Early transduction produces highly functional chimeric antigen receptor-modified virus-specific T-cells with central memory markers: A Production Assistant for Cell Therapy (PACT) translational application

Background: Virus-specific T-cells (VSTs) proliferate exponentially after adoptive transfer into hematopoietic stem cell transplant (HSCT) recipients, eliminate virus infections, then persist and provide long-term protection from viral disease. If VSTs behaved similarly when modified with tumor-specific chimeric antigen receptors (CARs), they should have potent anti-tumor activity. This theory was evaluated by Cruz et al. in a previous clinical trial with CD19.CAR-modified VSTs, but there was little apparent expansion of these cells in patients. In that study, VSTs were gene-modified on day 19 of culture and we hypothesized that by this time, sufficient T-cell differentiation may have occurred to limit the subsequent proliferative capacity of the transduced T-cells. To facilitate the clinical testing of this hypothesis in a project supported by the NHLBI-PACT mechanism, we developed and optimized a good manufacturing practices (GMP) compliant method for the early transduction of VSTs directed to Epstein-Barr virus (EBV), Adenovirus (AdV) and cytomegalovirus (CMV) using a CAR directed to the tumor-associated antigen disialoganglioside (GD2).Results: Ad-CMVpp65-transduced EBV-LCLs effectively stimulated VSTs directed to all three viruses (triVSTs). Transduction efficiency on day three was increased in the presence of cytokines and high-speed centrifugation of retroviral supernatant onto retronectin-coated plates, so that under optimal conditions up to 88% of tetramer-positive VSTs expressed the GD2.CAR. The average transduction efficiency of early-and late transduced VSTs was 55 ± 4% and 22 ± 5% respectively, and early-transduced VSTs maintained higher frequencies of T cells with central memory or intermediate memory phenotypes. Early-transduced VSTs also had higher proliferative capacity and produced higher levels of TH1 cytokines IL-2, TNF-α, IFN-γ, MIP-1α, MIP-1β and other cytokines in vitro.Conclusions: We developed a rapid and GMP compliant method for the early transduction of multivirus-specific T-cells that allowed stable expression of high levels of a tumor directed CAR. Since a proportion of early-transduced CAR-VSTs had a central memory phenotype, they should expand and persist in vivo, simultaneously protecting against infection and targeting residual malignancy. This manufacturing strategy is currently under clinical investigation in patients receiving allogeneic HSCT for relapsed neuroblastoma and B-cell malignancies (NCT01460901 using a GD2.CAR and NCT00840853 using a CD19.CAR)

Земство и мировой суд в России: организация публичной власти на местном уровне

The authors of the article reveal the essence and content of the Zemstvo and Judicial reforms (1864) in the general context of social relations and the domestic policy of the Russian Empire, showing their connection with the Peasant reform (1861). As a result of the reforms carried out by the government of Alexander II, local public authority was transformed. All-estate zemstvo institutions and courts independent from the administration appeared in the provinces and counties. The article describes the Zemstvo reform aimed at solving of social and economic problems of the citizens at local level, in its relationship with the Judicial reform, which had established courts of peace, guaranteed equal protection of personal and property rights of citizens, including peasants who had received freedom recently. The authors reveal the features of the new public order in the activities of zemstvo institutions and local courts; substantiate the organizational principles common to zemstvos and courts of peace (estate-inclusiveness, independence from administration, legality, etc.); characterize the structure and competence of zemstvo institutions and courts of peace in their interaction with each other, as well as with the government bodies. The self-government bodies and the courts of peace, introduced by the government of Alexander II, contradicted to the historically established social order and state system of the Russian Empire, which predetermined their future fate. During the implementation of the conservative course of the government of Alexander III the courts of peace were abolished and the institute of zemsky district commanders with administrative and judicial powers was established in rural areas in 1889.Авторы статьи раскрывают сущность и содержание земской и судебной реформ (1864 г.) в общем контексте социальных отношений и внутренней политики российского самодержавия, показывают их опосредованность крестьянской реформой (1861 г.). В результате проведенных правительством Александра II преобразований трансформировалась публичная власть на местах: в губерниях и уездах появились всесословные земские учреждения и независимые от администрации мировые суды. В статье дается характеристика земской реформы, нацеленной на решение социальных и хозяйственных проблем населения, в ее взаимосвязи с судебной реформой, учредившей мировые суды и гарантировавшей равную защиту личных и имущественных прав представителям всех сословий, включая вышедших из крепостной зависимости крестьян. Авторы выявляют черты нового публичного порядка в деятельности земских учреждений и мировых судов; обосновывают общие для земств и мировой юстиции принципы организации (всесословность, независимость от администрации, законность и др.); характеризуют структуру и компетенцию земских учреждений и мировых судов в их взаимодействии друг с другом, а также с органами государственного управления. Введенные правительством Александра II органы губернского и уездного земского самоуправления и мировые суды противоречили исторически сложившемуся общественному укладу и государственному строю Российской империи, что предопределило их дальнейшую судьбу. В ходе реализации консервативного курса правительства Александра III в 1889 г. были ликвидированы в сельской местности мировые суды и учрежден институт земских участковых начальников, наделенных административно-судебными полномочиями

Zemstvo and Court of peace in Russia: Public authority at local level [Земство и мировой суд в России: организация публичной власти на местном уровне]

The authors of the article reveal the essence and content of the Zemstvo and Judicial reforms (1864) in the general context of social relations and the domestic policy of the Russian Empire, showing their connection with the Peasant reform (1861). As a result of the reforms carried out by the government of Alexander II, local public authority was transformed. All-estate zemstvo institutions and courts independent from the administration appeared in the provinces and counties. The article describes the Zemstvo reform aimed at solving of social and economic problems of the citizens at local level, in its relationship with the Judicial reform, which had established courts of peace, guaranteed equal protection of personal and property rights of citizens, including peasants who had received freedom recently. The authors reveal the features of the new public order in the activities of zemstvo institutions and local courts; substantiate the organizational principles common to zemstvos and courts of peace (estate-inclusiveness, independence from administration, legality, etc.); characterize the structure and competence of zemstvo institutions and courts of peace in their interaction with each other, as well as with the government bodies. The self-government bodies and the courts of peace, introduced by the government of Alexander II, contradicted to the historically established social order and state system of the Russian Empire, which predetermined their future fate. During the implementation of the conservative course of the government of Alexander III the courts of peace were abolished and the institute of zemsky district commanders with administrative and judicial powers was established in rural areas in 1889. Copyright © 2019 by International Network Center for Fundamental and Applied Research