IL3-IGH fusion gene-positive pediatric acute lymphoblastic leukemia with hypereosinophilia as the first presentation: report of 1 case and review of literature / 白血病·淋巴瘤

Objective:To investigate the clinical characteristics and prognosis of IL3-IGH fusion gene-positive pediatric acute lymphoblastic leukemia (ALL) with hypereosinophilia as the first presentation.Methods:The clinical data of 1 pediatric IL3-IGH fusion gene-positive ALL patient with hypereosinophilia as the first presentation in January 2021 in Fujian Medical University Union Hospital was retrospectively analyzed and relevant literature was reviewed.Results:This 11-year-old male patient underwent bone marrow examination, and results showed that the proportion of eosinophils was increased; immunophenotyping disclosed that there were about 49.4% abnormal naive B lymphocytes in bone marrow; 43 leukemia fusion genes showed all negative; the whole transcriptome sequencing showed IL3-IGH fusion gene-positive. The patient was finally diagnosed as B-ALL with IL3-IGH fusion gene. According to the Chinese Children Cancer Group (CCCG)-ALL 2020 regimen, eosinophils returned to normal after induction therapy. Bone marrow examination on day 19 of induction showed that the proportion of promyelocytes was 0.005, the proportion of eosinophils was 0.05, and the minimal residual disease (MRD) was 23.02%. Bone marrow examination on day 46 of induction showed remission, and MRD was 0.18%. Consolidation chemotherapy used CAT (cyclophosphamide 1 g/m 2 once; cytarabine 50 mg/m 2, 12 h once, 7 days in total; mercaptopurine 40 mg/m 2, once per night, 7 days in total) regimen. Then the patient was added with lusotinib (75 mg 12 h once) orally and continued to receive high-dose methotrexate (5 g/m 2) regimen chemotherapy for 2 courses, the MRD was 0.20%. Chimeric antigen receptor T-cell (CAR-T) regimen was administered, followed by negative MRD. Conclusions:IL3-IGH fusion gene ALL is more frequently found in males, and more common in older children and young adults. It is prone to organ infiltration damage, and it has a high rate of induction failure and recurrence as well as poor prognosis.

Avaliação da interleucina 3 como marcador prognóstico na sepse / Role of interleukin-3 as a prognostic marker in septic patients

RESUMO Objetivo: Avaliar a acurácia dos níveis de interleucina 3 para predizer prognóstico em pacientes sépticos. Métodos: Conduzimos uma coorte prospectiva que incluiu pacientes adultos internados em unidade de terapia intensiva, que apresentassem sepse ou choque séptico iniciados há até 48 horas. Mediram-se os níveis séricos de interleucina 3 quando da inclusão (dia 1) e nos dias 3 e 7. O desfecho primário analisado foi a mortalidade hospitalar por qualquer causa. Resultados: Foram incluídos 120 pacientes. Os níveis séricos de interleucina 3 dosados à inclusão foram significativamente mais elevados em pacientes que faleceram em comparação aos que sobreviveram à internação hospitalar (91,2pg/mL versus 36pg/mL; p = 0,024). Em modelo de sobrevivência de Cox com inclusão de idade e valores sequenciais de SOFA, os níveis de interleucina 3 mensurados na inclusão mantiveram-se independentemente associados à mortalidade hospitalar (HR 1,032; IC95% 1,010 - 1,055; p = 0,005). Em curva Característica de Operação do Receptor construída para investigação adicional da acurácia da interleucina 3 na predição do prognóstico, encontrou-se área sob a curva de 0,62 (IC95% 0,51 - 0,73; p = 0,024) para mortalidade hospitalar. Valores iniciais de interleucina 3 acima de 127,5pg/mL mostraram-se significativamente associados à mortalidade hospitalar (p = 0,019; OR = 2,97; IC95% 1,27 - 6,97; p = 0,019), entretanto com baixo desempenho (especificidade de 82%, sensibilidade de 39%, valor preditivo positivo de 53%, valor preditivo negativo de 72%, razão de verossimilhança negativa de 0,73 e razão de verossimilhança positiva de 2,16). Conclusão: Níveis elevados de interleucina 3 mostraram-se independentemente associados à mortalidade hospitalar em pacientes sépticos, entretanto com baixo desempenho clínico.

ABSTRACT Objective: To evaluate the accuracy of IL-3 to predict the outcome of septic patients. Methods: Prospective cohort study with adult patients in an intensive care unit with sepsis or septic shock diagnosed within the previous 48 hours. Circulating IL-3 levels were measured upon inclusion (day 1) and on days 3 and 7. The primary outcome was hospital mortality. Results: One hundred and twenty patients were included. Serum levels of IL-3 on day 1 were significantly higher among patients who died than among patients who survived the hospital stay (91.2pg/mL versus 36pg/mL, p = 0.024). In a Cox survival model considering the IL-3 levels at inclusion, age and sequential SOFA, IL-3 values remained independently associated with mortality (HR 1.032; 95%CI 1.010 - 1.055; p = 0.005). An receiver operating characteristic curve was built to further investigate the accuracy of IL-3, with an area under the curve of 0.62 (95%CI 0.51 - 0.73; p = 0.024) for hospital mortality. A cutoff initial IL-3 value above 127.5pg/mL was associated with hospital mortality (OR 2.97; 95%CI: 1.27 - 6.97; p = 0.0019) but with a low performance (82% for specificity, 39% for sensibility, 53% for the positive predictive value, 72% for the negative predictive value, 0.73 for the negative likelihood and 2.16 for the positive likelihood ratio). Conclusion: Higher levels of IL-3 are shown to be independently associated with hospital mortality in septic patients but with poor clinical performance.

Cultivation and identification of mouse peritoneal and bone marrow-derived mast cells / 中华皮肤科杂志

Objective To explore the in vitro culture methods for oriented differentiation of peritoneal cells and bone marrow cells into high-purity mast cells,and to identify the function of these mast cells.Methods Peritoneal cells and bone marrow cells were isolated from the peritoneal cavity lavages and femur of C57BL/6 mice,and cultured with both interleukin-3 (IL-3) and stem cell factor for 2 and 4 weeks respectively.Light microscopy was performed to observe the morphology of these cells,toluidine blue staining to identify the degree of maturity of these mast cells,and flow cytometry to measure the expression of cell surface markers C D 117 and FceR Ⅰ α.After the stimulation with compound 48/80 at different concentrations,the degranulation rate of mast cells was counted under the microscope,and β-hexosaminidase release rate was measured by spectrophotometry.Results After 2-or 4-week culture,the mouse peritoneal and bone marrow cells all manifested as refractive suspension cells of uniform size.Toluidine blue staining showed violaceous metachromatic granules in the cytoplasm of the two kinds of cells.The proportions of CD117 or FcεR Ⅰ α single-positive peritoneal and bone marrow-derived mast cells were all more than 95％,and the proportions of CD117/FcεR Ⅰ α double-positive peritoneal and bone marrow-derived mast cells were 97.68％ ± 0.80％ and 96.12％ ± 0.76％ respectively.The degranulation rates of mast cells in the 100-and 1 000-mg/L compound 48/80 groups significantly differed from those in the blank control group (all P ＜ 0.01).Compared with the blank control group,the β-hexosaminidase release rates significantly increased in bone marrow-derived mast cells in the 100-mg/L compound 48/80 group and peritoneal mast cells in the 10-and 100-mg/L compound 48/80 groups (P ＜ 0.01 or 0.05).Conclusion IL-3 and stem cell factor can co-induce the directed differentiation and proliferation of mouse bone marrow stem cells and peritoneal cells,so as to harvest highnuritv mature degranulated mast cells,and lay a foundation for subsequent cell biology research.

Quantification of CD123+ BDCA-2+ plasmacytoid dendritic cells in skin lesions and peripheral blood of patients with psoriasis vulgaris / 中华皮肤科杂志

[Objective] To quantify CD123+ BDCA-2+ plasmacywid dendritic cells (PDCs) in skin lesions and peripheral blood of patients with psoriasis vulgaris,and to investigate their significance.[Methods] Skin tissue samples were resected from the lesions of 22 patients with psoriasis vulgaris and normal skin of 15healthy controls,and peripheral blood samples were also obtained from these subjects.Immunohistochemical technique (SP) was used to quantify PDCs in skin tissue samples,and flow cytometry to determine the proportion of PDCs in peripheral blood samples.[Results] Immunohistochemical study showed that the density of PDCs in the psoriatic lesions was significantly higher than that in the healthy controls (( 10.1 ± 2.1 )/mm2 vs.(0.4 + 0.6)/mm2,t =17.34,P ＜ 0.01 ).On the contrast,the proportion of PDCs in peripheral blood was 0.17％ ±0.07％ in the patients with psoriasis vulgaris,significantly lower than that in the healthy controls (0.33％ ±0.20％,t =4.48,P ＜ 0.01 ).[Conclusion]s The proportion of PDCs is reduced in the peripheral blood of patients with psoriasis vulgaris,which may be related to the initiation and progression of psoriasis vulgaris.

Preparation of a toxicity fusion protein targeting to CD123 on leukemia stem cell / 白血病·淋巴瘤

Objective To construct a fusion protein that used for treatment of resistance and palindromia in leukemia and studied its biological activity. Methods IL-3 and LP gene fragments were amplified by PCR. After enzymatic digestion and T4 ligation, the fusion gene was cloned into expression vector pAYZ. The product was purified by exchange chromatography and anti-Etag affinity chromatography. IL3-G4SLP fusion protein was analyzed by SDS-PAGE and Western blot. Protein biological activity was detected by FACS. Results The fusion protein was expressed as soluble protein by E.Coli 16C9. The protein expression level was about 1 mg/L, its purity was over 95 %, and the expression level was about 1 mg/L. The fusion protein can combined specificely with CD123 on leukemia stem cells. Conclusion Fusion protein IL-3-G4S-LP can target on leukemia stem cells and maybe as a potential drug used for treatment of resistance and palindromia in leukemia.

Relationship study between IL-3,IL-5 and IL-8 and Nasal polypus / 中国基层医药

Objective To explore the role of IL-3,IL-5 and IL-8 in the genesis and development of nasal polypus(NP) by testing IL-3 ,IL-5 and IL-8 Value in Nasal polypus issue. Methods 50 cases of nasal polyps and 30 casesof normal middle turbinate mucosa were involved in this study. Concentrations of IL-3, IL-5 and IL-8 were determined by enzyme linked immunosorbent assay(ELISA). Results The levels of IL-3 ,IL-5 and IL-8 in Nasal polypus issue were significantly higher than that in control group( P ＜ 0.05 ). Conclusion The levels of IL-3, IL-5 and IL-8 in Nasal polypus issue rised,They likely played a certain role in the genesis and development of NP.

Construction and prokaryotic expression of fusion gene IL3-PE38KDEL / 重庆医科大学学报

Objective:To construct a prokaryotic expression plasmid of PQE30-IL3-Linker-PE38KDEL and identify its recombinant protein expression.Methods:The IL3 and PE38KDEL gene were amplified by polymerase chain reaction(PCR) and cloned into the prokaryotic expression plasmid PQE30-Linker constructed after being sequenced.The recombinant vector confirmed by restriction endonucleases digestion,coenobium PCR,and DNA sequence analysis was transformed into E.coli SG13009.The expression of the protein was induced by IPTG.Relative molecular weight of the expression product was detected by SDS-PAGE.Finally,the fusion protein was examined by Western blot.Results:The results of restriction endonuclease digestion,coenobium PCR and DNA sequence analysis showed that the prokaryotic expression vector PQE30-IL3-Linker-PE38KDEL was constructed successfully.With induction of IPTG,the relative molecular weight of the expression product was identical to the expected value.The expressed 6?His-IL3-PE38KDEL fusion protein were identified at relative molecular mass of 57KD by Western blot with anti-His monoclonal antibody,showing the fusion protein expressed correctly.Conclusion:The fusion protein IL3-PE38KDEL is successfully constructed,which lays a solid foundation for the further research of protein purification and function.

Expression and activity detection of IL3-PE38KDEL / 重庆医科大学学报

Objective:To prepare a new type of anti-leukemia immunotoxin with killing activity.Methods:The method of cytotoxicity was used to study the activity of the immunotoxin after the induction of IPTG. Results:The expressed fusion proteins were detected mostly as inclusion bodies at high level.The result showed IL3-PE38KDEL had liable activity of toxicity. Conclusion:The fusion protein IL3-PE38KDEL has good biological activity,which paves way for the further study on its treatment of leukemia.

Effects of Intermittent Hypoxic Exposure on the Production of Erythrocyte and Some Haematopoietic Factor in Rats with Exercise-Induced Anemia / 中国运动医学杂志

0.05).Conclusion The hypoxic exposure can accelerate the production of erythrocyte, and treat the exercise-induced anemia effectively. Improvement of some haematopoietic factors and enhancement of hematopoiesis in marrow is thought to be the possible mechanism.

In Vitro Effect of Interleukin-11 (IL-11) on Megakaryopoiesis from Umbilical Cord Blood Cells

BACKGROUND: The megakaryopoiesis and platelet production is regulated by several hematopoietc factors such as thrombopoietin (TPO), interleukin-11 (IL-11) and interleukin-3 (IL-3). IL-11 is a potent stimulator of megakaryopoiesis in vivo, and acts primarily as a megakaryocyte maturation factor in vitro and it can act synergistically with IL-3 and TPO. We performed this study to investigate the effects of recombinant human IL-11 (rhIL-11) with other hematopoietic factors on megakaryocyte colony formation in vitro. METHODS: CD34+ cells were separated from umbilical cord blood and megakaryocyte colonies using MegaCult Assay Kit were cultured with rhIL-11, recombinant human IL-3 (rhIL-3), and recombinant human TPO (rhTPO) for 7 and 14 days. The number and percentage of CD34+ and CD41a+ cells were determined by flowcytometry. RESULTS: The number of CD41a+ cells were 0.54+/-0.05x10(4) (rhIL-11 100 ng/ml), 5.32+.-0.23x10(4) (rhIL-3 100 ng/ml), and 8.76+/-0.15x10(4) (rhTPO 50 ng/ml) of total expanded cells during the culture of the purified CD34+ cells in liquid phase for 7 days. The number of CD41a+ cells were increased to 7.47+/-0.69x10(4) (rhIL-3 rhIL-11), 11.92+/-0.19x10(4) (rhTPO rhIL-11) of total expanded cells, respectively, during the culture of the purified CD34+ cells in liquid phase for 7 days in the presence of rhIL-11 (100 ng/ml). When the purified CD34+ cells were cultured in semisolid media including various concentration of rhIL-11, the megakaryocyte colonies were not formed. When the purified CD34+ cells were cultured with rhIL-11 and rhTPO or with rhIL-11 and rhIL-3, the number of megakaryocyte colonies were increased compared with rhTPO or rhIL-3 alone. CONCLUSION: These results indicate that IL-11 exerts a potent proliferative activity to colony forming unit-megakaryocyte from human umbilical cord blood, and it acts with other hematopoietic factors synergistically

The expression and its clinical significances of interleukin-3 receptor alpha on Bone Marrow cells of the patients with acute myelocytic leukemia / 中国实用内科杂志

Objective To investigate the expression of interleukin-3 receptor alpha(CD123)on bone marrow cells in acute myelocytic leukemia(AML)and its clinical significances.Methods By means of Fluorescence-activated cell sorer(FACS)and semi-quantity reverse transcripition polymerase chain reaction(RT-PCR),the expression of IL-3R?(CD123+)protein on CD34+CD38-cells and mRNA in BMMNCs of 62 AML patients of Tianjin Medical University General Hospital from March 2008 to January 2009 and 12 normal controls were detected respectively;Then the correlation between IL-3R? and the clinical stages of AML were analyzed.Results CD34+CD38-CD123+/CD34+CD38-and IL-3R? mRNA in BMMNC of 33 deno-vo or relapsed AML patients were higher than those of control group(P

IL-3 Gene-Modified Bone Marrow Stromal Cells Accelerate Hematopoiesis Recovery of Mice Received High Dose Chemotherapy / 中国肿瘤生物治疗杂志

A main complication of chemotherapy in cancer patients is hematopoiesis suppression. Microenviroment transplantation using bone marrow stromal cells (BMSCs) has been demonstrated to be a potent method in recovery of hematopoiesis in animal models. Based on hematopoiesis-supportive ability of BMSCs and high potency of IL-3 in hematopoiesis stimulation, BMSCs were studied as a cellular delivery system for IL-3 gene transfection to promote hematopoiesis recovery of mice after high dose chemotherapy. BMSCs were transfected with recombinant adenovirous containing murine IL-3 gene(MOI = 10), the level of mIL-3 secreted by gene-modified BMSCs was 110U/ml/10~6 cells/ 24h in vitro. The mice were injected with high dose cyclophosphamide(200mg/kg) i.p. and after 24 hours the IL-3 gene-modified BMSCs(2 x 10~6/mouse) were transplanted intrasplenically. White blood cell counts in peripheral blood of mice received intrasplenic injection of IL-3-BMSCs were kept at a high level within two weeks after chemotherapy. The pathological sections of spleens and bone marrow showed significant recovery of hematopoiesis, compared with that of mice received chemotherapy only. The data indicated the feasibility of IL-3 gene-modified BMSCs transplantation in the acceleration of hematopoiesis recovery after chemotherapy.

Transfection of B16 Melanoma Cells with IL-3 Gene and Its in Vitro Proliferative Features / 中国肿瘤生物治疗杂志

IL-3 has effects on a wide variety of cell types, including immature cells of the immune cells, and mature immune cells such as granulocytes. In the purpose of studying the immunoregulatory function of IL-3 and its potential role in cancer therapy, we established a IL-3-secreting tumor model using gene transfection to deliver locally IL-3 to tumor site. First, we constructed IL-3 expression vector BMGNeo-mIL-3, then transfected it into B16 murine melanoma cells. By G418 resistant screening and limiting dilution, we obtained a transfectant that expressed high levels of IL-3 (806U / ml) . The IL-3 expression of the transfectant was confirmed by Northern blot analyses. Although the wild-type B16 cells and the B16-Neo cells transfected with BMGNeo did not express IL-3, the IL-3 expression in B16 cells had no obvious effect on the in vitro proliferation of the transfectant. These results showed we had successfully established a IL-3-secreting tumor cell clone which would enable us to further study its in vivo tumorigenicity and immune function.

Significance of the Changes of Interleukin-3,Interleukin-6 and Interleukin-8 Levels in Serum of Children with Acute Myeloid Leukemia / 实用儿科临床杂志

0.05).The IL-6 levels of children with M1 and M5 AML were remarkably higher than those of children with the other subtypes of AML(Pa

Experimental study on IL-3 gene expression induced by TSPG / 重庆医科大学学报

Objective:To explore the effect of TSPG (total saponins of panax ginseng) on gene expression of IL-3 which is one of the important hematopoietic growth factor at early stage of hematopoiesis.Methods:Techniques of RT-PCR and in situ hybridization were used.Results:A band corresponding to IL-3 was observed after RNA extraction from TSPG stimulated Jurkat cells were subjected to RT-PCR,while such band was not observed after RNA extraction from unstimulated Jurkat cells were subjected to RT-PCR.Comparing with the control group,both quantity and intensity of IL-3mRNA expression in fetus thymocytes and splenocytes induced by TSPG were obviously promoted.Conclusion:TSPG can induce human lymphocytes to express IL-3 mRNA,thus promote human hematopoiesis at early stage.

Siwutang enhancing production of IL-3 and IL-2 by splenocytes in Con A-primed mice / 第二军医大学学报

Objective:To investigate the effect of Siwutang on promoting hematopoietic function of mouse splenocytes. Methods:Effect of Siwutang on concanavalin A (Con A)-primed mice splenocytes production of IL-3 and IL-2 was studied by 3H-TdR incorporation and dot blot hybridization. Results: Siwutang significantly enhanced Con A-primed mice splenocytes production of IL-3 and IL-2(P

Construction of retroviral vector carrying human IL-3 cDNA under control of human AFP enhancer core sequence and SV40 early region promoter / 第二军医大学学报

To construct retroviral vector carrying human interleukin-3 c0mplementary DNA(HuIL-3 cDNA) under control of human a-fetoprotein gene enhancer core sequence and human SV4O pro-moter. Methods and Results: HuIL-3 cDNA was inserted into polylinker site of retroviral vector pMNSMto construct retr0viral vector pMNS-IL-3, in which the transcription of HuIL-3 cDNA was drived by SV40early region promoter. Human Q-fetoprotein gene enhancer core sequence was released from plasmidpGEM. 7Zf-AFPe and inserted into the polylinker site of pMNSM. Then human interleukin-3 cDNA wasinserted int0 p0lylinker site to construct retroviral vector pMNSA-IL-3, in which HuIL-3 cDNA transcrip-tion was drived by SV40 early region promoter and enhanced by human a-fetoprotein enhancer core se-quence- Conclusion: The vectors are of significance for hepatoma-specific gene therapy.

EXPERIMENTAL STUDY FOR THE EFFECT OF TSPG ON THE EXPRESSION OF IL-3 IN HEMATOPOIETIC STROMAL CELLS / 解剖学报

Objective Panax ginseng is a well-known Chinese traditional medicine functional in “invigorating qi”,and saponins are one of its main effective fractions.Our study investigates the effect of total saponins of panax ginseng(TSPG)on human hematopoiesis at early stage and its possible regulative mechanism to clarify the hematonic mechanism of Panax ginseng. Methods The techniques of culturing hematopoietic progenitor cells in vitro,bioassay of hematopoietic growth factor(HGF),immunocytochemistry and nucleic acid in situ hybridization were used to study the effect of TSPG on IL-3’s expression in hematopoietic stromal cell and its possible mechanism. Results TSPG directly added into culture system in vitro can markedly increase the colony forming yields of CFU-Mix;the different conditioned culture media prepared with TSPG can promote the proliferation and differentiation of CFU-Mix;the protein and mRNA expression of IL-3 in BMSC and cells of EcV304,THP induced by TSPG has been much intensified.Conclusion TSPG may regulate hematopoiesis at early stage by activating IL-3's gene expression in hematopoietic stromal cell,which is the most important element in hematopietic inductive microenvironment(HIM).