CD5 deletion enhances the antitumor activity of adoptive T cell therapies.

Most patients treated with US Food and Drug Administration (FDA)-approved chimeric antigen receptor (CAR) T cells eventually experience disease progression. Furthermore, CAR T cells have not been curative against solid cancers and several hematological malignancies such as T cell lymphomas, which have very poor prognoses. One of the main barriers to the clinical success of adoptive T cell immunotherapies is CAR T cell dysfunction and lack of expansion and/or persistence after infusion. In this study, we found that CD5 inhibits CAR T cell activation and that knockout (KO) of CD5 using CRISPR-Cas9 enhances the antitumor effect of CAR T cells in multiple hematological and solid cancer models. Mechanistically, CD5 KO drives increased T cell effector function with enhanced cytotoxicity, in vivo expansion, and persistence, without apparent toxicity in preclinical models. These findings indicate that CD5 is a critical inhibitor of T cell function and a potential clinical target for enhancing T cell therapies.

Noninvasive Monitoring of Mantle Cell Lymphoma by Immunoglobulin Gene Next-Generation Sequencing in a Phase 2 Study of Sequential Chemoradioimmunotherapy Followed by Autologous Stem-Cell Rescue.

BACKGROUND: Minimal residual disease (MRD) monitoring has been used to identify early molecular relapse and predict clinical relapse in mantle cell lymphoma (MCL). Few published data exist in MCL on the performance of next-generation sequencing-based assay of immunoglobulin gene rearrangements for MRD assessment. PATIENTS AND METHODS: In a prospective clinical trial (NCT01484093) with intensive induction chemotherapy and autologous stem-cell transplantation, posttreatment peripheral blood samples were collected from 16 MCL patients and analyzed with an earlier version of the Adaptive Biotechnologies MRD assay. RESULTS: Of the 7 patients whose disease remained in remission, the MRD test remained negative in 5 (71%). Of the 9 patients who experienced relapse, the MRD test was positive at least 3 months before relapse in 6 patients (67%) and positive at the time of relapse in 1 patient (11%). All patients with at least 2 positive MRD tests experienced relapse. CONCLUSION: The next-generation sequencing-based MRD assay identified early molecular relapse, and we observed more sensitivity in the cellular (circulating leukocytes) versus acellular (plasma cell-free DNA) compartment. This observation may be due to availability of tumor target or a limitation of the assay.

A meta-analysis for neurobehavioural effects due to electromagnetic field exposure emitted by GSM mobile phones.

BACKGROUND AND OBJECTIVE: Numerous studies have investigated the potential effects of electromagnetic fields (EMFs) emitted by GSM mobile phones ( approximately 900 MHz to approximately 1800 MHz) on cognitive functioning, but results have been equivocal. In order to try and clarify this issue, the current study carried out a meta-analysis on 19 experimental studies. DESIGN: Meta-analysis. METHODS: Nineteen studies were taken into consideration. Ten of them were included in the meta-analysis as they fulfilled several minimum requirements; for example, single-blind or double-blind experimental study design and documentation of means and standard deviation of the dependent variables. The meta-analysis compared exposed with non-exposed subjects assuming that there is a common population effect so that one single effect size could be calculated. When homogeneity for single effect sizes was not given, an own population effect for each study and a distribution of population effects was assumed. RESULTS: Attention measured by the subtraction task seems to be affected in regard to decreased reaction time. Working memory measured by the N-back test seems to be affected too: under condition 0-back target response time is lower under exposure, while under condition 2-back target response time increases. The number of errors under condition 2-back non-targets appears to be higher under exposure. CONCLUSION: Results of the meta-analysis suggest that EMFs may have a small impact on human attention and working memory.

Influence of hydrostatic pressure and sound amplitude on the ultrasound induced dispersion and de-agglomeration of nanoparticles.

In most applications, nanoparticles are required to be in a well-dispersed state prior to commercialisation. Conventional technology for dispersing particles into liquids, however, usually is not sufficient, since the nanoparticles tend to form very strong agglomerates requiring extremely high specific energy inputs in order to overcome the adhesive forces. Besides conventional systems as stirred media mills, ultrasound is one means to de-agglomerate nanoparticles in aqueous dispersions. In spite of several publications on ultrasound emulsification there is insufficient knowledge on the de-agglomeration of nanoparticulate systems in dispersions and their main parameters of influence. Aqueous suspensions of SiO2-particles were stressed up to specific energies EV of 10(4) kJ/m3 using ultrasound. Ultrasonic de-agglomeration of nanoparticles in aqueous solution is considered to be mainly a result of cavitation. Both hydrostatic pressure of the medium and the acoustic amplitude of the sound wave affect the intensity of cavitation. Furthermore, the presence of gas in the dispersion medium influences cavitation intensity and thus the effectiveness of the de-agglomeration process. In this contribution both, the influence of these parameters on the result of dispersion and the relation to the specific energy input are taken into account. For this, ultrasound experiments were carried out at different hydrostatic pressure levels (up to 10 bars) and amplitude values (64-123 microm). Depending on the optimisation target (time, energy input,...) different parameters limit the dispersion efficiency and result. All experimental results can be explained with the specific energy input that is a function of the primary input parameters of the process.

iNOS gene expression modulates microvascular responsiveness in endotoxin-challenged mice.

Septic shock is characterized by vasodilation and decreased responsiveness to vasoconstrictors. Recent studies suggest this results from nitric oxide (NO) overproduction after expression of the calcium-independent isoform of NO synthase (iNOS) in smooth muscle cells. However, direct evidence linking iNOS (NOS2) expression and decreased microvascular responsiveness after septic stimuli is lacking. In the present study, we determined the effect of bacterial lipopolysaccharide (LPS, 20 mg/kg, IP) on smooth muscle contraction and endothelial relaxation in mesenteric resistance arteries from wild-type and iNOS knockout mice. Four hours after challenge with LPS or saline in vivo, concentration-dependent responses to norepinephrine (NE) and acetylcholine (NE+ACh) were measured in cannulated, pressurized vessels ex vivo. In vessels from wild-type mice, NE-induced contraction was markedly impaired after LPS, and pretreatment with the iNOS inhibitor aminoguanidine (AG) partly restored the NE contraction. In contrast, NE contraction in microvessels from iNOS knockout mice was unaffected by LPS. ACh-induced relaxation was unaffected by LPS in vessels from either genotype. These data provide direct evidence that iNOS gene expression mediates the LPS-induced decrease in microvascular responsiveness to vasoconstrictors. Moreover, the observation that AG did not fully restore NE contraction after LPS, whereas iNOS gene deficiency did, suggests that iNOS expression plays a central role in the development of the NO-independent effect of LPS on microvascular responsiveness. Finally, our data indicate that LPS or iNOS expression has little effect on endothelium-dependent relaxation, and eNOS activity does not appear to play a role in the decreased smooth muscle responsiveness after LPS in this model. The full text of this article is available at http://www.circresaha.org.

Replication of an avian myxovirus in tumor cell cultures obtained from effusions of mammary carcinoma patients.

In view of the possible use of viruses for the immunotherapy of breast cancer, the replication of a strain of fowl plaque virus was studied in the tumor cells of 11 mammary carcinoma patients with malignant effusions. The tumor cells were obtained by centrifugation on iodamide solutions, then cultured in vitro, and infected by a fowl plaque virus previously adapted to grow in a mammary carcinoma cell line. Virus multiplication was observed in all cases, a prerequisite for the use of autologous viral oncolysates for immunotherapy in mammary carcinoma patients.

Efficacy and clinical cross-resistance of a new combination therapy (AMSA/VP16) in previously treated patients with acute nonlymphocytic leukemia.

We investigated the tolerance, efficacy, and clinical cross-resistance of a new combination chemotherapy in 38 patients with previously treated acute myeloblastic leukemia (AML). It consisted of 120 mg2/d 4'(9-acridinylamino) methanesulfon-m-Anisidide (m-AMSA) in a one-hour infusion and 80 mg/m2/d etoposide (VP-16) in a 24-hour infusion, both administered for 5 days. The first 27 patients also received vinblastine, 6 mg/m2 on day 8, but this therapy was discontinued because of intestinal complications. Thirteen of 23 patients (56%) at first or subsequent relapse and five of 15 patients (33%) who were primarily resistant to an anthracycline/cytarabine combination achieved a complete response (CR) (hemoglobin level not taken into account) with a median CR duration of 5 months and 2 months, respectively. The response rate was as high as 63% for patients at first or second relapse whether the remission was maintained or not. The median times to recovery of normal bone marrow cellularity, of blood granulocyte counts greater than 500/microL, and of platelets greater than 20,000/microL were 34, 27, and 22 days, respectively. Marked but reversible gastrointestinal toxicity was observed in 24% of the patients, and two patients died of infection during induction. The one-hour AMSA/continuous VP-16 combination is effective for patients with relapsing AML and shows no cross-resistance in a proportion of patients refractory to the standard anthracycline-cytarabine combination.

The free yeast aspartyl-tRNA synthetase differs from the tRNA(Asp)-complexed enzyme by structural changes in the catalytic site, hinge region, and anticodon-binding domain.

Aminoacyl-tRNA synthetases catalyze the specific charging of amino acid residues on tRNAs. Accurate recognition of a tRNA by its synthetase is achieved through sequence and structural signalling. It has been shown that tRNAs undergo large conformational changes upon binding to enzymes, but little is known about the conformational rearrangements in tRNA-bound synthetases. To address this issue the crystal structure of the dimeric class II aspartyl-tRNA synthetase (AspRS) from yeast was solved in its free form and compared to that of the protein associated to the cognate tRNA(Asp). The use of an enzyme truncated in N terminus improved the crystal quality and allowed us to solve and refine the structure of free AspRS at 2.3 A resolution. For the first time, snapshots are available for the different macromolecular states belonging to the same tRNA aminoacylation system, comprising the free forms for tRNA and enzyme, and their complex. Overall, the synthetase is less affected by the association than the tRNA, although significant local changes occur. They concern a rotation of the anticodon binding domain and a movement in the hinge region which connects the anticodon binding and active-site domains in the AspRS subunit. The most dramatic differences are observed in two evolutionary conserved loops. Both are in the neighborhood of the catalytic site and are of importance for ligand binding. The combination of this structural analysis with mutagenesis and enzymology data points to a tRNA binding process that starts by a recognition event between the tRNA anticodon loop and the synthetase anticodon binding module.

Routine bone marrow punctures during remission of acute myelogenous leukemia.

To determine whether serial examinations of the peripheral blood can replace regular bone marrow punctures in the diagnosis of a relapse of acute myelogenous leukemia (AML), the peripheral blood of 40 AML patients in remission undergoing regular bone marrow punctures was studied. Within three months prior to bone marrow examination proving relapse in 97% of the relapses, at least one of the following values of the peripheral blood was pathological: blasts (84%), neutrophil granulocytes (72%), thrombocytes (64%), and hemoglobin (58%). The simultaneous appearance of abnormalities in the peripheral blood and the bone marrow occurs in such high incidence that routine bone marrow punctures are rendered obsolete in the follow-up of AML patients. Needless pain and anxiety can therefore be avoided for many AML patients.

Hairy cell leukemia: an interferon deficient disease?

rIFN-alpha 2 is an effective substance in the treatment of HCL. Although the complete eradication of hairy cells from the bone marrow is rarely possible, a dramatic improvement of profound cytopenias can be obtained in most patients, 12 out of 13 in this study. The response can be seen after a median treatment duration of about 2 months and appears to be independent of the presence of the spleen. Once a stable improvement of the peripheral blood values is achieved, one dose of rIFN-alpha 2 every other week may be sufficient to maintain the cell counts. The mechanism(s) by which IFN acts remain unclear. We speculate, that HCL is an IFN deficient syndrome because monocytes which are the major source of IFN-alpha are severely depressed in this lymphoproliferative disorder. Following this hypothesis, the IFN therapy would be comparable to the substitution of insulin in diabetic patients.

Phase-II study of vindesine and hexamethylmelamine in patients with relapsing small cell carcinoma of the lung.

Twenty-five patients with measurable small cell lung cancer relapsing after first-line chemotherapy were treated with vindesine 3 mg/m2 IV on days 1 and 8 and hexamethylmelamine 100 mg/m2 PO on days 1-14, repeated every 3 weeks. Among 18 fully evaluable patients there was 1 partial remission lasting for 111 days. Two patients had disease stabilization for 127 and 152 days, respectively. Fifteen patients had disease progression. The treatment was well tolerated, myelosuppression being the major side-effect.

Influence of a new relapse treatment for acute myeloid leukemia (AML) on in vitro granulopoiesis.

Twelve AML patients in relapse were treated with cytosine arabinoside (Ara-C), VP 16-213, vincristine, and vinblastine (A-triple-V). For bone marrow (BM) evaluation, in vitro granulopoiesis by agar and liquid cultures was investigated. In 15 treatments, 12 complete remissions (CR) were observed. Three patients, treated with 2 A-triple-V cycles achieved CR twice. One to four months after treatment normal colony formation and cell differentiation was found in remission patients. Evidence of sustained recovery was obtained in sequential studies of BM cultures of patients in CR. These results indicate that-A-triple-V treatment does not irreversibly deplete normal myeloid progenitor cell population.

Malignant peritoneal mesothelioma after Thorotrast exposure.

A case of a malignant peritoneal mesothelioma in a 63-year-old male patient with a history of exposure to Thorotrast in 1945 is presented. There was no history of exposure to asbestos. The clinical manifestation was a serosal effusion, which required weekly ascites puncture until therapy with intraperitoneal bleomycin was initiated. The latter treatment led to a significant reduction of ascites without any influence on tumor progression. Unfortunately, intraperitoneal bleomycin was accompanied by pulmonary toxicity, but at a higher total dose than known for intravenous administration. Three years after diagnosis the patient is still alive, without relapse of ascites production after bleomycin had to be stopped. Considering the risk of pulmonary fibrosis with high-dose intraperitoneal bleomycin and the lack of efficacy on tumor reduction, bleomycin seems to offer no advantage with respect to cisplatin.

[Interdisciplinary follow-up in patients with tumors]. / Interdisziplinäre Kontrollen bei Tumorpatienten.

There are two main reasons for routine follow-up examinations after treatment of cancer patients: the assessment of treatment efficacy and detection of relapse and the rating of drug side-effects. Routine controls can only efficiently be performed with a profound knowledge of the biology of the tumor and of the therapeutic efficacy of the available treatments. The frequency and the type of the follow-up examinations depend mainly on the curative or the palliative treatment possibilities. Interdisciplinarity is important again only in case of new therapeutic considerations. Examples of useless controls are mentioned. Through the prevention of unnecessary examinations the primary-care physician could play an important role in the cut-down of healthcare costs.

Haploidentical hematopoietic SCT increases graft-versus-tumor effect against renal cell carcinoma.

Allogeneic hematopoietic SCT (HSCT) has been shown to be an effective treatment option for advanced renal cell cancer (RCC). However, tumor resistance/relapse remains as the main post transplant issue. Therefore, enhancing graft-versus-tumor (GVT) activity without increasing GVHD is critical for improving the outcome of HSCT. We explored the GVT effect of haploidentical-SCT (haplo-SCT) against RCC in murine models. Lethally irradiated CB6F1 (H2K(b/d)) recipients were transplanted with T-cell-depleted BM cells from B6CBAF1 (H2K(b/k)) mice. Haplo-SCT combined with a low-dose haploidentical (HI) T-cell infusion (1 × 10(5)) successfully provided GVT activity without incurring GVHD. This effect elicited murine RCC growth control and consequently displayed a comparative survival advantage of haplo-SCT recipients when compared with MHC-matched (B6D2F1CB6F1) and parent-F1 (B6CB6F1) transplant recipients. Recipients of haplo-SCT had an increase in donor-derived splenic T-cell numbers, T-cell proliferation and IFN-γ-secreting donor-derived T-cells, a critical aspect for anti-tumor activity. The splenocytes from B6CBAF1 mice had a higher cytotoxicity against RENCA cells than the splenocytes from B6 and B6D2F1 donors after tumor challenge. These findings suggest that haplo-SCT might be an innovative immunotherapeutic platform for solid tumors, particularly for renal cell carcinoma.

Interleukin-15 administration increases graft-versus-tumor activity in recipients of haploidentical hematopoietic SCT.

Utilizing a clinically relevant haploidentical (HI) murine transplant model, lethally irradiated B6D2F1 (H2K(b/d)) mice were transplanted with T cell-depleted (TCD) BM from B6CBAF1 (H2K(b/k)) mice. We found that administration of IL-15 significantly increases the numbers of CD8+ T and natural killer (NK) cells in spleen and BM after transplantion without GVHD. Graft-versus-tumor (GVT) potency of the graft was evaluated upon tumor challenge using P815 tumor cells (H2(d)). IL-15 administration without T-cell infusion did not result in any survival improvement. However, IL-15 in combination with very low-dose T-cell infusion (1 × 10(4)) significantly increased GVT activity and improved survival in recipients of HI hematopoietic SCT (HSCT). This effect was observed when IL-15 was given at a later time point, rather than immediately following transplantation. IL-15 administration also specifically increased slow-proliferative CD8+ T-cell proliferation and IFN-γ secretion in CD8+ T cells in recipients of CFSE (carboxyfluorescein succinimidyl ester)-labeled HI T-cell infusion, whereas there was no effect on CD4+ T-cell proliferation, suggesting the critical effect of IL-15 on CD8+ T-cell homeostasis in HI host. We conclude that IL-15 can be used for enhancing antileukemia effect of HI-HSCT, which requires presence of donor-derived T cells.

Produtividade de genótipos de trigo duplo propósito submetidos ao pastejo com vacas em lactação / Productivity of dual-purpose wheat genotypes under grazing with lactating cows

Esta pesquisa foi conduzida com o objetivo de avaliar a produtividade de dois genótipos de trigo de duplo propósito, BRS Tarumã e BRS Umbu, submetidos ao pastejo com vacas em lactação. O delineamento experimental foi o inteiramente ao acaso, com dois tratamentos (genótipos), três repetições (piquetes) e medidas repetidas no tempo (pastejos). Avaliaram-se a precocidade, a composição estrutural dos trigos, as produções de forragem e de biomassa de lâminas foliares, as taxas de acúmulo diário de forragem e de lâminas foliares, a taxa de lotação, as ofertas de forragem e de lâminas foliares, a eficiência de pastejo, o consumo aparente e a produção de grãos. O trigo mais precoce para produção de forragem foi o BRS Umbu. Houve diferença para a produção de forragem (3196 vs. 4143kg MS/ha) e de lâminas foliares (2281 vs. 3205kg MS/ha) para os genótipos BRS Umbu e BRS Tarumã, respectivamente. Valores similares foram encontrados para taxa de lotação (2,26UA/ha); eficiência de pastejo (52,26%), consumo aparente (2,91%) e produção de grãos (1716kg/ha). O genótipo BRS Tarumã é o mais indicado para o manejo de duplo propósito em condições de pastejo com vacas em lactação.(AU)

The objective of this research was to evaluate the productivity of two dual-purpose wheat genotypes BRS Tarumã and BRS Umbu under grazing with lactating cows. The experimental design was completely randomized, with two treatments, three replications (paddocks) and repeated measures (grazing cycles). Studied variables were early growth, the structural composition of wheat, forage production, leaf blade biomass, the stocking rate, the herbage and leaf blade allowance, the grazing efficiency, the herbage intake and grain yield. The earliest genotype for forage production was the BRS Umbu. Differences in herbage yield between BRS Umbu and BRS Tarumã genotypes (3196 vs. 4143kg DM/ha) and leaf blade production (2281 vs. 3205kg DM/ha), respectively, were detected. Similar values between cultivars were found in stocking rate (2,26AU/ha); grazing efficiency (52,26%), herbage intake (2,91 %) and grain yield (1716kg/ha). The BRS Tarumã genotype is the most suitable for dual-purpose under grazing with dairy cows.(AU)

Valor nutritivo de pastagens de Coastcross-1 em consórcio com diferentes leguminosas de ciclo hibernal / Nutritive value of coastcross-1 pastures mixed to different cool season legumes

Objetivou-se com esta pesquisa avaliar três sistemas forrageiros constituídos por Coastcross-1 (CC) + 100kg de N/ha/ano + ervilhaca comum; CC + 100kg de N/ha/ano + trevo vesiculoso; e CC + 200kg de N/ha/ano. Durante o período experimental (345 dias), foram realizados treze pastejos. O delineamento experimental utilizado foi o inteiramente ao acaso, com três tratamentos (sistemas forrageiros), três repetições (piquetes) em parcelas subdividas no tempo (valores médios dos pastejos em cada estação do ano). Para avaliação, foram utilizadas vacas em lactação da raça Holandesa. Amostras de simulação de pastejo foram coletadas para análise de proteína bruta (PB), fibra em detergente neutro (FDN) e ácido (FDA), digestibilidade in situ da matéria seca (DISMS) e da matéria orgânica (DISMO) e os nutrientes digestíveis totais (NDT). Os valores médios para PB, FDN, FDA, DISMS, DISMO e NDT foram de 18,1; 16,7 e 17,6%; 57,8; 58,9 e 58,7%; 26,5; 26,5 e 26,7%; 79,6; 78,9 e 80,6%; 79,8; 79,1 e 80,6%; 72,1; 71,4 e 72,7%, respectivamente. Melhores resultados de valor nutritivo foram obtidos no inverno, em especial para o consórcio de Coastcross-1 com ervilhaca.

The aim of this research was to evaluate three grazing systems with Coastcross-1 (CC) + 100kg N/ha/year + common vetch; CC + 100kg N/ha/year + arrowleaf clover; and CC + 200kg N/ha/year. Thirteen grazing cycles were performed during the experimental period (345 days). The experimental design was completely randomized with three treatments (grazing systems), three replicates (paddocks) in completely split-plot time (average values of grazing season). Lactating Holstein cows were used in the evaluation. Forage mass and botanical composition were evaluated. Samples from the hand-plucking method were collected to analyze crude protein (CP), neutral detergent fiber (NDF) and acid (ADF), in situ digestibility of dry matter (ISDMD) and organic matter (ISOMD) and total digestible nutrients (TDN). The averages of CP, NDF, ADF, ISDMD, ISOMD and TDN were 18.1, 16.7 and 17.6 %; 57.8, 58.9 and 58.7 %; 26.5, 26.5 and 26.7 %; 79.6, 78.9 and 80.6 %; 79.8, 79.1 and 80.6 %; 72.1, 71.4 and 72.7 %, respectively. Better results for nutritive value were found during winter, especially on Coastcross-1 mixed with common vetch.