Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production...Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.展开更多
Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to t...Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assayand Western blot. Cell growth kinetics analyses throughgrowth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth testwere performed to identify cells proliferation potential.Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormalappearance of M-CSF in nucleus could enhance cellproliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.展开更多
AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a ...AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a recipient of an organ graft with already impaired perfusion might cause further deterioration in the transplanted organ. This study evaluated whether G-CSF treatment worsens liver perfusion following liver transplantation in the rat model. METHODS： A non-arterialized rat liver transplantation model was employed to evaluate the effect of G-CSF treatment on the liver in a syngeneic and allogeneic strain combination. Study outcomes included survival time and liver damage as investigated by liver enzymes and liver histology. Observation times were 1 d, 1 wk and 12 wk. RESULTS： Rats treated with G-CSF had increased incidence and severity of biliary damage following liver transplantation. In these animals, hepatocellular necrosis was accentuated in the centrilobular region. These lesions are indicative of impaired perfusion in G-CSF treated animals. CONCLUSION： G-CSF should be used with caution in recipients of liver transplantation, as treatment might enhance preexisting, undetected perfusion problems and ultimately lead to ischemia induced biliary complications .展开更多
Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on e...Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on experience. The aim of this study is to evaluate the role of inflammatory markers in diagnosis of acute appendicitis. Material and Method: The study includes 77 cases with histopathologically proven acute appendicitis and 17 control cases. Blood samples were obtained from all cases and C-reactive protein (CRP), Granulocyte Colony Stimulating Factor (G-CSF) and Total Antioxidant Capacity (TAC) were measured. Findings: In cases with acute appendicitis, CRP and G-CSF levels were found to be related to acute appendicitis;however, TAC was not affected by the disease process. Moreover, CRP and G-CSF levels were correlated with the disease severity. Conclusion: Both CRP and G-CSF can be used in diagnosis of acute appendicitis. Furthermore, increased CRP level can be a marker to show advanced cases. However, G-CSF is not an effective marker to show disease severity.展开更多
Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal a...Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal antibody (McAb) of MAF-J6-1R RE2 and polyclonal antibody (PolyAb) of rhM-CSFR were prepared. The specificity of McAb RE2 to M-CSFR was confirmed by indirect ELISA, cross-neutralizing assay with J6-1 cell colony formation and neutralization test by ELISA. Results: the reactive activity of purified RE2 to M-CSFR was over 1: 16000. The inhibitory activity of M-CSFR and MAF-J6-1R could be blocked by RE2 and anti-M-CSFR antibody. The reactivity of RE2 to M-CSFR could be reduced by M-CSFR. Conclusion: The specificity of RE2 to M-CSFR was confirmed and the antigen association of MAF-J6-1R with M-CSFR was proved. It suggests that M-CSF and its receptor mediated auto-juxtacrine stimulation could be an operative mechanism in either leukemia or nonhematological malignancies.展开更多
The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This s...The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This study was undertaken to assess the effect ofgranulocyte-macrophage colony stimulating factor ( GM-CSF) on the repair of damaged iliac arteries.Twenty-four male New Zealand white rabbits undergoing primary iliac artery deendothelialization wererandomly divided into two groups ( GM-CSF group and control group) . The GM-CSF group received asubcutaneous injection of GM-CSF (10 μg ? kg^(-1) ? d^(-1) ) , and the control groupwas given a subcutaneous injection of equivalent saline. The iliac arteries of all animals weredamaged by balloon after 7 days. The levels of nitric oxide ( NO) were detected before, 1 week, 2weeks and 4 weeks after angioplasty. The repair and hyperplasia of the intima were observedmicroscopically and the indices of stenosis were evaluated by computerized planimetry after 4 weeksof angioplasty. The NO levels of the GM-CSF group were higher than those of the control group 2weeks and 4 weeks after angioplasty [91.92 +-11.57) μmol/L vs. (81. 67 +- 12. 18) μmol/L; (97. 67+- 10. 13 ) ( μmol/L vs. (83. 16 +-12. 64) μmol/L]. Four weeks after balloon damage,histological examination showed that neointima formation, vascular smooth muscle cells and fibroustissue of the GM-CSF group were less than those of the control group. The endothelium of the GM-CSFgroup was more integrated, and stenosis of lumen was slighter than that of the control group.Morphometry showed the lumen area of the GM-CSF group was larger than that of the control group[(1.27 +-0. 31) mm^2 vs. (0. 92 +- 0. 24) mm^2 ] , the neointimal area and percent of intimahyperplasia were significantly smaller than those of the control group [ (0. 85 +-0. 34) mm vs. (1.18 +-0. 38) mm^2; (40 +- 7)% vs. (55 +- 6)%]. GM-CSF could facilitate the repair of the intima,reduce neointima formation, better the function of the endothelium, and decrease the rate ofrestenosis.展开更多
Spinal cord injury represents a severe form of central nervous system trauma for which effective treatments remain limited.Microglia is the resident immune cells of the central nervous system,play a critical role in s...Spinal cord injury represents a severe form of central nervous system trauma for which effective treatments remain limited.Microglia is the resident immune cells of the central nervous system,play a critical role in spinal cord injury.Previous studies have shown that microglia can promote neuronal survival by phagocytosing dead cells and debris and by releasing neuroprotective and anti-inflammatory factors.However,excessive activation of microglia can lead to persistent inflammation and contribute to the formation of glial scars,which hinder axonal regeneration.Despite this,the precise role and mechanisms of microglia during the acute phase of spinal cord injury remain controversial and poorly understood.To elucidate the role of microglia in spinal cord injury,we employed the colony-stimulating factor 1 receptor inhibitor PLX5622 to deplete microglia.We observed that sustained depletion of microglia resulted in an expansion of the lesion area,downregulation of brain-derived neurotrophic factor,and impaired functional recovery after spinal cord injury.Next,we generated a transgenic mouse line with conditional overexpression of brain-derived neurotrophic factor specifically in microglia.We found that brain-derived neurotrophic factor overexpression in microglia increased angiogenesis and blood flow following spinal cord injury and facilitated the recovery of hindlimb motor function.Additionally,brain-derived neurotrophic factor overexpression in microglia reduced inflammation and neuronal apoptosis during the acute phase of spinal cord injury.Furthermore,through using specific transgenic mouse lines,TMEM119,and the colony-stimulating factor 1 receptor inhibitor PLX73086,we demonstrated that the neuroprotective effects were predominantly due to brain-derived neurotrophic factor overexpression in microglia rather than macrophages.In conclusion,our findings suggest the critical role of microglia in the formation of protective glial scars.Depleting microglia is detrimental to recovery of spinal cord injury,whereas targeting brain-derived neurotrophic factor overexpression in microglia represents a promising and novel therapeutic strategy to enhance motor function recovery in patients with spinal cord injury.展开更多
Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were tre...Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were treated with one course of CLAG from April 1st,2014 through December 9th,2015 in our hospital were retrospectively reviewed.Results Thirty-three展开更多
Radiotherapy(RT)serves as the primary treatment for solid tumors.Its potential to incite an immune response against tumors both locally and distally profoundly impacts clinical outcomes.However,RT may also promote the...Radiotherapy(RT)serves as the primary treatment for solid tumors.Its potential to incite an immune response against tumors both locally and distally profoundly impacts clinical outcomes.However,RT may also promote the accumulation of immunosuppressive cytokines and immunosuppressive cells,greatly impeding the activation of antitumor immune responses and substantially limiting the effectiveness of RT.Therefore,regulating post-RT immunosuppression to steer the immune milieu toward heightened activation potentially enhances RT's therapeutic potential.Cytokines,potent orchestrators of diverse cellular responses,play a pivotal role in regulating this immunosuppressive response.Identifying and promptly neutralizing early released immunosuppressive cytokines are a crucial development in augmenting RT's immunomodulatory effects.To this end,we conducted a screen of immunosuppressive cytokines following RT and identified macrophage colony-stimulating factor(MCSF)as an early up-regulated and persistent immune suppressor.Single-cell sequencing revealed that the main source of up-regulated McSF derived from tumor cells.Mechanistic exploration revealed that irradiation-dependent phosphorylation of the p65 protein facilitated its binding to the McSF gene promoter,enhancing transcription.Knockdown and chemical inhibitor experiments conclusively demonstrated that suppressing tumor cell-derived McSF amplifies RT's immune-activating effects,with optimal results achieved by early MCSF blockade after irradiation.Additionally,we validated that MCSF acted on macrophages,inducing the secretion of a large number of inhibitory cytokines.In summary,we propose a novel approach to enhance the immune activation effects of RT by blocking the MCSF-CSF1R signaling pathway early after irradiation.展开更多
To the Editor:After chimeric antigen receptor T(CAR-T)cell therapy,over 90%of patients experience either mild or severe cytopenia,with the prevalence of Grades 3–4 neutropenia ranging from 53%to 94.3%.[1,2]Granulocyt...To the Editor:After chimeric antigen receptor T(CAR-T)cell therapy,over 90%of patients experience either mild or severe cytopenia,with the prevalence of Grades 3–4 neutropenia ranging from 53%to 94.3%.[1,2]Granulocyte colony-stimulating factor(G-CSF),known for its ability to enhance the proliferation and differentiation of neutrophils,has been extensively employed for the prevention and treatment of neutropenia in various contexts.However,its application in patients with refractory/relapsing B-cell acute lymphoblastic leukemia(R/R B-ALL)after CAR-T cell therapy remains a topic of debate.[3]Several studies have suggested that G-CSF may exert anti-inflammatory effects by modulating the release of inflammatory factors,such as interleukin-12(IL-12),TNF-α,and IFN-γ,or by influencing the differentiation of T-cell subsets.[4]However,other researchers have posited that G-CSF could exacerbate cytokine release syndrome(CRS)and immune effector cell-associated neurotoxicity syndrome(ICANS)through the augmentation of antigen-presenting cell function.展开更多
Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Meth...Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Methods From November 1992 to March 1997, 45 patients wer e enrolled into our study and 15 were newly diagnosed All were treated with hi gh dose chemotherapy combined with rhGCSF Results Of 15 newly diagnosed patients, 13 achieved complete remission (CR) after one course of therapy and 2 achieved CR after two courses of therapy For newly diagnosed patients, the durations of absolute neutrophil counts (ANC ) <05109/L were 5 days and 10 days in rhGCSF group and control group res p ectively ( P <005) The incidences of infection of these two groups w ere 40% and 60% respectively ( P <005) As for patients who receive d intensive therapy, the durations of ANC <05109/L were 5 days and 8 days i n rhGCSF group and control group, respectively ( P <005), and the i ncidences of infection were 25% and 444% respectively ( P <005) Conclusions The application of rhGCSF in children with AML after chem otherapy may hasten the hematopoietic recovery The duration of neutropenia wa s shortened by 3-4 days, and the incidence of fatal infection was reduced rhG CSF does not stimulate AML growth in vivo展开更多
Severe alcoholic hepatitis(SAH)is associated with high short-term mortality.The SAH population exhibits extreme heterogeneity in disease severity,clinical presentation,decompensations,and outcomes.Nonetheless,improvin...Severe alcoholic hepatitis(SAH)is associated with high short-term mortality.The SAH population exhibits extreme heterogeneity in disease severity,clinical presentation,decompensations,and outcomes.Nonetheless,improving outcomes and preventing adverse events is a major challenge when selecting an appropriate treatment for alcoholic hepatitis.Currently,steroids are the standard of care for SAH with Maddrey’s discriminant function>32 and model for end stage liver disease>20;however,they have limited usage due to ineligibility in approx-imately two-third of such patients.Approximately 25%of patients do not respond to steroids and require alternative therapies.An array of evolving therapies,such as granulocyte colony-stimulating factors,plasma exchange,fecal microbiota transplantation,antibiotics,anti-cytokine therapies,and N-acetylcysteine,showing variable success,are emerging.Hence,it is also crucial to select appro-priate therapy.The present review discusses the standard of care,the existing therapies,risk stratification for outcomes,and the selection of appropriate therapy to improve survival in SAH patients.展开更多
Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B i...Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac) Methods A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 μg of GM CSF plus 10 μg of rHBvac injected intramuscularly at the same location (GM CSF group, 14 children) or 200 IU HBIG and 10 μg rHBvac in different muscles (HBIG group, 13 children) on a monthly four dose schedule HBV DNA quantification and other HBV serological markers were tested before and after the four dose therapy Results Twelve children in each group completed the study Of them, 3 children in the GM CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM CSF group and 10 in the HBIG group One from each group had an HBeAg/anti HBe seroconversion after the treatment The quantity of HBV DNA was significantly lower after the treatment ( P =0 023) in GM CSF group, but was not significantly reduced in HBIG group No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group Conclusion Combined GM CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis展开更多
Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acu...Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acute promyelocytic leukemia (APL) during the treatment with all trans retinoic acid (ATRA) Results The peak of increased serum G CSF level occurred on the 9th day, and WBC number was the highest on the 11th day After ATRA treatment, both serum G CSF level and WBC number increased in 68 1% of the cases In 19 2% of the cases treated, serum G CSF level was increased but without obvious change in WBC number, and the reverse was true in 12 7% of the cases Conclusion Serum G CSF level was statistically correlated to the number of WBC, promyelocytes and its late stage by Spearman's rank order correlation coefficient展开更多
The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated wit...The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.展开更多
Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cyt...Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cytokines that were rarely addressed during prion infection,the levels of 17 different cytokines in the brain homogenates of mice infected with different scrapie mouse-adapted agents were firstly screened with Luminex assay.Significant upregulations of interferon gamma-induced protein 10(IP10),keratinocyte chemoattractant(KC)and macrophage colony stimulating factor(M-CSF)were frequently detected in the brain lysates of many strains of scrapie infected mice.The upregulations of those three cytokines in the brains of scrapie infected mice were further validated by the individual specific ELISA and immunohistochemical assay.Increased specific mRNAs of IP10,M-CSF and KC in the brains of scrapie infected mice were also detected by the individual specific qRT-PCRs and IP10-specific digital PCR.Dynamic analyses of the brain samples collected at different time points post infection revealed the time-dependent increases of those three cytokines,particularly IP10 during the incubation period of scrapie infection.In addition,we also found that the levels of IP10 in cerebral spinalfluid(CSF)of 45 sporadic Creutzfeldt–Jakob disease(sCJD)patients were slightly but significantly higher than those of the cases who were excluded the diagnosis of prion diseases.These data give us a better understanding of inflammatory reaction during prion infection and progression of prion disease.展开更多
Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage col...Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor （GM-CSF） transfection in mice ex vivo. Methods： The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c＋ cells were cultured with cytokines. For adenoviral （Ad）-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of multiplicity of infection （MOI） to determine the optimal gene transfection conditions, and detecting the expression of GM-CSF after transfection. The variation of GM-CSF gene-modified DCs were analyzed by morphological observation, phenotype analysis, and mixed lymphocyte reaction （MLR）. DCs were loaded with gastric cancer antigen obtained by frozen and thawed method. The stimulated DCs vaccination induced T lymphocytes, and the killing effect of T cells to gastric cancer cells was assayed by MTT. INF-γ production was determined with the INF-γ ELISA kit. Results： B220–CD11c＋ cells numbers increased after CCL3 injection. ELISA results showed that after GM-CSF gene modification, DC could produce high level of GM-CSF. When DCs were transferred AdGM-CSF gene at MOI equal to 1：100, GM-CSF level in culture supernatants reached saturation [（130.00 ± 12.61） pg/mL]. After GM-CSF gene-modification, DCs tended to more maturated through morphological observation and were phenotypically identical to typical DC and gained the capacity to stimulate allogeneic T cells. T lymphocytes stimulated with DC transduced with GM-CSF gene showed the specific killing effect on gastric carcinoma cells and produced high level of INF-γ [（1245.00 ± 13.75） pg/mL]. Conclusion： CCL3-recruited DCs modified by adenovirus-transducted GM-CSF could produce high level of GM-CSF, which tended to more maturated, and the capacity of activating allogeneic T lymphocytes proliferation was enhanced greatly. Moreover, they could stimulate specific cytotoxic T lymphocyte （CTL） to gastric cancer ex vivo.展开更多
AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-C...AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-CSF administration on virological response. METHODS:Sixty-eight patients undergoing antiviral treatment for post-liver transplantation(OLT)HCV recurrence were enrolled.All patients developing neutropenia received G-CSF. RESULTS:Twenty three(34%)received G-CSF.Mean neutrophil count at the onset of neutropenia was 700/mmc(range 400-750/mmc);after 1 mo of G-CSF it increased to 1210/mmc(range 300-5590/mmc) (P<0.0001).Three patients did not respond to G-CSF. Treatment duration was similar in neutropenic and non-neutropenic patients.No differences in the rate of discontinuation,infections or virological response were observed between the two groups.G-CSF was protective for the onset of de novo autoimmune hepatitis(P<0.003). CONCLUSION:G-CSF administration is effective in the case of Peg-IFN induced neutropenia increasingneutrophil count,prolonging treatment and leading to sustained virological response(SVR)rates comparable to non-neutropenic patients.It prevents the occurrence of de novo autoimmune hepatitis.展开更多
Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1...Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1-GM-CSF plasmid which was controlled by the CMV promoter was transferred into CHO cell by lipofectamine, selected by G418 and the positive clones was got. The recombinant vector which was rejoined into the groups of DNA of CHO was identified by PCR. Results: The results showed that the protein of rhGM-CSF was about 28 KD by using ELISA, SDS-PAGE and Western blot. Conclusion: rhGM-CSF was expressed steadily and highly. The rhGM-CSF will be of more use value.展开更多
Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiat...Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiation.Methods DCs from peripheral blood mononuclear cells (PBMC) that maintain the antigen capturing and processing capacity charateristicof immature cells have been established in vitro, using granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then, in cholangiocarcinoma cells apoptosis was induced by J-irradiation. The experimental groups were as follows: (1) coculture ofDCs and apoptotic cancer cells and T cells; (2) coculture of DCs and necrotic cancer cells and T cells; (3) coculture of DCs, culturedcancer cell and T cells. They are cocultured for 7 days. DCs and T cells were riched, isolated and their antitumor response was tested.Results The cells had typical dendritic morphology, expressed high levels of GDI a and B7, acquired antigen from apoptotic cells causedby y-irradiation and induced an increased T cell stimulatory capacity in mixed lymphocyte reactions (MLR) .Conclusion DCs obtained from PBMCs using GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by y-irradiation and efficiently induce T cells. This strategy, therefore, may present an effective approach to transduce DCs with antigen.展开更多
文摘Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.
基金This work was supported by a grant from Tianjin Science and Technology Development Project (No. 003119311).
文摘Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assayand Western blot. Cell growth kinetics analyses throughgrowth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth testwere performed to identify cells proliferation potential.Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormalappearance of M-CSF in nucleus could enhance cellproliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.
基金Supported by the Deutsche Forschungsgemeinschaft (KFO 117/1) and the IFORES Research Program, University Hospital Essen
文摘AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a recipient of an organ graft with already impaired perfusion might cause further deterioration in the transplanted organ. This study evaluated whether G-CSF treatment worsens liver perfusion following liver transplantation in the rat model. METHODS： A non-arterialized rat liver transplantation model was employed to evaluate the effect of G-CSF treatment on the liver in a syngeneic and allogeneic strain combination. Study outcomes included survival time and liver damage as investigated by liver enzymes and liver histology. Observation times were 1 d, 1 wk and 12 wk. RESULTS： Rats treated with G-CSF had increased incidence and severity of biliary damage following liver transplantation. In these animals, hepatocellular necrosis was accentuated in the centrilobular region. These lesions are indicative of impaired perfusion in G-CSF treated animals. CONCLUSION： G-CSF should be used with caution in recipients of liver transplantation, as treatment might enhance preexisting, undetected perfusion problems and ultimately lead to ischemia induced biliary complications .
文摘Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on experience. The aim of this study is to evaluate the role of inflammatory markers in diagnosis of acute appendicitis. Material and Method: The study includes 77 cases with histopathologically proven acute appendicitis and 17 control cases. Blood samples were obtained from all cases and C-reactive protein (CRP), Granulocyte Colony Stimulating Factor (G-CSF) and Total Antioxidant Capacity (TAC) were measured. Findings: In cases with acute appendicitis, CRP and G-CSF levels were found to be related to acute appendicitis;however, TAC was not affected by the disease process. Moreover, CRP and G-CSF levels were correlated with the disease severity. Conclusion: Both CRP and G-CSF can be used in diagnosis of acute appendicitis. Furthermore, increased CRP level can be a marker to show advanced cases. However, G-CSF is not an effective marker to show disease severity.
文摘Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal antibody (McAb) of MAF-J6-1R RE2 and polyclonal antibody (PolyAb) of rhM-CSFR were prepared. The specificity of McAb RE2 to M-CSFR was confirmed by indirect ELISA, cross-neutralizing assay with J6-1 cell colony formation and neutralization test by ELISA. Results: the reactive activity of purified RE2 to M-CSFR was over 1: 16000. The inhibitory activity of M-CSFR and MAF-J6-1R could be blocked by RE2 and anti-M-CSFR antibody. The reactivity of RE2 to M-CSFR could be reduced by M-CSFR. Conclusion: The specificity of RE2 to M-CSFR was confirmed and the antigen association of MAF-J6-1R with M-CSFR was proved. It suggests that M-CSF and its receptor mediated auto-juxtacrine stimulation could be an operative mechanism in either leukemia or nonhematological malignancies.
文摘The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This study was undertaken to assess the effect ofgranulocyte-macrophage colony stimulating factor ( GM-CSF) on the repair of damaged iliac arteries.Twenty-four male New Zealand white rabbits undergoing primary iliac artery deendothelialization wererandomly divided into two groups ( GM-CSF group and control group) . The GM-CSF group received asubcutaneous injection of GM-CSF (10 μg ? kg^(-1) ? d^(-1) ) , and the control groupwas given a subcutaneous injection of equivalent saline. The iliac arteries of all animals weredamaged by balloon after 7 days. The levels of nitric oxide ( NO) were detected before, 1 week, 2weeks and 4 weeks after angioplasty. The repair and hyperplasia of the intima were observedmicroscopically and the indices of stenosis were evaluated by computerized planimetry after 4 weeksof angioplasty. The NO levels of the GM-CSF group were higher than those of the control group 2weeks and 4 weeks after angioplasty [91.92 +-11.57) μmol/L vs. (81. 67 +- 12. 18) μmol/L; (97. 67+- 10. 13 ) ( μmol/L vs. (83. 16 +-12. 64) μmol/L]. Four weeks after balloon damage,histological examination showed that neointima formation, vascular smooth muscle cells and fibroustissue of the GM-CSF group were less than those of the control group. The endothelium of the GM-CSFgroup was more integrated, and stenosis of lumen was slighter than that of the control group.Morphometry showed the lumen area of the GM-CSF group was larger than that of the control group[(1.27 +-0. 31) mm^2 vs. (0. 92 +- 0. 24) mm^2 ] , the neointimal area and percent of intimahyperplasia were significantly smaller than those of the control group [ (0. 85 +-0. 34) mm vs. (1.18 +-0. 38) mm^2; (40 +- 7)% vs. (55 +- 6)%]. GM-CSF could facilitate the repair of the intima,reduce neointima formation, better the function of the endothelium, and decrease the rate ofrestenosis.
基金supported by the National Natural Science Foundation of China,Nos.82072165 and 82272256(both to XM)the Key Project of Xiangyang Central Hospital,No.2023YZ03(to RM)。
文摘Spinal cord injury represents a severe form of central nervous system trauma for which effective treatments remain limited.Microglia is the resident immune cells of the central nervous system,play a critical role in spinal cord injury.Previous studies have shown that microglia can promote neuronal survival by phagocytosing dead cells and debris and by releasing neuroprotective and anti-inflammatory factors.However,excessive activation of microglia can lead to persistent inflammation and contribute to the formation of glial scars,which hinder axonal regeneration.Despite this,the precise role and mechanisms of microglia during the acute phase of spinal cord injury remain controversial and poorly understood.To elucidate the role of microglia in spinal cord injury,we employed the colony-stimulating factor 1 receptor inhibitor PLX5622 to deplete microglia.We observed that sustained depletion of microglia resulted in an expansion of the lesion area,downregulation of brain-derived neurotrophic factor,and impaired functional recovery after spinal cord injury.Next,we generated a transgenic mouse line with conditional overexpression of brain-derived neurotrophic factor specifically in microglia.We found that brain-derived neurotrophic factor overexpression in microglia increased angiogenesis and blood flow following spinal cord injury and facilitated the recovery of hindlimb motor function.Additionally,brain-derived neurotrophic factor overexpression in microglia reduced inflammation and neuronal apoptosis during the acute phase of spinal cord injury.Furthermore,through using specific transgenic mouse lines,TMEM119,and the colony-stimulating factor 1 receptor inhibitor PLX73086,we demonstrated that the neuroprotective effects were predominantly due to brain-derived neurotrophic factor overexpression in microglia rather than macrophages.In conclusion,our findings suggest the critical role of microglia in the formation of protective glial scars.Depleting microglia is detrimental to recovery of spinal cord injury,whereas targeting brain-derived neurotrophic factor overexpression in microglia represents a promising and novel therapeutic strategy to enhance motor function recovery in patients with spinal cord injury.
文摘Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were treated with one course of CLAG from April 1st,2014 through December 9th,2015 in our hospital were retrospectively reviewed.Results Thirty-three
基金supported by the National Natural Science Foundation of China(grant nos.82102931,82330085,82073354,and 82102201)Hubei Provincial Key Laboratory Open Project Fund for Precision Radiotherapy in Oncology(grant no.jzfs001).
文摘Radiotherapy(RT)serves as the primary treatment for solid tumors.Its potential to incite an immune response against tumors both locally and distally profoundly impacts clinical outcomes.However,RT may also promote the accumulation of immunosuppressive cytokines and immunosuppressive cells,greatly impeding the activation of antitumor immune responses and substantially limiting the effectiveness of RT.Therefore,regulating post-RT immunosuppression to steer the immune milieu toward heightened activation potentially enhances RT's therapeutic potential.Cytokines,potent orchestrators of diverse cellular responses,play a pivotal role in regulating this immunosuppressive response.Identifying and promptly neutralizing early released immunosuppressive cytokines are a crucial development in augmenting RT's immunomodulatory effects.To this end,we conducted a screen of immunosuppressive cytokines following RT and identified macrophage colony-stimulating factor(MCSF)as an early up-regulated and persistent immune suppressor.Single-cell sequencing revealed that the main source of up-regulated McSF derived from tumor cells.Mechanistic exploration revealed that irradiation-dependent phosphorylation of the p65 protein facilitated its binding to the McSF gene promoter,enhancing transcription.Knockdown and chemical inhibitor experiments conclusively demonstrated that suppressing tumor cell-derived McSF amplifies RT's immune-activating effects,with optimal results achieved by early MCSF blockade after irradiation.Additionally,we validated that MCSF acted on macrophages,inducing the secretion of a large number of inhibitory cytokines.In summary,we propose a novel approach to enhance the immune activation effects of RT by blocking the MCSF-CSF1R signaling pathway early after irradiation.
基金supported by grants from the General Project of the National Natural Science Foundation of China(No.81970180)the Science and Technology Project of Tianjin Municipal Health Committee(No.TJWJ2022QN030)+7 种基金the Key projects of Tianjin Applied Basic Research and Multi-Investment Fund(No.21JCZDJC01240)the Science and Technology Project of Tianjin Municipal Health Committee(No.TJWJ2022XK018)Tianjin Key Medical Discipline(Specialty)Construction Project(No.TJYXZDXK-056B)Tianjin Municipal Natural Science Foundation(No.22JCQNJC00820)Tianjin Health Research Project(No.TJWJ2023QN027)Tianjin Health Bureau Project(No.ZC20074)Tianjin Key Medical Discipline(Specialty)Construction Project(No.TJWJ2023XK010)Henan Provincial Youth and Middle-aged Health Science and Technology Talents Excellent Youth Program(No.20230273).
文摘To the Editor:After chimeric antigen receptor T(CAR-T)cell therapy,over 90%of patients experience either mild or severe cytopenia,with the prevalence of Grades 3–4 neutropenia ranging from 53%to 94.3%.[1,2]Granulocyte colony-stimulating factor(G-CSF),known for its ability to enhance the proliferation and differentiation of neutrophils,has been extensively employed for the prevention and treatment of neutropenia in various contexts.However,its application in patients with refractory/relapsing B-cell acute lymphoblastic leukemia(R/R B-ALL)after CAR-T cell therapy remains a topic of debate.[3]Several studies have suggested that G-CSF may exert anti-inflammatory effects by modulating the release of inflammatory factors,such as interleukin-12(IL-12),TNF-α,and IFN-γ,or by influencing the differentiation of T-cell subsets.[4]However,other researchers have posited that G-CSF could exacerbate cytokine release syndrome(CRS)and immune effector cell-associated neurotoxicity syndrome(ICANS)through the augmentation of antigen-presenting cell function.
文摘Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Methods From November 1992 to March 1997, 45 patients wer e enrolled into our study and 15 were newly diagnosed All were treated with hi gh dose chemotherapy combined with rhGCSF Results Of 15 newly diagnosed patients, 13 achieved complete remission (CR) after one course of therapy and 2 achieved CR after two courses of therapy For newly diagnosed patients, the durations of absolute neutrophil counts (ANC ) <05109/L were 5 days and 10 days in rhGCSF group and control group res p ectively ( P <005) The incidences of infection of these two groups w ere 40% and 60% respectively ( P <005) As for patients who receive d intensive therapy, the durations of ANC <05109/L were 5 days and 8 days i n rhGCSF group and control group, respectively ( P <005), and the i ncidences of infection were 25% and 444% respectively ( P <005) Conclusions The application of rhGCSF in children with AML after chem otherapy may hasten the hematopoietic recovery The duration of neutropenia wa s shortened by 3-4 days, and the incidence of fatal infection was reduced rhG CSF does not stimulate AML growth in vivo
文摘Severe alcoholic hepatitis(SAH)is associated with high short-term mortality.The SAH population exhibits extreme heterogeneity in disease severity,clinical presentation,decompensations,and outcomes.Nonetheless,improving outcomes and preventing adverse events is a major challenge when selecting an appropriate treatment for alcoholic hepatitis.Currently,steroids are the standard of care for SAH with Maddrey’s discriminant function>32 and model for end stage liver disease>20;however,they have limited usage due to ineligibility in approx-imately two-third of such patients.Approximately 25%of patients do not respond to steroids and require alternative therapies.An array of evolving therapies,such as granulocyte colony-stimulating factors,plasma exchange,fecal microbiota transplantation,antibiotics,anti-cytokine therapies,and N-acetylcysteine,showing variable success,are emerging.Hence,it is also crucial to select appro-priate therapy.The present review discusses the standard of care,the existing therapies,risk stratification for outcomes,and the selection of appropriate therapy to improve survival in SAH patients.
基金ThisstudywassupportedbyagrantfromtheNationalPublicHealthMinistry (No97030223)andagrantfromtheNationalNaturalScienceFoundationofChina (No 39670 667)
文摘Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac) Methods A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 μg of GM CSF plus 10 μg of rHBvac injected intramuscularly at the same location (GM CSF group, 14 children) or 200 IU HBIG and 10 μg rHBvac in different muscles (HBIG group, 13 children) on a monthly four dose schedule HBV DNA quantification and other HBV serological markers were tested before and after the four dose therapy Results Twelve children in each group completed the study Of them, 3 children in the GM CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM CSF group and 10 in the HBIG group One from each group had an HBeAg/anti HBe seroconversion after the treatment The quantity of HBV DNA was significantly lower after the treatment ( P =0 023) in GM CSF group, but was not significantly reduced in HBIG group No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group Conclusion Combined GM CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis
文摘Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acute promyelocytic leukemia (APL) during the treatment with all trans retinoic acid (ATRA) Results The peak of increased serum G CSF level occurred on the 9th day, and WBC number was the highest on the 11th day After ATRA treatment, both serum G CSF level and WBC number increased in 68 1% of the cases In 19 2% of the cases treated, serum G CSF level was increased but without obvious change in WBC number, and the reverse was true in 12 7% of the cases Conclusion Serum G CSF level was statistically correlated to the number of WBC, promyelocytes and its late stage by Spearman's rank order correlation coefficient
文摘The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.
基金supported by the National Natural Science Foundation of China(81772197,81401670 and 81630062)the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(2018RC330004)+3 种基金National Key R&D Program of China(2018YFC1200305Y)National Science and Technology Major Project of China(2018ZX10102001)SKLID Development Grant(2019SKLID401 and 2016SKLID603)the Natural Science Foundation of Heilongjiang Province(C2018044)
文摘Activation of inflammatory cells and upregulations of a number of cytokines in the central nervous system(CNS)of patients with prion diseases are frequently observed.To evaluate the potential changes of some brain cytokines that were rarely addressed during prion infection,the levels of 17 different cytokines in the brain homogenates of mice infected with different scrapie mouse-adapted agents were firstly screened with Luminex assay.Significant upregulations of interferon gamma-induced protein 10(IP10),keratinocyte chemoattractant(KC)and macrophage colony stimulating factor(M-CSF)were frequently detected in the brain lysates of many strains of scrapie infected mice.The upregulations of those three cytokines in the brains of scrapie infected mice were further validated by the individual specific ELISA and immunohistochemical assay.Increased specific mRNAs of IP10,M-CSF and KC in the brains of scrapie infected mice were also detected by the individual specific qRT-PCRs and IP10-specific digital PCR.Dynamic analyses of the brain samples collected at different time points post infection revealed the time-dependent increases of those three cytokines,particularly IP10 during the incubation period of scrapie infection.In addition,we also found that the levels of IP10 in cerebral spinalfluid(CSF)of 45 sporadic Creutzfeldt–Jakob disease(sCJD)patients were slightly but significantly higher than those of the cases who were excluded the diagnosis of prion diseases.These data give us a better understanding of inflammatory reaction during prion infection and progression of prion disease.
基金Supported by grants of Medical Science and Technology Development Foundation, Jiangsu Province Department of Health (No. H201013)the Program for Postgraduate Research Innovation in University of Jiangsu Province (No. CX10B_054Z)the Project of Youth Foundation in Science and Education of Department of Public Health of Suzhou (2010, No. 4)
文摘Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor （GM-CSF） transfection in mice ex vivo. Methods： The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c＋ cells were cultured with cytokines. For adenoviral （Ad）-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of multiplicity of infection （MOI） to determine the optimal gene transfection conditions, and detecting the expression of GM-CSF after transfection. The variation of GM-CSF gene-modified DCs were analyzed by morphological observation, phenotype analysis, and mixed lymphocyte reaction （MLR）. DCs were loaded with gastric cancer antigen obtained by frozen and thawed method. The stimulated DCs vaccination induced T lymphocytes, and the killing effect of T cells to gastric cancer cells was assayed by MTT. INF-γ production was determined with the INF-γ ELISA kit. Results： B220–CD11c＋ cells numbers increased after CCL3 injection. ELISA results showed that after GM-CSF gene modification, DC could produce high level of GM-CSF. When DCs were transferred AdGM-CSF gene at MOI equal to 1：100, GM-CSF level in culture supernatants reached saturation [（130.00 ± 12.61） pg/mL]. After GM-CSF gene-modification, DCs tended to more maturated through morphological observation and were phenotypically identical to typical DC and gained the capacity to stimulate allogeneic T cells. T lymphocytes stimulated with DC transduced with GM-CSF gene showed the specific killing effect on gastric carcinoma cells and produced high level of INF-γ [（1245.00 ± 13.75） pg/mL]. Conclusion： CCL3-recruited DCs modified by adenovirus-transducted GM-CSF could produce high level of GM-CSF, which tended to more maturated, and the capacity of activating allogeneic T lymphocytes proliferation was enhanced greatly. Moreover, they could stimulate specific cytotoxic T lymphocyte （CTL） to gastric cancer ex vivo.
文摘AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-CSF administration on virological response. METHODS:Sixty-eight patients undergoing antiviral treatment for post-liver transplantation(OLT)HCV recurrence were enrolled.All patients developing neutropenia received G-CSF. RESULTS:Twenty three(34%)received G-CSF.Mean neutrophil count at the onset of neutropenia was 700/mmc(range 400-750/mmc);after 1 mo of G-CSF it increased to 1210/mmc(range 300-5590/mmc) (P<0.0001).Three patients did not respond to G-CSF. Treatment duration was similar in neutropenic and non-neutropenic patients.No differences in the rate of discontinuation,infections or virological response were observed between the two groups.G-CSF was protective for the onset of de novo autoimmune hepatitis(P<0.003). CONCLUSION:G-CSF administration is effective in the case of Peg-IFN induced neutropenia increasingneutrophil count,prolonging treatment and leading to sustained virological response(SVR)rates comparable to non-neutropenic patients.It prevents the occurrence of de novo autoimmune hepatitis.
基金This work was supported by the Natural Science foundation of Fujian Province, China (No. C97067).
文摘Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1-GM-CSF plasmid which was controlled by the CMV promoter was transferred into CHO cell by lipofectamine, selected by G418 and the positive clones was got. The recombinant vector which was rejoined into the groups of DNA of CHO was identified by PCR. Results: The results showed that the protein of rhGM-CSF was about 28 KD by using ELISA, SDS-PAGE and Western blot. Conclusion: rhGM-CSF was expressed steadily and highly. The rhGM-CSF will be of more use value.
文摘Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiation.Methods DCs from peripheral blood mononuclear cells (PBMC) that maintain the antigen capturing and processing capacity charateristicof immature cells have been established in vitro, using granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then, in cholangiocarcinoma cells apoptosis was induced by J-irradiation. The experimental groups were as follows: (1) coculture ofDCs and apoptotic cancer cells and T cells; (2) coculture of DCs and necrotic cancer cells and T cells; (3) coculture of DCs, culturedcancer cell and T cells. They are cocultured for 7 days. DCs and T cells were riched, isolated and their antitumor response was tested.Results The cells had typical dendritic morphology, expressed high levels of GDI a and B7, acquired antigen from apoptotic cells causedby y-irradiation and induced an increased T cell stimulatory capacity in mixed lymphocyte reactions (MLR) .Conclusion DCs obtained from PBMCs using GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by y-irradiation and efficiently induce T cells. This strategy, therefore, may present an effective approach to transduce DCs with antigen.
