The immune injury effects and clinical significance of eosinophils in auto-immune-related hematocytopenia.

Eosinophils (EOS) quantity, active state, peroxidase activity (POX), and HLA-DR expression in bone marrow of 176 Auto-Immune-Related Hematocytopenia (AIRH) patients were analyzed. Immunofluorescent staining (IF) is performed to observe the expression of immunizing molecules on EOS. In serum of AIRH patients the levels of IL-4, IL-5, IL-6, IL-12, IL-17, and IFN- γ were increased but there was no significance on IL-2 level. In marrow of AIRH, activated EOS expressed POX, and other molecules, it played various cell-mediated immunity injury roles to hemocyte. EOS might be possessed with multiple immunological fuctions, it playes an important immune effect in AIRH autoimmune pathological processes.

[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.