Download the Free Unbound MEDLINE PubMed App to your smartphone or tablet.
Available for iPhone, iPad, iPod touch, and Android.
T cell growth factor [keywords]
- Retrospective Analysis of the Safety and Efficacy of High-dose Interleukin-2 After Prior Tyrosine Kinase Inhibitor Therapy in Patients With Advanced Renal Cell Carcinoma. [JOURNAL ARTICLE]
- J Immunother 2014 Jul 29.
Although tyrosine kinase inhibitors (TKI) are the most common first-line therapy for metastatic renal cell carcinoma, high-dose interleukin-2 (HD-IL2) remains the only agent that provides durable complete responses. The optimal sequence of these agents remains uncertain. This retrospective multi-institutional study examined the safety and efficacy of HD-IL2 following TKI therapy. After IRB approval at 7 HD-IL2 centers, data relating to patient, disease, and treatment characteristics among 40 consecutive patients with metastatic renal cell carcinoma who were treated with HD-IL2 after at least 1 prior TKI therapy were retrospectively collected. The most common cardiac adverse events were grade 3 hypotension and vascular leak syndrome. Six patients (15%) experienced other grade ≥3 cardiac adverse events. There were 2 treatment-related deaths due to congestive heart failure, occurring in 1 patient with short TKI to HD-IL2 interval and another patient with an abnormal baseline cardiac stress test. Best responses included 2 CRs (5%, duration 40+ and 62+ mo), 3 PRs (8%, duration 6, 11, and 24 mo), 13 SD (32%, median duration 12 mo), 20 PD (50%), and 2 not evaluable patients. Median overall survival was 22 months. Administration of HD-IL2 could be safe and effective after TKI therapy; however, careful selection of patients is critical. We recommend baseline cardiac risk factor assessment, screening with both cardiac stress test and echocardiogram, and allowing a TKI to HD-IL2 interval of at least 2 months.
- An Epithelial Cell Adhesion Molecule- and CD3-Bispecific Antibody Plus Activated T-Cells Can Eradicate Chemoresistant Cancer Stem-like Pancreatic Carcinoma Cells In Vitro. [Journal Article]
- Anticancer Res 2014 Aug; 34(8):4509-19.
Cancer stem-like properties of various types of cancer, including pancreatic cancer, one of the most aggressive types, correlate with metastasis, invasion, and therapeutic resistance. More importantly, chemoresistance in cancer stem-like cells (CSLCs) is a critical problem for eradication of pancreatic cancer. Several cell surface markers, such as CD44 and epithelial cell adhesion molecule (EpCAM), are molecular targets on CSLCs of pancreatic carcinoma. In this study, we investigated whether catumaxomab, a clinical-grade bi-specific antibody that binds to both EpCAM on tumor cells and CD3 on T-cells, combined with activated T-cells can eliminate chemoresistant pancreatic CSLCs in vitro. Firstly, we established a CSLC line (MU-PK1) from human pancreatic carcinoma cells derived from a patient with chemoresistant and disseminated pancreatic cancer. These CSLCs were almost completely resistant to gemcitabine-mediated cytotoxicity up to a concentration of 10 μg/ml. The cells expressed high levels of CSLC markers (CD44 and EpCAM) and had significantly higher capacities for sphere formation, invasion, and aldehyde dehydrogenase-1 expression, which are associated with cancer stemness properties. We found that pre-treatment with catumaxomab and subsequent addition of interleukin-2/OKT3 activated autologous T-cells eliminated CSLCs during a short incubation period. Moreover, when MU-PK1 cells were cultured under hypoxic conditions, the CSLCs became more aggressive. However, the combination of cytokine-activated killer T-cells with catumaxomab successfully lysed almost all these cells. In conclusion, catumaxomab combined with activated T-cells may be a potent therapeutic modality to eradicate chemoresistant pancreatic CSLCs.
- High Expression of Fusion Proteins Consisting of a Single-chain Variable Fragment Antibody Against a Tumor-associated Antigen and Interleukin-2 in Escherichia coli. [Journal Article]
- Anticancer Res 2014 Aug; 34(8):3937-46.
The aim of this study was to establish a strategy for high-level production of single-chain variable fragment (scFv) antibodies fused with interleukin-2 (IL-2) in Escherichia coli.We constructed two fusion sequences consisting of a scFv gene derived from a mouse monoclonal antibody against a tumor-associated antigen (MK-1) and human Interleukin-2(IL-2) gene, ligated the fusions into pET15b and transformed into three different E. coli strains. The effects of temperature, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration and duration of IPTG induction were investigated.Employing E. coli strain Rosetta-gami B, which has an oxidizing cytoplasm that facilitates cytoplasmic disulfide bond formation, improved the level of soluble protein expression. Under optimal conditions, the highest levels of fusion protein expression and high percentages of the proteins were found in their soluble form. Specifically, 89.29% (0.28 g/l) of one fusion protein was soluble after a 10-h induction and 84.61% (0.26 g/l) of the other fusion protein was soluble after a separate 10-h induction. When analyzed by enzyme-linked immunosorbent assay, the partially-purified fusion proteins retained a specific binding activity to the cell lysate of Chinese hamster ovary (CHO) cells expressing MK-1.Taken together, the methods described herein permit the production of substantial amounts of the fusion proteins for conducting functional studies on the biological role of these fusion proteins.
- Optimal Production of a Fusion Protein Consisting of a Single-chain Variable Fragment Antibody Against a Tumor-associated Antigen and Interleukin-2 in Fed-batch Culture of Pichia pastoris. [Journal Article]
- Anticancer Res 2014 Aug; 34(8):3925-35.
The aim of the present study was to establish the strategy for producing a single-chain variable fragment (scFv) antibody fused with interleulin-2 (IL2) by Pichia pastoris and to optimize production during fed-batch cultivation in a 5-l fermenter.We constructed a fusion sequence consisting of an scFv gene derived from a mouse monoclonal antibody against a tumor-associated antigen (designated MK-1 antigen) and human interleulin-2 (IL-2) gene, ligated the sequences to expression vector pPICZα-A and separately transformed the constructs into Pichia pastoris strains GS115 and KM71H.The highest concentration of secreted fusion protein, 738±44 mg/l, was obtained after a 60-h induction. To investigate the specific binding activity of the partially purified fusion protein, we used an enzyme-linked immunosorbent assay and antigen from a whole-cell lysate. Student's t-test showed that the specific binding activity of the partially-purified fusion protein to the lysate of Chinese hamster ovary cell lines expressing the MK-1 antigen was significantly higher than that of the lysate of CHO cell lines that do not express MK-1.The method described here permits the production of substantial amounts of the fusion protein for conducting functional studies on the biological role of these fusion proteins.
- Modulation of both activator protein-1 and nuclear factor-kappa B signal transduction of human T cells by amiodarone. [JOURNAL ARTICLE]
- Exp Biol Med (Maywood) 2014 Jul 29.
Amiodarone, a common and effective antiarrhythmic drug, has been reported to have anti-inflammatory effects such as reducing the activation and movement of neutrophils. However, its effects on human T cells remain unclear. The aim of this study was to elucidate the effects and possible underlying mechanisms of amiodarone on human T cells. We isolated human primary T cells from the peripheral blood of healthy volunteers and performed enzyme-linked immunosorbent assay (ELISA), flow cytometry, electrophoretic mobility shift assay, luciferase assay, and Western blotting to evaluate the modulatory effects of amiodarone on human T cells. We found that amiodarone dose dependently inhibited the production of cytokines, including interleukin-2 (IL-2), IL-4, tumor necrosis factor-alpha, and interferon-gamma in activated human T cells. By flow cytometry, we demonstrated that amiodarone suppressed the expression of IL-2 receptor-alpha (CD25) and CD69, the cell surface markers of activated T cells. Moreover, molecular investigations revealed that amiodarone down-regulated activator protein-1 (AP-1) and nuclear factor kappa-B (NF-κB) DNA-binding activities in activated human T cells and also inhibited DNA binding and transcriptional activities of both AP-1 and NF-κB in Jurkat cells. Finally, by Western blotting, we showed that amiodarone reduced the activation of c-Jun NH2-terminal protein kinase and P38 mitogen-activated protein kinase, and suppressed stimuli-induced I-kappa B-alpha degradation in activated human T cells. Through regulation of AP-1 and NF-κB signaling, amiodarone inhibits cytokine production and T cell activation. These results show the pleiotropic effects of amiodarone on human T cells and suggest its therapeutic potential in inflammation-related cardiovascular disorders.
- [Immunogenicity of allogeneic freezing periosteum and bone marrow]. [English Abstract, Journal Article]
- Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2014 May; 28(5):649-53.
To investigate the immunogenicity of freezing periosteum and bone marrow during allogeneic joint transplantation, and to explore proper pretreatment of allogeneic joint.The allogeneic periosteum and bone marrow were harvested from knee joints of 5 New Zealand white rabbits (aged, 6 months; weighing, 2.6-3.0 kg). After gradient cooling, the tissue was cryopreserved for 1 month. The freezing periosteum and bone marrow were grinded to pieces after rewarming to prepare the suspension of periosteum and bone marrow. Eighteen Chinchilla rabbits (aged, 6 months; weighing, 2.1-2.8 kg) were divided into 3 groups randomly: normal saline injection group (group A, n = 6), periosteum injection group (group B, n = 6), and bone marrow injection group (group C, n=6). The normal saline, periosteum suspension, and bone marrow suspension were injected into the peritoneal cavity in groups A, B, and C, respectively. The concentrations of interleukin 2 (IL-2), IL-6, and tumor necrosis factor alpha (TNF-alpha) in serum and the ratio of CD4' T cell/CD8+ T cell in venous blood were measured before injection, at 1 week and 2 weeks after injection.There was no significant difference in the concentration of IL-2 between before and after injection in the same group (P=0.241), and between groups (P = 0.055). The concentration of IL-6 after injection was significantly lower than that before injection in the same group (P = 0.040), but no significant difference was found between groups (P = 0.357). The concentration of TNF-a showed no significant difference between before and after injection in the same group (P = 0.925), but the concentration of TNF-a in group B was significantly higher than that in groups A and C (P < 0.05). The ratio of CD4+ T cell/CD8+ T cell of venous blood had no significant difference between before and after operation in the same group (P = 0.248), and between groups (P=0.646).The freezing periosteum and bone marrow are lowly immunogenic. In order to decrease the immunogenicity of the joint, preserving the periosteum and removing the marrow cavity are recommended.
- Current Systemic Therapies for Melanoma. [JOURNAL ARTICLE]
- Dermatol Surg 2014 Jul 28.
Systemic agents are used in melanoma for adjuvant therapy and to treat metastatic disease. Currently, interferon-α is the only agent approved for adjuvant therapy. Six drugs are FDA approved for metastatic disease: dacarbazine, interleukin-2 (IL-2), vemurafenib, ipilimumab, dabrafenib, and trametinib. Vemurafenib and ipilimumab were approved in 2011, whereas dabrafenib and trametinib were approved in 2013.This review will update the practicing dermatologist on the differences in efficacy, adverse events, and cost of systemic therapies available for the treatment of melanoma.This article is a review of the current literature on systemic therapies for advanced melanoma. Key search words included "advanced melanoma," "systemic therapy," and "adjuvant therapy" with particular focus on the past 20 years.Before 2011, dacarbazine and IL-2 were the only FDA approved therapies for metastatic melanoma, and IFN-α is the only approved agent for adjuvant therapy. The new agents vemurafenib, ipilimumab, dabrafenib, and trametinib are the first to have improved overall survival in Phase III studies in comparison with other systemic therapies.Despite new developments, there remains a significant need for better therapies with improved long-term efficacy and decreased toxicity.
- Cytokine pathway disruption in a mouse model of schizophrenia induced by Munc18-1a overexpression in the brain. [JOURNAL ARTICLE]
- J Neuroinflammation 2014 Jul 29; 11(1):128.
An accumulating body of evidence points to the significance of neuroinflammation and immunogenetics in schizophrenia, and an imbalance of cytokines in the central nervous system (CNS) has been suggested to be associated with the disorder. Munc18- overexpressing mice (Munc18-OE) have provided a model for the study of the alterations that may underlie the symptoms of subjects with schizophrenia. The aim of the present study was to elucidate the involvement of neuroinflammation and cytokine imbalance in this model.Cytokines were evaluated in the cortex and the striatum of Munc18-OE and wild-type (WT) mice by enzyme-linked immunosorbent assay (ELISA). Protein levels of specific microglia and macrophage, astrocytic and neuroinflammation markers were quantified by western blot in the cortex and the striatum of Munc18-OE and WT mice.Each cytokine evaluated (Interferon-gamma (IFN-gamma), Tumor Necrosis Factor-alpha (TNF-alpha), Interleukin-2 (IL-2) and CCL2 chemokine) was present at higher levels in the striatum of Munc18-OE mice than WT. Cortical TNF-alpha and IL-2 levels were significantly lower in Munc18-OE mice than WT mice. The microglia and macrophage marker CD11b was lower in the cortexes of Munc18-OE mice than WT, but no differences were observed in the striatum. Glial Fibrillary Acidic Protein (GFAP) and Nuclear Factor-kappaB (NF-kappaB)p65 levels were not different between the groups. Interleukin-1beta (IL-1beta) and IL-6 levels were beneath detection limits.The disrupted levels of cytokines detected in the brain of Munc18-OE mice was found to be similar to clinical reports and endorses study of this type for analysis of this aspect of the disorder. The lower CD11b expression in the cortex but not in the striatum of the Munc18-OE mice may reflect differences in physiological activity. The cytokine expression pattern observed in Munc18-OE mice is similar to a previously published model of schizophrenia caused by maternal immune activation. Together, these data suggest a possible role for an immune imbalance in this disorder.
- Mechanical removal of dendritic cell-generating non-classical monocytes via ex vivo lung perfusion. [Journal Article]
- J Heart Lung Transplant 2014 Aug; 33(8):864-9.
Ex vivo lung perfusion (EVLP) is a novel procedure designed to rapidly assess and recondition unusable donor lungs for transplantation (LTx). EVLP may reduce graft immunogenicity and allorecognition via removal of passenger leukocytes. We aimed to explore this hypothesis using human EVLP and in vitro analysis.Explanted human lungs (n = 7) underwent standard EVLP. Perfusate samples and the leukocyte filter were collected, and cells characterized via flow cytometry. Isolated alveolar monocytes (from post-LTx bronchoalveolar lavage) were differentiated to dendritic cells and characterized (n = 10). An in vitro (air epithelial-liquid endothelial) lung model was utilized to evaluate monocyte migration and differentiation within the lung.Non-classical monocytes (NCM, normally <1% of total white blood cell repertoire) mobilized within 30 minutes of EVLP and represented 80.04% of the passenger leukocyte population. This subset readily differentiated to dendritic cells and secreted pro-inflammatory cytokines (interferon-γ and interleukin-2) after stimulation. NCM rapidly diapedesed from the vascular bed to the alveolus and, when cultured on the alveolus, differentiated to dendritic cells with inflammatory phenotypes.The lung possesses a reservoir of NCM, which can readily diapedese to the alveolus or mobilize in the circulation. After activation, NCM differentiate to inflammatory dendritic cells with T-cell co-stimulatory capacity. EVLP may impart additional benefits after LTx via the removal of passenger monocytes, which may represent a previously unidentified beneficial mechanism of action.
- Evaluation of Prolonged Fatigue Post-West Nile Virus Infection and Association of Fatigue with Elevated Antiviral and Proinflammatory Cytokines. [JOURNAL ARTICLE]
- Viral Immunol 2014 Jul 25.
Abstract This study aimed to characterize fatigue postinfection among study participants with a history of West Nile virus (WNV) infection and determine whether antiviral and pro-inflammatory cytokines were significantly elevated in those reporting prolonged fatigue. We found that 31% (44/140) of study participants experienced prolonged (more than 6 months) fatigue postinfection, with an average length of fatigue duration of 5 years. Females, those younger than 50 years of age, and those with symptomatic clinical WNV disease were significantly more likely to report fatigue. Pro-inflammatory and antiviral cytokines (granulocyte macrophage colony stimulating factor, interferon-γ, interferon-γ inducing protein 10, interleukin 2, interleukin 6, and interleukin 12p70) were significantly elevated in those reporting fatigue postinfection compared to those not reporting fatigue. Clinicians should consider history of WNV infection as a possible factor when evaluating causes of prolonged fatigue following a febrile viral illness in their patients.