[The immunological effect of anti-leukemic tumor induced by eosinophilic granulocyte].

OBJECTIVE: To investigate the biological effect of anti-leukemic cells induced by eosinophilic granulocyte (EOS) in bone marrow of patients with chronic myelogenous leukemia (CML). METHODS: The BCR-ABL fusion gene as well as the expression of IL-12 and IL-17 mRNA were performed by RT-PCR. The serum concentrations of cytokine IL-12 and IL-17 were determined by enzyme-linked immuno sorbent assay (ELISA). Immunochemistry staining and cytochemistry staining were used to observe the peroxidase (POX) and human leukocyte antigen (HLA)-DR expression of EOS in bone marrow. Immunofluorescence staining was used to observe mannose receptor (MR), IL-12, IL-17A and IL-17 receptor A (IL-17RA) expression of EOS. The results between the CML patients and the healthy controls were compared. RESULT: Serum levels of IL-12 and IL-17 were higher in the 60 CML patients [(196.33 ± 21.79) ng/L and (36.55 ± 3.01) ng/L] than those in the controls [(96.60 ± 4.92) ng/L and (23.74 ± 1.36) ng/L]. In the 32 patients with activated EOS, the levels of IL-12 and IL-17 were (273.12 ± 17.16) ng/L and (40.11 ± 6.13) ng/L, which were significantly higher than those in the non-activated EOS [(126.16 ± 14.27) ng/L and (28.14 ± 5.29) ng/L] (P values < 0.01). IL-12 and IL-17 mRNA were expressed in activated EOS, while BCR-ABL fusion gene was not found. The amounts of EOS were increased abnormally in the bone marrow and peripheral blood of the CML patients with POX positive staining in the cytoplasm and weakly positive HLA-DR staining. It was observed easily by a microscope that EOS could attack leukemic cells in bone marrow through adhesion, capture and phagocytosis. Activated EOS could express IL-12, IL-17A and MR, which was related with the serum levels of these cytokines. CONCLUSIONS: Activated EOS in bone marrow of CML patients could express IL-12 and IL-17. Activated EOS could induce coup injury to leukemic cell by releasing POX and expressing IL-12 and IL-17. It can also capture or swallow target cells via the expression of MR on the membrane. EOS may play an important role in the anti-tumor immunologic function in bone marrow of CML patients.

[Pathological effects and clinical significance of eosinophils in autoimmune-related hematocytopenia].

OBJECTIVE: To observe the pathological effects induced by eosinophils (EOS) in the process of cellular damage in immune-related hematocytopenia (IRH) and elucidate the immunologic mechanism and clinical significance of EOS. METHODS: Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum concentrations of interleukins (IL)-2, IL-4, IL-5, IL-6, IL-12 and IL-17 in 117 IRH patients from February 2008 to February 2012 in our hospital. Their quantity, activity, peroxidase (POX) and HLA-DR expression of EOS were observed and analyzed. Immunofluorescent staining was used for detecting anti-human immunoglobulin (IgG) on the surface of hematopoietic cell and the expressions of intercellular adhesion molecule-1 (ICAM-1), Fcγ receptor II (FcγRII), mannose receptor (MR), IL-5 receptor (IL-5R), IL-12, IL-12 receptor (IL-12R), IL-17A and IL-17 A receptor (IL-17RA) on EOS. TdR incorporation method was employed to determine the capability of antigen presentation and IL-17 mRNA expression examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The serum levels of IL-4, IL-5, IL-6, IL-12 and IL-17 were (170.9 ± 14.7), (112.9 ± 8.2), (131.8 ± 13.8), (339.2 ± 26.1) and (42.5 ± 2.2) ng/L in patient group versus (60.3 ± 11.0), (34.1 ± 2.2), (91.0 ± 12.3), (94.0 ± 3.3) and (20.0 ± 1.1) ng/L in control group respectively. All incremental percentages of IL-5, IL-12 and IL-17 were 100% (117/117) and they were correlated with disease. Diastrophic EOS could be found in marrow of patient with decreased leucocyte with HLA-B27(+). Hematopoietic cells were adhered, captured and phagocytized actively by activated EOS. Activated EOS showed strongly positive POX while POX of neutrophils attacked by EOS became weakened. Activated EOS could express HLA-DR and played a role in antigen presentation. EOS could secrete cytokine IL-17 through a transcription of IL-17 mRNA. And EOS could also express ICAM-1, FcγRII, MR, IL-5R and IL-12, IL-17A, IL-17RA. CONCLUSIONS: Capable of expressing or secreting various cytokines and molecules, EOS have immunological functions of adhering, capturing and phagocytizing pathological hemocytes. It also participates in pathogenic process of IRH. Thus EOS is probably an important immune effector cell in the process of in situ marrow damage.

[Clinical significance of telomerase activity detection of myeloid element in immune related hematocytopenia].

OBJECTIVE: To observe the telomerase activity (TA) in bone marrow mononuclear cell (BMMNC) at pre-therapy and post-therapy in patients with immune related hematocytopenia and immune related pancytopenia (IRH/IRP) so as to explore the relationship between TA and disease process. METHODS: TA in BMMNC of 108 IRH/IRP patients was detected with telomeric repeat amplification protocol (TRAP)-PCR-ELISA. The expressions of anti-human immunoglobulin IgG, FcγII receptor (FcγIIR), mannose receptor (MR) and interleukin 17A (IL-17A) located on membrane surface were observed by immunofluorescence (IF). A total of 30 iron deficiency anemia patients were selected as case control. RESULTS: The TA of 108 IRH/IRP patients were 0.251 ± 0.021. And it increased as compared with the control group of 0.062 ± 0.031 at pre-therapy (P < 0.01). The HLA-B27(+) patients among them was 0.270 ± 0.020 while HLA-B27(-) patients 0.181 ± 0.013 (P < 0.05). And HLA-B27(+)IgG elevated IRP was 0.341 ± 0.016 and HLA-B27(-)IgG elevated 0.183 ± 0.010, the difference of TA was significant between two groups (P < 0.01). IgG antibody positive immature hemocyte could be observed in marrow of IRH/IRP patients, and FcγIIR was expressed highly on those activated macrophage (Mφ) in bone marrow. Both IL-17A and MR were expressed on activated dendritic cells and Mφ in marrow of HLA-B27(+) patients. After the therapy of glucocorticoids associated with cyclosporine A and other drugs, the patient's hemogram recovered and the telomerase in BMMNC were re-devitalized. CONCLUSIONS: The TA of the BMMNC in marrow of IRH/IRP patients is increased, and it shows a close relation between apoptotic degree of hematopoietic cell and the activation of immunocytes. In IRH/IRP, synchronism and equilibrium exist between BMMNC apoptosis and TA.

[Nitrogen Removal Characteristics and Analysis of Microbial Community Structure in an IEM-UF Simultaneous Separation and Denitrification System].

A system that combines an ion exchange membrane and ultrafiltration membrane (IEM-UF) to form a simultaneous separation and denitrification system was proposed for domestic sewage with a low carbon/nitrogen ratio. The removal of nitrogen and COD in the system was studied under a three phase operating condition. The characteristics of the microbial community in each reactor were analyzed using metagenomics. The results show that, the average rate of ammonia nitrogen enrichment in the separator reached above 116.1% when the current intensity was 0.2 A. When the system was at C/N 2.80 and operating well, the average removal rates of COD and TN reached above 90% and 50%, respectively. The maximum removal rate of TN was above 65.4%. The results of metagenomics showed a genus of phylum Nitrospirae (Nitrospira) and a genus of phylum Proteobacteria (Nitrosomonas), with the proportions of 12.23% and 2.31%, respectively. In the denitrifying reactor, Dechloromonas, Thauera, and Azospira were detected in the proportions 4.57%, 1.76%, and 1.03%, respectively. These proportions were far larger than those of other bacteria in this reactor. Meanwhile, the presence of iron autotrophic denitrifying bacteria increased the denitrification efficiency of the system.

[Analysis of Low C/N Wastewater Treatment and Structure by the CEM-UF Combined Membrane-Nitrification/Denitrification System].

In this study, a CEM-UF composite membrane with ammonia nitrogen enrichment and separation characteristics was combined with nitrification/denitrification to treat low C/N wastewater. The denitrification characteristics of low C/N wastewater at different flow ratios were investigated, and the structural characteristics of functional microbial communities in nitrifying and denitrifying activated sludge were analyzed by 16Sr DNA high-throughput sequencing. The results showed that influent TN was 60 mg·L-1, COD/TN was 2.65, the nitrification effect of each flow rate was good, and the average ammonia nitrogen removal rate was 98.7%. When the flow ratio increased from 1:2 to 1:6, the m(COD)/m(NO3--N) of denitrification was increased, and the removal of average nitrate nitrogen reached its highest level at 1:6, which was 86.28%, and the removal of total nitrogen increased from 22.56% to 46.8%. An analysis of Illumina sequencing showed that nitrogen fixing bacteria Proteobacteria accounted for 30.9%, and the important nitrite oxidizing bacteria, Nitrospirae, accounted for 3.06%. At the genus level, Nitrosomonas and Nitrosospira, belonging to the ammonia oxidizing bacteria (AOB) category and Nitrospira and Nitrobacter, belonging to the nitrite oxidizing bacteria (NOB) category were detected. The ratio of AOB and NOB bacteria was high, which is consistent with good nitrification in the nitrification reactor. The dominant bacteria in denitrification sludge were Proteobacteria (53.13%), followed by Bacteroidetes (10.93%). A variety of bacteria related to denitrification were detected at the genus level, such as Dechloromonas, Thauera, Castellaniella, Alicycliphilus, Azospira, Comamonas, Caldilinea, and Saccharibacteria. The proportion of denitrifying bacteria was 25.91% as denitrifying bacteria microbial species were rich in the denitrifying sludge, giving a good denitrification effect.