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Objective:To investigate the correlation of CD8 positive tumor-infiltrating lymphocytes (CD8 + TIL) density and programmed-death receptor ligand 1 (PD-L1) expression in rectal cancer with clinicopathological characteristics and prognosis of patients after neoadjuvant chemoradiotherapy. Methods:The clinicopathological data of 166 patients with locally advanced rectal cancer (LARC) who received neoadjuvant therapy before surgery in the Beijing Chao-Yang Hospital, Capital Medical University from January 2015 to December 2018 were retrospectively analyzed. CD8 + TIL density and PD-L1 expression were detected by using immunohistochemistry. The correlation of CD8 + TIL density and PD-L1 expression with clinicopathological characteristics of patients after neoadjuvant chemoradiotherapy was analyzed. Kaplan-Meier method was used to analyze the disease-free survival (DFS) and Cox regression risk model was used to make univariate and multivariate analysis of the influencing factors for DFS. Results:Among 166 LARC patients, 81 cases (48.8%) had high density of CD8 + TIL, 85 cases (51.2%) had low density of CD8 + TIL; 63 cases (38.0%) had PD-L1 expression, and 103 cases (62.0%) had non-expression of CD8 + TIL. The expression rate of PD-L1 in CD8 + TIL high density group was higher than that in CD8 + TIL low density group [50.6% (41/81) vs. 25.9%(22/85), χ2 = 10.78, P < 0.001]. According to the density of CD8 + TIL and PD-L1 expression, immunophenotype was divided among 4 groups; the 3-year DFS rate of the CD8 + TIL high density /PD-L1 expression group was 87.1%, which was higher than that of the other groups (CD8 + TIL low density /PD-L1 expression group was 72.8%, CD8 + TIL high density /PD-L1 non-expression group was 67.0%, CD8 + TIL low density /PD-L1 non-expression group was 64.3%), and the difference was statistically significant ( P < 0.05). Univariate analysis showed that tumor differentiation degree, TNM stage, CD8 + TIL density, PD-L1 expression and CD8 + TIL density /PD-L1 expression were correlated with the DFS of patients (all P < 0.05). Multivariate analysis results showed that CD8 + TIL high density /PD-L1 expression was an independent protective factor for DFS ( HR = 0.049, 95% CI 0.005-0.497, P = 0.011), while TNM stage 3 was an independent risk factor for DFS ( HR = 2.752,95% CI 1.300-5.825, P = 0.008). Conclusions:In LARC after neoadjuvant therapy, CD8 + TIL density is positively correlated with the expression of PD-L1, and the high density of CD8 + TIL/PD-L1 expression is an independent influencing factor for good prognosis, suggesting that these patients may benefit from the immunotherapy.
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Objective:To develop the anti-CD30 monoclonal antibody 64Cu-1, 4, 7-trizacyclononane-1, 4, 7-triacetic acid (NOTA)-CD30 and visualize CD30 expression in lymphoma non-invasively. Methods:The CD30 expression levels of 5 cell lines (Karpas299, Raji, Daudi, Ramos, and U266) were assessed by Western blot. Cell lines with high and low CD30 expression were selected for flow cytometry to evaluate the specific binding affinity of anti-CD30 monoclonal antibody. Thirteen NSG mice were used to established CD30 positive and negative subcutaneous xenograft models. 64Cu-NOTA-CD30 was obtained and 64Cu-NOTA-immunoglobulin (Ig)G was used as the control. ImmunoPET imaging was performed 2, 24, and 48 h after the injection of 64Cu-NOTA-CD30 or 64Cu-NOTA-IgG. Finally, the biodistribution studies were conducted. Repeated-measures analysis of variance and Bonferroni test were conducted for comparison. Results:Karpas299 showed the highest CD30 expression, while Raji showed the lowest. Flow cytometry showed specific binding affinity of the anti-CD30 monoclonal antibody to the Karpas299 cell line. The radiochemical purities of the probes were both higher than 95%. In microPET, the 64Cu-NOTA-CD30 uptake of Karpas299 xenograft tumors increased over time, with (11.46±0.58), (17.60±1.16) and (19.46±0.99) percentage activity of injection dose per gram of tissue (%ID/g) at 2, 24 and 48 h respectively. The contrast to normal tissue was good at 48 h, with the tumor/heart (blood) ratio of 2.20±0.22. The uptake of 64Cu-NOTA-CD30 in Karpas299 tumor at 48 h after injection was significantly higher than that in Raji tumor ((6.10±1.03) %ID/g) and 64Cu-NOTA-IgG in Karpas299 tumor ((5.12±0.89) %ID/g; F=290.99, t values: 19.65 and 22.25, all P<0.001). The uptake of 64Cu-NOTA-CD30 and the control probe in the heart and liver decreased over time in all groups. Ex vivo biodistribution at 48 h was mainly consistent with the results of microPET in vivo. Conclusions:64Cu-NOTA-CD30 is able to visualize the expression level and distribution of CD30 non-invasively. It is promising to be applied for screening the beneficial groups and evaluating efficacy for CD30-targeted immunotherapy.
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Objective:To explore the efficacy and safety of blinatumomab in treatment of relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL).Methods:The data of 8 patients with relapsed/refractory B-ALL treated with blinatumomab in Shanghai Zhaxin Traditional Chinese and Western Medicine Hospital from September 2020 to December 2021 were retrospectively analyzed, and their clinical characteristics, overall survival, lymphocyte subsets, cytokines, tandem transplantation and adverse reactions were analyzed.Results:The median follow-up time of 8 patients was 143 d (range: 41-534 d). Five of the 8 patients were alive; among them, 4 of 6 patients assessed to be in minimal residual disease (MRD)-negative complete remission (CR) and 1 of 2 patients assessed to be in non-remission at the time of belintuzumab discontinuation were alive. The median duration of treatment with belintuzumab was 28 d (10-56 d), and it was 23 d (10-56 d) for patients with MRD-positive at baseline and 28 d (25-31 d) for the 4 non-remission patients. Six patients achieved MRD-negative CR after treatment, of which 4 were assessed as MRD-positive at baseline and 2 were assessed as non-remission at baseline. All 4 patients with MRD-positive CR achieved MRD-negative CR after treatment with belintuzumab, including 1 patient with Philadelphia chromosome-positive (Ph +) ALL bridged to autologous hematopoietic stem cell transplantation, and 1 patient with Ph + ALL and 1 patient with Ph - ALL received sequential allogeneic hematopoietic stem cell transplantation and had persistent MRD-negative CR. Two of the 4 non-remission patients achieved MRD-negative CR after treatment with belintuzumab, including 1 patient with Ph + ALL bridged to autologous hematopoietic stem cell transplantation, and 1 patient with Ph - ALL received sequential allogeneic hematopoietic stem cell transplantation, and the 2 patients had persistent MRD-negative CR. Leukocyte counts and neutrophils decreased in both MRD-positive CR and non-remission patients after receiving belintumomab. The proportion and absolute number of CD3 + T and CD3 + CD8 + T lymphocytes in patients with MRD-positive CR were higher than those in patients without remission, and both decreased after drug administration. Median interleukin-6 (46.23, 1.42 pg/ml), interleukin-8 (17.85, 2.10 pg/ml), interleukin-10 (7.43, 1.49 pg/ml) and interferon-γ (11.82, 0.39 pg/ml) levels were elevated in MRD-positive CR and non-remission patients at week 3 of treatment. Grade 1 cytokine release syndrome occurred in 1 case with clinical manifestations of fever, which improved after drug suspension. Three cases developed infections, 2 of which were pulmonary and 1 of which was upper respiratory tract infection. No immune effector cell-associated neurotoxic syndrome was observed. Conclusions:Belintumomab is effective for MRD clearance in relapsed/refractory B-ALL with manageable adverse reactions, providing an effective therapeutic option for bridging hematopoietic stem cell transplantation to prolong the survival of patients.
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Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.
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Objective:To investigate clinicopathological features and prognosis of transformed mycosis fungoides (TMF) .Methods:A retrospective analysis was performed on clinicopathological data collected from 24 patients with TMF, as well as on flow cytometry results of 16 peripheral blood samples obtained from 11 of the 24 patients, who visited Hospital of Dermatology, Chinese Academy of Medical Sciences between 2014 and 2020.Results:Among the 24 patients, 11 were males and 13 were females. Their average age at diagnosis of TMF was 50.0 years (range: 18 - 77 years), and patients with early-stage TMF (9 cases) and tumor-stage TMF (15 cases) were aged 44.8 and 52.6 years on average, respectively. The average time interval from diagnosis of MF to large cell transformation was 3.7 years, and 8 patients were diagnosed with TMF at the initial visit. Histopathologically, large cells infiltrated in a diffuse pattern in 20 cases, as well as in a multifocal pattern in 4, and the proportion of large cells in 7 cases was greater than 75%. Immunohistochemically, 18 patients showed positive staining for CD30, and the proportion of CD30-positive large cells was greater than 75% in 9; negative staining for CD30 was observed in 6. Flow cytometry of 16 peripheral blood samples showed the presence of cell subsets expressing clonal T cell receptor (TCR) -vβ in 2 of 4 patients with early-stage TMF and 10 of 12 with tumor-stage TMF, and tumor cells with higher forward scatter than normal lymphocytes were detected in 16 samples. During the follow-up, among the patients with early-stage TMF, 3 progressed to tumor-stage TMF 3.3 years on average after large cell transformation, 1 progressed to erythrodermic MF in stage IIIA, and the other 4 still showed an indolent course; among the patients with tumor-stage TMF, 1 progressed to stage-IV TMF, and 5 died 3.3 (1.5 - 6) years after large cell transformation.Conclusion:Large cell transformation may occur in patients with MF in any stage, some patients have poor prognosis, so close follow-up is needed for patients with TMF.
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Objective:To prepare 89Zr labeled Daratumumab and evaluate its feasibility in the imaging diagnosis of multiple myeloma (MM). Methods:According to the principle of 89Y (p, n) 89Zr nuclear reaction, 89Zr was produced by cyclotron solid target system (30 μA, 1.5 h) and automatic purification module. The radionuclide purity, half-life and impurity metal ion concentration were detected. Desferrioxamine (DFO) was coupled with Daratumumab and then chelated with 89Zr to prepare 89Zr-DFO-Daratumumab. The quality control analyses of three consecutive batches were carried out. Pharmacokinetic evaluation and 89Zr-DFO-Daratumumab microPET/CT imaging were performed in normal rabbits and orthotopic myeloma mouse models, respectively. The SUV in situ myeloma and that in normal bone were compared by independent-sample t test. Results:About 560 MBq of 89Zr was obtained, and there were only two characteristic energy peaks of 89Zr (909 keV and 511 keV) by γ spectrometer. The half-life of 89Zr was 78.2 h, and the content of metal impurities was small. 89Zr-DFO-Daratumumab was prepared with pH of 7.2, radiochemical purity of more than 99%, good stability in vitro, and sterility and endotoxin tests were passed. Pharmacokinetic studies in rabbits showed that 89Zr-DFO-Daratumumab was gradually distributed from blood to liver, spleen, kidney and bone joints over time and metabolism. The results of microPET/CT imaging in orthotopic myeloma mouse models showed that the SUVs of 89Zr-DFO-daratumumab in situ myeloma were significantly higher than those in normal bone (2 h: 0.22±0.02 vs 0.06±0.00; 1 d: 0.38±0.01 vs 0.08±0.00; t values: 8.89, 21.90, both P=0.001). Conclusion:89Zr and 89Zr-DFO-daratumumab are successfully prepared, and relevant quality control and biological evaluation in vivo and in vitro are completed, which verify the feasibility of 89Zr-DFO-Daratumumab in the imaging diagnosis of MM, thus laying a foundation for clinical transformation.
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Doenças autoimunes são doenças universais, e os diagnósticos e tratamentos primários são habitualmente iniciados por clínicos em enfermarias ou ambulatórios, antes de serem encaminhados a especialistas. Além disso, pacientes em uso de biológicos internados em hospitais gerais têm sido cada vez mais frequentes na prática clínica. Conhecer o perfil de segurança, as indicações e os efeitos colaterais dessas drogas deve ser preocupação dos clínicos. Neste trabalho, foi realizada revisão de literatura sobre terapia biológica com rituximabe no tratamento das principais doenças autoimunes sistêmicas da prática clínica: artrite reumatoide, lúpus eritematoso sistêmico, vasculites relacionadas aos anticorpos anticitoplasma de neutrófilo, púrpura trombocitopênica imune e espondilite anquilosante. (AU)
AutoimmunAutoimmune diseases are universal diseases and primary diagnosis and treatment are typically initiated by internists in wards or outpatient clinics before being referred to specialists. In addition, patients on use of biologicals hospitalized in general hospitals have been increasingly common in clinical practice. Knowing the safety profile, the indications, and the side effects of these drugs should be a concern for the internists as well. In this study, the literature review was performed on biological therapy with Rituximab for treating the main systemic autoimmune diseases of clinical practice: rheumatoid arthritis, systemic lupus erythematosus, anti-neutrophil cytoplasmic antibody-associated vasculitides, immune thrombocytopenic purpura, and ankylosing spondylitis. (AU)
Subject(s)
Humans , Autoimmune Diseases/drug therapy , Rituximab/therapeutic use , Immunologic Factors/therapeutic use , Antibodies, Monoclonal/therapeutic use , Spondylitis, Ankylosing/drug therapy , Immunoglobulins/drug effects , B-Lymphocytes/drug effects , Antigens, CD20/drug effects , Rituximab/pharmacologyABSTRACT
Chimeric antigen receptor T (CAR-T) cell has achieved excellent efficacy in hematological tumors, especially for lymphoma. Many products have been approved to market all over the world, and 2 products targeting CD19 have been approved to treat relapsed and refractory large B-cell lymphoma in China. The current experiences of using CAR-T cells come from previous clinical studies. How to use CAR-T cells in a standardized and rationalized way is still a challenge faced by our clinicians. Based on the CAR-T cell treatment experiences from Peking University Cancer Hospital and the latest research progresses in CAR-T in China and abroad, this article will elaborate on patient screening, peripheral blood mononuclear cell collection, bridging treatment, lymphocyte depletion chemotherapy, CAR-T cell infusion, the monitoring and treatment of adverse events after infusion, and long-term follow-up after infusion, in order to guide clinicians to better use CAR-T cell and to bring maximum benefits to patients.
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Objective:To investigate the expressions of CD200 and inducible costimulator (ICOS) protein in angioimmunoblastic T-cell lymphoma (AITL) and the relationship with prognosis as well as their significances in the differential diagnosis of AITL.Methods:A total of 39 AITL patients in the First People's Hospital of Chenzhou, the Fourth People's Hospital of Chenzhou, Xiangnan College Affiliated Hospital and Chenzhou 3rd People's Hospital from June 2012 to December 2019, and 10 patients with classic Hodgkin lymphoma (CHL) and 10 patients with peripheral T cell lymphoma, not otherwise specified (PTCL-NOS) from August 2016 to July 2019 in the First People's Hospital of Chenzhou were selected. Immunohistochemistry was used to detect the expressions of CD200, ICOS, CD10, programmed death 1 (PD-1), bcl-6 and CXC chemokine receptor-13 (CXCL13) proteins, and the correlation of CD200 and ICOS with clinicopathological features and prognosis of AITL patients was analyzed, and the diagnostic significance of both in differentiating AITL from PTCL-NOS and CHL was also analyzed.Results:The positive rates of CD200 and ICOS in 39 AITL patients were 71.79% (28/39) and 61.54% (24/39), respectively. There were 7 cases of CD200 weak and moderate positive in 10 CHL patients, and ICOS proteins were all negative. Among 10 PTCL-NOS patients, 4 patients had CD200 positive and 1 patient had ICOS positive. The differences in positive rates of ICOS protein between AITL patients and CHL, PTCL-NOS patients were statistically significant (all P < 0.05); the differences in positive rates of CD200 protein between AITL patients and CHL, PTCL-NOS patients were not statistically significant ( χ2=0.013, P=0.911; χ2=3.551, P=0.060). The positive rate of CD200 in AITL patients with elevated lactate dehydrogenase (LDH) and international prognostic index (IPI) score of 3-4 was higher than that in AITL patients with normal LDH and IPI score of 0-2 (both P < 0.05); The positive rate of ICOS in AITL patients with elevated LDH and PD-1 positive was higher than that in AITL patients with normal LDH and PD-1 negative (both P < 0.05). CD200 negative AITL patients had better 3-year overall survival (OS) rate (4.2% vs. 66.7%) and 3-year progression-free survival (PFS) rate (5.3% vs. 77.1%) compared with those in CD200 positive AITL patients, and the differences between both groups were statistically significant (both P < 0.01); there was a statistically significant difference in 3-year OS rate between ICOS positive AITL patients and ICOS negative AITL patients (15.3% vs. 38.6%, P=0.011), while there was no statistically significant difference in 3-year PFS rate of both groups (18.6% vs. 41.5%, P=0.059). Multivariate analysis showed CD200 ( HR=0.076, 95% CI 1.555-79.497, P=0.001), extranodal involvement or not ( HR=11.117, 95% CI 1.555-79.497, P=0.016) and LDH ( HR=2.147, 95% CI 0.844-5.459, P=0.109) were independent influencing factors of OS in AITL patients; CD200 ( HR=0.075, 95% CI 0.016-0.357, P=0.001) and LDH ( HR=2.335, 95% CI 0.929-5.870, P=0.071) were independent influencing factors of PFS in AITL patients. Conclusions:CD200 and ICOS can be used as immunohistochemical indicators to assist the diagnosis of AITL patients. ICOS protein helps to differentiate AITL from CHL and PTCL-NOS; CD200 can be used as indicators to judge the prognosis and deterioration of AITL patients.
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Objective:To investigate the expression of programmed cell death protein 1 (PD-1), programmed cell death ligand 1(PD-L1), and lymphocyte-activation gene 3(LAG-3) in different subsets of lymphocytes and their relationship with the immunologic imbalance in preeclampsia.Methods:We enrolled 25 cases of singleton pregnant women with preeclampsia who were delivered by cesarean section in the Shandong Provincial Hospital Affiliated to Shandong First Medical University from May 2019 to January 2020 as the preeclampsia group. According to the allocation ratio of 1∶1 matched for the date of cesarean section and pregnancy week at delivery, another 25 healthy singleton pregnant women underwent elective cesarean section were selected as the normal group. The decidua tissue was obtained during cesarean section. The expression levels of PD-1, PD-L1, and LAG-3 on decidual T cells, natural killer (NK), and natural killer T (NKT) cells were measured by flow cytometry and compared between the two groups using two independent samples- t test. Results:(1) The expression of PD-1 on decidual T cells and NK cells of the preeclampsia group were lower than those of the normal group (37.84±3.82 vs 57.02±3.89, t=3.529, P<0.001; 3.28±0.48 vs 5.69±0.99, t=2.184, P=0.034), but did not differ significantly in the expression on decidual NKT cells ( P=0.461). PD-L1 expression on decidual NK cells of preeclampsia group was lower than that of the normal group (0.60±0.11 vs 1.32±0.19, t=3.319, P=0.002), but showed no significant difference in the expression level on T cells and NKT cells (both P>0.05). The preeclampsia group was noted for a lower expression of LAG-3 on decidual T cells and NKT cells compared with the normal group (2.32±0.36 vs 4.09±0.67, t=2.335, P=0.024; 35.40±4.97 vs 56.27±4.49, t=3.282, P=0.002), while showed no significant difference in the expression level of NK cells ( P=0.112). Conclusions:The decreased expression of PD-1, PD-L1, and LAG-3 in the decidual lymphocyte subsets may be involved in the immunologic imbalance of preeclampsia through the over-activation of immunocytes at the maternal-fetal interface.
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Objective:To investigate the efficacy and safety of infliximab in the treatment of severe plaque psoriasis and its effect on the expression of programmed cell death-1 (PD-1) and programmed cell death ligand-1 (PD-L1) in psoriatic lesions.Methods:A total of 17 patients with severe plaque psoriasis were enrolled from Shanghai Skin Disease Hospital from February 2019 to April 2019, and were treated with intravenous drips of infliximab at a dose of 5 mg/kg at weeks 0, 2, 6, 14, 22, 30, 38 and 46. Efficacy was evaluated by using the psoriasis area and severity index (PASI) score at weeks 2, 6, 10, 14, 22, 30, 38, 46 and 52, and adverse events were recorded during the trial. Real-time PCR was performed to determine the expression of PD-1 and PD-L1 in skin tissues of 8 volunteer controls, as well as in skin lesions of 14 patients with plaque psoriasis before treatment and 5 patients with plaque psoriasis after 10-week treatment, and immunofluorescence assay to measure the expression of PD-1 and PD-L1 in skin tissues of 5 volunteers and 5 patients with psoriasis. The independent sample t-test was used to compare the expression of PD-1 and PD-L1 in skin tissues between the patients with plaque psoriasis and controls, and paired t-test to compare the expression of PD-1 and PD-L1 in the skin lesions of patients before and after infliximab treatment. Results:After 2, 6, 10, 14, 22, 30, 38, 46 and 52 weeks of infliximab treatment, the proportion of patients with plaque psoriasis achieving PASI75 was 1/17, 6/16, 9/16, 10/16, 15/15, 14/15, 13/14, 11/13 and 10/11, respectively. Antinuclear antibody staining turned positive in 12 patients, which was the most common adverse reaction, and 1 patient experienced an infusion reaction, which was the most severe adverse reaction. Before the treatment, the expression of PD-1 and PD-L1 (1.111 ± 0.391, 0.902 ± 0.169, respectively) was significantly higher in the skin lesions of patients with psoriasis than in the skin tissues of controls (0.620 ± 0.225, t=3.116, P=0.007; 0.474 ± 0.360, t=3.208, P=0.006, respectively) ; after infliximab treatment, the expression of PD-1 and PD-L1 (0.570 ± 0.230, 0.150 ± 0.050, respectively) in the improved skin lesions was significantly lower than that in the corresponding lesions before the treatment (1.238 ± 0.414, t=3.107, P=0.036; 0.966 ± 0.184, t=8.423, P=0.001, respectively) . Conclusions:Infliximab is effective and safe for the treatment of plaque psoriasis, but monitoring is necessary during treatment. The expression of PD-1 and PD-L1 is aberrantly upregulated in plaque psoriasis lesions, and decreased after infliximab treatment, suggesting that PD-1/PD-L1 may be involved in inflammation regulation in psoriasis.
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Objective:To prepare a 68Ga labeled probe targeting integrin alpha M(CD11b) receptor, namely 68Ga-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid-Glycine-Arginine-Glutamate-Arginine-Glutamate-polyethylene glycol 11-1, 2, 4, 5-terazine/CD11b antibody-F(ab′) 2-trans-cyclooctene ( 68Ga-NOTA-Polypeptide-PEG 11-Tz/anti-CD11b-F(ab′) 2-TCO), and to explore its feasibility as a molecular probe for CD11b receptor through microPET imaging. Methods:Immunofluorescence was used to detect the expression of CD11b on the surface of RAW264.7 cell. CD11b specific monoclonal antibody (M1/70) was conjugated with TCO, and anti-CD11b-F(ab′) 2-TCO fragment was obtained. The ligand NOTA-Polypeptide-PEG 11-Tz was labeled with 68Ga, and its specific activity and radiochemical purity were detected. Pre-targeted cell binding experiment was conducted to evaluate the binding ability of molecular probe. CT26 colon cancer bearing mouse models were established, and then pre-targeted biodistribution and imaging experiments were performed. Immunohistochemical experiment was used to verify the expression of CD11b receptor in tumor. The one-way analysis of variance was used to compare the data. Results:The results of immunofluorescence demonstrated CD11b receptor was highly expressed on the surface of RAW264.7 cell. Anti-CD11b-F(ab′) 2-TCO fragment was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). 68Ga-NOTA-Polypeptide-PEG 11-Tz was successfully synthesized, with the labeling efficiency of 94.6%. The specific activity was 7.0-7.4 MBq/μg, and the radiochemical purity was higher than 95%. Pre-targeted cell binding experiment confirmed that the molecular probe bound to the CD11b receptor. The biodistribution and imaging experiments showed that the kidney radioactivity uptake was high at pre-targeted 4, 12 and 24 h intervals, which proved that probe was excreted through the urinary system. In addition, molecular probe had higher radioactive uptake at the tumor site, with the tumor/muscle ratios of 9.23±1.45, 12.53±1.36 and 10.74±1.11 ( F=848.8, P<0.05). When the radioligand was injected 1 h after the pre-positioned 12 h interval, the images contrast was the best, with the standardized uptake value (SUV) in tumor and muscle of 0.67±0.12, 0.09±0.04, respectively. Immunohistochemistry verified the highly expression of CD11b receptor in tumor. Conclusions:The pre-targeted molecular probe 68Ga-NOTA-Polypeptide-PEG 11-Tz/anti-CD11b-F(ab′) 2-TCO is successfully synthesized. The molecular probe has targeting ability for CD11b + colon cancer, and is expected to be used as a tracer targeting CD11b receptor in vivo.
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ObjectiveTo investigate the main clinical influencing factors for the rate of active immune response to hepatitis B vaccine in adult hepatitis B recipients after liver transplantation. MethodsAnalysis was performed for the clinical follow-up data of 15 hepatitis B recipients after liver transplantation who received hepatitis B vaccine in Peking University International Hospital from May 2019 to November 2020, and all patients received liver transplantation at least 3 years before and had a CD4 level of 300-800 cells/u before vaccination. Each dose of vaccination was 40 μg recombinant hepatitis B vaccine (Saccharomyces cerevisiae), with a total of 4 injections at 0, 1, 6, and 8 months. Anti-HBs ≥100 mIU/L after four injections which lasted for 12 weeks without attenuation was considered successful response. Pearson correlation analysis and Kendall correlation analysis were used to investigate the correlation between CD4 level before vaccination and vaccine response rate; a linear regression analysis was used to investigate whether CD4 level before vaccination could predict the titer of anti-HBs after active vaccine immunization; a logistic regression analysis was used to investigate whether CD4 level before vaccination could predict vaccine response. ResultsOf all patients at week 12 of monitoring, 6 patients had response, among whom 1 had an anti-HBs level of ＞1000 mIU/L and 5 had an anti-HBs level of ≥100 mIU/L, and the antibody titer did not attenuate till week 16; the response rate of hepatitis B vaccine was 40%. The 6 patients with response had a mean CD4 level of ≥592 cells/u before vaccination, while the 9 patients without response had a mean CD4 level of ≤500 cells/u before vaccination. CD4 level before vaccination was strongly correlated with the response rate of hepatitis B vaccine (Pearson correlation analysis: r=0.767, P=0.001; Kendall correlation: r=0.717, P=0.001). ConclusionCD4 level before vaccination is a key clinical factor affecting the response rate of hepatitis B vaccine after liver transplantation.
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Abstract: Inflammatory periapical lesions are characterized by infiltration of different immune cell types, the functions of which depend on an effective vascular network. This study aimed to evaluate the mast cells density (MCD) in inflamatory odontogenic cysts capsules concerning microvascular density (MVD), microvascular area (MVA), and microvascular perimeter (MVP), and correlate such findings with the type of lesion, intensity of the inflammatory infiltrate, and thickness of the epithelial lining. Twenty inflamatory dentigerous cysts (IDCs), twenty radicular cysts (RCs), and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using anti-tryptase and anti-CD34 antibodies. RCs exhibited the highest MCD, MVD, MVA, and MVP indexes (p = < 0.001, p = 0.008, p = 0.003 and p = < 0.001, respectively), and lesions with inflammatory infiltrate grade III showed the highest MVD (p = 0.044). Considering epithelial thickness, a higher MVP index was identified in lesions with hyperplastic epithelium (p = 0.018). In IDCs, RCs, and RRCs, a strong positive correlation was observed between MVA and MVP (r = 0.950 and p = < 0.001; r = 0.914 and p = < 0.001; r = 0.713 and p = < 0.001, respectively). In IDCs, a moderate correlation was observed between MCD and both MVA and MVP (r = 0.660 and p = 0.002; r = 0.634 and p = 0.003, respectively). These results suggest that tryptase-positive mast cells might play an important role in the angiogenic activity of IDCs, while RCs had the highest indexes. Our findings also confirmed that the intensity of the inflammatory infiltrate and epithelial thickness influence angiogenesis.
Subject(s)
Humans , Odontogenic Cysts , Radicular Cyst , Epithelium , Tryptases , Mast CellsABSTRACT
Propósito de la revisión: el objetivo de la revisión es delinear la fisiopatología de los linfomas de estirpe B.Buscamos reportes endonde se incluyela descripción del origen de los Linfomas B para una mejor comprensión de esta patología, a la luz de los avances en las diferentes áreas. Recientes hallazgos: El Grupo Euroflow ha publicado una lista de paneles de Expresión de Antígenos de Superficie en Linfoma no Hodgkin, cuya lista se presenta en este artículo. Extracto: Las neoplasias hematológicas han tenido grandes avances en los últimos años en varios campos, evolucionando desde la identificación citológica, pasando por su caracterización inmunofenotípica por medio de la Citometría de Flujo e Inmunohistoquímica y llegando a la caracterización molecular, iniciando por Técnicas de Cariotipo Convencional, continuando por técnicas de Inmunohibridación in situ y actualmente con la identificación molecular por medio de la Secuenciación de Nueva Generación. Esta es la razón por la que los sistemas de estadificación han ido evolucionando también, siendo el que está al momento en vigencia el propuesto por la Organización Mundial de la Salud en el año 2016.Los linfomas constituyen un grupo heterogéneo de neoplasias hematológicas con un amplio espectro de presentación clínica, cuyo origen se encuentra en los precursores de linfoides y que afectan a los diversos órganos linfoides. De estos, los linfomas dela línea B son los más comunes, motivo de esta revisión
Purpose of the review: the objective of the review is to delineate the pathophysiology of B-line lymphomas. We are looking for reports that include a description of the origin of B-lymphomas for a better understanding of this pathology, in light of advances in the different areas. Recent Findings: The Euroflow Group has published a list of Surface Antigen Expression panels in Non-Hodgkin Lymphoma, the list of which is presented in this article. Extract: Hematological neoplasms have had great advances in recent years in several fields, evolving from cytological identification, passing through their immunophenotypic characterization through Flow Cytometry and Immunohistochemistry and reaching molecular characterization, starting with Conventional Karyotype Techniques , continuing with in situ Immunohybridization techniques and currently with molecular identification through New Generation Sequencing. This is the reason why staging systems have also evolved, the one currently in force being the one proposed by the World Health Organization in 2016. Lymphomas constitute a heterogeneous group of hematological neoplasms with a wide spectrum of clinical presentation, originating from lymphoid precursors and affecting the various lymphoid organs. Of these, line B lymphomas are the most common, which is the reason for this review
Subject(s)
Humans , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma , Antigens, CD20 , Flow Cytometry , Review , High-Throughput Nucleotide Sequencing , KaryotypeABSTRACT
O fibromixoma acral superficial é um tumor mesenquimal raro e benigno. Acomete principalmente homens de meia-idade; entretanto, pode ocorrer em qualquer sexo e faixa etária. Apresenta crescimento lento, com predileção por áreas ungueais e periungueais.
Superficial acral fibromyxoma is a rare and benign mesenchymal tumor. It mainly affects middle-aged men; however, it can occur in any gender and age group. It has a slow growth, with a preference for nail and periungual areas
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ObjectiveTo investigate the anti-liver fibrosis effect and mechanism of action of Kangxian Ruangan prescription based on hepatic sinusoidal capillarization. MethodsA total of 16 male Wistar rats were divided into Kangxian Ruangan prescription group and control group, with 8 rats in each group. The rats in the Kangxian Ruangan prescription group were given crude drug 40 g/kg body weight by gavage, and those in the control group were given normal saline by gavage. Serum containing Kangxian Ruangan prescription was prepared, and then primary liver sinusoidal endothelial cells were treated with the serum containing Kangxian Ruangan prescription or the control serum. Western blot was used to measure the protein expression of the vascular endothelial markers cluster of differentiation 31 (CD31) and von Willebrand factor (vWF), and immunocytochemistry was used to measure the expression of CD31 and vWF in cells. Scanning electron microscopy was used to observe the change in the fenestrae of liver sinusoidal endothelial cells. The t-test was used for comparison of continuous data between two groups. ResultsKangxian Ruangan prescription had no marked influence on the morphology of liver sinusoidal endothelial cells. The liver sinusoidal endothelial cells in the control group had a small number of narrow fenestrae with an occluded structure, while those in the Kangxian Ruangan prescription group had an increased number of large fenestrae with a clear structure. The results of immunocytochemistry showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the expression of CD31 (0.069±0015 vs 0.115±0.011, t=4.257, P＜0.05) and vWF (0.092±0.009 vs 0.132±0.014, t=4.274, P＜0.05). The results of Western blot showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the protein expression of CD31 (0.334±0.029 vs 0.448±0.013, t=6.245, P＜0.01) and vWF (0.560±0.047 vs 0709±0.045, t=3.966, P＜0.05). ConclusionKangxian Ruangan prescription can significantly resist hepatic sinusoid capillarization and thus prevent the progression of liver fibrosis.
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Objective@#To observe the change and clinical significance of the peripheral blood T-lymphocytes CD45RA+ and CD45RO+ in children with Henoch-Schonlein purpura (HSP) under different clinical classification.@*Methods@#From October 2015 to July 2017, the clinical data of 80 children with HSP in the Affiliated Hospital of North Sichuan Medical College were retrospectively analyzed.According to the clinical classification, they were divided into three groups: skin involvement group (35 cases), abdominal type group (36 cases), and renal type group (9 cases). Another 80 healthy children were selected as the control group.The changes and clinical significance of peripheral blood T-lymphocytes CD45RA+ , CD45RO+ and CD45RA+ /CD45RO+ ratio were analyzed.@*Results@#Among the 80 children, 40 cases were male and 40 cases were female, with age of (7.2±2.3)years old.The CD45RA+ and CD45RO+ rates in the HSP group were (13.19±7.09)%, (12.07±3.46)%, respectively, which were significantly lower than those in the control group [(23.26±6.01)%, (21.74±3.46)%], the differences were statistically significant (t=9.69, 16.42, all P<0.05). The CD45RA+ ratio, CD45RA+ count and CD45RA+ /CD45RO+ ratio of the kidney type group were (8.02±3.63)%, (2.19±0.33)/μL, (-0.28±0.19), respectively, which were significantly lower than those of the skin group [(15.74±7.71)%, (2.55±0.33)/μL, (0.27±0.12)], the differences were statistically significant(t=3.085, 2.709, 4.013, all P<0.05). The ratio of CD45RA+ /CD45RO+ in the abdominal group was significantly lower than that in the skin group[(-0.07±0.27)vs.(0.27±0.12), t=2.989, P<0.05].@*Conclusion@#Peripheral blood T lymphocyte subsets CD45RA+ and CD45RO+ may play a role in the generation and development of immune function changes in children with HSP, and the decrease of CD45RA+ T cells may be related to the risk of HSP nephritis.
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Objective To assess the role of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in regulation of interleukin(IL)-12 secretion by CD14+ peripheral blood monocytes from patients with psoriasis vulgaris.Methods From November 2017 to March 2018,a total of 47 patients with psoriasis vulgaris (psoriasis group) and 19 healthy volunteers (control group) were enrolled from Shanxi Provincial People's Hospital.Peripheral blood mononuclear cells (PBMC) were isolated,and stimulated with Toll-like receptor (TLR) ligands.TIM-3 expression and IL-12 secretion by CD14+ monocytes were measured by flow cytometry.After blockade of TIM-3 pathway by anti-TIM-3 neutralizing antibody,changes in the downstream signaling pathway molecules in and IL-12 secretion by CD14+ monocytes were investigated.Two independent samples t-test was used for comparison between two groups,and Pearson correlation test for correlation analysis.Results Under the unstimulated condition,the level of IL-12 secreted by CD14+ monocytes was very low,and the proportion of CD14+TIM-3+ cells was significantly higher in the psoriasis group (12.20％ ± 2.83％) than in the control group (9.91％ ± 1.77％,t =3.270,P =0.001 7).After the stimulation with TLR ligands,the proportion of CD14+IL-12+ cells was significantly lower in the psoriasis group (13.49％ ± 2.80％) than in the control group (28.97％ ± 8.97％,t =10.71,P ＜0.000 1),but the proportion of CD14+TIM-3+ cells was still higher in the psoriasis group (6.80％ ± 1.11％)than in the control group (4.85％ ± 1.37％,t =6.064,P ＜ 0.000 1).The proportion of CD14+TIM-3+ cells was negatively correlated with that of CD14+IL-12+ cells in both the control group and psoriasis group (r =-0.473,-0.371 respectively,both P ＜ 0.05).The TIM-3 pathway blockade could induce the decrease of suppressor of cytokine signaling 1 in CD14+ monocytes,increase the phosphorylation of signaling transducers and activators of transcription 1,and promote IL-12 secretion by CD 14+ monocytes induced by keratinocytes isolated from psoriatic skin lesions.Conclusion TIM-3 plays a crucial role in the negative regulation of CD 14+ monocyte-induced innate immune response in psoriasis.
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Objective To investigated the protective effect and immunology mechanism of Astragaloside Ⅳ(Ast) on rat renal ischemia-reperfusion injury model. Methods The SD rats were randomly divided into four group as the control, sham-operation, renal ischemia-reperfusion injury, and Ast treatment groups. Rat serum and urine were collected and detective for kidney function and interleukin cytokines. The kidney tissue was collected for histology exam. The rats in astragaloside group were intraperitoneally injected with 100 mg/kg astragaloside, and the other three groups were intraperitoneally injected with an equal volume of normal saline. The models of renal ischemia-reperfusion injury were prepared to generate in model and astragaloside groups, after 30 minutes of astragaloside injection. The rats with renal ischemia-reperfusion injury model were sacrificed after 24 hours, and the level of blood-urine creatinine, neutrophil gelatinase-associated lipocalin and Kidney damage molecule-1 were determined. The level of Th1 type cytokines (TNF-α, IFN-γ, IL-2) and Th2 type cytokines (IL-4, IL-5, IL-10) in serum were measured by using ELISA. The protein and gene expression of TNF-α, IFN-γ, IL-2, IL-4, IL-5, IL-10 in renal tissue were tested by western blot and PCR, respectively. The pathological changes and apoptosis of renal tissue in each group were detected by HE staining and TUNEL staining, respectively. The expression of CD20 protein in renal tissue was determined by immunohistochemistry. Results Compared with the model group, Ast treatment reduced serum creatinine (58.74 ± 9.44 μmol/L vs. 85.03 ± 23.48 μmol/L), increased creatinine clearance rate (0.81 ± 0.13 ml/min vs. 0.37 ± 0.08 ml/min), and reduce urine neutrophil gelatinase-associated lipocalin (NGAL) (579.34 ± 11.70 pg/ml vs. 827.60 ± 14.48 pg/ml), kidney injury molecule-1 (KIM-1) (105.06 ± 2.10 pg/ml vs. 151.67 ± 3.06 pg/ml) (P<0.05). Compared with the model group, Ast treatment alleviated renal tubular epithelial cell injury and significantly decreased the apoptosis (14.36 ± 1.36% vs. 28.63 ± 2.03%) (P<0.05), and significantly decreased the serum TNF-α (361.44 ± 9.66 pg/ml vs. 515.93 ± 10.61 pg/ml), IFN-γ (64.11 ± 1.21 pg/ml vs. 93.51 ± 2.15 pg/ml), IL-2 (388.33 ± 1.21 pg/ml vs. 557.82 ± 15.29 pg/ml), IL-4 (60.89 ± 1.21 pg/ml vs. 95.56 ± 2.75 pg/ml), IL-5 (94.02 ± 2.81 pg/ml vs. 147.07 ± 3.50 pg/ml), and IL-10 (52.62 ± 2.51 pg/ml vs. 78.22 ± 3.24 pg/ml) (P<0.01). Compared with the model group, Ast treatment significantly decreased the kidney TNF-α mRNA (1.89 ± 0.59 vs. 2.87 ± 0.97), IFN-γ mRNA (3.11 ± 1.02 vs. 5.98 ± 1.52), IL-2 mRNA (1.68 ± 0.44 vs. 4.09 ± 1.65), IL-4 mRNA (2.41 ± 0.81 vs. 4.69 ± 1.62), IL-5 mRNA (1.56 ± 0.19 vs. 2.92 ± 0.55), IL-10 mRNA (1.45 ± 0.14 vs. 2.85 ± 0.32) (P<0.01). The ratio of IL-4 to IFN-γ was basically restored to the level of sham operation group (1.05 ± 0.02 vs. 1.02 ± 0.06) (P<0.01), and CD20 cells in renal tissue was reduced. Conclusions The Th1 and B lymphocytes play an important role in renal ischemia reperfusion injury, and Th2 cells play a protective role. Astragaloside can regulate the imbalance of Th1/Th2 in the early stage after acute renal injury, and alleviate renal tubular injury.