Specific immunotherapy of experimental myasthenia gravis in vitro and in vivo: the Guided Missile strategy.

Current immunotherapy of myasthenia gravis (MG) is often effective, but entails risks of infection and neoplasia. The "Guided Missile" strategy described here is designed to target and eliminate the individual's unique AChR-specific T cell repertoire, without otherwise interfering with the immune system. We genetically engineered dendritic cells to present AChR epitopes and simultaneously express Fas ligand in an ongoing EAMG model. In both in vitro and in vivo experiments, these engineered cells specifically killed AChR-responsive T cells without otherwise damaging the immune system. AChR antibodies were markedly reduced in the treated mice. Translation of this method to treat human MG is possible.

[Orthovoltage roentgenotherapy in the treatment of lymphorrhea after reconstructive operations on the lower extremities arteries].

After performance of 728 reconstructive-restoration operations on the lower extremities arteries in 58 (7.9%) patients a lymphorrhea from the wound have had occurred. While roentgenotherapy application for postoperative lymphorrhea treatment in 67% patients a good result was achieved. The number of the ray therapy procedures was determined in accordance with the clinical effect obtained.

Granzyme B: evidence for a role in the origin of myasthenia gravis.

PURPOSE OF RESEARCH: Although the pathogenesis of myasthenia gravis (MG) as an antibody mediated disorder of acetylcholine receptors (AChRs) at neuromuscular junctions is well understood, the origin of the autoimmune response is unclear. The thymus is intimately involved in initiation of the autoimmune response; the antigen, AChR, is present in the thymus, but how the autoimmune response is triggered is not known. Granzyme B (GrB), a proteolytic enzyme present in cytolytic T cells and natural killer (NK) cells, selectively cleaves many potential autoantigens (but few non-autoantigens), generating novel fragments that trigger autoreactive responses. This protease has been strongly implicated in the pathogenesis of several autoimmune diseases including lupus, rheumatoid arthritis, dermatomyositis, and others. In the studies described in this manuscript, we examined the ability of GrB to cleave the AChR subunits, and performed biochemical, immunohistochemical and molecular studies on thymus glands from myasthenic patients and controls to assess GrB expression. MAIN RESULTS: GrB efficiently and specifically cleaves subunits of AChR, especially the epsilon subunit. GrB is present in thymus glands from myasthenia patients, but is absent in control thymuses. CONCLUSIONS: Our results provide evidence supporting a potential role for GrB in the process of initiation of MG, and are consistent with the concept of an immunodominant epsilon epitope.

Specific immunotherapy of experimental myasthenia by genetically engineered APCs: the "guided missile" strategy.

Although treatment of MG with general immunosuppressive agents is often effective, it has important drawbacks, including suppression of the immune system as a whole, with the risks of infection and neoplasia, and numerous other adverse side effects. Ideally, treatment of MG should eliminate the specific pathogenic autoimmune response to AChR, without otherwise suppressing the immune system or producing other adverse side effects. Although antibodies to AChR are directly responsible for the loss of AChRs at neuromuscular junctions in MG, the AChR antibody response is T cell-dependent, and immunotherapy directed at T cells can abrogate the autoantibody response, with resulting benefit. As in other autoimmune diseases, the T cell response in MG is highly heterogeneous. The design of specific immunotherapy must take this heterogeneity into account and target the entire repertoire of AChR-specific T cells. We describe our investigation of a novel strategy for specific immunotherapy of MG, involving gene transfer to convert antigen-presenting cells (APCs) to "guided missiles" that target AChR-specific T cells, and that induce apoptosis and elimination of those T cells. This strategy uses the ability of APCs from a given individual to present the entire spectrum of AChR epitopes unique for that individual, and thereby to target the entire repertoire of antigen-specific T cells of the same individual. Using viral vectors, we have genetically engineered the APCs to process and present the most important domain of the AChR molecule, and to express a "warhead" of Fas ligand (FasL) to eliminate the activated AChR-specific T cells with which they interact. Our results show that the APCs express the appropriate gene products, and effectively and specifically eliminate AChR-specific T cells by the Fas/FasL pathway, while sparing T cells of other specificities.

[Correcting effect of the locus coeruleus on ophthalmic hypertension of the hypothalamic genesis]. / Korrigiruiushchee vliianie sinego piatna mozga na oftal'mogipertenziiu gipotalamicheskogo geneza.

We studied the intraocular pressure and the humor aquosus volume changes after a chronic emotional stress produced by a prolonged electrical stimulation of the multiple ventromedial (VMHN) hypothalamic nuclei and the locus coeruleus. The VMHN stimulation caused an increase in the intraocular pressure and production of the humor aquosus, as well as a decrease in the outflow ratio. Stimulation of the locus coeruleus increased the intraocular pressure to a lesser extent. A combined effect of both types of stimulation normalised the intraocular pressure due to a decrease in the humor aquosus production, the outflow ratio remaining unchanged.

Endogenous regulation of a therapeutic transgene restores homeostasis in arthritic joints.

The treatment of chronic inflammatory diseases is complicated by their unpredictable, relapsing clinical course. Here, we describe a new strategy in which an inflammation-regulated therapeutic transgene is introduced into the joints to prevent recurrence of arthritis. To this end, we designed a recombinant adenoviral vector containing a two-component, inflammation-inducible promoter controlling the expression of human IL-10 (hIL-10) cDNA. When tested in vitro, this system had a low-level basal activity and was activated four to five orders of magnitude by various inflammatory stimuli, including TNF-alpha, IL-1 beta, IL-6, and LPS. When introduced in joints of rats with recurrent streptococcal cell wall-induced arthritis, the IL-10 transgene was induced in parallel with disease recurrence and effectively prevented the influx of inflammatory cells and the associated swelling of the joints. Levels of inflammation-inducible hIL-10 protein within the joints correlated closely with the severity of recurrence. An endogenously regulated therapeutic transgene can thus establish negative feedback and restore homeostasis in vivo while minimizing host exposure to the recombinant drug.

Oncogenic mutants of RON and MET receptor tyrosine kinases cause activation of the beta-catenin pathway.

beta-Catenin is an oncogenic protein involved in regulation of cell-cell adhesion and gene expression. Accumulation of cellular beta-catenin occurs in many types of human cancers. Four mechanisms are known to cause increases in beta-catenin: mutations of beta-catenin, adenomatous polyposis coli, or axin genes and activation of Wnt signaling. We report a new cause of beta-catenin accumulation involving oncogenic mutants of RON and MET receptor tyrosine kinases (RTKs). Cells transfected with oncogenic RON or MET were characterized by beta-catenin tyrosine phosphorylation and accumulation; constitutive activation of a Tcf transcriptional factor; and increased levels of beta-catenin/Tcf target oncogene proteins c-myc and cyclin D1. Interference with the beta-catenin pathway reduced the transforming potential of mutated RON and MET. Activation of beta-catenin by oncogenic RON and MET constitutes a new pathway, which might lead to cell transformation by these and other mutant growth factor RTKs.

Specific immunotherapy of experimental myasthenia gravis in vitro: the "guided missile" strategy.

We describe a strategy for specific immunotherapy of myasthenia gravis (MG) based on genetic engineering of antigen presenting cells (APCs) to present the autoantigen acetylcholine receptor (AChR) and express the "warhead" Fas ligand (FasL). For transduction of APCs we prepared recombinant attenuated vaccinia virus vectors carrying the following three gene constructs: (i) AChR fused to LAMP1 to present AChR and target AChR-specific T cells; (ii) FasL to eliminate the targeted T cells; and (iii) truncated FADD to protect APCs from self-destruction by FasL. The engineered APCs effectively expressed the genes of interest and killed AChR-specific T cells in culture by the Fas/FasL pathway. T cells specific for an unrelated antigen were spared. Our in vitro demonstration that engineered APCs target and kill antigen-specific T cells represents a promising novel strategy for specific immunotherapy of MG and other autoimmune diseases.

Specific immunotherapy by genetically engineered APCs: the "guided missile" strategy.

We tested the hypothesis that APCs genetically engineered to present an Ag and to express Fas ligand (FasL) simultaneously can target and eliminate Ag-specific T cells. Transgenic T cells specific for influenza hemagglutinin (HA) were used as targets. We prepared recombinant vaccinia virus vectors (VVV) to transfer the gene constructs individually or simultaneously into APCs. We prevented unwanted viral replication by attenuating the VVVs with psoralen-UV light treatment. For presentation of the HA Ag, APCs were transduced with cDNA for HA flanked by sequences of the lysosome-associated membrane protein that direct efficient processing and presentation of the Ag by APCs. As a "warhead" for the APCs, we transduced them with the gene for FasL, which induces apoptosis of Fas-expressing activated T cells. To protect the transduced APCs from self-destruction by FasL, we transferred cDNA for a truncated form of Fas-associated death domain, which inhibits Fas-mediated cell death. Our results show that the engineered APCs effectively expressed the genes of interest. APCs transduced with VVV carrying all three gene constructs specifically killed HA-transgenic T cells in culture. Coculture with T cells specific for an unrelated Ag (OVA) had no significant effect. Our in vitro findings show that APCs can be genetically engineered to target and kill Ag-specific T cells and represent a promising novel strategy for the specific treatment of autoimmune diseases.

NF-kappaB activation provides the potential link between inflammation and hyperplasia in the arthritic joint.

The transcription factor NF-kappaB is a pivotal regulator of inflammatory responses. While the activation of NF-kappaB in the arthritic joint has been associated with rheumatoid arthritis (RA), its significance is poorly understood. Here, we examine the role of NF-kappaB in animal models of RA. We demonstrate that in vitro, NF-kappaB controlled expression of numerous inflammatory molecules in synoviocytes and protected cells against tumor necrosis factor alpha (TNFalpha) and Fas ligand (FasL) cytotoxicity. Similar to that observed in human RA, NF-kappaB was found to be activated in the synovium of rats with streptococcal cell wall (SCW)-induced arthritis. In vivo suppression of NF-kappaB by either proteasomal inhibitors or intraarticular adenoviral gene transfer of super-repressor IkappaBalpha profoundly enhanced apoptosis in the synovium of rats with SCW- and pristane-induced arthritis. This indicated that the activation of NF-kappaB protected the cells in the synovium against apoptosis and thus provided the potential link between inflammation and hyperplasia. Intraarticular administration of NF-kB decoys prevented the recurrence of SCW arthritis in treated joints. Unexpectedly, the severity of arthritis also was inhibited significantly in the contralateral, untreated joints, indicating beneficial systemic effects of local suppression of NF-kappaB. These results establish a mechanism regulating apoptosis in the arthritic joint and indicate the feasibility of therapeutic approaches to RA based on the specific suppression of NF-kappaB.

[Infectious complications in patients in the terminal stages of chronic renal insufficiency]. / Infektsionnye oslozhneniia u bol'nykh s terminal'noi stadiei khronicheskoi pochechnoi nedostatochnosti.

Characteristics of infectious complications were investigated in 112 patients at the terminal stage of renal insufficiency treated by programme hemodialysis. High frequency of sepsis (32.4 per cent), urinary tract infections (27.7 per cent), respiratory tract infections (26.4 per cent) and tuberculosis (5.6 per cent) was stated. The lethal outcomes of the infectious complications averaged 20.3 per cent.

[Lomefloxacin pharmacokinetics in patients in the terminal stage of chronic renal insufficiency, undergoing treatment with programmed hemodialysis]. / Farmakokinetika lomefloksatsina u bol'nykh v terminal'noi pochechnoi nedostatochnosti, nakhodiashchikhsia na lechenii programmnym gemodializom.

The most frequent complications in patients with the terminal stage of chronic renal insufficiency are infections of various severity. The problem of the antibacterial therapy choice is especially urgent because of a high frequency of antibiotic resistant strains and the necessity to correct the treatment regimens in regard to the severity of the renal failure. The pharmacokinetics of lomefloxacin, a new fluoroquinolone, was studied in the treatment of patients with the terminal stage of chronic renal insufficiency treated by programmed hemodialysis. Lomefloxacin was administered orally in a dose of 400 mg at an interval of 48 hours 24 hours prior to the hemodialysis application. There was observed a decrease in the maximum serum concentration of the drug by comparison to that in healthy persons which could be due to slow absorption of the drug and hyperhydration in the patients because of anuria. In the treatment of such patients it is necessary to provide high serum concentrations of lomefloxacin attainable by using higher single doses of the drug.

[Pulmonary hemodynamics and external respiratory function in patients with chronic obstructive lung diseases. The diagnostic value of complex radiologic study]. / Legochnaia gemodinamika i funktsiia vneshnego dykhaniia u bol'nykh khronicheskimi obstruktuvnymi zabolevaniiami legkikh. Diagnosticheskaia tsennost' kompleksnogo luchevogo issledovaniia.

A study made of the external respiration function and pulmonary hemodynamics in 43 patients with chronic obstructive pulmonary diseases by fluorography and tomography disclosed highly significant signs of impairement of the external respiration function and pulmonary hemodynamics at early stages in the course of these conditions' development. The above findings are confirmed by a series of non-invasive techniques of investigation, such as radiopulmonography, jugular phlebography, tetrapolar pulmonary rheoplethismography.

[Biochemical study of the A2F4 antigen associated with human pulmonary adenocarcinoma]. / Biokhimicheskoe izuchenie antigena A2F4, assotsiirovannogo s adenokartsinomoi legkogo cheloveka.

The tissue-specific antigen associated with human lung adenocarcinoma had been investigated using immunological and biochemical methods. The antigen, which represents a new tissue-specific marker, has a molecular weight of 400 kDa. Purification of the antigen was achieved by gel chromatography. Antibody binding to the antigen was studied using enzyme-linked immunoassay after preincubation with enzymes or treatment with periodate. The results obtained testify to the proteinaceous nature of the antigenic determinant and the glycoprotein nature of the antigen.

[Intensive therapy of DIC-syndrome in patients with acute renal insufficiency caused by long-term crush syndrome]. / Intensivnaia terapiia DVS-sindroma u bol'nykh s ostroi pochechnoi nedostatoch-nost'iu, obuslovlennoi sindromom dlitel'nogo sdavleniia.

Impairment of parenchymatous organs, primarily kidneys, responsible for their dysfunction in crush syndrome results in many respects from disseminated intravascular coagulation (DIC). It is also associated with hemorrhagic complications. It is demonstrated that treatment modalities aimed at arrest of DIC syndrome (plasmapheresis, heparin, dysaggregation drugs, transfusions of large amounts of fresh frozen plasma) stopped bleeding and septic shock in 12 patients with crush syndrome following the earthquake in Armenia (1988).

[The results of using pseudoplasmapheresis in bronchial asthma (a preliminary report)]. / Rezul'taty primeneniia lozhnogo plazmafereza pri bronkhial'noi astme (predvaritel'noe soobshchenie).

The authors provide the results of the treatment of 15 patients with infectious bronchial asthma by plasmapheresis (PA). All the patients received PA in accordance with the techniques developed by the authors. These techniques simulate standard procedures using a plasticized container and a refrigerator centrifuge. As a result of the studies carried out, it has been concluded that the clinical efficacy of false PA is similar to that of routine PA as regards the clinical manifestations of bronchial asthma. Apparently, the elimination effect and the effect of red blood cells plasma withdrawal are of no material importance in the mechanism by which PA influences bronchial asthma. The psychosomatic causes, effects of temporary blood loss, blood contact with polymeric materials, and the influence of temporary blood cooling may be under discussion.

[Relation between arterial hypertension and disorder of erythrocyte deformability in patients with chronic renal failure]. / O sviazi mezhdu arterial'noi gipertoniei i narusheniem deformiruemosti éritrotsitov u bol'nykh s khronicheskoi pochechnoi nedostatochnost'iu.

A total of 29 patients with chronic renal insufficiency (CRI) were investigated. Noticeable disorder of erythrocytic deformability (ED) was detected in half of them. The expression of arterial hypertension showed correlation with a degree of ED disorder. A possibility of the main role of ED disorder in the development and progression of arterial hypertension was discussed. ED disorder was found to correlate with a degree of expression of laboratory signs of the DIC-syndrome and was practically unassociated with the blood level of creatinine.