Lymphocyte Stimulation Research Articles

Loss of DIAPH1 causes SCBMS, combined immunodeficiency, and mitochondrial dysfunction

Homozygous loss of DIAPH1 results in seizures, cortical blindness, and microcephaly syndrome (SCBMS). We studied 5 Finnish and 2 Omani patients with loss of DIAPH1 presenting with SCBMS, mitochondrial dysfunction, and immunodeficiency. We sought to further characterize phenotypes and disease mechanisms associated with loss of DIAPH1. Exome sequencing, genotyping and haplotype analysis, B- and T-cell phenotyping, invitro lymphocyte stimulation assays, analyses of mitochondrial function, immunofluorescence staining for cytoskeletal proteins and mitochondria, and CRISPR-Cas9 DIAPH1 knockout in heathy donor PBMCs were used. Genetic analyses found all Finnish patients homozygous for a rare DIAPH1 splice-variant (NM_005219:c.684+1G>A) enriched in the Finnish population, and Omani patients homozygous for a previously described pathogenic DIAPH1 frameshift-variant (NM_005219:c.2769delT;p.F923fs). In addition to microcephaly, epilepsy, and cortical blindness characteristic to SCBMS, the patients presented with infection susceptibility due to defective lymphocyte maturation and 3 patients developed B-cell lymphoma. Patients' immunophenotype was characterized by poor lymphocyte activation and proliferation, defective B-cell maturation, and lack of naive T cells. CRISPR-Cas9 knockout of DIAPH1 in PBMCs from healthy donors replicated the T-cell activation defect. Patient-derived peripheral blood T cells exhibited impaired adhesion and inefficient microtubule-organizing center repositioning to the immunologic synapse. The clinical symptoms and laboratory tests also suggested mitochondrial dysfunction. Experiments with immortalized, patient-derived fibroblasts indicated that DIAPH1 affects the amount of complex IV of the mitochondrial respiratory chain. Our data demonstrate that individuals with SCBMS can have combined immune deficiency and implicate defective cytoskeletal organization and mitochondrial dysfunction in SCBMS pathogenesis.

Weaning Induces Stress-Dependent DNA Methylation and Transcriptional Changes in Piglet PBMCs.

Changes to the epigenome, including those to DNA methylation, have been proposed as mechanisms by which stress can induce long-term physiological changes in livestock species. Pig weaning is associated with dietary and social stress, both of which elicit an immune response and changes to the hypothalamic–pituitary–adrenal (HPA) axis. While differential methylation following stress has been assessed in model organisms, it remains poorly understood how the pig methylome is altered by stressors in production settings. We quantified changes in CpG methylation and transcript abundance in piglet peripheral blood mononuclear cells (PBMCs) following weaning and also assessed differential patterns in pigs exhibiting high and low stress response as measured by cortisol concentration and lesion scores. Blood was collected from nine gilt piglets 24 h before and after weaning, and whole-genome bisulfite sequencing (WGBS) and RNA-sequencing were performed on six and nine animals, respectively, at both time points. We identified 2,674 differentially methylated regions (DMRs) that were enriched within promoters of genes associated with lymphocyte stimulation and transcriptional regulation. Stress groups displayed unique differential methylation and expression patterns associated with activation and suppression of T cell immunity in low and high stress animals, respectively. Differential methylation was strongly associated with differential expression; specifically, upregulated genes were enriched among hypomethylated genes. We observed post-weaning hypermethylation of the glucocorticoid receptor (NR3C1) promoter and a significant decrease in NR3C1 expression (n = 9, p = 6.1 × 10–3). Our results indicate that weaning-associated stress elicits genome-wide methylation changes associated with differential gene expression, reduced T cell activation, and an altered HPA axis response.

Sustained hyperbilirubinemia following clarithromycin administration

Abstract Rationale: Hyperbilirubinemia is observed in drug-induced liver injury of cholestatic type, but direct inhibition of transporter proteins which facilitates bilirubin transport also causes hyperbilirubinemia. Here we present a case of sustained hyperbilirubinemia after the administration of clarithromycin. Patient concerns: A 52-year-old man was referred to our hospital because of sustained hyperbilirubinemia. Both contrast-enhanced computed tomography and Magnetic resonance cholangiopancreatography did not reveal any abnormality. But his liver was not enhanced in the hepatocellular phase of Gd-EOB-DTPA MRI. Indocyanine Green (ICG) retention rate at 15 minutes (ICG(R15)) was 58%, urinary coproporphyrin was 324 μg/g Cr, and serum bile acid was 283 nmol/mL. Technetium-99m galactosyl human serum albumin (99mTc-GSA) liver scintigram revealed that his liver function was normal (blood clearance ratio (HH15), 0.572; and hepatic uptake ratio (LHL15), 0.948) Diagnoses: These findings suggested that hyperbilirubinemia in the present case was caused by impaired organic anion-transporting polypeptide (OATP) 1B1 and 1B3 activity. Drug-induced lymphocyte stimulation test (DLST) for clarithromycin was positive. Among the drugs prescribed before the presentation of hyperbilirubinemia, clarithromycin is the only agent that has been reported to reduce the OATP1B1/1B3 transporter activity. Interventions: The patient continued taking oral UDCA, but hyperbilirubinemia did not improve. Bilirubin absorption therapy was performed twice. Outcomes: Following bilirubin absorption therapy, hyperbilirubinemia resolved and the bilirubin levels became normal. With the decrease in serum bilirubin, the liver was normally enhanced on the hepatocellular phase of Gd-EOB-DTPA MRI. ICG(R15). Urinary coproporphyrin, and bile acid levels returned to normal range (ICG(R15) of 9.2%, urinary coproporphyrin of 58 μg/g Cr, and bile acid of 13.5 nmol/mL). Lessons: The present case indicates that uptake of Gd-EOB-DTPA in the liver, ICG retention test, serum bile acid levels combined with urinary coproporphyrin can be a potential marker of OATP transporter activity and help in differential diagnosis of sustained hyperbilirubinemia.

B lymphocyte stimulating factor of Trachinotus ovatus (TroBLyS) promotes antibacterial immune defense

B lymphocyte stimulating factor (BLyS) belongs to the tumor necrosis family (TNF), which is a key cytokine in promoting the survival, proliferation, differentiation and apoptosis of B cells, and regulating the functions of T cells. However, functions of BLyS in teleosts are not well known. In the present study, the functions of BLyS in golden pompano (Trachinotus ovatus) (TroBLyS) were conducted through in vivo and in vitro experiments. Gene expression analysis suggested that TroBLyS was ubiquitously expressed in different tissues. Particularly, TroBLyS was most expressed in head kidney, followed by spleen, and liver. After Vibrio harveyi stimulation, the expressions of TroBLyS in head kidney, spleen, and liver were significantly upregulated. Recombinant TroBLyS (rTroBLyS) in vitro could promote lymphocytes proliferation and survival. Moreover, overexpression of TroBLyS in vivo in fish significantly promoted the activation of macrophages, increased the secretion of cytokines and chemokines, and reduced V. harveyi colonization in tissues. When the expression of TroBLyS was inhibited in T. ovatus, the bacterial count of V. harveyi was significantly increased than that in the control fish. Furthermore, the mortality of fish which was administrated of pTroBLyS for two weeks, was significantly reduced after V. harveyi infection, suggesting that TroBLyS had the potential as an immune enhancer. Our study indicated that TroBLyS might be involved in the antibacterial immune responses and has the potential as an immune system enhancer in T. ovatus.

Non-IgE- or Mixed IgE/Non-IgE-Mediated Gastrointestinal Food Allergies in the First Years of Life: Old and New Tools for Diagnosis.

non-IgE and mixed gastrointestinal food allergies present various specific, well-characterized clinical pictures such as food protein-induced allergic proctocolitis, food protein-induced enterocolitis and food protein-induced enteropathy syndrome as well as eosinophilic gastrointestinal disorders such as eosinophilic esophagitis, allergic eosinophilic gastroenteritis and eosinophilic colitis. The aim of this article is to provide an updated review of their different clinical presentations, to suggest a correct approach to their diagnosis and to discuss the usefulness of both old and new diagnostic tools, including fecal biomarkers, atopy patch tests, endoscopy, specific IgG and IgG4 testing, allergen-specific lymphocyte stimulation test (ALST) and clinical score (CoMiss).

Pharmacokinetics analysis based on target-mediated drug distribution for RC18, a novel BLyS/APRIL fusion protein to treat systemic lupus erythematosus and rheumatoid arthritis

Background and PurposeRC18 is a novel recombinant fusion protein targeting on B lymphocyte stimulator (BLyS). We aimed to develop and qualify a population pharmacokinetics (PopPK) model for RC18 in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) patients, taking into account the mechanistic target-mediated drug disposition (TMDD) process. MethodsA TMDD model of RC18 was developed using data from two phase I clinical trial (n = 23). The TMDD structural model was developed by simultaneous fitting of the serum free RC18 and serum RC18-BLyS complex. Potential covariates were screened using stepwise method, and predictive performance was qualified using a prediction-corrected visual predictive check (pcVPC) and bootstrap. ResultsA two compartment TMDD model with first order absorption for subcutaneous administration was built. The final model included a significant relationship between distribution volume of the central compartment and body weight. And the baseline of immunoglobulin IgG had significant effect on the baseline of target BLyS. The plots from goodness-of-fit and pcVPC confirmed good predictive performance of this TMDDmodel. ConclusionsThis mechanistic TMDD model integrated the interaction of RC18 with its target BLyS and accurately predicts both RC18 and RC18-BLyS complex profiles in RA and SLE patients. Simulated target change profiles can be used to help guide rational dose regimen selection and used as a biomarker for efficacy evaluation.

Exploring the effects of formula feeding on infant immune development in China: A prospective cohort study.

The worldwide exclusive breastfeeding rate is suboptimal and this study aims to evaluate effects on infant immune development of formula feeding. A prospective study including 221 infants fed with breast milk or formula was conducted. At 3-month and 9-month, the concentrations of total immunoglobulin (Ig)G, IgM, IgA, IgG1, IgG2, interleukin (IL)-4, interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were measured by using enzyme-linked immunosorbent assay (ELISA). Natural killer (NK) cell activity and lymphocyte transformation testing were conducted. Furthermore, the occurrence of infantile diarrhea, respiratory infections and allergic diseases were questioned. The levels of total IgG (Z=-3.21, p=0.001), IgG1 (Z=-2.12, p=0.034), IFN-γ (t=-2.09, p=0.039) and NK cell activity (t=-2.14, p=0.034) were significant higher in formula-fed infants compared to breast-fed after 3 months. At 9-month, the levels of total IgG (Z=-4.34, p<0.001), IgA (Z=-2.05, p=0.041) and TNF-α (t=-2.10, p=0.037) of formula-fed infants were higher, but the lymphocyte stimulation index (t=2.76, p=0.007) was lower than breast-fed infants. While, no significant differences were found in the incidences of diarrhea and respiratory tract infection (p>0.05). This investigation suggested that formula- and breast-feeding have different contributions to infant immune development, but the formula feeding would not cause significantly increase of diarrhea and respiratory infections.

CIPLUKAN (PHYSALIS ANGULATA LINN.) EXTRACT AS A NATURAL ADJUVANT REMEDIES FOR SCLERODERMA

Scleroderma or systemic sclerosis is a rare connective tissue autoimmune disorder.1 Its prevalence in Indonesia is not well recorded but scleroderma is listed as the third most common disease in rheumatology outpatient clinic in Hasan Sadikin Hospital, Bandung, West Java, Indonesia.2,3 Scleroderma is characterized by skin fibrosis, small vessel vasculopathy, and immune system abnormality. The cutaneous lesion of scleroderma is presenting with excessive collagen type-1 production and deposition, as well as exaggerated inflammatory process. There is no specific diagnosis test is available, but in most cases, there are some laboratory findings which support the diagnosis of scleroderma, such as elevated erythrocyte sedimentation rate (ESR), B lymphocyte stimulator (BAFF), C-Reactive Protein (CRP), Soluble Cd40 Ligand (sCD40L), and Procollagen Type-1 N-Terminal Propeptide (P1NP).3-6&#x0D; The treatment of this autoimmune disease remains a challenge for clinicians. The current treatment for scleroderma is expensive and some side effects as well as drug resistance are reported. Some new medicines are still under research, not accessible, and too expensive for most Indonesian communities. Hence, new therapeutic options are constantly sought.7 &#x0D; Ciplukan herb (Physalis angulate Linn) is a medicinal plant which grows in Indonesia. It has been known for its efficacy and safety as long-term anti-inflammatory, anti-proliferative, and anti-angiogenesis agent.7 Dewi et al conducted a double –blind, randomized clinical trial using ciplukan herb extract as an adjuvant treatment for scleroderma. The clinical trial was performed in Jakarta and Bandung, Indonesia for 2 years. There are 59 scleroderma patients, age 15-60 years old, had stable treatment for at least 3 months, who randomly divided into treatment group (29 people) and control group (30 people). The treatment group received 250 mg ciplukan herb extract three times a day and the control group received 250 mg amylum three times a day for 12 weeks. A 35,9% skin thickness reduction was found in treatment group. It is greater than control group which only gained 6,3% reduction (p

Clinical effects and applications of the gut microbiome in hematologic malignancies

The gut microbiome and its effects on host immunity have exciting implications for cancer prognosis and therapy. Examples in allogeneic hematopoietic stem cell transplantation (allo-SCT) demonstrate the role of the gut microbiome as a biomarker for clinical outcomes, and animal models demonstrate how microbiota manipulation may augment therapeutic responses. There are multiple mechanisms that gut microbiota may have in affecting distant tumor environments, including control of cytokine release, dendritic cell activation, and T-cell lymphocyte stimulation. Recently, there has been a marked interest in understanding interactions between host and microbiome in hematologic malignancies. This review summarizes the current understanding of the gut microbiome and its impact on leukemia, lymphoma, multiple myeloma, and allo-SCT and highlights several broad methods for targeting the gut microbiome in therapeutic trials.

Cytokines mediated hyperinflammation in SARS-CoV2: An Overview

Hyperinflammation induced by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV2) is a major cause of disease severity and mortality in infected patients. The immunopathogenesis of SARS-CoV2 infection is similar to the previous Middle East Respiratory Syndrome-related coronavirus (MERS-CoV) and SARS-CoV coronavirus with severe inflammatory responses. Therefore, severity of this viral infection is not only associated with the virus but also due to host immune responses. Hyperinflammatory responses due to cytokine storm are a centerpiece of SARS-CoV2 pathogenesis with overwhelming consequences for the host. Virus infected monocyte derived macrophages produce cytokines and this contributes to damage of lymphoid tissue and limits the lymphocyte responses. Blocking the deadly cytokine storm and T lymphocyte stimulation is a vital defense for treating SARS-CoV2. Here, we will describe the role of hyperinflamation and the involvement of cytokines in severe SARS-CoV2 infection.

B cell activating factor (BAFF): Structure, functions, autoimmunity and clinical implications in Systemic Lupus Erythematosus (SLE).

The B cell activating factor (BAFF), or B lymphocyte stimulator (BLyS), is a B cell survival factor which supports autoreactive B cells and prevents their deletion. BAFF expression is closely linked with autoimmunity and is enhanced by genetic alterations and viral infections. Furthermore, BAFF seems to be involved in adipogenesis, atherosclerosis, neuro-inflammatory processes and ischemia reperfusion (I/R) injury. BAFF is commonly overexpressed in Systemic Lupus Erythematosus (SLE) and strongly involved in the pathogenesis of the disease. The relationship between BAFF levels, disease activity and damage accrual in SLE is controversial, but growing evidence is emerging on its role in renal involvement. Belimumab, a biologic BAFF inhibitor, has been the first biologic agent licensed for SLE therapy so far. As Rituximab (RTX) has been shown to increase BAFF levels following B cell depletion, the combination therapy of RTX plus belimumab (being evaluated in two RCT) seems to be a valuable option for several clinical scenarios. In this review we will highlight the growing body of evidence of immune and non-immune related BAFF expression in experimental and clinical settings.

Mesalazine allergy and an attempt at desensitization therapy in patients with inflammatory bowel disease

Mesalazine is a key drug used for remission induction and maintenance therapy in inflammatory bowel disease (IBD). We sometimes encounter patients who develop allergic reactions to the drug and inevitably discontinue treatment. Of 692 patients who received mesalazine for IBD between 2014 and March 2020, 33 diagnosed with mesalazine allergy (43 episodes) were included, and their clinical manifestations were evaluated. For ten patients undergoing desensitization therapy, therapeutic outcomes were evaluated. The incidence of mesalazine allergy was 4.8%. The time from the start of oral medication to allergy onset was 10 ± 5 days for the first allergic attack and 2 ± 1 days for the second and subsequent allergic attacks. The observed clinical symptoms included fever (93%), diarrhea (26%), abdominal pain (23%), and bloody stool (12%). Drug-induced lymphocyte stimulation test was performed in 85% of the patients (28/33), and the sensitivity was 51%. Desensitization therapy with a time-dependent mesalazine granule formulation was successful in nine of the ten patients (90%), allowing them to receive 2000 mg or more of the drug. Fever was a common allergic symptom, and its presence appeared to be useful for distinguishing mesalazine allergy from exacerbation of the primary disease. Desensitization therapy was useful in patients with mesalazine allergy.

Antigenic Potency of LY6E in Stimulating Dendritic Cells to Elicit Tumor-Specific Responses Against Human Colorectal and Gastric Cancer Cell Lines

Early-stage gastrointestinal (GI) carcinomas are amenable malignancies, however, due to late diagnosis or lack of proper medication, alternative treatment necessitates new approaches such as dendritic cell (DC) therapy. Our previous microarray study indicated Lymphocyte antigen-6 (LY6E) as a commonly overexpressed biomarker in three lethal GI cancers, colon, gastric, and pancreatic. Therefore, we examined the antigenic potency of LY6E in stimulating DCs to elicit tumor-specific responses against human colorectal cancer (CRC) and gastric cancer (GC) cell lines HT-29 and AGS, respectively. LY6E peptide-pulsed DCs stimulated lymphocytes up to 55.9% in comparison with mature DCs (48.3%). Also, flow cytometry analysis of lymphocyte proliferation illustrated that the populations CD4+ and CD8+ were increased after treating by peptide-pulsed DCs (62.9% and 48.7% respectively). Furthermore, the cytotoxicity assay demonstrated that the 40:1 ratio of stimulated lymphocytes on AGS and HT29 cell lines was 65.1% and 66.2%, respectively. The research exposed that LY6E loaded DCs had substantial impact stimulation, proliferation, and lineage differentiation of lymphocytes. Besides, co-cultured of primed lymphocytes with AGS and HT29 cell lines exhibited cytotoxic activity. These data suggest LY6E as a potential candidate in developing DC therapy against CRC and AGS.

Insulin resistance and obesity affect monocyte-derived dendritic cell phenotype and function

AimCardiovascular disease (CVD) is prevalent in women after menopause, which may be associated with obesity, insulin resistance and metaflammation. Despite the recognized role of immunological mechanisms in vascular remodeling, the role of dendritic cells (DCs) is still unclear. The aim was to characterize monocyte-derived DCs (Mo-DC) in post-menopausal patients with type 2 diabetes (T2DM) and obese woman, without clinical manifestations of atherosclerosis. MethodsObese post-menopausal women with or without T2DM were enrolled and were compared to age-matched healthy women. DCs obtained from patients were phenotypically and functionally characterized by flow cytometry and mixed lymphocyte reaction. MRNA integrins expression was assessed by real time RT-PCR; circulating fetuin-A and adiponectin levels were measured by ELISA. ResultsPhenotypic dysregulation of Mo-DC reported was related to a defective allogenic lymphocyte stimulation and to an increased mRNA of CD11c, CD18 and DC-SIGN/CD209 which regulate their adhesion to vascular wall cells. Fetuin-A and adiponectin levels were significantly altered and negatively correlated. Hyperglycaemia significantly impaired CD14+ transdifferentiation into Mo-DC. ConclusionsThese data show a dysfunction of Mo-DCs obtained from precursors isolated from T2DM obese post-menopausal woman without any documented clinical CV event. Association of obesity to diabetes seems to worsen DC's phenotype and function and increase vascular inflammation.

Belimumab in systemic lupus erythematosus: a profile of its use

Belimumab (Benlysta®), a human immunoglobulin (Ig) G1λ monoclonal antibody targeting B lymphocyte stimulator, is effective as an add-on to standard therapy in patients with active, autoantibody-positive systemic lupus erythematosus (SLE). Belimumab was first approved in 2011 as an intravenous (IV) treatment for adults with SLE. Since then, a subcutaneous formulation of belimumab has been approved in adults and, most recently, IV belimumab has been approved in children aged ≥ 5 years. In adults with active, autoantibody-positive SLE, add-on belimumab improves overall disease activity and flare rates. Belimumab appears to have a steroid-sparing effect and disease control is maintained over long-term use. The clinical benefits of IV belimumab in children with SLE are consistent with those in adults. Belimumab is generally well tolerated in both adults and children.

Immunopathological effects of experimental T-2 mycotoxicosis in Wistar rats.

It is well known that T-2 toxin has cytotoxic radiomimetic like effects on the immune system. Because of scant research data demonstrating the chronic effects of low doses of the T-2 toxin on humoral and cellular responses in rats, the present experiment was undertaken. The animals were divided into four groups, namely, group I (0.5 ppm), group II (0.75 ppm) and group III (1.0 ppm) and group IV (control) were given toxin-free diet for 12 weeks and eight animals each were sacrificed at 2, 4, 6, 8, 10, and 12-week of the experimental period. The humoral immune response was evaluated based on hemagglutination test (HA), and levels of serum immunoglobulins (IgA, IgG, IgM) while the cell-mediated immune response was evaluated by delayed-type hypersensitivity (DTH) response to ovalbumin, lymphocyte stimulation index, analyses of CD4+ and CD8+ T lymphocytes and mRNA expression levels of selected cytokines like IL-2, IFN-γ, IL-4 and IL-10 by quantitative Real-time PCR in experimental groups. T-2 treatment caused suppression in both humoral and cell-mediated immune responses as evidenced by a decrease in all these parameters in toxin fed animals compared to the control in the dose and duration-dependent manner. This dose-dependent effect on the immune system has been further reflected largely by the depletion of lymphocytes from lymphoid organs as observed histopathologically in the spleen, thymus, and Peyer's patches in the present study.

Lymphocyte stimulation test in the diagnostics of adverse reactions to antituberculouse line 1 and 2 drugs.

Adverse drug reactions to anti-TB drugs (ADR) are found in 6-20% of patients and have various clinical manifestations and are detected in the lymphocyte stimulation test (LST), recorded by the incorporation on H3 thymidine, but nowadays it has significant limitations. We used LST with WST-1 reagent to detect ADR to the main 1-st and 2-nd line antituberculosis drugs in 11 tuberculosis patients who had ADR (6 - hepatotoxic reaction, 3 - blood eosinophilia and 2 - with joint pain syndrome). 6 people with tuberculosis contacts made up the control group. LST evaluation with WST-1 showed that in patients with a hepatotoxic reaction, the SI index was>2 and exceeded the values in the control group (3.28±0.59, 95% CI-1.16 and 0, 74±0.16, 95% CI - 0.31, respectively) upon stimulation of cell cultures with rifampicin alone but not with other drugs. Cell cultures stimulated with the PHA mitogen have SI >2 in ADR patients (mainly with hepatotoxic reactions). Control group SI was <2 (4,93±0.53, 95% CI - 1, 04 and 1.97±0.3, 95% CI - 0.59, respectively). We have not detected PPD-L cell cultures stimulation with WST-1 reagent both in the group of patients with ADR and the control group. In patients with eosinophilia and joint pain syndrome SI was low for all studied drugs and did not differ from the control group. The sensitivity of the LST test with WST-1 reagent is not sufficient to determine ADR to anti-TB drugs.

Safety assessment of a newly synthesized copolymer for micellar delivery of hydrophobic caffeic acid phenethyl ester

Caffeic acid phenethyl ester (CAPE), a major pharmacologically active component of poplar type propolis, is known for its proapoptotic, anti-inflammatory, antioxidant, antiviral, and enzyme inhibiting activities. The aim of this study was to perform an in vitro and in vivo safety assessment of a micellar system based on a newly synthesized copolymer, consisting of polyglycidol and poly(allyl glycidyl ether) (C12-PAGE-PG) as a drug delivery platform for CAPE. The in vitro studies on HepG2 and L929 cells by MTT and LDH assays after treatment with the empty and CAPE-loaded micelles showed no cytotoxic effects of the empty micelles and retained cytotoxic activity of CAPE loaded in the micelles. No hemolysis or stimulation of mouse lymphocytes or macrophages was observed in vitro. In vivo hematological, biochemical, and histological assays on rats, treated with the empty (2580 and 5160 µg/kg) or CAPE-loaded (375 and 750 µg CAPE/kg) micelles did not reveal pathological changes of any of the parameters assayed after 14-days’ treatment. In conclusion, initial toxicological data characterize C12-PAGE-PG as a non-toxic and promising copolymer for development of micellar drug delivery systems, particularly for a hydrophobic active substance as CAPE.

Methicillin resistant staphylococcus aureus（MRSA）多発膿瘍経過中に発症し，ステロイドが著効したDrug reaction with eosinophilia and systemic symptoms（DRESS）の1例(A case report: Corticosteroid effective Drug reaction with eosinophilia and systemic symptoms (DRESS) due to antibiotics drugs)

要旨 症例は83歳の男性。発熱，呼吸困難，血圧低下で救急搬送となり，化膿性椎間板炎および腸腰筋膿瘍，敗血症性ショックの診断で，腸腰筋膿瘍に対して経皮的穿刺ドレナージ後にICU入室となった。血液培養からはMethicillin resistant staphylococcus aureus（MRSA）が検出され，全身に多発膿瘍形成を認めた。バンコマイシンを長期間投与する方針とし，洗浄ドレナージを追加した。第9病日に血液培養は陰性化したが，第18病日に発熱，拡大する紅斑，好酸球増多，腎機能障害，肝機能障害が出現し，循環動態の悪化および炎症所見が増悪した。敗血症の二次感染を疑い加療したが状態は改善せず，RegiSCARスコアよりDrug reaction with eosinophilia and systemic symptoms（DRESS）と診断した。被疑薬である抗菌薬を中止し，ステロイド投与により循環動態は安定し，発熱，紅斑，好酸球増多，腎機能障害，肝機能障害は改善した。本症例は敗血症の二次感染との鑑別に難渋し，敗血症診療のピットフォールであると考えられた。

Efficient Colorectal Cancer Gene Therapy with IL-15 mRNA Nanoformulation.

Immunogene therapy is a novel method for the treatment of colorectal cancer. Cytokine IL-15 has exhibited therapeutic anticancer potential due to its immune-stimulation property. However, conventional IL-15-based cancer gene therapy studies have been performed using the plasmid DNA form, which has potential shortcomings including weak delivery efficiency and backbone effect. In this study, an IL-15 immunogene therapy study for colon cancer using in vitro transcript mRNA is described. A protamine/liposome system (CLPP) is developed to provide efficient condensation and delivery capacity for in vivo mRNA transportation. They demonstrated that the prepared CLPP system could deliver the IL-15-encoding mRNA into C26 cells with high efficacy. The secretory expressed IL-15 cytokine by the C26 cells successfully produced lymphocyte stimulation and triggered anticancer cytotoxicity upon cancer cells in vitro. Local or systemic administration of the CLPP/mIL-15 complex exhibited obvious inhibition effects on multiple C26 murine colon cancer models with inhibition rates of up to 70% in the C26 abdominal cavity metastasis tumor model, 55% in the subcutaneous model, and 69% in the pulmonary metastasis model, demonstrating high efficacy and safety. These results successfully demonstrated the high therapeutic potential of the CLPP/mIL-15 complex for colorectal cancer immunogene therapy.