Frequency analysis of cytolytic T lymphocyte precursors (CTL-P) generated in vivo during lethal rabies infection of mice. II. Rabies virus genus specificity of CTL-P

Cytolytic T lymphocyte precursors (CTL-P) were sensitized in vivo by intraplantar infection of C57BL/6 mice with a lethal dose of rabies virus, strain ERA (ERA). As a result of sensitization CTL-P matured to interleukin-receptive CTL-P (IL-CTL-P) that could be expanded in vitro to Thy-1+, Lyt-2+ CTL clones in the presence of IL without subjection to antigen-driven selection. After infection with ERA, IL-CTL-P-derived CTL lysed fibroblasts infected with rabies virus but not those infected with another rhabdovirus, the vesicular stomatitis virus. These CTL, however, did not discriminate between fibroblasts infected with the serologically closely related laboratory strains of classic rabies virus, ERA and HEP-Flury, and the serologically distinct rabies-related African isolate Mokola. This finding implies that in vivo sensitized IL-CTL-P recognize common genus-specific determinants expressed on cells infected with members of the lyssavirus genus

Satisfiability Games for Branching-Time Logics

The satisfiability problem for branching-time temporal logics like CTL*, CTL and CTL+ has important applications in program specification and verification. Their computational complexities are known: CTL* and CTL+ are complete for doubly exponential time, CTL is complete for single exponential time. Some decision procedures for these logics are known; they use tree automata, tableaux or axiom systems. In this paper we present a uniform game-theoretic framework for the satisfiability problem of these branching-time temporal logics. We define satisfiability games for the full branching-time temporal logic CTL* using a high-level definition of winning condition that captures the essence of well-foundedness of least fixpoint unfoldings. These winning conditions form formal languages of \omega-words. We analyse which kinds of deterministic {\omega}-automata are needed in which case in order to recognise these languages. We then obtain a reduction to the problem of solving parity or B\"uchi games. The worst-case complexity of the obtained algorithms matches the known lower bounds for these logics. This approach provides a uniform, yet complexity-theoretically optimal treatment of satisfiability for branching-time temporal logics. It separates the use of temporal logic machinery from the use of automata thus preserving a syntactical relationship between the input formula and the object that represents satisfiability, i.e. a winning strategy in a parity or B\"uchi game. The games presented here work on a Fischer-Ladner closure of the input formula only. Last but not least, the games presented here come with an attempt at providing tool support for the satisfiability problem of complex branching-time logics like CTL* and CTL+

Frequency analysis of cytolytic T cell precursors (CTL-P) generated in vivo during lethal rabies infection of mice. I. Distinction of CTL-P with different interleukin 2 sensitivity

The aim of this study was to determine the number and state of activity of cytolytic T lymphocytes (CTL) and their precursors (CTL-P) present in vivo during the early stages of viral infection. The local response to lethal infection with rabies virus was used as a model system that is not accessible to analysis by secondary activation in vitro. The local response to alloantigen served as a control. Experimental protocols were established that allow frequency estimates of in vivo antigen-triggered CTL-P. Data allow a distinction between CTL-P activated in vivo by alloantigen and viral antigen with respect to their different capacity to utilize T cell growth factors (inter-leukins). In vivo alloantigen-primed CTL-P generate, in vitro, an active effector progeny in the presence of interleukins of xenogeneic origin, whereas the majority of virus-specific CTL-P, in spite of considerable expansion in vivo, fail to generate CTL in vitro unless antigen is added

Alloreactive cytotoxic T lymphocytes generated in the presence of viral- derived peptides show exquisite peptide and MHC specificity

The nature of alloreactivity to MHC molecules has been enigmatic, primarily because of the observation that allogeneic responses are considerably stronger than syngeneic responses. To better determine the specificity potential of allogeneic responses, we have generated alloreactive CTL specific for exogenous, viral-derived peptide ligands. This approach allowed us to critically evaluate both the peptide- and MHC-specificity of these alloreactive T cells. Exploiting the accessibility of the H-2Ld class I molecule for exogenous peptide ligands, alloreactive CTL were generated that are specific for either murine cytomegalovirus (MCMV) or lymphocytic choriomeningitis virus (LCMV) peptides bound by Ld alloantigens. Peptide specificity was initially observed in bulk cultures of alloreactive CTL only when tested on peptide-sensitized T2.Ld target cells that have defective presentation of endogenous peptides. Subsequent cloning of bulk alloreactive CTL lines generated to MCMV yielded CTL clones that had exquisitely specific MCMV peptide recognition requirement. All of the MCMV/Ld alloreactive CTL clones were also exquisitely MHC-specific in that none of the CTL clones lysed targets expressing MCMV/Lq complexes, even though Lq differs from Ld by only six amino acid residues and Lq also binds the MCMV peptide. This observation clearly demonstrates that alloreactive CTL are capable of the same degree of specificity for target cell recognition as are syngeneic CTL in MHC-restricted responses

Model Checking CTL is Almost Always Inherently Sequential

The model checking problem for CTL is known to be P-complete (Clarke, Emerson, and Sistla (1986), see Schnoebelen (2002)). We consider fragments of CTL obtained by restricting the use of temporal modalities or the use of negations—restrictions already studied for LTL by Sistla and Clarke (1985) and Markey (2004). For all these fragments, except for the trivial case without any temporal operator, we systematically prove model checking to be either inherently sequential (P-complete) or very efficiently parallelizable (LOGCFL-complete). For most fragments, however, model checking for CTL is already P-complete. Hence our results indicate that in most applications, approaching CTL model checking by parallelism will not result in the desired speed up. We also completely determine the complexity of the model checking problem for all fragments of the extensions ECTL, CTL +, and ECTL +

Shared determinants between virus-infected and trinitrophenyl-conjugated H-2-identical target cells detected in cell-mediated lympholysis

Infection of H-2-identical mice with either lymphocytic choriomeningitis (LCM) virus, vaccinia virus, or paramyxo (Sendai) virus resulted in the generation of specifically sensitized cytotoxic T lymphocytes (CTL). CTL generated in vitro against 2,4,6-trinitrophenyl (TNP)-conjugated syngeneic stimulator cells were specifically cytotoxic for TNP-conjugated H-2K(D) region identical targets. Both LCM and vaccinia-induced CTL, however, were found to be strongly cytotoxic towards TNP-conjugated, H-2K(D) region-identical target cells. In contrast, Sendai virus-induced CTL did not lyse TNP-conjugated, syngeneic target cells. Inhibition experiments using cold targets suggested that shared antigenic determinants can be detected on either LCM virus-infected and TNP-conjugated targets, which are not present on the cell surface 6f normal target cells

The cytolytic T lymphocyte response to the murine cytomegalovirus

Limiting dilution (LD) analysis with two modifications, the expansion and the restimulation LD assay, led to the detection and quantification of two distinct in vivo maturation stages within the lineage of virus- specific self-restricted CTL after infection of mice with the murine cytomegalovirus (MCMV). A low frequency set, representing an average of 15% of the specifically activated CTL-P in a draining lymph node, generated virus-specific lytic activity in the absence of antigen, solely under expansion conditions provided by growth and differentiation interleukins. These cells were considered to be active and were denoted antigen-independent or interleukin-receptive CTL-P (IL- CTL-P). A high frequency set required additional antigen in vitro to generate functionally active clones, and therefore the cells were termed antigen-dependent. Both sets are present in vivo simultaneously at the peak of the acute immune response and represent antigen- activated cells because their existence strictly depends on a preceding priming event. IL-CTL-P disappear quickly after acute infection and are absent during the memory state. It is proposed that the isolation of IL- CTL-P could serve to detect viral antigen expression during persistent and/or recurrent herpes virus infections

Model Checking CTL is Almost Always Inherently Sequential

The model checking problem for CTL is known to be P-complete (Clarke, Emerson, and Sistla (1986), see Schnoebelen (2002)). We consider fragments of CTL obtained by restricting the use of temporal modalities or the use of negations---restrictions already studied for LTL by Sistla and Clarke (1985) and Markey (2004). For all these fragments, except for the trivial case without any temporal operator, we systematically prove model checking to be either inherently sequential (P-complete) or very efficiently parallelizable (LOGCFL-complete). For most fragments, however, model checking for CTL is already P-complete. Hence our results indicate that, in cases where the combined complexity is of relevance, approaching CTL model checking by parallelism cannot be expected to result in any significant speedup. We also completely determine the complexity of the model checking problem for all fragments of the extensions ECTL, CTL+, and ECTL+