ABSTRACT
Objective:To investigate the significance of 12 inflammatory cytokines in early detection and treatment guidance of hematologic malignant patients with Cytokine release syndrome (CRS) after Chimeric antigen receptor (CAR)-T cell immunotherapy.Methods:A total of 12 patients, including 6 males and 6 females, aged 53.0 (49.8, 62.5) years old, were treated with CAR-T cell immunotherapy in the First Affiliated Hospital of Nanjing Medical University from 2017 to 2020. Cytometric bead array was used to detect the serum levels of IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, IFN-α, IFN-γ, and TNF-α at different time points after cell infusion in all patients receiving CAR-T cell immunotherapy. C-reactive protein (CRP), D-dimer (D-D), serum ferritin (SF), and lactate dehydrogenase (LDH) were measured at the corresponding period. CRS was classified into four grades according to the diagnostic criteria, from 0 to 3. The differences of the above mentioned parameters between the four groups were compared. The Speedman correlation coefficient was used to analyze the correlation between inflammatory cytokine expression levels and CRS grades. Plot the subject′s receiver operating characterist (ROC) curve to determine the sensitivity and specificity of inflammatory cytokines to predict CRS.Results:CRS grading was performed on day 1, 4, 7, and 11 after CAR-T cell infusion in 12 patients. There are 48 cases in total, including 25 cases of CRS grade 0, 6 cases of CRS grade 1, 9 cases of CRS grade 2, and 8 cases of CRS grade 3. The correlation analysis of 48 cases showed that the expression levels of IL-4, IL-6, IL-10, TNF-α, IFN-γ, IL-1β, and IL-8 were positively correlated with CRS grade ( P<0.05). The correlation coefficients were 0.384, 0.730, 0.632, 0.341, 0.681, 0.319, and 0.622, respectively. 7 inflammatory cytokines (IL-6, IL-10, IFN-γ, IL-8, IL-2, TNF-α, and IFN-α) were elevated in 12 patients, and the average time to start the rise was 3.4, 5.3, 6.1, 2.9, 4.3, 6.0 and 5.8 days, respectively. The time for CRP, D-D, SF, and LDH to begin to rise were 6.6, 7.6, 8.3 and 7.6 days, which were higher than that of the 7 inflammatory cytokines. After effective treatment, except for IL-6, the remaining 6 inflammatory cytokines (IL-10, IFN-γ, IL-8, IL-2, TNF-α and IFN-α) had their recovery times as 7.8, 3.9, 5.1, 8.0, 6.0, and 2.5 day,respectively, which were lower than that of CRP, D-D, SF, and LDH(9.7, 9.2, 13.7, and 13.8 days, respectively). The ROC showed that IL-6, IL-10, IFN-γ, and IL-8 can serve as biomarkers for diagnosis of CRS with high sensitivity and specificity. Conclusion:The monitoring of 12 inflammatory cytokines play an important role in CRS grading after CAR-T cell immunotherapy, which contributes to the early diagnosis of CRS and the prediction of clinical outcome.
ABSTRACT
Objective:To explore expressions of interleukin 6 (IL-6), interleukin 8 (IL-8) and interleukin 10 (IL-10) in the peripheral blood of patients with diffuse large B-cell lymphoma (DLBCL) and their clinical significances.Methods:The clinical data of 78 newly diagnosed patients with DLBCL from March 2018 to March 2021 in the First Affiliated Hospital of Xiamen University were retrospectively analyzed, and 58 healthy people receiving physical examination during the same period were taken as the healthy controls. The expressions levels of IL-6, IL-8 and IL-10 in peripheral blood were tested by using cytometric bead array (CBA), and the association of the levels of IL-6, IL-8 and IL-10 with clinical characteristics, disease staging and prognosis was analyzed.Results:The expression levels of IL-6, IL-8 and IL-10 in DLBCL group were higher than those in the healthy control group [(171.81±70.91) pg/ml vs. (2.71±0.28) pg/ml, (47.95±13.04) pg/ml vs. (3.69±0.47) pg/ml, (38.02±10.35) pg/ml vs. (1.77±0.23) pg/ml], and differences were statistically significant ( t values were 2.38, 3.39, 3.50, all P<0.05). In DLBCL group, the expression levels of IL-6, IL-8 and IL-10 in patient with bone marrow invasion, international prognostic index (IPI) 3-5 scores and clinical staging Ⅲ-Ⅳ were higher than those in patients with bone marrow non-invasion, IPI 1-2 scores and clinical stagingⅠ-Ⅱ(all P<0.05). There was a relationship between the expression levels of IL-6 and IL-8, IL-6 and IL-10, IL-8 and IL-10 in peripheral blood of DLBCL patients ( r2 value was 0.93, 0.89, 0.89, respectively; all P < 0.05). Among patients with high expressions of IL-6, IL-8 and IL-10, the proportion of patients not receiving remission after 6 cycles of treatment in clinical staging Ⅲ-Ⅳ was higher than that of patients with high expressions of IL-6, IL-8 and IL-10 alone or any two of them, and differences were statistically significant (all P < 0.05). Conclusions:There is a high correlation of IL-6, IL-8 and IL-10 expression levels; the increasing expression levels of them may predict the later disease stage and poor prognosis for DLBCL patients.
ABSTRACT
Background: Psoriasis is a chronic inflammatory skin disease characterized by hyperproliferation and incomplete differentiation of epidermis, and accumulation of neutrophils and proinflammatory T cells in epidermis and dermis. Chemokines are believed to be the main players mediating the chemotaxis of leucocytes to the lesional site. Previous studies have established the role of various chemokine ligands and receptors at the lesional site in psoriasis. Aims: In this study, we have compared the serum levels of various chemokines, namely, inducible protein-10 (IP-10) (CXCL10), MCP-1 (CCL-2), monokine induced by gamma interferon (MIG) (CXCL-9), RANTES (CCL5), interleukin (IL)-8, and eotaxin in patients with chronic plaque psoriasis with that of healthy controls. We also studied whether the chemokine levels varied within different patient groups based on various clinical and demographic parameters, and if any of these chemokines correlated with disease activity. Methods: We studied 40 patients with chronic plaque psoriasis from a single center. Their clinical and demographic details were recorded in predesigned prforma. Patients with unstable forms of psoriasis like guttate, erythrodermic, or pustular psoriasis were excluded. The serum chemokine levels were measured by flow cytometry–based bead array set system. The serum levels of the patients were compared with that of 25 healthy controls. A subgroup analysis was also done to study the correlation of chemokine levels with age, sex, duration, and severity of disease. Results: We observed a significant decrease in serum level of all these chemokines in patients, when compared with that of healthy controls. We also found that MIG levels showed a positive correlation with disease severity based on Psoriasis Area and Severity Index. Limitations: The major limitation of the study is lack of data on the lesional chemokine levels compared to serum chemokines. Conclusion: The inflammatory process in psoriasis is orchestrated through chemokines. MIG is a potential serum biomarker for assessing disease severity.
ABSTRACT
Objective: To investigate the effect of melatonin(MLT) on the expression of type Thl/Th2/Thl7 cytokines such as interferon(IFN)-Î3, tumor necrosis factor(TNF), interleukin(IL)-2,IL-4,IL-6,IL-10,IL-17a and so on of gastric cancer in vitro and in vivo. Methods: 1. Model of gastric cancer-bearing mice was established. Then 32 male 615 mice were all inoculated with murine foregastric carcinoma cells and randomly divided into 4 groups, and injected intraperitoneally with melatonin at doses of 0, 25, 50 and 100 mg/kg, and measureed long and short diameter of tumor, respectively. After a week of intervention, peripheral blood was taken and the tumor tissue was removed for weighing and measurement. 2. Murine foregastric carcinoma (MFC) cells were inoculated into a six-well cell culture plate and routinely cultured. After 24 hours of adherence, they were treated with melatonin at different concentrations of 0, 2, 4, 6, 8 and 10 mmol/L. After 24 hours again, the cells morpology were observed and the corresponding supematants were collected. 3. The expressions of melatonin concentrations in peripheral blood serum was detected by enzyme -linked imm unosorbent assay (ELISA). The expressions of Thl/Th2/Thl7 cytokines in peripheral blood serum and cell supematants were detected by cytometric bead array (CBA) respectively. Results: 1. Tumor-bearing mice models were successfully established. Compared with the negative control group, the melatonin concentration in peripheral blood serum of the middle and high dose MLT group which increased significantly, and the tumor volume significantly decreased. Compared with the negative control group, the concentration of 1L-10 in the middle dose group increased significantly. And the concentration of IFN-y, IL-2 and IL-10 in the high dose group increased significantly too. 2. Compared with the 0 mmol/L MLT group, the concentrations of IFN-Î3 in the 6 and 10 mmol/L MLT group were significantly decreased; the concentrations of IL-6 in the 4, 6, 8 and 10 mmol/L MLT group were significantly decreased, and the concentration of IL-10 in 6 mmol/L MLT group was significantly increased. All the difference were statistically significant (P<0. 05). Conclusion: Melatonin inhibits the proliferation of murine foregastric carcinoma cell MFC both in vitro and in vivo, and may enhance tumor immunity by adjust the expression of IFN-Î3, IL-2, IL-6 and IL-10 cytokines of type Thl/Th2/Thl7 cells.
ABSTRACT
Objective To appraise the analytical capability of flow cytometric bead array for lung cancer markers through the tests of limit of detection,relative standard deviation,specificity,methods comparation and linearity rang.Methods The limit of detection,relative standard deviation,specificity and linearity rang in detection of Carcinoembryonic antigen (CEA),cytokeratin 19 (Cyfra21-1) and neuron specific enolase (NSE) in serum were evaluated by flow cytometer.Western blotting method was ultilized to validate the specificity of antibody-antigen recognization.The interference of hemoglobin,three acyl glycerol and bilirubin on the detection of CEA,Cyfra21-1 and NSE was tested.Compared to electrochemiluminescence immunoassay,the relative error for flow cytometric bead array was assessed.Results Flow cytometric bead array demonstrated that the limit of detection was 1.71 pg/mL for CEA,3.97 pg/mL for cyfra21-1,and 2.27 pg/mL for NSE.The relative standard deviation for intra-assay and inter-assay were below 10％ and 15％,respectively.The pair of antibodies can defferentially recognize antigens.The measurement for CEACAM6,CK18,NSE appeared that there was no significant cross-talking reaction.Three acyl glycerol and bilirubin did not significantly interfere with the detection for serum samples.Hemoglobin of 500 ng/mL can significantly interfere with the detection of Cyfra21-1 (P ＜ 0.05) and NSE (P ＜ 0.05).The correlation coefficient between flow cytometric array and electrochemiluminescence immunoassay was 0.984 2 for serum CEA,0.962 2 for serum cyfra 21-1 and 0.982 0 for serum NSE.The linearity ranged from 355.76 pg/mL to 367.74 ng/mL for CEA,from 87.89 pg/mL to 107.8 ng/mL for cyfra21-1,and from 90.12 pg/mL to 86.07 ng/mL for NSE.Conclusion Flow cytometric array for lung cancer markers may be of use in clinical detection.
ABSTRACT
OBJECTIVETo study the role of interleukin 17 (IL-17) in the pathogenesis of chronic otitis media with effusion (COME).METHODSThe expression of IL-17 in middle ear effusion (MEE) and blood plasma were measured in 30 patients (48 ears) by means of Cytometric Bead Array (CBA), as well as in 20 normal volunteers.RESULTSCompared with the control group, the level of IL-17 significantly increased in the peripheral blood of COME patients (P<0.05). What was more, the level of IL-17 in the MEE was higher than that in peripheral blood of COME patients (P<0.05).CONCLUSIONIL-17, as an important immunoregulatory mediator, may play an important role in chronic course of COME.
ABSTRACT
Objective: To explore the significance of Th17 in hepatocellular carcinoma, expecially with HBV infection.Methods:Cytometric bead array(CBA) was employed to detect 5 cytokines(IL-2,IL-4,IL-6,IFN-γ,IL-17A)from 39 tumor and non-tumor tissues of HCC and combined clinical data for comparative statistic analysis.Results:The expression of IL-2,IL-4,IFN-γin liver cancer tissue[(4.61±0.28),(3.37±0.58),(3.08±1.08)pg/ml,respectively] was significant lower than non-cancer tissue [(5.57±0.59),(3.77±0.70),(3.69±1.20)pg/ml,respectively].Otherwise,the expression of IL-6,IL-17A in cancer tissue [(280.09±254.68), (2.66±1.66) pg/ml, respectively] was higher than non-cancer [(6.58 ±1.92), (1.49 ±0.98) pg/ml, respectively].And,whatever cancer or non-cancer tissue,the expression of IL-17A in tissue[(3.45±1.86)pg/ml] with high HBV load (>1 000 U/ml) was significant higher than tissue with low HBV load[(1.97±1.16)pg/ml].Conclusion: IL-17A was highly expressed in HCC,and IL-2,IL-4,IFN-γmay inhibit its expression,and IL-6 may promote it.Hepatitis B virus infection may promote Th17 expression,thereby reducing patient′s prognosis.
ABSTRACT
Objective To explore the changes of inflammatory factors in patients with different stage of rheumatoid arthritis (RA) ,and to study the changes of immune microenvironment in patients with RA .Methods The serum levels of tumor necrosis factor(TNF) ,interleukin‐10(IL‐10) ,interleukin‐6(IL‐6) ,interleukin‐1β(IL‐1β) and interleukin‐4(IL‐4) in patients with RA on ac‐tive stage(36 cases) ,patients with RA on remitting stage and healthy individuals (30 cases)were detected by using cytometric bead array .Results The serum levels of IL‐6 ,IL‐1βand TNF in active stage RA group were higher than those in control group and re‐mitting stage RA group ,while serum levels of IL‐4 and IL‐10 were lower than those in control group and remitting stage RA group , and the differences were statistically significant (P0 .05) .As the disease develops ,except for IL‐6 which tended to be stable , the serum levels of IL‐4 and IL‐10 had a rising trend ,while serum levels of IL‐1βand TNF had a downward trend with the progres‐sion of RA .Conclusion There is an imbalance between Th1 and Th2 cytokines in patients with RA on active stage ,in which Th1 cytokines are dominantly expressed .Periodic detection of inflammatory factors according to the course of RA could provide a relia‐ble basis for the assessment of disease activity .
ABSTRACT
Objective To study the effects of purified rabies vaccine for human use (RV) on specific Th1/Th2 cytokines in human. Methods Twenty cases were injected intramuscularly with 5 full doses of RV. PBMCs were isolated from the blood sample collected at day 0, 14, 45 after the RV inoculation. Neutralizing antibody was determined by ELISA, and the proliferation of lymphocyte by in vitro test. The levels of RV specific IFN-γ, TNF, IL-2,IL-4, IL-5, IL-10 in the culture supernatants were detected by cytometric bead array (CBA). Results The neutralizing antibody was tested positive in 19 cases 45 days after inoculation and 1 case after 60 days, with the positive rate reaching 100%. After stimulation with RV, the lymphocyte transformation index at day 14, 45 in cases were significantly higher than those day of 0 (P< 0.05), and similar results were confirmed with IFN-γ, IL-2, IL-4, IL-5 tested by CBA (P<0.05). Condusion The RV could induce humoral and antigen-specific cellular immune responses in human, tested by showing good protective effect on rabies virus.
ABSTRACT
Chemokines recruit and activate leukocytes, assisting granuloma formation. Herein, we evaluated plasma chemokines in patients with active tuberculosis (ATB) and after completing treatment (TTB) and compared them to BCG-vaccinated healthy controls (HC). Levels of chemokines were measured by cytometric bead array. Levels of CXCL8, CXCL9 and CXCL10 were higher in ATB patients compared to HC, but they decreased in TTB. Levels of CCL2 and CCL5 in ATB patients were similar to those observed in HC. Thus, the high levels of CXC-chemokines detected during ATB, which can modulate the trafficking of immune cells from the periphery to the site of infection, were reversed by anti-mycobacterial treatment.
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Antibiotics, Antitubercular/therapeutic use , Chemokines, CXC/blood , Tuberculosis, Pulmonary/blood , Tuberculosis, Pulmonary/drug therapy , BCG Vaccine , Case-Control Studies , Chemokines, CXC/analysis , Flow Cytometry/methods , Statistics, Nonparametric , Time Factors , Treatment Outcome , Young AdultABSTRACT
Objective To detect the level of Tnl and T.2 type cytokines in the patients with lym-phoma in order to find out the laboratory evidence of tumor immunotherapy. Methods The levels of serum IFN-γ, TNF-α, IL-2, IL-4, IL-5 and IL-10 were measured by cytometric bead array (CBA) in 92 patients with lymphoma, and 70 normal sera as control. Results The levels of TH1 type cytokincs in 92 patients with lymphoma were: IFN-γ (34.26 ± 33.48) pg/ml, TNF-α (8.17 ± 10. 09) pg/ml, IL-2 (3.74 ±1.72) pg/ml; and the levels of TH2 type cytokines were: IL-10 (6. 28±8.56) pg/ml, IL-5 (3.53 ±3.20) pg/ml, IL-4 (6.22±7.13) pg/ml. The levels of TH1 and TH2 cytokines in lymphoma patients were significantly higher than those in controls(P ＜ 0.01) except TNF-α. And the rate of IL-2/IL-4 was signifi-cantly decreased in lymphoma patients(P ＜0.01). The level of IL-10 in Ⅲ/Ⅳ stage lymphoma patients was much higher than that in Ⅰ/Ⅱ stage patients(P ＜0.01). The level of IFN-γ/was significantly decreased in aged patients with lymphoma (P ＜0. 05). Conclusion TH 1/TH2 is imbalance in lymphoma, which may provide clinical index for the evaluation of progrossien and prognosis. In the lymphoma patients TH1/TH2 shifts to TH2, which may be the mechanism of tumor arising and transferring by immune escape from immu-nosurveillance.