Number Of CD34 Cells Research Articles

A Comparison of Cyclophosphamide, Bortezomib, and Dexamethasone Versus Bortezomib and Dexamethasone in Transplant Eligible Patients with Newly Diagnosed Multiple Myeloma

▪IntroductionCyclophosphamide, bortezomib, and dexamethasone (CyBor-D) is considered a standard induction regimen for transplant eligible patients with newly diagnosed multiple myeloma (MM) based primarily on phase 2 data. It has not been directly compared to bortezomib and dexamethasone (Bor-Dex) alone. Therefore, we sought to compare the efficacy CyBor-D and Bor-Dex induction in transplant eligible patients treated at a tertiary care center.MethodsA retrospective observational study was performed from a single tertiary care center between January 1, 2008 and April 30, 2016. All transplant eligible patients with MM who received induction chemotherapy with CyBor-D or Bor-Dex and were included. The primary outcome was progression free survival (PFS). Secondary outcomes were response to induction, toxicity, stem cell collection yield and failures, and response to autologous stem cell transplant (autoSCT). Continuous variables were reported as median and compared using Mann-Whitney test. Categorical variables were compared using Pearson's chi-square test. PFS was estimated with Kaplan-Meier.ResultsOne hundred and fifty nine patients were included, 82 (51.6%) treated with CyBor-D and 77 (48.4%) treated with Bor-Dex. The relative proportion of patients treated with CyBor-D increased annually: 1/23 (4%) in 2012, 20/30 (66%) in 2013, 33/34 (97%) in 2014, and 28/28 (100%) in 2015. Patient demographics and disease characteristics were similar between all groups for sex, subtype of MM and stage by the International Staging System. The Bor-Dex group was significantly younger than the CyBor-D group (mean age 56 vs. 61 years, p < 0.001). Median number of cycles was the same for both groups (4, p= 0.73); however, the Bor-Dex group was more likely to have treatment held, reduced or discontinued than the CyBor-D group (19.7% vs. 6.4%, p=0.017). The overall response rate (ORR) was similar between both groups, 88.5% for CyBor-D and 82.7% for Bor-Dex (p=0.36), but patients treated with CyBor-D had deeper responses to induction with 57.5% achieving ≥VGPR whereas 38.7% in the Bor-Dex group achieved ≥ VGPR (p=0.02). The stem cell mobilization regimen was predominantly Cyclophosphamide and GCSF for both groups. Plerixafor use was more common in the CyBor-D cohort, 14.6% vs. 1.3% in Bor-Dex (p=0.002), and there were more collection failures in the CyBor-D group compared to Bor-Dex (6.1% vs. 0%, p=0.03). The median number of CD34 cells collected was higher in the Bor-Dex compared to CyBor-D (9.88 x 106 cells/kg vs. 7.66 x 106cells/kg, p=0.004), however median collection days was similar (1 in both groups, p=0.175). The mobilization toxicity was similar between both groups with a trend towards increased hospitalizations in the CyBor-D group (26% vs. 14%, p=0.075). The conditioning regimen was the same in both groups, predominantly Melphalan 200 mg/m2. Day 100 response was similar, with 76.7% of CyBor-D patients achieving ≥ VGPR vs. 71.21% for Bor-Dex (p= 0.25). The median PFS was 36.5 months in the Bor-Dex group and not yet reached in the CyBor-D group, due to shorter follow up time. However, the percentage of patients free from relapse was similar at 1 year (92.8% for CyBorD vs 98.7% for Bor-Dex) and at 2 years (69.4% for CyBor-D vs 72.9% for Bor-Dex).ConclusionsCyBor-D and Bor-Dex appear to have similar overall efficacy for newly diagnosed transplant eligible patients with multiple myeloma in our cohort. CyBor-D was associated with deeper response to induction, but after autoSCT there was no difference in the depth of response between the two cohorts. Longer follow up is required to detect if the deeper response to induction with CyBor-D leads to differences in PFS. When used with Cyclophosphamide-GCSF mobilization, CyBor-D appears to result in more collection failures and a trend towards increased hospitalizations. Further prospective studies are required to examine this relationship. DisclosuresKew:Celgene: Honoraria. McCurdy:Celgene: Honoraria.

The clinical effects of low-dose splenic irradiation combined with chest three-dimensional conformal radiotherapy on patients with locally advanced non-small-cell lung cancer: a randomized clinical trial.

ObjectiveThe objective of this study was to explore the clinical effects of low-dose splenic irradiation on locally advanced non-small-cell lung cancer (NSCLC) patients.MethodsThirty-eight patients with stage III NSCLC were randomly divided into a control group and a combined treatment group. The control group only received chest three-dimensional conformal radiotherapy, while the combined treatment group received low-dose splenic irradiation followed by chest three-dimensional conformal radiotherapy after 6 hours. T lymphocyte subsets of the blood cells were tested before, during, and after treatment once a week. The side effects induced by radiation were observed, and a follow-up was done to observe the survival statistics.ResultsThe ratio differences in CD4+ cells, CD8+ cells, and CD4+/CD8+ before and after treatment were not statistically significant (P>0.05) in both the groups. The immune indexes were also not statistically significant (P>0.05) before and after radiotherapy in the combined treatment group. However, the numbers of CD4+ cells and CD4+/CD8+ ratios before radiotherapy were higher than after radiotherapy in the control group. There were no differences in the incidence of radiation toxicities between the two groups; however, the incidence of grade III or IV radiation toxicities was lower, and the dose at which the radiation toxicities appeared was higher in the combined treatment group. The total response rate was 63.16% (12/19) in the combined treatment group vs 42.11% (8/19) in the control group. The median 2-year progression-free survival (15 months in the combined treatment group vs 10 months in the control group) was statistically significant (P<0.05). The median 2-year overall survival (17.1 months in the combined treatment group vs 15.8 months in the control group) was not statistically significant (P>0.05).ConclusionLow-dose radiation can alleviate the radiation toxicities, improve the short-term efficacy of radiotherapy, and improve the survival of locally advanced NSCLC patients.

Immune phenotypes predict survival in patients with glioblastoma multiforme.

BackgroundGlioblastoma multiforme (GBM), a common primary malignant brain tumor, rarely disseminates beyond the central nervous system and has a very bad prognosis. The current study aimed at the analysis of immunological control in individual patients with GBM.MethodsImmune phenotypes and plasma biomarkers of GBM patients were determined at the time of diagnosis using flow cytometry and ELISA, respectively.ResultsUsing descriptive statistics, we found that immune anomalies were distinct in individual patients. Defined marker profiles proved highly relevant for survival. A remarkable relation between activated NK cells and improved survival in GBM patients was in contrast to increased CD39 and IL-10 in patients with a detrimental course and very short survival. Recursive partitioning analysis (RPA) and Cox proportional hazards models substantiated the relevance of absolute numbers of CD8 cells and low numbers of CD39 cells for better survival.ConclusionsDefined alterations of the immune system may guide the course of disease in patients with GBM and may be prognostically valuable for longitudinal studies or can be applied for immune intervention.Electronic supplementary materialThe online version of this article (doi:10.1186/s13045-016-0272-3) contains supplementary material, which is available to authorized users.

The Relationship between Obesity, Prostate Tumor Infiltrating Lymphocytes and Macrophages, and Biochemical Failure.

BackgroundObesity reflects a chronic inflammatory environment that may contribute to prostate cancer progression and poor treatment outcomes. However, it is not clear which mechanisms drive this association within the tumor microenvironment. The aim of this pilot study was to examine prostatic inflammation via tumor infiltrating lymphocytes and macrophages characterized by obesity and cancer severity.MethodsWe studied paraffin-embedded prostatectomy tissue from 99 participants (63 non-obese and 36 obese) from the Study of Clinical Outcomes, Risk and Ethnicity (University of Pennsylvania). Pathologists analyzed the tissue for type and count of lymphocytes and macrophages, including CD3, CD8, FOXP3, and CD68. Pathology data were linked to clinical and demographic variables. Statistical analyses included frequency tables, Kruskal-Wallis tests, Spearman correlations, and multivariable models.ResultsWe observed positive univariate associations between the number of CD68 cells and tumor grade (p = 0.019). In multivariable analysis, CD8 counts were associated with time to biochemical failure (HR = 1.09, 95% CI = 1.004–1.192, p-value = 0.041.) There were no differences in lymphocytes or macrophages by obesity status or BMI.ConclusionsThe number of lymphocytes and macrophages in the tumor microenvironment did not differ by obesity status. However, these inflammation markers were associated with poor prostate cancer outcomes. Further examination of underlying mechanisms that influence obesity-related effects on prostate cancer outcomes is warranted. Such research will guide immunotherapy protocols and weight management as they apply to diverse patient populations and phenotypes.

Abstract 739: The role of obesity on inflammation markers in the prostate tumor microenvironment

Abstract Background: Obesity reflects a chronic inflammatory environment that may contribute to prostate cancer progression and poor treatment outcomes. However, it is not clear which mechanisms drive this association within the tumor microenvironment. The aim of this pilot study was to examine prostatic inflammation via tumor infiltrating lymphocytes and macrophages characterized by obesity and cancer severity. Methods: We studied paraffin-embedded prostatectomy tissue from 99 participants (63 non-obese and 36 obese) from the Study of Clinical Outcomes, Risk and Ethnicity (University of Pennsylvania) Pathologists analyzed the tissue for type and count of lymphocytes and macrophages, including CD3, CD8, FOXP3, and CD68. Pathology data were linked to clinical and demographic variables. Statistical analyses included frequency tables, Kruskal-Wallis tests, Spearman correlations, and multivariable models. Results: We observed positive univariate associations between the number of CD68 cells and tumor grade (p = 0.019), as well as biochemical failure (p = 0.033). In multivariable analysis, CD3 and CD8 counts were associated with time to biochemical failure (HR = 0.93, 95% CI = 0.88-0.99; HR = 1.08, 95% CI = 1.01-1.17, respectively.) Preliminary results suggested differences in lymphocytes by race. There were no differences in lymphocytes or macrophages by obesity status or BMI. Conclusions: The number of lymphocytes and macrophages in the tumor microenvironment did not differ by obesity status. However, these inflammation markers were associated with poor prostate cancer outcomes. Further examination of underlying mechanisms that influence obesity-related effects on prostate cancer outcomes is warranted. Such research will guide immunotherapy protocols and weight management as they apply to diverse patient populations and phenotypes. Citation Format: Charnita M. Zeigler-Johnson, Knashawn H. Morales, Priti Lal, Timothy R. Rebbeck, Michael Feldman. The role of obesity on inflammation markers in the prostate tumor microenvironment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 739.

Early syphilis treatment in HIV-infected patients: single dose vs. three doses of benzathine penicillin G.

Current treatment guidelines for early stages of syphilis are the same regardless of HIV serostatus. There is still controversy about the best treatment for syphilis in HIV patients and the current recommendations are based on limited data. The primary goal of this study was to compare the serological response rates to a single dose vs. three weekly doses of benzathine penicillin G (BPG) in HIV-infected patients with early syphilis and to assess the adequacy of current recommendations. Clinical and laboratory data of HIV patients with early syphilis treated in Sexually Transmitted Disease Clinic between January 2000 and December 2014 were recorded. A good serological response was defined as a ≥4-fold decline in Venereal Disease Research Laboratory (VDRL) titre within 12 months after treatment. Serological failure was defined as a lack of at least fourfold decrease in VDRL titres within 12 months after treatment. After applying inclusion and exclusion criteria, 60 patients were enrolled in the study. Seventeen (28.3%) patients were treated with a single dose of BPG, while in 43 (71.7%) patients, three weekly doses were used. Fifty eight (96.7%) had a good serological response at 12 months and seroconversion was confirmed in 29 (48.3%) patients. There was no statistically significant difference between the two treatment groups regarding serological response, seroconversion rate and the time needed to obtain a good serological response. Furthermore, treatment response was not affected by the number of CD4 cells. The results of our study support the current international treatment guidelines, recommending early syphilis treatment with a single dose of BPG in HIV patients.

Immunological changes in patients with community-acquired pneumonia with concomitant coronary artery disease during treatment with Simvastatin

Introduction. Immunological changes in the elderly people with concomitant diseases affect all major parts of the immune status (cellular and humoral immunity, mucosal immunity, the complement system, and so on). With features of immune system in the elderly people related some of clinical problems: hypersensitivity to non- and intra-hospital infections, subacute infection that increases concomitant diseases, especially cardiovascular. Aim of this work was determination of the characteristics of immunological changes in patients with community-acquired pneumonia and concomitant coronary heart disease and their dynamics during treatment with Simvastatin. Material and methods. 50 patients with community acquired pneumonia the third group and 30 healthy individuals were included in an open prospective study. Immunogram definition was made by lymphocytes phenotyping tests rosette of particles coated with monoclonal antibodies. Immunoglobulin G, A, M were determined by radial immunodiffusion Agar gel by Mancini using monospecific sera. Also absorption capacity of the neutrophil count phagocytic index, phagocytic index, neutrophil bacterial activity by a test restore them of nitroblue tetrazolium were defined. Results. Subanalysis in patients with pneumonia with concomitant coronary artery disease, who were randomly assigned to receiving simvastatin, after treatment showed a statistically significant increase of IgG to 18.6% (p=0.02) of 9.57±4.64 to 11.36±3.7. The rest of the performance was not likely changed. After treatment of pneumonia with concomitant coronary artery disease treated and not treated with Simvastatin group the following immunogram indicators significantly differed: the relative number of lymphocytes, the number of CD8 relative and absolute number of CD22 cells, complement activity, completion of the index of phagocytosis. Thus in patients receiving Simvastatin after treatment were lower rates relative number of CD8 cells by 38% (p=0.037) – 18.2±7.38 vs 25.1±8.0; complement activity by 33% (p=0.03) – 66.9±14.8 vs 89.0±17.8; the absolute number of CD22 cells by 26.7% (p=0.005) – 17.68±7.03 vs 24.11±5.87; completion phagocytosis index by 6.3% (p=0.01) – 0.89±0.10 vs 0.95±0.11; and was higher by 36% (p=0.02), relative number of lymphocytes (27.07±5.94 vs 17.32±7.26), than patients who did not receive simvastatin. Conclusions. The Simvastatin use in the treatment of patients with community acquired pneumonia with concomitant coronary artery disease is accompanied by the following positive changes in the immunogram: reducing the number of stick-nuclear leukocytes by 52% and 36% of ESR, increasing content of IgG 18.6%. However, patients with coronary artery disease who did not use Simvastatin in the treatment of pneumonia, had further growth in immuno-reactivity index by 4% and reduction of IgG 12.6%. Immunogram in patients with coronary artery disease who received Simvastatin After a course of treatment was characterized by significantly lower performance of the relative number of CD8 cells by 38% (p=0.037); complement activity by 33% (p=0.03); the absolute number of CD22 cells by 26.7% (p=0.005); completion phagocytosis index by 6.3% (p=0.01); and higher by 36% (p=0.02) relative quantity of the number of lymphocytes than in patients who did not receive Simvastatin. The article presents data of changes in immune status in patients with community acquired pneumonia. There are positive changes in the cellular link of immunity in these patients. In pneumonia patients with concomitant coronary artery disease indicators of immuno- cellular reactivity after Simvastatin treatment were not changed significantly, but humoral immunity has been changed towards in growing of Ig G and M content.

‘Role of IL-28A in Herpes Stromal Keretitis’.

Abstract Ocular infection with herpes simplex virus (HSV) can result in a chronic immune-inflammatory lesion that is a significant cause of human blindness. A key to controlling SK lesion severity is to identify cellular and molecular events responsible for tissue damage. Interferons are key cytokines in the establishment of a multifaceted antiviral response. Although all IFNs are important mediators of antiviral protection, their roles in antiviral defence vary. Interferon lambda which is a type III cytokine is capable of inducing an antiviral response both in vitro as well as in vivo. This family consists of three structurally related interferon lambda subtypes called IFN-λ (IL-29), IFN-λ2 (IL-28A) and IFN-λ3 (IL-28B). Here, we show that IFN-λ2 (IL-28A) has a prominent role in HSK. The binding of IFN-λ to their cognate receptors induces a common signaling cascade that results in the activation of STAT1 and STAT2, which promotes MX1, OAS1 and IRF7 which in turn mediate the antiviral activity of the type-III IFNs. Here we show that IL-28A appears to specifically target neutrophil function and cause a major reduction in the total number of CD4 and γδT cells and also in TH17 cells in DLN. Also, we found that the serum levels of IL-1β were reduced at the early stage of treatment. Administration of IL-28A effectively reduces the HSK lesions and can prove to be a very reliable therapeutic agent.

How frequently are predicted peptides actually recognized by CD8 cells?

Detection of antigen-specific CD8 cells frequently relies on the use of peptides that are predicted to bind to HLA Class I molecules or have been shown to induce immune responses. There is extensive knowledge on individual HLA alleles’ peptide-binding requirements, and immunogenic peptides for many antigens have been defined. The 32 individual peptides that comprise the CEF peptide pool represent such well-defined peptide determinants for Cytomegalo-, Epstein–barr-, and Influenza virus. We tested the accuracy of these peptide recognition predictions on 42 healthy human donors that have been high-resolution HLA-typed. According to the predictions, 241 recall responses should have been detected in these donors. Actual testing showed that 36 (15 %) of the predicted CD8 cell responses occurred in the high frequency range, 41 (17 %) in mid-frequencies, and 45 (19 %) were at the detection limit. In 119 instances (49 %), the predicted peptides were not targeted by CD8 cells detectably. The individual CEF peptides were recognized in an unpredicted fashion in 57 test cases. Moreover, the frequency of CD8 cells responding to a single peptide did not reflect on the number of CD8 cells targeting other determinants on the same antigen. Thus, reliance on one or a few predicted peptides provides a rather inaccurate assessment of antigen-specific CD8 cell immunity, strongly arguing for the use of peptide pools for immune monitoring.

Clinical and immunologic phenotype associated with activated phosphoinositide 3-kinase δ syndrome 2: A cohort study

Activated phosphoinositide 3-kinase δ syndrome (APDS) 2 (p110δ-activating mutations causing senescent Tcells, lymphadenopathy, and immunodeficiency [PASLI]-R1), a recently described primary immunodeficiency, results from autosomal dominant mutations in PIK3R1, the gene encoding the regulatory subunit (p85α, p55α, and p50α) of class IA phosphoinositide 3-kinases. We sought to review the clinical, immunologic, and histopathologic phenotypes of APDS2 in a genetically defined international patient cohort. The medical and biological records of 36 patients with genetically diagnosed APDS2 were collected and reviewed. Mutations within splice acceptor and donor sites of exon 11 of the PIK3R1 gene lead to APDS2. Recurrent upper respiratory tract infections (100%), pneumonitis (71%), and chronic lymphoproliferation (89%, including adenopathy [75%], splenomegaly [43%], and upper respiratory tract lymphoid hyperplasia [48%]) were the most common features. Growth retardation was frequently noticed (45%). Other complications were mild neurodevelopmental delay (31%); malignant diseases (28%), most of them being B-cell lymphomas; autoimmunity (17%); bronchiectasis (18%); and chronic diarrhea (24%). Decreased serum IgA and IgG levels (87%), increased IgM levels (58%), B-cell lymphopenia (88%) associated with an increased frequency of transitional Bcells (93%), and decreased numbers of naive CD4 and naive CD8 cells but increased numbers of CD8 effector/memory Tcells were predominant immunologic features. The majority of patients (89%) received immunoglobulin replacement; 3 patients were treated with rituximab, and 6 were treated with rapamycin initiated after diagnosis of APDS2. Five patients died from APDS2-related complications. APDS2 is a combined immunodeficiency with a variable clinical phenotype. Complications are frequent, suchassevere bacterial and viral infections, lymphoproliferation, and lymphoma similar to APDS1/PASLI-CD. Immunoglobulin replacement therapy, rapamycin, and, likely in the near future, selective phosphoinositide 3-kinase δ inhibitors are possible treatment options.

MxA expression is associated with tumor-infiltrating lymphocytes and is a prognostic factor in triple-negative breast cancer.

Interferons (IFNs) play an important role in tumor-immune system interactions. As one of the main mediators of IFNs, myxovirus resistance A (MxA) is upregulated in various cancers. However, the exact role of MxA in breast cancer is not fully understood. As part of the immune response to tumors, tumor-infiltrating lymphocytes (TILs) have prognostic significance in breast cancer. The aim of our present study was to examine the relationship between MxA and immune system components, including the amount of TILs and human leukocyte antigen (HLA) expression, in breast cancer. TILs, MxA expression, HLA intensity, and clinicopathological factors were retrospectively analyzed in 688 patients with primary breast cancer between 1993 and 1998 and in 705 patients with triple-negative breast cancer (TNBC) between 2004 and 2011. MxA expression was higher in TNBC tumors than in other subtypes. High MxA levels were associated with a higher histologic grade, abundant TILs, and stronger HLA-ABC expression in both the TNBC subtype within the consecutive breast cancer cohort and the validation TNBC cohort. MxA expression showed a significant positive correlation with TILs, the number of CD8(+) cells, and the number of CD69(+) cells in the validation TNBC cohort. High MxA levels and abundant TILs were found to be independent prognostic factors for disease-free survival in patients with TNBC. These results indicate that MxA expression is closely related to TILs in TNBC and, along with TILs, provides prognostic information after chemotherapy in patients with TNBC.

Computer-Assisted Inflammation Analysis Of Kidney-Graft Biopsy To Improve Risk Stratification In Allograft Rejection

Introduction/ Background Kidney graft biopsy plays a key role in diagnosis of antibody-mediated rejection (AMR), the major cause of renal graft failure. The diagnosis of AMR requires the presence of i) donor specific antibodies (DSA), and ii) microvascular inflammatory lesions on kidney graft biopsy. Aims Histological assessment relies onBanffclassification [1] that has quantitative and qualitative limitations and faces in terms of diagnostic accuracy and risk prediction: 1) this grading is categorical with risks of threshold effect; 2) the nature of inflammatory cells is not considered. Hence we propose a new method of computerized image analysis in order to finely characterize the quality and intensity of graft inflammation. Methods Data 57 kidney recipients fulfilled theBanffcriteria for AMR between 2004 and 2012 at the Lyon Hospitals. Double immunohistological stainings were performed with CD31 (capillaries) and respectively CD68 (macrophages), CD3 (T lymphocytes), CD66b (granulocytes), CD20 (B lymphocytes). 288 glass slides were scanned (MiraxScan, 20x, NA=0.8). Algorithms The goal is to quantify the number of immune cells in the different parts of the kidney cortex. Due to the biopsy preparation such as biopsy slicing and image quality staining many variations within the observed object could happen and result to incorrect image segmentation. Hence we combine color component images to extract Regions of Interest (ROI) based on the use of their contextual data information in order to correctly extract the capillaries and immune cells. The algorithms are implemented in the Icy software ( http://icy.bioimageanal ysis.org) [2]. In the workflow: 1) color deconvolution [3], 2) pre-processing step to segment the pixel staining, 3) extraction of stained objects by combined information. The color deconvolution separates the initial image into 3 component-staining images: blue component for nuclei, brown component for capillaries and the purple one for immune cells. These component images are first preprocessed, by gaussian filtering and then by the k-means classification to segment the images. We combine the segmented ROI and their spatial relation to extract the objects of interest. Results 34 patients had C3d+ DSA and 23 had C3d- DSA. Al- though allograft survival was lower in the C3d+ group (p<0,001 by log-rank),Banffscores for AMR were similar in the 2 groups (3.4±1.1 vs 3.5±1.2, p=0.65). In contrast, our approach revealed notable differences in graft inflammation between the two groups. The number of CD68 cells in the capillaries and in the interstitium allows identifying patients with a risk of graft loss (HR=3.18, p<0.01 et HR=2.62, p=0.01 respectively). The combining C3d test and quantification of monocytes in interstitium allow clustering patients into 3 groups of renal prognosis: C3d-, C3d+/CD68 low and C3d+/CD68 high (p<0.0001 by log rank; C3d+/CD68 high vs C3d+/ CD68 low: HR=2.43, p=0.04; C3d+/CD68 low vs C3d-: HR=4.99, p=0.006). The isolated C3d test has an excellent value negative prediction (89,5% for the 1 year graft loss) but perfectible positive prediction (52,9%) [4] . The monocyte quantification allows to accurate the prognosis of patients in the C3d+ group. Using this novel reproducible approach for topological quantification of inflammation, we observed that histopathological features of complement-binding DSA are different from that of non-complement. The computer-assisted analysis of graft inflammation improves the risk stratification of graft loss.

CD4细胞检测在艾滋病诊疗中的意义

艾滋病是由HIV病毒感染机体CD4细胞后，损伤患者的免疫系统所引发的一种疾病。艾滋病具有传染性强、危害大、不可治愈的特点，给患者的家庭和社会都造成极大的负担。及早的发现病情，并积极的接受治疗可显著提升病人生活质量，降低传染风险。艾滋病的病程与患者的CD4阳性T细胞数量紧密相连，所以监测CD4细胞对于用药指导和药效评价都具有重大意义。 AIDS (acquired immunodeficiency syndrome) is caused by the invasion of HIV viruses and resultant damages to the immune-system. AIDS is a disease characterized by high infectivity, severe harm. So far, there is no cure for AIDS. Thus, it brings great burden to patients’ families and the society. Early detection and treatment of AIDS can significantly improve the quality of patients’ life and reduce the risk of disease expansion. The disease course of AIDS is closely linked with the number of CD4 positive T cells in the peripheral blood. Thus, monitoring the number of CD4 cells is of great significance for the guidance of drug use and the evaluation of drug efficacy.

Investigating Lymphocytic Involvement in Minimal Change Nephrotic Syndrome

Minimal change nephrotic syndrome, also known as minimal change disease (MCD) or minimal change nephrosis, is a subcategory of non-inflammatory glomerulopathy – a degenerative disorder of the main functional unit of the kidney. Neither the cause nor the pathogenesis of minimal change nephrotic syndrome is known. A destructive epithelial disorder would explain the clinical signs of MCD such as proteinuria (excessive protein in the urine), hypoalbuminemia (low serum albumin), and hyperlipidemia (high serum lipids). As a result of the association between the diagnosis of minimal change disease (MCD) and a history of heightened immune responses, it is believed that MCD may be caused by a disorder of T lymphocytes. It is suspected that T-lymphocytic deregulation may release lymphokines toxic to the glomerular basement membrane, which in turn leads to increased ultrastructural permeability (thus leading to symptoms of MCD). We investigated the hypothesis that minimal change nephrotic syndrome is caused by abnormal T lymphocytes, specifically CD4 T cells, as they are known to be involved in many microscopic degenerative kidney disorders involving increased membranous permeability [1]. To test this hypothesis, we measured T-cell counts in peripheral blood samples by flow cytometry. We found that eight of ten patients had decreased CD4:CD8 ratio, thus strengthening the immune involvement postulate of MCD. More specifically, we found the number of CD4 cells to be abnormally low; the results in this study can be used to further investigate whether or not MCD is a true autoimmune disorder. Understanding T-lymphocyte abnormalities in MCD can lead to further research efforts in finding a cause for MCD.

Interferon-Gamma Signaling in the Pathogenesis of Idiopathic Pneumonia Syndrome Following Allogeneic Bone Marrow Transplantation

Idiopathic pneumonia syndrome (IPS) is a life-threatening acute noninfectious diffuse lung injury occurring after allogeneic bone marrow transplantation (BMT). We have recently shown that absence of STAT1 signaling in recipient APC leads to accelerated Graft Versus Host Disease (GVHD) and Transplant-associated mortality and morbidity after lethal irradiation in fully MHC-mismatched allogeneic or in non-irradiated P→F1 BMT models. We now addressed the impact of recipient Type II interferon signaling on the development of lung injury after allogeneic BMT using a non-irradiated P→F1 model. 1x107 BMCs plus 2.5x107 SPCs from C57BL/6 (B6) mice were i.v. injected into IFNγR-deficient CB6F1 (F1.GRKO) or CB6F1 wild type (WT) mice. F1.GRKO recipients had significantly reduced survival (MST 32 days post-BMT vs. not reached p<0.001, Log-rank test). Pulmonary function tests were performed 3 weeks post-BMT on anaesthetized mice that were tracheotomized and mechanically ventilated using a computer-controlled animal ventilator (Flexivent, SCIREQ). Static compliance was derived from plateau pressure measurements between total lung capacity and functional residual capacity, and broadband forced oscillations were used to determine airway resistance at baseline and following increasing doses of methacholine (3.1, 12.5 and 50 mg/ml). Static compliance and inspiratory capacity were reduced in F1.GRKO recipients when compared with WT mice (p<0.01). In addition, airway resistance, both at baseline and in response to methacholine, was increased in the KO recipient mice (p<0.05). Histopathology was evaluated on H&E, Trichrome and PAS stained lung sections to assess for inflammation, fibrosis and mucus hyperplasia, respectively. Lungs from the KO mice showed increased interstitial inflammation involving bronchioles, vasculature and parenchyma (pathology index 10 vs 1; p<0.001), fibrosis (fibrosis score 2.8 vs 0.15; p<0.05) and a trend for mucus hyperplasia/metaplasia. To further evaluate the cellular mediators of the observed lung injury, we assessed the cellular composition of the bronchioalveolar lavage fluid (BAL) at day 30 post-BMT. We observed a 10-fold increase in the total cell number of the BAL in F1.GRKO recipient mice compared to WT mice (4x105 vs.4x104, p<0.01); the majority of the cells in the BAL from wild type mice were macrophages/monocytes, while neutrophils predominated in the F1.GRKO mice. Importantly, more than 50% of the CD45+ BAL cells were donor derived in the F1.GRKO recipients, compared to only 10% in the wild type recipients. Furthermore, the absolute cell number of donor CD4+ and CD8+ cells in the knockout mice significantly outnumbered that in their wild type counterparts (p<0.01). Thus, our results suggest that absence of recipient IFN-γR signaling promotes accumulation and/or trafficking of donor neutrophils and T cells to the lungs. To better understand the underlying mechanisms of enhanced infiltration of donor cells in the lungs of IFNγR-/- mice, the activation, proliferation, and differentiation status of donor lymphocytes were assessed by Flowcytometry after BMT. In line with the lung infiltration, there was significantly increased donor cell engraftment in PBMC, spleen, lymph nodes, and bone marrow in F1.GRKO recipients (all with p<0.01); Moreover, there were also significantly increased activation (measured as CD44high CD62Llow cells), Th1 and Tc1 differentiation of donor CD4 and CD8 cells (p<0.01). Furthermore, enhanced Th17 differentiation, but reduced Foxp3+ Treg cell differentiation was noticed. Interestigly, we observed significantly lower PD-L1 expression on recipient APC in the F1.GRKO mice compared to wild type mice, suggesting that absence of PD-L1 expression may contribute to reduced contraction of activated donor T cells. Interestingly, in line with these findings we also observed significantly reduced PD-L1 expression when single cell suspension from lung tissues were analyzed by flow. Reduced PD-L1 expression was observed on both recipient CD45+, and CD45- cells in F1.GRKO when tested on day 22 post-BMT, meanwhile there was no difference in PD-L1 expression pattern in the cells isolated from small intestine. These results suggest that IFN-γ induced PD-L1 expression on hematopoietic and non-hematopoietic cells in the lung may be of significance in protecting recipients against acute donor-cell mediated lung injury and IPS. DisclosuresLentzsch:Celgene: Membership on an entity's Board of Directors or advisory committees; Axiom: Speakers Bureau; Novartis: Membership on an entity's Board of Directors or advisory committees; BMS: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees.

A Pilot Study of Peripheral Blood Hematopoietic Stem Cells Mobilization with the Combination of Bortezomib and G-CSF in Multiple Myeloma and Non-Hodgkin's Lymphoma Patients.

Background: Bortezomib has become an integral part of front-line therapy of multiple myeloma in a large majority of patients. There are preliminary reports which show that addition of bortezomib can augment the peripheral blood CD34 count during stem cell mobilization. In this single center prospective trial we added bortezomib to G-CSF to evaluate the effects of bortezomib on peripheral CD34 counts and collection.Methods: Patients aged 18-70 years with diagnosis of multiple myeloma (MM) or non-hodgkin's lymphoma (NHL) who were eligible for autologous stem cell transplantation (ASCT) and had received no more than three prior chemotherapeutic regimens were eligible for the study. Patients were enrolled in two groups. Group A (N=3) received G-CSF 16mcg/kg for 5 days and proceeded to stem cell collection on D5 and then received bortezomib 1.3mg/m2 on D5 after stem cell collection and G-CSF 16mcg/kg on D6, 7, 8 and repeat stem cell collection on D6, 7, 8 till the goal was achieved. Group B (N=17) received G-CSF 16mg/kg on D1-5 and received bortezomib 1.3mg/m2 on D4 and proceeded to stem cell collection on D5. If the patient was not able to collect the predefined goal CD34, G-CSF was continued on D 6, 7, 8 and a second dose of bortezomib 1.3mg/m2 was given on D7. Mobilization procedure was stopped once the predefined goal CD34 collection (4 x 106/kg for MM and 2 x 106/kg for NHL) had been collected.Primary objectives of the study was to determine if addition of bortezomib to G-CSF will result in an increase in PBSCs by > 2-fold and to achieve median neutrophil engraftment 12 days post ASCT. Secondary objectiveswere to evaluate the collected product for co-mobilization of lymphoma or myeloma cells and to determine if the use of bortezomib increases the mobilization of immune-stimulatory Dendritic cell (DC) -1 subsets.Results: A total of 23 patients were enrolled and 20 were evaluable for the results. Only one patient with NHL was enrolled and rest had MM. Median age of pts was 57 years, M/F 8/12, median number of previous chemotherapy regimens was 1 (range 1-3).The median peripheral blood CD34 count pre and post bortezomib in all patients were 28.8 x 106/kg and 37 x 106/kg respectively. All three patients in group A had drop in peripheral blood CD34 counts on D6 post bortezomib as they had undergone stem cell collection on day 5. In part B (N=17), 15 patients had increase in peripheral blood CD 34+ve cell counts with 4 patients achieved doubling while 11 pts had less than doubling of peripheral blood CD34 count after receiving bortezomib. Two patients had minimal drop in the peripheral blood CD34 counts post bortezomib. Median number of CD34 cells collected in15 patients (part B) were 5.06 x 106 CD34 cells/kg (range 4-15.1). 18 patients proceeded to ASCT and median time to neutrophil engraftment (ANC ≥500/cumm) post transplant was 12 days (range 11-16) and platelet engraftment (Plt count ≥ 20,000/cumm) was 18 days (range 15-27). There was no significant change in DC1/DC2 ratio in both groups following treatment with bortezomib and G-CSF (Figure 1).In group A all three patients collected goal CD34 count on day 5 and 2/3 patients collected >4 x106 CD34 cells/kg on D6 post bortezomib and1/3 patients collected 2.6 x 106 on D6 post bortezomib. In group B (n=17), 2 patients were unable to collect because of low CD34 counts on D4 and D5, 11 pts collected the goal in one day (D 5) and 4 pts required two days of apheresis (D 5 and 6). None of the patients received D7 bortezomib.Conclusion: Use of bortezomib during autologous stem cell collection was safe and well tolerated. Majority of patients had increase in peripheral blood CD34 counts post bortezomib administration on D4. Future trials should explore bortezomib as an alternate strategy to chemo-mobilization in combination with growth factors. [Display omitted] [Display omitted] DisclosuresOff Label Use: Bortezomib for stem cell mobilization. Lum:Karyopharm Therapeutics Inc: Equity Ownership; Transtarget.Inc: Equity Ownership. Deol:Bristol meyer squibb: Research Funding. Abidi:celgene: Speakers Bureau; Millenium: Research Funding.

Stimulation of Adrenergic Activity By Desipramine Enhances Hematopoietic Stem and Progenitor Cell Mobilization Along with G-CSF in Multiple Myeloma - a Pilot Study of Safety and Efficacy

Background:Hematopoietic stem cell release is regulated by the sympathetic nervous system through the β (3) adrenergic receptor [Mendez-Ferrer et al. Nature 2008]. Peripheral sympathetic nerve neurons express the G-CSF receptor and stimulation of peripheral sympathetic nerve neurons with G-CSF reduced norepinephrine (NE) reuptake significantly, suggesting that G-CSF potentiates the sympathetic tone by increasing NE availability [Lucas et al Blood 2012]. Based on preclinical data, we investigated the NE reuptake inhibitor desipramine in HSC mobilization.Despite augmentation with Plerixafor (CXCR4 inhibitor), 20% of all patients fail to mobilize 6*10^6 CD34 cells/kg in myeloma and the collection rate with G-CSF alone is 51.1% [Diperiso et al Blood 2012]. The cost of upfront plerixafor is $9,081 per patient while desipramine costs $40. We undertook a feasibility study of adult patients with MM undergoing autologous transplantation (ASCT) to study safety and efficacy of mobilization with desipramine and G-CSF.Patients & Methods:From 2013- 2014, 10 patients between the ages of 18-70, eligible for ASCT were enrolled. Desipramine 100mg daily was administered for 7 days, starting 4 days prior to starting G-CSF (D-3) and continue along with G-CSF for a total of 7 days. CBC and CD34 counts were determined on Day+5. If CD34 counts were > 10/ul, stem cell collection was commenced and if < 10/ul, plerixafor was added as salvage therapy. The endpoints were safety and efficacy in mobilizing CD34 cells for ASCT in patients with multiple myeloma. This trial was registered at clinicaltrials.gov as NCT01899326.ResultsSix of ten patients enrolled completed the protocol and underwent stem cell transplantation. Reasons for not completing were 1. Lack of insurance coverage 2. Non-compliance with study treatment 3. Disease relapse prior to ASCT. Five patients did not have any grade 3 or 4 adverse events and 1 had disease-related Grade 4 hypercalcemia and Grade 2 AKI at the time of stem cell mobilization. No patients had significant treatment related adverse effects. All 6 patients who completed the protocol achieved the target collection of 5*10^6 CD34 cells/kg. Four patients achieved 6*10^6 CD34 cells/kg or more and the remaining 2 patients achieved 5.52 and 5.92 *10^6 CD34 cells/kg respectively. Among the 6 patients, 2 patients received salvage plerixafor. The median time to achieve WBC >1000/ul, ANC >500/ul and platelets>20k was 11.5, 11, 13.5 daysTable 1AgeInd. RegimneDisease statusPPB CD34/ulCD34 collected *10^6 / kgTotal CD34/kg collectedEngraftment (Days to)Adverse effects from desipramineD1D2D3D4D2D3D4ANC >0.5Platelets> 20kG1,G2G3,G4162 Free λVRDVGPRN45.866.07.017.011213nonenone250 Free λVRDVGPRN88.0143.512.2212.221212nonenone358 IgAVCDVGPRN38.067.731.64.222.756.971317nonenone470 IgAκVRDVGPRY2.4040.216.64.311.615.921214nonenone556 IgGκVCDVGPRY8.7011.937.119.41.334.571.617.511112nonenone670 IgGλVD RDRelapseN76.297.15.545.541120AKIhypercalcemiaP-Plerixafor; V-Velcade; R-Lenalidomide; D-Dexamethasone; C-CyclophosphamideConclusionsOverall G-CSF + Desipramine combination appears to be safe, well tolerated and shows signs of efficacy. G-CSF and desipramine was successful in 4/6 (66%) and all achieved the stem cell collection in 2 days or less. Desipramine, GCSF and Plerixafor was successful in all (6/6) patients to achieve a target of 5*10^6 CD34 cells/kg. The mean number of CD34 cells collected in the desipramine+ G-CSF mobilisers was 7.24*10^6 CD34 cells/kg which, based on historical data, is higher than what would be expected with G-CSF alone even though 3/4 of these patients had lenalidomide as induction therapy. Based on these results, a phase II clinical study evaluating the efficacy of G-CSF with desipramine with or without salvage plerixafor in multiple myeloma and lymphoma will be initiated. DisclosuresBarta:Seattle Genetics: Research Funding.

호마자 추출물이 NC/Nga 생쥐의 아토피 피부염에 미치는 영향

Objectives The purpose of this study is to investigate the effects of Sesamum indicum extracted (SEI) on atopic dermatitis in an in-vitro and in-vivo experiment using a MC/9 murine mast cells and a NC/Nga mouse. Methods In-vitro experiment, IL-4, IL-5, IL-6, IL-13, TNF-<TEX>${\alpha}$</TEX> and GM-CSF mRNA expression were evaluated by Real-time PCR, IL-13, MIP-<TEX>$1{\alpha}$</TEX> production by ELISA and manifestations of NFAT-1, NFAT-2, c-jun, c-fos, NF-<TEX>${\kappa}B$</TEX> p65 transcription factors by western blotting. In-vivo experiment, we measured WBC, Eosinophil, Neutrophil, and serum IL-5, IL-13 in NC/Nga atopic dermatitis mouse, IL-5, IL-13, IFN-<TEX>${\gamma}$</TEX>, IL-4 in the spleenocyte culture supernatant by ELISA, the absolute cell numbers of CD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the axillary lymph node (ALN), peripheral blood mononuclear cells (PBMCs) and dorsal skin tissue, IL-5, IL-13 by Real-time PCR, the distribution of tissue inflammation and cellular infiltration by H&E and toluidine blue. Results SEI decreased IL-4, IL-5, IL-6, IL-13, GM-CSF, TNF-<TEX>${\alpha}$</TEX> mRNA expression, IL-13, MIP-<TEX>$1{\alpha}$</TEX> production and the expression of transcription factors including NFAT-1, c-jun, NF-<TEX>${\kappa}B$</TEX> p65 in MC/9 murine mast cells. SEI orally administration decreased cell number of WBC, Eosinophil, the level of serum IgE, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, B220+CD23+ in the ALN. SEI orally administration also increased absolute cell number of CD8+/CD3+ and decreased Gr-1+/CD11b+ in PBMCs, decreased CD4+ in dorsal skin tissue, inhibited IL-5, IL-13 mRNA expression. Infiltration levels of inflammatory immune cells, mast cells and thickness of epidermis decreased in dorsal skin tissue. Conclusions SEI can regulate allergic inflammatory response suppressed the gene expression and production of cytokines that mediate allergic reactions, and will be able to be effectively utilized in the treatment of atopic dermatitis future.

PDL1 Regulation by p53 via miR-34.

Background:Although clinical studies have shown promise for targeting PD1/PDL1 signaling in non–small cell lung cancer (NSCLC), the regulation of PDL1 expression is poorly understood. Here, we show that PDL1 is regulated by p53 via miR-34.Methods:p53 wild-type and p53-deficient cell lines (p53–/– and p53+/+ HCT116, p53-inducible H1299, and p53-knockdown H460) were used to determine if p53 regulates PDL1 via miR-34. PDL1 and miR-34a expression were analyzed in samples from patients with NSCLC and mutated p53 vs wild-type p53 tumors from The Cancer Genome Atlas for Lung Adenocarcinoma (TCGA LUAD). We confirmed that PDL1 is a direct target of miR-34 with western blotting and luciferase assays and used a p53R172HΔg/+K-rasLA1/+ syngeneic mouse model (n = 12) to deliver miR-34a–loaded liposomes (MRX34) plus radiotherapy (XRT) and assessed PDL1 expression and tumor-infiltrating lymphocytes (TILs). A two-sided t test was applied to compare the mean between different treatments.Results:We found that p53 regulates PDL1 via miR-34, which directly binds to the PDL1 3’ untranslated region in models of NSCLC (fold-change luciferase activity to control group, mean for miR-34a = 0.50, SD = 0.2, P < .001; mean for miR-34b = 0.52, SD = 0.2, P = .006; and mean for miR-34c = 0.59, SD = 0.14, and P = .006). Therapeutic delivery of MRX34, currently the subject of a phase I clinical trial, promoted TILs (mean of CD8 expression percentage of control group = 22.5%, SD = 1.9%; mean of CD8 expression percentage of MRX34 = 30.1%, SD = 3.7%, P = .016, n = 4) and reduced CD8+PD1+ cells in vivo (mean of CD8/PD1 expression percentage of control group = 40.2%, SD = 6.2%; mean of CD8/PD1 expression percentage of MRX34 = 20.3%, SD = 5.1%, P = .001, n = 4). Further, MRX34 plus XRT increased CD8+ cell numbers more than either therapy alone (mean of CD8 expression percentage of MRX34 plus XRT to control group = 44.2%, SD = 8.7%, P = .004, n = 4). Finally, miR-34a delivery reduced the numbers of radiation-induced macrophages (mean of F4-80 expression percentage of control group = 52.4%, SD = 1.7%; mean of F4-80 expression percentage of MRX34 = 40.1%, SD = 3.5%, P = .008, n = 4) and T-regulatory cells.Conclusions:We identified a novel mechanism by which tumor immune evasion is regulated by p53/miR-34/PDL1 axis. Our results suggest that delivery of miRNAs with standard therapies, such as XRT, may represent a novel therapeutic approach for lung cancer.

Indirubin Increases CD4+CD25+Foxp3+ Regulatory T Cells to Prevent Immune Thrombocytopenia in Mice.

Indirubin, a traditional Chinese medicine, is used to treat autoimmune diseases in clinics. However, the effects of indirubin on the immunosuppressive CD4+CD25+Foxp3+ regulatory T cells (Treg) have not been addressed. Thus, we aimed to investigate the effects of indirubin on CD4+CD25+Treg cells in immune thrombocytopenia (ITP) CBA mice, which were established by immunization with Wistar rat platelets. 50 mg/kg indirubin treatment daily for 4 weeks significantly decreased anti-platelet antibody production and prevented the decrease of platelets caused by immunization in ITP mice. Consistently, indirubin significantly enhanced the percentage and cell number of CD4+CD25+Foxp3+Treg cells in the peripheral blood, spleen and lymph nodes. We also observed a significant increase of the frequency and cell number of CD4+CD25+Foxp3+Treg cells in the thymus upon indirubin treatment. Furthermore, CD4+CD25+Treg cells from indirubin-treated mice showed similar immunosuppression on T effector cells as compared to those from control mice. Altogether, indirubin ameliorates ITP by enhancing CD4+CD25+Foxp3+Treg cell level with preserving immunosuppressive function.