Serum Granulocyte-macrophage Colony-stimulating Factor Research Articles

Repeated inhalation of GM-CSF by nonhuman primates induces bronchus-associated lymphoid tissue along the lower respiratory tract

BackgroundRepeated inhalation of granulocyte-macrophage colony-stimulating factor (GM-CSF) was recently approved in Japan as a treatment for autoimmune pulmonary alveolar proteinosis. However, the detailed physiological and pathological effects of repeated inhalation in the long term, especially at increasing doses, remain unclear.MethodsIn this chronic safety study, we administered 24 cynomolgus monkeys (Macaca fascicularis) aged 2–3 years with aerosolized sargramostim (a yeast-derived recombinant human GM-CSF [rhGM-CSF]) biweekly for 26 weeks across four dosing groups (0, 5, 100, and 500 µg/kg/day). We measured the serum GM-CSF antibody (GM-Ab) concentration by an ELISA and assessed the neutralizing capacity of GM-Ab using the GM-CSF-dependent cell line TF-1. We subjected lung tissue samples taken from all monkeys at 27 weeks to histopathological assessment using a sargramostim-specific monoclonal antibody to detect localization of residual sargramostim.ResultsAll the animals maintained good body condition and showed steady weight gain throughout the study. The pathological analyses of the lung revealed the formation of induced bronchus-associated lymphoid tissue (iBALT) in the lower respiratory tract, even at the clinical dose of 5 µg/kg/day. There was a relationship between the number or size of BALT and sargramostim dose or the serum GM-Ab levels. Immunohistochemical analyses revealed GM-Ab–producing cells in the follicular region of iBALT, with residual sargramostim in the follicles. Leucocyte counts were inversely correlated with GM-Ab levels in the high-dose groups. Additionally, serum GM-Ab from the treated animals significantly suppressed the alveolar macrophage proliferation activity of both Cynomolgus recombinant and rhGM-CSF in vitro.ConclusionLong-term repeated inhalation of sargramostim led to iBALT formation in the lower respiratory tract, even at the clinical dose of 5 µg/kg/day, with the extent of iBALT formation increasing in a dose-dependent manner. Inhaled sargramostim was localized to the follicular region of iBALT nodules, which may induce the production of GM-Ab.

Serum GM-CSF level is a predictor of treatment response to tocilizumab in rheumatoid arthritis patients: a prospective observational cohort study

BackgroundThe aim of this prospective observational cohort study was to unveil the predictors of treatment response to tocilizumab (TCZ) therapy in rheumatoid arthritis (RA) patients, in terms of clinical characteristics and serum proinflammatory cytokines, especially to explore the predictive value of granulocyte macrophage-colony stimulating factor (GM-CSF).MethodsActive adult RA patients with inadequate response to MTX intending to receive TCZ therapy were recruited prospectively in the study. A total of 174 severe RA patients were included for the identification of the associations between treatment response and the following characteristic features: demographics, medications, disease activity, serum proinflammatory cytokines and so on.ResultsDisease duration (OR = 0.996), tender joint count (TJC)/68 (OR = 0.943), neutrophil ratio (W4/baseline) (OR = 0.224), the high level of GM-CSF > 5 ng/ml (OR = 0.414) at baseline were the independent adverse predictors of good response assessed by clinical disease activity index (CDAI) at week 24 (W24) for TCZ therapy in RA patients. Moreover, DAS28-ESR (OR = 2.951, P = 0.002) and the high level of GM-CSF > 10 ng/ml at baseline (OR = 5.419, P = 0.002) were independent predictors of poor response, but not the high level of GM-CSF > 5 ng/ml (OR = 2.713, P = 0.054). The patients in the high GM-CSF group had significantly higher DAS28-ESR and serum levels of cytokines (IL-17A, IL-1β, IL-6, TNF-α) at baseline, as well as significantly higher rate of non-good response (62.8% vs. 39.4%, P = 0.010) and poor response (27.9% vs. 9.1%, P = 0.004) than the low GM-CSF group at W24. In addition, poor responders had significantly higher levels of GM-CSF with concomitant increase in the serum levels of IL-17A and IL-1β at baseline than those in moderate and good response groups, while serum levels of IL-6 and TNF-α at baseline were not significantly different in three response groups.ConclusionThe high levels of GM-CSF (> 5 ng/ml and > 10 ng/ml) at baseline were the independent predictors of non-good response and poor response to TCZ at W24 respectively. The high level of GM-CSF at baseline is a marker of high disease activity and a predictor of poor response to TCZ in severe RA patients, which may facilitate the development of individualized treatment strategies for refractory RA.

Pathogenesis-driven treatment of primary pulmonary alveolar proteinosis.

Pulmonary alveolar proteinosis (PAP) is a syndrome that results from the accumulation of lipoproteinaceous material in the alveolar space. According to the underlying pathogenetic mechanisms, three different forms have been identified, namely primary, secondary and congenital. Primary PAP is caused by disruption of granulocyte-macrophage colony-stimulating factor (GM-CSF) signalling due to the presence of neutralising autoantibodies (autoimmune PAP) or GM-CSF receptor genetic defects (hereditary PAP), which results in dysfunctional alveolar macrophages with reduced phagocytic clearance of particles, cholesterol and surfactant. The serum level of GM-CSF autoantibody is the only disease-specific biomarker of autoimmune PAP, although it does not correlate with disease severity. In PAP patients with normal serum GM-CSF autoantibody levels, elevated serum GM-CSF levels is highly suspicious for hereditary PAP. Several biomarkers have been correlated with disease severity, although they are not specific for PAP. These include lactate dehydrogenase, cytokeratin 19 fragment 21.1, carcinoembryonic antigen, neuron-specific enolase, surfactant proteins, Krebs von Lungen 6, chitinase-3-like protein 1 and monocyte chemotactic proteins. Finally, increased awareness of the disease mechanisms has led to the development of pathogenesis-based treatments, such as GM-CSF augmentation and cholesterol-targeting therapies.

73 Effects of blend of organic acids and monoglycerides on immune responses of weaned pigs experimentally infected with enterotoxigenic Escherichia coli F18

Abstract Our previous research implies that dietary supplementation of organic acids blend, monoglycerides blend, and the combination of both can mitigate diarrhea of weaned pigs infected with Escherichia coli (E. coli) F18. The objective of this study was to investigate the effects of dietary supplementation with a blend of organic acids, a blend of monoglycerides, or the combination of both on immune responses of weaned pigs experimentally infected with E. coli F18. Forty pigs [average body weight (BW) = 7.81 ± 0.84 kg] were individually housed and randomly assigned to one of four treatment groups with 10 replicate pigs per treatment. The four dietary treatments were: () Control: nursery basal diet, 2) Acids: basal diet supplemented with a blend of organic acids (formic acid, lactic acid, and sodium formate) at 0.3%, 3) Monoglycerides: basal diet supplemented with a blend of monoglycerides of fatty acids at 0.3%, or 4) Combination of acids and monoglycerides: basal diet added with the acids blend at 0.2% and the monoglycerides blend at 0.2%. The experiment lasted 28 d, with 7 d before and 21 d after the first inoculation (d 0). All pigs were orally inoculated with E. coli F18 (1010 CFU/3 mL) for 3 consecutive days from d 0. Serum samples were collected on d 0, 2, 5, and 14 post-inoculation (PI) to measure the concentration of C-reactive protein (CRP) and haptoglobin using ELISA. Porcine Cytokine/Chemokine13-Plex Discovery Assay (Eve Technology, Canada) was also conducted with serum samples from d 0, 5, and 14 PI. Data were analyzed by ANOVA using PROC MIXED of SAS with a randomized complete block design. The concentrations of serum CRP and haptoglobin were increased (P < 0.0001) on d 2 PI and then gradually decreased on d 5 and 14 PI. However, no differences were observed in serum CRP and haptoglobin concentrations among all treatment groups throughout the experiment. Pigs fed with monoglycerides tended (P < 0.10) to have decreased serum granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 1-alpha (IL-1α), and tumor necrosis factor-alpha (TNF-α) concentrations on d 0 PI compared with other treatments. Pigs supplemented with monoglycerides also tended (P < 0.10) to have decreased GM-CSF on d 5 PI and less IL-1α on d 14 PI among all treatments. Supplementation of the combination tended (P < 0.10) to have greater GM-CSF concentration on d 5 PI compared with other treatments. In conclusion, dietary supplementation of monoglycerides blend or the combination of organic acids and monoglycerides alleviates systemic inflammation of weaned pigs during E. coli F18 infection by modulating related cytokines. However, additional research is needed to understand the modulation mechanisms.

Can serum granulocyte-macrophage colony-stimulating factor and CCL17 levels be a marker of disease activation in spondyloarthritis?

The aim of this cross-sectional study was to investigate if serum levels of granulocytemacrophage colony-stimulating factor (GM-CSF) and CC chemokine ligand 17 (CCL17) correlate with disease activity in axial spondyloarthritis (axSpA) and peripheral spondyloarthritis (pSpA) patients. The cross-sectional study was conducted with 80 individuals (48 females, 32 males; mean age: 47.7±11.5) between March 2021 and September 2021. Of the participants, 20 were axSpA, 20 were pSpA, and 20 were active rheumatoid arthritis patients, and the remaining 20 were healthy controls. Age, sex, body mass index, disease duration, comorbid diseases, smoking status, medical treatments, C-reactive protein (CRP) level and human leukocyte antigen B27 (HLA-B27) positivity were recorded. Serum GM-CSF and CCL17 levels were analyzed by ELISA. Ankylosing Spondylitis Disease Activity Score with CRP (ASDAS-CRP) was used to evaluate the disease activity of patients with spondyloarthritis. Functional status of spondyloarthritis patients was evaluated by Bath Ankylosing Spondylitis Functional Index (BASFI). While the serum GM-CSF levels were similar in the axSpA and pSpA groups, they were significantly higher than the healthy control group (p=0.021 and p=0.009, respectively). There was a significant correlation between GM-CSF levels and ASDAS-CRP (r=0.545, p=0.013) and BASFI (r=0.546, p=0.013) in the axSpA group. In active axSpA patients, the cut-off value for GM-CSF was 15.89 pg/mL (sensitivity 50%, specificity 100%). No differences were detected in serum CCL17 levels among the groups. The results suggest that serum GM-CSF levels may be used as a new marker for the evaluation of disease activity in axSpA, and GM-CSF might be a therapeutic target.

Association of serum levels of inflammatory cytokines with retinopathy of prematurity in preterm infants.

Retinopathy of prematurity (ROP) is a retinal vascular developmental disease associated with risks factors such as supplementary oxygen use or low birth weight/early gestational age. Multiple studies have reported associations between ROP and systemic inflammation. In this study, we investigated serum cytokines associated with ROP development and severity and assessed their applicability as potential biomarkers of ROP. This prospective study was conducted at an institutional referral center between 2019 and 2021. To measure the serum levels of 40 inflammatory cytokines in eligible premature patients, we collected their serum samples during the enrollment of patients or the intravitreal injection of anti-vascular endothelial growth factor (VEGF) agents and after 2 and 4 weeks. Fifty patients were enrolled. In patients with type 1 ROP who received anti-VEGF agents (n = 22), the levels of serum intercellular adhesion molecule-1 decreased significantly (p < 0.05) at 4 weeks compared with the baseline level, whereas those of serum granulocyte-macrophage colony-stimulating factor increased significantly (p < 0.05). In patients with ROP who did not require any treatment (n = 14), no significant change was noted in the level of any of the 40 inflammatory cytokines. In control infants without ROP (n = 14), the serum levels of tumor necrosis factor-α, interleukin (IL)-15, and IL-12p40 increased significantly (p < 0.05) at 4 weeks. The changes in the levels of serum inflammatory cytokines did not vary significantly among the aforementioned three groups. A generalized estimating equation indicated that zone 1 ROP, stage 3 ROP, older postmenstrual age, respiratory distress syndrome, necrotizing enterocolitis, and sepsis were associated with the changes in serum cytokine levels. Although significant changes (compared with baseline) were observed in the serum levels of certain inflammatory cytokines in patients with type 1 ROP and infants without ROP, no significant difference in cytokine level fluctuations were noted among the three groups. Changes in serum inflammatory cytokine levels may not predict ROP development or severity. Additional comprehensive studies are warranted to establish their definitive role and significance in ROP, emphasizing the need for continued research in this area.

Efficacy of Heshouwu () on gut mircobiota in mice with autoimmune encephalomyelitis.

To learn the mechanisms between gut microbiome and the autoimmunity benefits on Traditional Chinese Medicine (TCM) in central nervous system (CNS), we investigated the neuro-protection effects and gut mircobiota changes of Heshouwu () on experimental autoimmune encepha-lomyelitis (EAE), an animal model of multiple sclerosis (MS). Mice were randomly divided into four groups: EAE mice (control phosphate-buffered saline group), 50 mg·kg·d Heshouwu ()-treated EAE mice, 100 mg·kg·d Heshouwu ()-treated EAE mice, and 200 mg·kg·d Heshouwu ()-treated EAE mice. The spinal cords were stained with hematoxylin and eosin (HE) and luxol fast blue for evaluating inflammatory infiltration and demyelination. The percentages of granulocyte macrophage-colony stimulating factor (GM-CSF)+CD4+, interleukin 17 (IL-17)+CD4+, Foxp3 CD4+, and interferon-γ (IFN-γ)+CD4+ T cells in the inguinal lymph nodes (LNs) and brain were determined by flow cytometry analysis. 16S rRNA gene sequencing was employed to analyze the changes in gut microbiota. We found that Heshouwu () alleviated the disease severity and neuropathology of EAE as evaluated by clinical and histopathologyical scores. Heshouwu () increased the diversity and abundance of the gut microbiota, and decreased / ratio (F/B ratio). Heshouwu () also decreased the concentrations of IL-10, and IL-21 and increase the levels of GM-CSF, IL-17A, IL-17F and IL-22 in serum of EAE mice. Moreover, Heshouwu () modulated the T cell responses by inhibiting Th17 cells and restoring Treg cells in the small intestine lymphoid tissues and inguinal lymph nodes. Microbiota-depleted mice receiving Heshouwu ()-treated fecal microbiota trans-plantation had lower disease severity, neuropathology scores and alleviation of Th17/Treg imbalance compared to ad libitum group. Our findings suggested that the vital neuro-protection role of Heshouwu () (TCM) in immunomodulation effects partly by regulations of gut microbiome.

Serum Level of Granulocyte-macrophage Colony-stimulating Factor in Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis

BACKGROUND: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe cutaneous adverse drug reactions. Activated T-cells secrete high amounts of cytokines that increase the expression and activity of keratinocytes, including granulocyte-macrophage colony-stimulating factor (GM-CSF). AIMS: The aims of this study were to evaluate the serum level of GM-CSF in SJS and TEN as well as the relationship between it and the progress of SJS and TEN. METHODS: This was a sectional descriptive study conducted at the National Hospital of Dermatology and Venereology, in Hanoi, Vietnam, from October 2017 to September 2019. Forty-eight SJS/TEN patients, 43 erythema multiforme (EM) patients, and 20 healthy controls (HCs) participated. GM-CSF levels were measured using the fluorescence covalent microbead immunosorbent assay (ProcartaPlex Immunoassay Panels kit, Thermo Fisher Scientific, USA). The Mann–Whitney U-test was used to compare serum SJS/TEN levels of the two groups. The Wilcoxon tests were used to compare quantitative variables before and after the treatment. Differences were considered to be statistically significant at p &lt; 0.05. RESULTS: There were 19 SJS patients (39.5%) and 29 TEN patients (60.5%). The mean age was 49.3 years, range of 19–77 years. The male patients were 47.9%. The most common causative drugs were traditional medicine (29.1%), followed by carbamazepine (12.5%), and allopurinol (12.5%). On the day of hospitalization, the mean serum level of GM-CSF in the SJS/TEN group was 10.6 pg/mL, which was significantly higher than that of the EM group (p &lt; 0.05) but not higher than that of the HCs group and was higher than that on the day of re-epithelialization (3.6 pg/mL) and the difference was statistically significant with p &lt; 0.05. CONCLUSION: Serum GM-CSF level can be a good biomarker to evaluate the progress of SJS/TEN.

Role of IL4 and GMCSF in Predicting Survival in Esophageal Cancer.

Esophageal cancer (EC) originates in the setting of chronic inflammation. Although previous studies have sought to understand the role of inflammatory signaling in EC, the effect of these immunologic changes on patient outcomes remains understudied. This study's objective was to identify relationships between cytokine levels and prognosis in a mixed cohort of esophageal adenocarcinoma (EAC) and esophageal squamous cell carcinoma (ESCC) patients. A total of 37 serum cytokines were profiled at the time of resection using multiplex ELISA in 47 patients (42 esophageal adenocarcinoma, 5 esophageal squamous cell carcinoma). Cytokine levels were median-binarized and assessed using Cox regression models. Findings were validated at the RNA level using The Cancer Genome Atlas EC cohort (81 esophageal adenocarcinoma, 81 esophageal squamous cell carcinoma). Univariable analysis revealed high serum interleukin 4 (IL4) and granulocyte-macrophage colony-stimulating factor (GMCSF) were negatively associated with overall survival (p = 0.046, p = 0.040). Multivariable analysis determined both high serum IL4 or high serum GMCSF were negatively associated with survival independent of important clinical factors (hazard ratio [HR] 7.55, p < 0.001; HR 5.24, p = 0.001). These findings were validated at the RNA level in The Cancer Genome Atlas EC cohort, where multivariable analysis identified high IL4 expression, high CSF2 expression (encodes GMCSF), and advanced pathologic stage as independent negative predictors of survival when controlled for clinical factors (HR 2.35, p = 0.012; HR 1.97, p = 0.040). These results show that high IL4/GMCSF levels are negatively associated with survival in EC. These relationships are independent of pathologic stage and are identified across modalities, histologic subtypes, and the presence/absence of neoadjuvant therapy.

Multiple Cytokine Analysis of Th1/Th2/Th9/Th17/Th22/Treg Cytokine Pathway for Individual Immune Profile Assessment in Patients with Psoriasis.

BACKGROUND Psoriasis is an autoimmune and autoinflammatory disorder that has a significant impact on patient quality of life. The aim of the study was to assess the immune profiles of patients with psoriasis with multiple cytokine analysis. MATERIAL AND METHODS Fifty-two male psoriatic patients and 24 healthy male volunteers were recruited. Granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-gamma), interleukin (IL)-1 beta, IL-2, Il-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17A, IL-18, IL-21, IL-22, IL-23, IL-27, and tumor necrosis factor (TNF)-alpha were measured in patients' serum with a Th1/Th2/Th9/Th17/Th22/Treg Cytokine 18-Plex Human ProcartaPlex Panel, based on Luminex xMAP technology. RESULTS The median fluorescence intensities of serum GM-CSF, IL-2, IL-5, IL-10, IL-13, IL-17A, IL-21, and IL-22 were not intensive enough to calculate the cytokine concentration. We observed elevated levels of IL-6 (P=0.001) and IL-9 (P=0.003) in patients, compared with the control group. The levels of IL-1beta (P=0.008) and IL-27 (P=0.006) were decreased. In patients with psoriatic arthritis, we noticed a decreased level of IL-9 compared with that in patients without arthritis (P=0.034). The levels of IL-12 (P<0.05) and IL-18 (P<0.05) correlated positively with the Psoriasis Area and Severity Index. We found negative correlations of IL-9 (P<0.05), IL-12 (P<0.05), and IL-23 (P<0.05) with the age of psoriatic patients; IL-12 (P<0.05) and IL-23 (P<0.05) with psoriasis duration; and IL-6 (P<0.05) and IL-9 (P<0.05) with the Nail Psoriasis Severity Index. CONCLUSIONS Multiple cytokine analysis seems to be an important form of individual immune profile assessment before treatment selection.

Secondary Pulmonary Alveolar Proteinosis Due to Sirolimus

Introduction Pulmonary alveolar proteinosis (PAP) is a rare, progressive lung disease characterized by the accumulation of lipoproteinaceous material in alveoli due to impaired clearance by alveolar macrophages. Although most cases are due to the disruption of granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling, secondary PAP from immunodeficiency is described in solid organ recipients. We present a treatment conundrum in a heart transplant (OHT) patient with a history of graft rejection who develops secondary PAP while on sirolimus. Case Report A 63-year-old male 6 years post re-do OHT, initially transplanted for ischemic cardiomyopathy then re-do for antibody mediated rejection presented with new hypoxia requiring 2L O2. His immunosuppression regimen included myfortic, prednisone, sirolimus, and tacrolimus. A CT scan demonstrated central and basilar ground glass opacities with “crazy paving”. He underwent a bronchoscopy with bronchoalveolar lavage (BAL) showing 90% macrophages, negative cultures, and cytology with hemosiderin pigment and proteinaceous material. He was empirically placed on levofloxacin and weaned to room air. At 5-month follow-up a repeat CT re-demonstrated similar findings. Repeat BAL, performed for concern for PAP, was Periodic acid-Schiff (PAS) stain positive for proteinaceous material. Serum GM-CSF antibodies were absent and the patient was diagnosed with secondary PAP due to sirolimus. Given the patient's prior history of graft failure, his sirolimus dose was unchanged and he was treated with inhaled sargramostim. Summary PAP is a rare disease caused by disorders in surfactant homeostasis further divided into three distinct etiologies: autoimmune (90%), secondary (5-10%), and congenital (2%). Secondary PAP is due to functional impairment in alveolar macrophages from immunodeficiency and is well-described in kidney and lung transplant recipients. To our knowledge, this is the first case described in a heart transplant recipient. The prognosis of transplant-associated secondary PAP is difficult to establish due to the limited number of published cases, though resolution is typically observed 2-4 months after removal of the offending agent. We present a case treated with GM-CSF due to difficulties reducing immunosuppression. Pulmonary alveolar proteinosis (PAP) is a rare, progressive lung disease characterized by the accumulation of lipoproteinaceous material in alveoli due to impaired clearance by alveolar macrophages. Although most cases are due to the disruption of granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling, secondary PAP from immunodeficiency is described in solid organ recipients. We present a treatment conundrum in a heart transplant (OHT) patient with a history of graft rejection who develops secondary PAP while on sirolimus. A 63-year-old male 6 years post re-do OHT, initially transplanted for ischemic cardiomyopathy then re-do for antibody mediated rejection presented with new hypoxia requiring 2L O2. His immunosuppression regimen included myfortic, prednisone, sirolimus, and tacrolimus. A CT scan demonstrated central and basilar ground glass opacities with “crazy paving”. He underwent a bronchoscopy with bronchoalveolar lavage (BAL) showing 90% macrophages, negative cultures, and cytology with hemosiderin pigment and proteinaceous material. He was empirically placed on levofloxacin and weaned to room air. At 5-month follow-up a repeat CT re-demonstrated similar findings. Repeat BAL, performed for concern for PAP, was Periodic acid-Schiff (PAS) stain positive for proteinaceous material. Serum GM-CSF antibodies were absent and the patient was diagnosed with secondary PAP due to sirolimus. Given the patient's prior history of graft failure, his sirolimus dose was unchanged and he was treated with inhaled sargramostim. PAP is a rare disease caused by disorders in surfactant homeostasis further divided into three distinct etiologies: autoimmune (90%), secondary (5-10%), and congenital (2%). Secondary PAP is due to functional impairment in alveolar macrophages from immunodeficiency and is well-described in kidney and lung transplant recipients. To our knowledge, this is the first case described in a heart transplant recipient. The prognosis of transplant-associated secondary PAP is difficult to establish due to the limited number of published cases, though resolution is typically observed 2-4 months after removal of the offending agent. We present a case treated with GM-CSF due to difficulties reducing immunosuppression.

Serum granulocyte-macrophage colony-stimulating factor (GM-CSF) is increased in patients with active radiographic axial spondyloarthritis and persists despite anti-TNF treatment

BackgroundAccumulating evidence supports the role of monocytes and neutrophils in radiographic axSpA (r-axSpA). Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a growth factor for both leukocyte lineages and a pro-inflammatory cytokine activating myeloid cells and promoting osteoclastogenesis. It acts through the JAK-STAT pathway. We measured serum GM-CSF and markers of bone metabolism in patients with r-axSpA before and after anti-TNF treatment.MethodsPatients with active r-axSpA despite treatment with NSAIDs, all eligible for treatment with a biologic agent, were recruited. Healthy donors were sampled as controls. Serum was collected before (baseline) and after 4–6 months (follow-up) of anti-TNF treatment and the following molecules were measured with ELISA: GM-CSF, sclerostin (SOST), and dickkopf-1 (Dkk-1).ResultsTwelve r-axSpA patients (7 males, 5 females, median age 37 years) with a median disease duration of 1 year and 16 age- and sex-matched controls were included. At baseline, patients had mean BASDAI 6.3±2 and ASDAS 3.2±0.7, which decreased to 4.1±1.7 and 2.2±0.6 at follow-up, respectively. At baseline, r-axSpA patients had significantly higher mean serum levels of GM-CSF (150 vs 62pg/ml, p=0.049), significantly lower Dkk-1 (1228 vs 3052pg/ml, p=0.001), but similar levels of SOST (369 vs 544pg/ml, p=0.144) compared to controls. Anti-TNF treatment did not affect GM-CSF, Dkk-1, or SOST levels. Spearman correlation analysis showed that GM-CSF correlated positively with ASDAS at baseline (r=0.61, p=0.039), while no correlations were identified between bone markers (Dkk-1, SOST) on one hand and GM-CSF or disease activity indices on the other.ConclusionsGM-CSF is increased in patients with active AS and strongly correlates with disease activity. TNF inhibition does not affect GM-SCF levels, despite improving disease activity. GM-CSF may represent an important pathway responsible for residual inflammation during TNF blockade, but also a potential target of JAK inhibitors, explaining their efficacy in r-axSpA.

Granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-\u03b1 in combination is a useful diagnostic biomarker to distinguish familial Mediterranean fever from sepsis

ObjectiveTo identify potential biomarkers to distinguish familial Mediterranean fever (FMF) from sepsis.MethodWe recruited 28 patients diagnosed with typical FMF (according to the Tel Hashomer criteria), 22 patients with sepsis, and 118 age-matched controls. Serum levels of 40 cytokines were analyzed using multi-suspension cytokine array. We performed a cluster analysis of each cytokine in the FMF and sepsis groups in order to identify specific molecular networks. Multivariate classification (random forest analysis) and logistic regression analysis were used to rank the cytokines by importance and determine specific biomarkers for distinguishing FMF from sepsis.ResultsFifteen of the 40 cytokines were found to be suitable for further analysis. Levels of serum granulocyte-macrophage colony-stimulating factor (GM-CSF), fibroblast growth factor 2, vascular endothelial growth factor, macrophage inflammatory protein-1b, and interleukin-17 were significantly elevated, whereas tumor necrosis factor-α (TNF-α) was significantly lower in patients with FMF compared with those with sepsis. Cytokine clustering patterns differed between the two groups. Multivariate classification followed by logistic regression analysis revealed that measurement of both GM-CSF and TNF-α could distinguish FMF from sepsis with high accuracy (cut-off values for GM-CSF = 8.3 pg/mL; TNF-α = 16.3 pg/mL; sensitivity, 92.9%; specificity, 94.4%; accuracy, 93.4%).ConclusionDetermination of GM-CSF and TNF-α levels in combination may represent a biomarker for the differential diagnosis of FMF from sepsis, based on measurement of multiple cytokines.

Initial serum GM-CSF levels are associated with the severity of cerebral small vessel disease in microscopic polyangiitis patients

Serum cytokine levels were comprehensively measured, and the association with cerebrovascular lesions on brain magnetic resonance imaging (MRI) in microscopic polyangiitis (MPA) patients was investigated. The initial serum granulocyte-macrophage colony-stimulating factor (GM-CSF) levels were significantly higher in the high-grade white matter hyperintensities (WMH) group than those in the low-grade WMH group. In multivariate analyses, high serum levels of GM-CSF were independently associated with high-grade WMH. The initial serum GM-CSF levels correlated positively with the Birmingham Vasculitis Activity Score and semi-quantitative scales of WMH. The initial serum GM-CSF levels were associated with the severity of WMH in MPA patients.

Assessment of molecular response after tyrosine kinase inhibitors & serum granulocyte-macrophage colony-stimulating factor levels in Egyptian patients with chronic myelogenous leukemia in chronic phase

BackgroundIn chronic myelogenous leukemia (CML), one of the most important & documented benefits of the profitable use of tyrosine kinase inhibitors (TKIs) is achieving major molecular response (MMR) & undoubtedly raising the life expectancy. We evaluate the molecular response after one year of 32 newly diagnosed Egyptian CML patients in chronic phase receiving TKIs (imatinib vs nilotinib). Quantitative assessment of BCR-ABL1 mRNA transcripts using real-time reverse transcription polymerase chain reaction was done. Also, baseline quantitative detection of serum GM-CSF level was done using enzyme linked immunosorbent assays technique together with correlation to molecular response. ResultsThirty-two patients were included in the study, but the molecular response was assessed in only 17 patients; 6 patients were under imatinib treatment and 11 patients received nilotinib. Overall within 1 year, 7/17 (41.2%) patients achieved major molecular response (MMR) with median BCR-ABL1 level of 0.06% on the international scale (IS). Five patients (29.4%) reached MR4 with median BCR-ABL1 level 0.007% on IS. Regarding TKIs therapy, each MMR and MR4 was noticed in 2/6 (33.3%) patients who received imatinib treatment with median BCR-ABL1 level of 0.01% & 0.005% on the IS, respectively. However, MMR and MR4 were detected in 5/11 (45.4%) & 3/11 (27.3%) patients who received nilotinib treatment, respectively with median BCR-ABL1 level 0.06% & 0.008% on the IS, respectively. Median serum GM-CSF level was higher in patients who reached MR4 than patients who achieved MMR but it did not reach significant difference (p value = 0.7). ConclusionThis study demonstrates the molecular profile of Egyptian CML patients after TKIs treatment. Also, it assesses the baseline serum GM-CSF levels to predict therapeutic outcome in CML patients.

Polysaccharide of atractylodes macrocephala koidz activated T lymphocytes to alleviate cyclophosphamide-induced immunosuppression of geese through novel_mir2/CD28/AP-1 signal pathway

Polysaccharide Of Atractylodes Macrocephala Koidz (PAMK) has been proved to have anti-cancer, antitumor, anti-inflammation function and improve the immune level of the organism. The miRNA plays a very important role in regulating the immune function by negatively regulate the expression of target genes. To explore the molecular mechanism of PAMK active the lymphocytes, thirty 61-d-old geese were randomly divided into 4 groups (C, CTX, PAMK, PAMK+CTX). The thymus morphology, the level of serum granulocyte-macrophage colony-stimulating factor (GMC-SF), IL-1β, IL-3, IL-5, the relative mRNA expression of CD25, novel_mir2, CTLA4 and CD28 signal pathway were measured. Further more, the lymphocytes was extracted from thymus to measure the relative mRNA expression of CD28 signal pathway. The results showed that PAMK could significantly maintain normal cell morphology of thymus, alleviate the decrease level of GMC-SF, IL-1β, IL-5, IL-6, TGF-β, the increase level of IL-4, IL-10, and the decrease relative mRNA expression of novel_mir2, CD25 and CD28 signal pathway in thymus and lymphocytes induced by cyclophosphamide (CTX). In conclusion, PAMK alleviated the decreased T lymphocytes activation levels induced by CTX through novel_mir2/CTLA4/CD28/AP-1 signal pathway.

Mechanism of nano-indium-tin oxide inducing pulmonary alveolar proteinosis in Sprague-Dawley rats

Objective: To investigate the pathogenesis of pulmonary alveolar proteinosis in rats induced by nano-indium-tin oxide exposure, and to provide a basis for further determining the limit of occupational exposure to indium and developing related protection measures. Methods: In August 2018, a total of 40 specific pathogen-free Sprague-Dawley rats, with an age of 6-8 weeks and a body weight of (200±10) g, were randomly divided into control group, low-dose group (1.2 mg/kg) , middle-dose group (3 mg/kg) , and high-dose group (6 mg/kg) , with 10 rats in each group. After 1 week of routine feeding, the rats were given non-exposed intratracheal instillation twice every week, with an interval of 3 days, for 12 consecutive weeks. Body weight was measured every week during exposure to observe the change in body weight; The rats were anesthetized and sacrificed by chloral hydrate after the exposure ended, and lung tissue and serum were collected; Hematoxylin-eosin staining and periodic acid-Schiff (PAS) staining were performed for lung tissue to observe pathological results; Inductively coupled plasma mass spectrometry was used to measure the serum level of indium; ELISA was used to measure the levels of surfactant protein A (SP-A) , surfactant protein D (SP-D) , and the type II alveolar cell surface antigen Krebs von den Lungen-6 (KL-6) in lung tissue and the serum level of granulocyte-macrophage colony-stimulating factor (GM-CSF) . Results: The pathological results showed that the rats in the control group had basically complete alveolar structure, and after intratracheal instillation of nano indium-tin oxide, uniform, eosinophilic, and unstructured granular substances were observed in the alveolar space of the low-, middle-, and high-dose exposure groups, with macrophage proliferation and an increase in macrophages, especially in the high-dose group. Negative PAS staining was observed in the control group, while substances with positive PAS staining were observed in lung tissue in each exposure group. The three exposure groups had a significantly higher serum level of indium than the control group (P<0.05) . Compared with the control group, the three exposure groups had significant increases in SP-A, SP-D, and KL-6 in lung tissue and a significant reduction in GM-CSF in serum (P<0.05) . Conclusion: Pulmonary alveolar proteinosis in rats may be associated with the destruction of alveolar macrophages caused by nano-indium-tin oxide and the aggregation of pulmonary surfactants due to disorders in the metabolism and clearance of pulmonary surfactants by macrophages.

Pulmonary alveolar proteinosis: from classification to therapy.

Pulmonary alveolar proteinosis (PAP) is a rare respiratory syndrome characterised by the accumulation of surfactant lipoproteins within the alveoli. According to various pathogenetic mechanisms and aetiologies, PAP is classified as primary, secondary or congenital. Primary PAP is led by a granulocyte–macrophage colony-stimulating factor (GM-CSF) signalling disruption; the autoimmune form is driven by the presence of anti GM-CSF autoantibodies and represents 90% of all the PAP cases; and the hereditary form is the result of mutations in genes encoding GM-CSF receptor. Secondary PAP is associated with various diseases causing a reduction in function and/or number of alveolar macrophages. Congenital PAP emerges as a consequence of corrupted surfactant production, due to mutations in surfactant proteins or lipid transporter, or mutations affecting lung development. The clinical manifestations are various, ranging from insidious onset to acute or progressive respiratory failure, including premature death within the first days of life in neonates with congenital surfactant production disorders. The diagnostic workup includes clinical and radiological assessment (respiratory function test, high-resolution chest computed tomography), laboratory tests (anti-GM-CSF autoantibodies dosage, GM-CSF serum level and GM-CSF signalling test), and genetic tests. Whole-lung lavage is the current gold standard of care of PAP; however, the therapeutic approach depends on the pathogenic form and disease severity, including GM-CSF augmentation strategies in autoimmune PAP and other promising new treatments.Educational aimsTo update knowledge about a rare respiratory syndrome, pulmonary alveolar proteinosis, in order to promote early diagnosis and correct management.To highlight recent treatment options based on pathogenesis and disease severity.

Early predicting of preeclampsia in pregnant women after assisted reproductive technologies

Aim: to identify new prognostic criteria of potential preeclampsia (PE) in pregnant women after assisted reproductive technologies (ART) for timely PE prophylaxis.Materials and methods. A prospective study of 85 patients who entered the program of ART was conducted. All patients were examined for possible hemostatic abnormalities (genetic thrombophilia and chronic hypercoagulation) and also for granulocytemacrophage colony-stimulating factor (GM-CSF) in the serum during the most critical periods (4–6, 12–14, 22–24 и and 30–32 weeks) of the fetoplacental complex formation.Results. The lowest level of GM-CSF was observed in patients with hemostatic disorders. Thus, in pregnant women who later developed PE, there was a decrease in GM-CSF level below the physiological: in those diagnosed with genetic thrombophilia – by 79.4 %, and those with hypercoagulation – by 63.6 %.Conclusion. The determination of serum GM-CSF and identification of hemostatic abnormalities in pregnant women after ART has a prognostic importance for potential PE. This result is significant for the understanding of the pathogenesis of PE and also has a practical value: it allows the doctor to attribute the patient to a high risk group from the first trimester of her pregnancy and start preventive therapy rather early.

Synergistic and toxicity-attenuating effects of Periplaneta americana extract CII-3 combined with cisplatin on lewis lung cancer-bearing mice

Objective: The objective was to study the synergistic and attenuating effects of CII-3 combined with cisplatin. Materials and Methods: A Lewis tumor-bearing mouse model was established. After 15 days of continuous administration of CII-3 and cisplatin, the pathological changes in the tumor, liver, lung, and femur tissues were observed; the life prolongation rate, tumor inhibition rate, Q value, organ indices, spleen T- and B-lymphocyte proliferation activities, NK cell killing activity, the bone marrow cell proliferation rate and cell cycle phase, and the number of peripheral blood cells and bone marrow nucleated cells were measured. The expression of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in mouse serum and bone marrow tissue was measured. Results: The combination of CII-3 and cisplatin could enhance the ability of cisplatin to inhibit tumor cell proliferation and protects the liver damage and femoral injury and could significantly increase the life prolongation rate; tumor inhibition rate of cisplatin; the liver, spleen, lung, and thymus indices; the T- and B-lymphocyte proliferation activity; the NK cell killing activity; and the number of peripheral blood cells and bone marrow nucleated cells. The combination of drugs can stimulate the transformation of bone marrow cells from S phase to G2/M phase, significantly increase the proliferation rate of bone marrow cells and the contents of G-CSF and GM-CSF in mouse serum, and downregulate the mRNA and protein levels of them in bone marrow tissue. Conclusion: These results suggest that CII-3 combined with cisplatin can significantly enhance the antitumor effect and reduce the toxicity and side effects of cisplatin in Lewis tumor-bearing mice.