Targeting lentiviral vectors to antigen-specific immunoglobulins

Gene transfer into B cells by lentivectors can provide an alternative approach to managing B lymphocyte malignancies and autoreactive B cell-mediated autoimmune diseases. These pathogenic B cell Populations can be distinguished by their surface expression of monospecific immunoglobulin. Development of a novel vector system to deliver genes to these specific B cells could improve the safety and efficacy of gene therapy. We have developed an efficient rnethod to target lentivectors to monospecific immunoglobulin-expressing cells in vitro and hi vivo. We were able to incorporate a model antigen CD20 and a fusogenic protein derived from the Sindbis virus as two distinct molecules into the lentiviral Surface. This engineered vector could specifically bind to cells expressing Surface immunoglobulin recognizing CD20 (αCD20), resulting in efficient transduction of target cells in a cognate antigen-dependent manner in vitro, and in vivo in a xenografted tumor model. Tumor suppression was observed in vivo, using the engineered lentivector to deliver a suicide gene to a xenografted tumor expressing αCD20. These results show the feasibility of engineering lentivectors to target immunoglobulin-specific cells to deliver a therapeutic effect. Such targeting lentivectors also Could potentially be used to genetically mark antigen-specific B cells in vivo to study their B cell biology

Somatodendritic secretion in oxytocin neurons is upregulated during the female reproductive cycle

During the female reproductive cycle, hypothalamic oxytocin (OT) neurons undergo sharp changes in excitability. In lactating mammals, bursts of electrical activity of OT neurons result in the release of large amounts of OT in the bloodstream, which causes milk ejection. One hypothesis is that OT neurons regulate their own firing activity and that of nearby OT neurons by somatodendritic release of OT. In this study, we show that OT neuron activity strongly reduces inhibitory synaptic transmission to these neurons. This effect is blocked by antagonists of both adenosine and OT receptors and is mimicked by OT application. Inhibition of soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex formation by tetanus toxin completely blocked the stimulation-induced reduction in inhibitory input, as did the calcium chelator BAPTA. During lactation, the readily releasable pool of secretory vesicles in OT cell bodies was doubled, and calcium currents were upregulated. This resulted in an increased inhibition of GABAergic synaptic transmission by somatodendritic release during lactation compared with the adult virgin stage. These results demonstrate that somatodendritic release is augmented during lactation, which is a novel form of plasticity to change the strength of synaptic transmission

Generation of Functional CLL-Specific Cord Blood CTL Using CD40-Ligated CLL APC

PMCID: PMC3526610This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

A Comparative Study of Different Methodologies for Fault Diagnosis in Multivariate Quality Control

Different methodologies for fault diagnosis in multivariate quality control have been proposed in recent years. These methods work in the space of the original measured variables and have performed reasonably well when there is a reduced number of mildly correlated quality and/or process variables with a well-conditioned covariance matrix. These approaches have been introduced by emphasizing their positive or negative virtues, generally on an individual basis, so it is not clear for the practitioner the best method to be used. This paper provides a comprehensive study of the performance of diverse methodological approaches when tested on a large number of distinct simulated scenarios. Our primary aim is to highlight key weaknesses and strengths in these methods as well as clarifying their relationships and the requirements for their implementation in practice.Vidal Puig, S.; Ferrer, A. (2014). A Comparative Study of Different Methodologies for Fault Diagnosis in Multivariate Quality Control. Communications in Statistics - Simulation and Computation. 43(5):986-1005. doi:10.1080/03610918.2012.720745S9861005435Arteaga, F., & Ferrer, A. (2010). How to simulate normal data sets with the desired correlation structure. Chemometrics and Intelligent Laboratory Systems, 101(1), 38-42. doi:10.1016/j.chemolab.2009.12.003Doganaksoy, N., Faltin, F. W., & Tucker, W. T. (1991). Identification of out of control quality characteristics in a multivariate manufacturing environment. Communications in Statistics - Theory and Methods, 20(9), 2775-2790. doi:10.1080/03610929108830667Fuchs, C., & Benjamini, Y. (1994). Multivariate Profile Charts for Statistical Process Control. Technometrics, 36(2), 182-195. doi:10.1080/00401706.1994.10485765Hawkins, D. M. (1991). Multivariate Quality Control Based on Regression-Adiusted Variables. Technometrics, 33(1), 61-75. doi:10.1080/00401706.1991.10484770Editorial Board. (2007). Computational Statistics & Data Analysis, 51(8), iii-v. doi:10.1016/s0167-9473(07)00125-9Hayter, A. J., & Tsui, K.-L. (1994). Identification and Quantification in Multivariate Quality Control Problems. Journal of Quality Technology, 26(3), 197-208. doi:10.1080/00224065.1994.11979526HOCHBERG, Y. (1988). A sharper Bonferroni procedure for multiple tests of significance. Biometrika, 75(4), 800-802. doi:10.1093/biomet/75.4.800HOMMEL, G. (1988). A stagewise rejective multiple test procedure based on a modified Bonferroni test. Biometrika, 75(2), 383-386. doi:10.1093/biomet/75.2.383Kourti, T., & MacGregor, J. F. (1996). Multivariate SPC Methods for Process and Product Monitoring. Journal of Quality Technology, 28(4), 409-428. doi:10.1080/00224065.1996.11979699Li, J., Jin, J., & Shi, J. (2008). Causation-BasedT2Decomposition for Multivariate Process Monitoring and Diagnosis. Journal of Quality Technology, 40(1), 46-58. doi:10.1080/00224065.2008.11917712Mason, R. L., Tracy, N. D., & Young, J. C. (1995). Decomposition ofT2 for Multivariate Control Chart Interpretation. Journal of Quality Technology, 27(2), 99-108. doi:10.1080/00224065.1995.11979573Mason, R. L., Tracy, N. D., & Young, J. C. (1997). A Practical Approach for Interpreting Multivariate T2 Control Chart Signals. Journal of Quality Technology, 29(4), 396-406. doi:10.1080/00224065.1997.11979791Murphy, B. J. (1987). Selecting Out of Control Variables With the T 2 Multivariate Quality Control Procedure. The Statistician, 36(5), 571. doi:10.2307/2348668Rencher, A. C. (1993). The Contribution of Individual Variables to Hotelling’s T 2 , Wilks’ Λ, and R 2. Biometrics, 49(2), 479. doi:10.2307/2532560Roy, J. (1958). Step-Down Procedure in Multivariate Analysis. The Annals of Mathematical Statistics, 29(4), 1177-1187. doi:10.1214/aoms/1177706449Runger, G. C., Alt, F. B., & Montgomery, D. C. (1996). Contributors to a multivariate statistical process control chart signal. Communications in Statistics - Theory and Methods, 25(10), 2203-2213. doi:10.1080/03610929608831832Sankoh, A. J., Huque, M. F., & Dubey, S. D. (1997). Some comments on frequently used multiple endpoint adjustment methods in clinical trials. Statistics in Medicine, 16(22), 2529-2542. doi:10.1002/(sici)1097-0258(19971130)16:223.0.co;2-jTukey, J. W., Ciminera, J. L., & Heyse, J. F. (1985). Testing the Statistical Certainty of a Response to Increasing Doses of a Drug. Biometrics, 41(1), 295. doi:10.2307/253066

A Two-Gene Signature, SKI and SLAMF1, Predicts Time-to-Treatment in Previously Untreated Patients with Chronic Lymphocytic Leukemia

We developed and validated a two-gene signature that predicts prognosis in previously-untreated chronic lymphocytic leukemia (CLL) patients. Using a 65 sample training set, from a cohort of 131 patients, we identified the best clinical models to predict time-to-treatment (TTT) and overall survival (OS). To identify individual genes or combinations in the training set with expression related to prognosis, we cross-validated univariate and multivariate models to predict TTT. We identified four gene sets (5, 6, 12, or 13 genes) to construct multivariate prognostic models. By optimizing each gene set on the training set, we constructed 11 models to predict the time from diagnosis to treatment. Each model also predicted OS and added value to the best clinical models. To determine which contributed the most value when added to clinical variables, we applied the Akaike Information Criterion. Two genes were consistently retained in the models with clinical variables: SKI (v-SKI avian sarcoma viral oncogene homolog) and SLAMF1 (signaling lymphocytic activation molecule family member 1; CD150). We optimized a two-gene model and validated it on an independent test set of 66 samples. This two-gene model predicted prognosis better on the test set than any of the known predictors, including ZAP70 and serum β2-microglobulin

Rituximab in B-Cell Hematologic Malignancies: A Review of 20 Years of Clinical Experience

Rituximab is a human/murine, chimeric anti-CD20 monoclonal antibody with established efficacy, and a favorable and well-defined safety profile in patients with various CD20-expressing lymphoid malignancies, including indolent and aggressive forms of B-cell non-Hodgkin lymphoma. Since its first approval 20 years ago, intravenously administered rituximab has revolutionized the treatment of B-cell malignancies and has become a standard component of care for follicular lymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, and mantle cell lymphoma. For all of these diseases, clinical trials have demonstrated that rituximab not only prolongs the time to disease progression but also extends overall survival. Efficacy benefits have also been shown in patients with marginal zone lymphoma and in more aggressive diseases such as Burkitt lymphoma. Although the proven clinical efficacy and success of rituximab has led to the development of other anti-CD20 monoclonal antibodies in recent years (e.g., obinutuzumab, ofatumumab, veltuzumab, and ocrelizumab), rituximab is likely to maintain a position within the therapeutic armamentarium because it is well established with a long history of successful clinical use. Furthermore, a subcutaneous formulation of the drug has been approved both in the EU and in the USA for the treatment of B-cell malignancies. Using the wealth of data published on rituximab during the last two decades, we review the preclinical development of rituximab and the clinical experience gained in the treatment of hematologic B-cell malignancies, with a focus on the well-established intravenous route of administration. This article is a companion paper to A. Davies, et al., which is also published in this issue

IGHV gene mutational status and 17p deletion are independent molecular predictors in a comprehensive clinical-biological prognostic model for overall survival prediction in chronic lymphocytic leukemia

Prognostic index for survival estimation by clinical-demographic variables were previously proposed in chronic lymphocytic leukemia (CLL) patients. Our objective was to test in a large retrospective cohort of CLL patients the prognostic power of biological and clinical-demographic variable in a comprehensive multivariate model. A new prognostic index was proposed