3.394, 95% CI 1.093-10.539, P=0.035).@*CONCLUSION@#High RDW and high Fib associate with poor efficacy in DLBCL patients.

Prognostic Value of Immune Status Changes after Chemotherapy in Patients with Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the prognostic significance of immune changes in patients with newly diagnosed multiple myeloma(MM) after chemothrapy.@*METHODS@#The clinical data of 99 patients with multiple myeloma received treatment in Department of Hematology, Lanzhou University Second Hospital from April 2011 to December 2017 were collected and retrospectively analyzed. The change of immune status was defined by changes of lymphocyte/monocyte ratio(LMR) level. The prognosis value of age, sex, typing, hemoglobin (Hb), β2-microglobulin (β2-MG), lactate dehydrogenase (LDH), albumin (albumin, ALB) and LMR changes were investigated in patients with newly diagnosed MM, and the relationship between above inentioned factors and changes of LMR was also explored. Overall survival rate between different subgroups was compared by using Kaplan-Meier curves and detected by Log-rank tests. Univariate and multivariate analysis of prognosis was performed by using the COX proportional hazards regression model. Paired samples Wilcoxon test were used to compare changes in ALC, AMC and LMR before and after chemotherapy, and logistic regression was used to investigate the clinical factors that affect the changes of LMR.@*RESULTS@#The median value of ALC increased from 1.25 (0.84-1.81)×10/L to 1.39 (1.02-1.9)×10/L (P=0.029) after treated for 1 month; the median value of AMC decreased from 0.37 (0.23-0.47) ×10/L to 0.29 (0.2-0.44)×10/L (P=0.026), and the median value of LMR increased from 3.552 (2.405-5.208) to 5.138 (3.22-6.471) (P=0.002). Multivariate survival analysis showed that increasing of LMR (HR 0.459, 95% CI 0.241-0.875, P=0.018) and LDH (HR 2.368, 95% CI 1.123-4.995, P=0.024) were considered to be the independent factors affecting the prognosis of MM patients.@*CONCLUSION@#The increasing of LMR level after treatment indicates a longer survival time of newly prognostic MM patients. Combination with LMR can not only reflect the effect of treatment on the immune status, but also predict the prognosis of MM patients much better.

Effect of plasmacytoid dendritic cells activited by bacteria on spontaneous remission of leukemia / 中国实验血液学杂志

Spontaneous remission (SR) of leukemia is a rare event in clinic, which possibly correlated with severe infection and sepsis, but its exact mechanism has not been confirmed. Plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) play a key role in innate and adaptive immunity respectively. A patient with severe infection of staphylococcus aureus acquired completely spontaneous remission (SR), moreover a increased number of pDC were observed, suggesting that bacteria-activated pDC may play an important role in SR. This study was purposed to explore if the bacteria can stimulate pDC successfully and get a functional pDC. Both pDC and mDC were isolated from freshly collected, leukocyte-rich buffy coats from healthy blood donor and leukemic patient with SR by using MACS and FACS. The pDC were cultured in RPMI 1640 medium and were stimulated with different kinds of bacteria and the expression of CD40, CD86 and HLA-DR on the cell surface was analyzed by flow cytometry. The cytokine (IFN-α, IL-12, IFN-γ, IL-2, IL-4, IL-10) production was measured by using ELISA kits. The results showed that the stimulation with staphylococcus aureus and pseudomonas aeruginosa resulted in the maturation of pDC, which secrete a large number of IFN-α and promote the differentiation of naive CD4⁺ T cells to Th1 cells. The activated pDC expressed high level of CD40 and CD86 and showed higher T cell stimulatory capacities. It is concluded that staphylococcus aureus and pseudomonas aeruginosa can activate pDC, the activated pDC secrete high quantity of IFN-α. This result suggests that bacteria stimulated pDC may play a key role in SR of leukemia following severe infections.

Immuno-effect of plasmacytoid dendritic cells on bacteria infection induced spontaneous remission of leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the immuno-effect of plasmacytoid dendritic cells (pDC) on bacteria infection induced spontaneous remission (SR) of leukemia.</p><p><b>METHODS</b>Both pDC and myeloid dendritic cells (mDC) were isolated and purified from leukemic patient with SR and healthy donor by combination of immunomagnetic beads and flow cytometry. pDC were cultured in RPMI1640 medium and stimulated with different bacteria. The T cells proliferation was detected by MTT, and cytokine production by ELISA kits.</p><p><b>RESULTS</b>The human bacterial pathogen Staphylococcus aureus and Pseudomonas aeruginosa stimulation for 48 h resulted in the maturation of pDC with production of high quantity of IFN-α at (15.34 ± 2.91) ng/ml and (10.38 ± 1.41) ng/ml, respectively, comparing with that of negative group at (1.36 ± 0.13) ng/ml (P<0.01). Activated pDC could promote the differentiation of naive CD4⁺ T cells to Th1 cells with secretion of IFN-γ at (2.16 ± 0.37) ng/ml and (2.73 ± 1.11) ng/ml, respectively, comparing with that of positive control at (2.55 ± 0.23) ng/ml (P > 0.05). Activated pDC showed higher T cell stimulatory capacities [proliferation index (PI) was 4.36 and 4.05, respectively] than that of non-activated pDC (PI was 1.23 and 0.13, respectively) (P < 0.01).</p><p><b>CONCLUSION</b>Staphylococcus aureus and Pseudomonas aeruginosa activated pDC may play a key role in SR of leukemia following severe infections.</p>

Effect of high dose dexamethasone on function and TLR-9 mRNA expression of plasmacytoid dendritic cells in patients with immune thrombocytopenic purpura / 中国实验血液学杂志

This study was purposed to investigate the effect of high-dose dexamethasone (DXM) on function and Toll like receptor 9 (TLR-9) expression of plasmacytoid dendritic cells (pDC) in peripheral blood of patients with immune thrombocytopenic purpura (ITP). 15 newly diagnosed patients with ITP received high dose DXM at single daily doses of 40 mg for 4 consecutive days. The peripheral blood plasmacytoid dendritic cells from 13 remission patients and 15 normal controls were separated by immunomagnetic beads and then induced by CpG-OND2216. 24 h later, the levels of IFN-α, IL-6 and TNF-α in the supernatant were detected by enzyme linked immunosorbent assay (ELISA). The expression of TLR9 mRNA of pDC was detected by real-time quantitative PCR. The results indicated that the levels of IFN-α, IL-6 and TNF-α produced by pDC in ITP patients were significantly higher than those in normal controls (P < 0.05). After high dose DXM treatment, the levels of IFN-α, IL-6 and TNF-α decreased without significant difference compared with normal controls (P > 0.05). The expression of TLR9 mRNA in pDC of untreated patients was significantly higher than that in control group (P < 0.05), and significantly reduced after treatment without difference from that in control group (P > 0.05). It is concluded that pDC may play an important role in ITP by their TLR9 and secreted cytokines; dexamethasone may down regulate the expression of TLR9, inhibit pDC function, and thus play a therapeutic role.

The prevalence of Th17 cells in patients with acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the proportion of Th17 cells in the peripheral blood of the patients with acute myeloid leukemia (AML) and evaluate the potential association of Th17 cells with AML.</p><p><b>METHODS</b>The cytokines IL-17 and TGF-β1 in the peripheral blood of AML patients before therapy (group 1), AML patients in complete remission (AML-CR, group 2) and healthy donors (group 3) were measured by enzyme-linked immunosorbent assay (ELISA). The proportion of Th17 cells of each group was evaluated by flow cytometry. The level of IL-17 mRNA of each group was examined by reverse transcription-PCR (RT-PCR).</p><p><b>RESULTS</b>The percentage of Th17 cells and the level of IL-17, IL-17 mRNA in group 1 \[(10.502 ± 1.071) ng/L, (0.935 ± 0.140)% and 0.262 ± 0.510\] and group 2 \[(11.345 ± 0.987) ng/L, (1.091 ± 0.159)% and 0.307 ± 0.031\] was significantly lower than that in group 3 \[(16.852 ± 1.198) ng/L, (2.586 ± 0.235)% and 0.501 ± 0.060\]. The percentage of Th17 cells and the level of IL-17, IL-17 mRNA in group 1 was lower than that in the group 2. But the level of TGF-β1 in the group 1 (29.963 ± 1.588) ng/L and the group 2 (25.163 ± 1.848) ng/L was significantly higher than that in group 3 (13.366 ± 1.565) ng/L. However, the level of TGF-β1 in the group 3 was higher than that of the group 2.</p><p><b>CONCLUSION</b>Th17 cells might be negatively correlated with the AML development. The overexpression of TGF-β1 in AML patients might suppress the differentiation of Th17 cells.</p>

Clinical characteristic of primary myelofibrosis with positive JAK2 V617F terminating in acute lymphoblastic leukemia / 白血病·淋巴瘤

Objective To report clinical and laboratory features of a rare case who was primary myelofibrosis (PMF) with positive JAK2 V617F terminating in T-acute lymphoblastic leukemia (T-ALL).Methods A rare case of PMF terminating in ALL was found, the quantity and quality of JAK2 gene mutation were defined by PCR methods, collected the clinical dates. Results The JAK2 V617F gene mutations were all positive pre and post-blast, the mutant rates respectively were 71.21% and 83.56 %. The interval from PMF to leukemia was 22 months, and from leukemia to death was 27 days. Conclusion PMF with positive JAK2 V617F terminating in ALL is a rare case, clinical prognosis is poor and survival time is short.

Killing activity of co-cultured cytokine-induced killer cells and dendritic cells against multi-drug resistant tumor cell lines / 中华肿瘤杂志

<p><b>OBJECTIVE</b>A lot of studies have suggested that a certain amount of T cells may be involved among cytokine-induced killer (CIK) cells. The aim of the present study was to prove whether an antigen-specific killing effect on tumor cells is involved during the CIKs-induced killing process.</p><p><b>METHODS</b>Bone marrow mononuclear cells (BMMNCs) derived from healthy subjects were separately cultured to generate dendritic cells (DC) and CIKs. A human mammary cancer cell line MCF-7/ADR, expressing P-gp antigen, was frozen-thawed and the lysate including P-gp antigen was obtained. The DC pulsed with or without tumor antigen lysate was co-cultured with CIK (pulsed-DC + CIK and DC + CIK), and CIK cultured alone was used as control. The cell phenotype of DC and CIK was analyzed by flow cytometry. The secretion of IL-12 and IFN-gamma was assayed by ELSA. The antitumor effect of the three CIK groups targeted at MCF-7/ADR cells expressing P-gp antigen and MCF-7 cells was detected by MTT.</p><p><b>RESULTS</b>Pulsed-DC + CIK group and DC + CIK group showed a higher expression level of DC mature phenotypes than those before co-culture with CIK, with a significant difference (P = 0.003, P = 0.001, respectively). The phenotypes (CD3, CD8, CD56) of CIK in pulsed-DC + CIK group and DC + CIK group was higher than those in CIK group (P = 0.003, P = 0.011, respectively). Among the three CIK groups, pulsed-DC + CIK group had the highest phenotypes on CD3+ CD56 (pulsed-DC + CIK vs. DC + CIK, P = 0.001; pulsed-DC + CIK vs. CIK, P < 0.001) and CD3 CD8 (P = 0.002, P = 0.002, respectively). Among the three groups, the pulsed-DC + CIK group showed the lowest CD45RA phenotype (pulsed-DC + CIK vs. DC + CIK, P < 0.001; pulsed-DC + CIK vs. CIK, P = 0.004). Among the three groups the secretion of IL-12 and IFN-gamma had the highest level in pulsed-DC + CIK group, with a value of 254 +/- 14.5 pg/ml and 3100 +/- 286 pg/ml, respectively. The antitumor killing effect on MCF-7/ADR cells had a significant difference between any two groups (pulsed-DC + CIK VS. DC + CIK, P = 0.039; pulsed-DC + CIK VS. CIK, P = 0.002; DC + CIK vs. CIK, P = 0.049). The highest was in pulsed-DC + CIK group and the lowest was in CIK group. The CIK group showed a significantly lower antitumor effect on MCF-7 cells than the other two groups (pulsed-DC + CIK vs. CIK, P = 0.007; DC + CIK vs. CIK, P = 0.048), but no significant difference between the pulsed-DC + CIK and DC + CIK groups.</p><p><b>CONCLUSION</b>In the present study, DC and CIK cells have been successfully obtained and cultured from bone marrow mononuclear cells. After their co-culture, not only both their specific phenotypes were increased, but also the associated cytokines were secreted. An improved antitumor killing effect and some possible specific immunocytotoxicity were observed. Our findings provided a basis for experimental and clinical research on bio-immunotherapy targeted at multi-drug resistant tumor cells.</p>

In vitro cytokines-induced differentiation in mononuclear cell derived dendritic cells from chronic myeloid leukemia / 中国实验血液学杂志

The purpose of this study was to investigate the mechanism of effects of interferon-alpha (IFN-alpha) on chronic myeloid leukemia (CML). Bone marrow mononuclear cells (BMMNC) were obtained from heparinized blood of CML patients by Ficoll-Paque density gradient centrifugation. The expressions of CD1a, CD83, CD86, HLA-ABC, HLA-DR and CD54 on DC induced by IFN-alpha + GM-CSF, IFN-alpha + GM-CSF+IL-4 and IL-4 + GM-CSF for 7 days in vitro were assayed by flow cytometry. The morphologic features were observed by transmission and optical microscopy. The mixed lymphocyte reactions (MLR) with DC were evaluated by MTT assay. The results showed that the DC cultured in different cytokine combinations expressed significantly higher levels of CD1a, HLA-ABC, HLA-DR, CD86, CD54, and CD83 than those in the precultured. The DC growing with IFN-alpha + GM-CSF expressed significantly higher levels of HLA-ABC, HLA-DR than those in GM-CSF + IL-4. The CD86 expression and MLR levels in IFN-alpha + GM-CSF + IL-4 increased significantly. The expression rate of DC antigens and MLR in the IFN resistant group significantly lower than those in the newly diagnosed and the effectively treated groups after at least 6 months of IFN-alpha treatment (P < 0.05). The DC from the IFN resistant group did not express significantly CD86 and MLR in IFN-alpha + GM-CSF + IL-4 groups compared to those in the newly diagnosed and IFN effective treated groups. It is concluded that the BMMNC from CML cultured in combination with IFN-alpha and other cytokines can be induced into DC with typical morphologic and immunophenotypic characteristics. Addition of IFN-alpha + GM-CSF + IL-4 to DC cultures can significantly up-regulate the expression of major histocompatibility complex molecules, co-stimulatory molecules and various adhesion molecules. The deficiency of DC differentiation and function may play a role in the development of clinical resistance to IFN-alpha.

Induction of anti-leukemic immunity of dendritic cells derived from multidrug resistant leukemia K562/A02 cells with high expression of P-glycoprotein and sensitive K562 cells / 中国实验血液学杂志

This study was aimed to investigate and compare the anti-leukemic effect mediated by dendritic cells (DC) derived from multidrug resistant (MDR) leukemia K562/A02 cells with high expression of p-glycoprotein (P-gp) and sensitive K562 cells. Multidrug resistant K562/A02 cell line and sensitive K562 cell line from chronic myeloid leukemia (CML) were induced for differentiating to DC in complete RPMI 1640 culture medium supplemented with GM-CSF (1 000 U/ml), IL-4 (500 U/ml) and TNF-alpha (100 ng/ml) for 14 days. The morphologic features of DC were observed by means of optical microscopy and the phenotype of DC was detected by flow cytometry. T-cell stimulating activity was determined by allogeneic lymphocyte reaction (allo-MLR). Cytotoxic activity was measured by MTT assay. The results indicated that DC derived from K562/A02 cells and K562 cells similarly showed the typical morphology of dendritic cell and expressed the surface differentiation antigens and costimulatory molecules CD1a, CD83, HLA-DR, CD80 and CD86 of DC. In allo-MLR, K562/A02-DC had a higher capacity to induce lymphocyte proliferation, compared with K562-DC (P < 0.05). K562/A02-DC and K562-DC could similarly generate specific cytotoxic activity against K562/A02 cells or K562 cells respectively, but low reactivity against HL-60 cells. More importantly, the cytotoxic activity mediated by K562/A02-DC was stronger than that by K562-DC against K562/A02 cells or HL-60/VCR cells (P < 0.01, respectively). It is concluded that functional DC can be differentiated from multidrug resistant leukemia K562/A02 cells as well as sensitive K562 cells in the presence of GM-CSF, IL-4 and TNF-alpha. Especially, DC derived from K562/A02 cells can induced a p-glycoprotein specific anti-leukemic immunity.

Study on karyotype of 306 cases of myelodysplastic syndrome / 中国实验血液学杂志

The purpose of this study was to explore the significance of abnormal karyotype in diagnosis and prognosis estimation of myelodysplastic syndrome (MDS). Chromosome analysis were performed in 306 cases of MDS using the short-term culture of bone marrow cell and G-banding technique, and in partial cases FISH technique was used for this analysis. 93 out of 306 cases were followed up. The results showed that 144 cases (47.1%) had clonal chromosome aberrations. The most common chromosomal aberrations included +8, translocation, complex or high complex karyotype, -7/7q-, 20q-/-20, trisomy 1 or partial trisomy 1, +11/+11q-, -9/9q-, +9/9q+, -Y, dup(1q), +21. The rate of abnormal karyotype in refractory anemia with erythroblasts (RAEB) and refractory anemia with erythroblasts-transformation (RAEBT) were much higher than in refractory anemia (RA) and refractory anemia with sideroblasts (RAS) (P < 0.05). The rate of abnormal karyotype among those cases with mutagen contact history were higher than those in cases without mutagen contact history. The patients with abnormal karyotype had a mean survival time much shorter than patients with normal karyotype (P < 0.005) and had a higher risk transforming into acute leukemia (P < 0.05). The worst outcome was observed in those patients with a complex or high complex karyotype, -7/7q- and trisomy 11. In conclusion, MDS is highly heterogeneous disorders and karyotype analysis is helpful for its diagnosis, treatment selection and prognosis estimation.