[State of collective immunity against poliomyelitis in some regions of Russia].

AIM: Study the state of collective immunity against poliomyelitis in 7 regions of Russia in the last 3 years. MATERIALS AND METHODS: 2579 sera were studied for antibodies against poliomyelitis virus. Antibodies (AT) against 3 types of viruses were determined in neutralization reaction in RD cell culture, the state of collective immunity in the examined individuals was evaluated by the percent of individuals with AT against a type of poliovirus and geometric mean AT titer. The circulation of wild polioviruses was judged by the presence of strain specific AT against wild and vaccine viruses in the examined children (311 sera were studied). RESULTS: The indicators of collective immunity against poliomyelitis in both select examined regions and select age groups were generally high. The data obtained allow to make a conclusion that the quality of vaccine prophylaxis in the examined regions is good. Introduction of wild poliovirus type 1 from Tajikistan in 2010 caused disease in 7 residents of Russia whereas an epidemic that had affected more than 700 individuals emerged in Tajikistan. CONCLUSION: The studies carried out confirmed the necessity to continue qualitative poliomyelitis vaccine prophylaxis in the country despite the lack of circulation of wild polioviruses that can be introduced at any time.

Immunomodulatory effects of mixed hematopoietic chimerism: immune tolerance in canine model of lung transplantation.

Long-term survival after lung transplantation is limited by acute and chronic graft rejection. Induction of immune tolerance by first establishing mixed hematopoietic chimerism (MC) is a promising strategy to improve outcomes. In a preclinical canine model, stable MC was established in recipients after reduced-intensity conditioning and hematopoietic cell transplantation from a DLA-identical donor. Delayed lung transplantation was performed from the stem cell donor without pharmacological immunosuppression. Lung graft survival without loss of function was prolonged in chimeric (n = 5) vs. nonchimeric (n = 7) recipients (p < or = 0.05, Fisher's test). There were histological changes consistent with low-grade rejection in 3/5 of the lung grafts in chimeric recipients at > or =1 year. Chimeric recipients after lung transplantation had a normal immune response to a T-dependent antigen. Compared to normal dogs, there were significant increases of CD4+INFgamma+, CD4+IL-4+ and CD8+ INFgamma+ T-cell subsets in the blood (p < 0.0001 for each of the three T-cell subsets). Markers for regulatory T-cell subsets including foxP3, IL10 and TGFbeta were also increased in CD3+ T cells from the blood and peripheral tissues of chimeric recipients after lung transplantation. Establishing MC is immunomodulatory and observed changes were consistent with activation of both the effector and regulatory immune response.

[Evolution of tick-borne encephalitis and a problem of evolution of its causative agent].

The evolution of tick-borne encephalitis (TBE) is marked by the expanded nosological area, the transformation of landscapes, the formation of anthropurgic foci, the change of environmental systems, the increase of mortality rate mainly among urban dwellers, as well as pathomorphism. The evolution of natural TBE virus (TBEV) populations was studied in Eastern and Western Siberia, Middle Urals, and the European part of the nosological area. The paper first describes the types of evolutionary transformations of viral populations under the conditions of a varying environmental and epidemiological situation. These include: 1) the change of TBEV subtypes over 50-60 years; substitution of the Far-Eastern subtype for its Siberian subtype (the Sverdlovsk and Kemerovo regions); 2) the steady-state circulation of one Siberian subtype with mutanttypes being accumulated (the Vologda region); 3) co-existence of the Far-Eastern and Siberian subtypes with the common vector Ixodes persulcatus (the Yaroslavl and Irkutsk regions, etc.); 4) original mixed TBEV strains including the gene sites of proteins E and NSI of two subtypes. There is new evidence that the Siberian subtype is able to induce focal TBE forms, leading to death.

Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection.

Several clinical studies of gene-modified T cells have shown limited in vivo function of the cells, immunogenicity of the transgene, and lack of a selective advantage for gene-modified T cells. To address these problems, we developed a lentiviral vector (LV) that provides a selectable, proliferative advantage and potentially decreases immunogenicity for transduced T cells. The bicistronic vector expressed two genes linked with an internal ribosomal entry site. One gene is a variant of the inosine monophosphate dehydrogenase 2, inosine monophosphate dehydrogenase (IMPDH(IY)), conferring resistance to the immunosuppressive drug mycophenolate mofetil (MMF). The other is a suicide gene, herpes simplex virus thymidine kinase (HSV-TK), rendering proliferating cells sensitive to ablation with ganciclovir, fused to the selectable transmembrane marker DeltaCD34 (DeltaCD34/TK). Cells transduced with LV-DeltaCD34/TK.IMPDH(IY) were efficiently enriched by immunomagnetic selection for CD34, proliferated in 0.5-5 microM MMF, and were killed by 0.5-25 microg ml(-1) ganciclovir. We demonstrate efficient selection and killing of gene-modified cells and suggest LV-DeltaCD34/TK.IMPDH(IY)-transduced T cells could be used to facilitate allogeneic hematopoietic cell engraftment. The expression of IMPDH(IY) would allow in vivo selection with MMF, and DeltaCD34/TK expression would allow rapid and safe elimination of transduced T cells if graft-versus-host disease developed.

Protection of human and murine hepatocytes against Fas-induced death by transferrin and iron.

Recent studies in a murine model show that transferrin (Tf) interferes with Fas-mediated hepatocyte death and liver failure by decreasing pro-apoptotic and increasing anti-apoptotic signals. We show here in vitro in murine and human hepatocyte cell lines and in vivo in mice that Fas-induced apoptosis is modulated by exogenous Tf and iron. The results obtained with iron-free Tf (ApoTf), iron-saturated Tf (FeTf), and the iron chelator salicylaldehyde isonicotinoyl hydrazone (SIH) in its iron-free and iron-saturated (FeSIH) forms indicate that apoptosis-modulating effects of Tf are not mediated by iron alone. Both the Tf molecule and iron affect multiple aspects of cell death, and the route of iron delivery to the cell may be critical for the final outcome of cellular Fas signaling. Survival of hepatocytes 'stressed' by Fas signals can be manipulated by Tf and iron and may be a target for prophylactic and therapeutic interventions.

Pro-apoptotic and anti-apoptotic effects of transferrin and transferrin-derived glycans on hematopoietic cells and lymphocytes.

The aim of this study was to test the hypothesis that transferrin (Tf) has anti-apoptotic properties and thereby exerts a cytoprotective effect against tissue damage induced by irradiation and other cytotoxic modalities. This hypothesis was tested in several models, including in vitro human short-term marrow cultures, subpopulations of marrow cells, particularly, CD56(+) natural killer cells (and natural killer cell lines), and in vivo radioprotection of murine marrow cells. Reverse transcriptase polymerase chain reaction analysis was used for determination of cytokine mRNA. Preincubation of human marrow with Tf protected cells (except for a CD56(+) subpopulation) against cell death induced by gamma-irradiation, tumor necrosis factor-alpha (TNF-alpha), and agonistic anti-Fas monoclonal antibody. Deglycosylation of Tf abrogated this action of Tf; conversely, Tf-derived glycans (Tf-Gly) (but not glycans isolated from other proteins) mimicked the effects of the intact Tf molecule on apoptosis. Antibodies specific for the Tf receptor (CD71) did not block the effects of Tf or Tf-Gly on apoptosis. Determination of cytokine mRNA in the course of Fas-mediated apoptosis in the presence of Tf or Tf-Gly showed upregulation of mRNA for Fas ligand and TNF-alpha in CD56(+) and downregulation of these transcripts along with upregulation of mRNA for interleukin-10 in CD3(+) marrow cells. Under these conditions, a distinct increase in Fas-associated phosphatase-1 message was observed in CD3(+) cells that were protected by Tf or Tf-Gly against apoptosis. The in vitro data were confirmed in a murine in vivo model in which pretreatment of mice with Tf protected marrow cells against gamma-irradiation-induced cell death. These data suggest a role for Tf and particularly Tf-Gly in the regulation of programmed cell death, apparently via alterations in cytokine expression, and provide a basis for additional studies on the use of Tf in cytoprotective protocols.

Upregulation of interleukin-10 and inhibition of alloantigen responses by transferrin and transferrin-derived glycans.

Previous studies have shown that critically timed administration of transferrin (Tf) facilitates induction of immunologic unresponsiveness. Here, we determined in mixed leukocyte culture (MLC) and in concanavalin A (ConA)-driven cultures the effect of exogenous Tf and Tf-derived glycans (Tf-Gly) on lymphocyte proliferation. In cultures of human blood lymphocytes, Tf inhibited selectively alloantigen-driven proliferation in MLC, but not ConA-stimulated lymphocyte proliferation. Deglycosylation of Tf abrogated the inhibitory effect of Tf on alloantigen-induced lymphocyte proliferation, and, consistent with a role for glycans, an effect qualitatively and quantitatively similar to Tf was exerted by purified Tf-Gly. Glycans isolated from other proteins, for example, immunoglobulin G (IgG) or fibrinogen, failed to inhibit alloantigen-induced proliferation selectively. Rather, they suppressed lymphocyte proliferation in a non-specific manner. Determination of cytokines in MLC supernatant showed a downregulation of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), IL-2, and IL-12 (p40), along with an upregulation of IL-10, a pattern entirely consistent with the observed effects of Tf and Tf-Gly on alloantigen-induced lymphocyte proliferation. The effect of Tf on MLC was directly IL-10-dependent. IL-10 levels were inversely correlated with lymphocyte proliferation and CD86 expression. Neutralization of IL-10 by anti-IL-10 monoclonal antibody (mAb) blocked the effect of Tf. The MLC-modulating effect of Tf (or Tf-Gly) was not dependent upon the Tf receptor CD71 but appeared to be mediated by a Gly-responsive receptor. These data suggest a role of Tf, and, in particular, Tf-Gly, in allo-interactions that is independent from the role of Tf in iron metabolism, and appears to involve co-stimulatory signals.

Negative regulators of hemopoiesis and stroma function in patients with myelodysplastic syndrome.

The mechanism that leads to hemopoietic failure in patients with myelodysplastic syndrome (MDS) is not well understood. There is evidence, however, that regulatory molecules such as tumor necrosis factor (TNF)-alpha, Fas (CD95), and Fas-ligand, which negatively affect hemopoiesis by way of apoptosis are upregulated. Here we analyzed marrow samples from 80 patients with MDS in regard to TNF-alpha and Fas-ligand levels and a possible correlation with various disease parameters and risk factors. TNF-alpha levels were elevated in comparison to samples from normal marrow donors, however, no significant correlation with FAB subtype, cytogenetic risk group or score by the International Prognostic Scoring System (IPSS) was observed. However, there was an inverse correlation between the cytogenetic risk category (low, intermediate, high) and levels of soluble Fas-ligand. The major source of TNF-alpha were mononuclear (non-stromal) cells which appeared to produce TNF-alpha at maximum levels. Limiting dilution analysis of CD34+ precursor cells showed that individually assayed cells, removed from companion cells that presumably provided negative signals such as TNF-alpha or Fas-ligand, were able to generate progressively increasing numbers of colonies. Stromal layers derived from MDS marrow did not have an inhibitory effect. In fact, higher colony numbers were obtained from both normal and MDS marrow derived hemopoietic precursors propagated on irradiated stromal layers from MDS marrow than on stromal layers from normal marrow. These results show that substantial numbers of normal hemopoietic precursors persist in MDS marrow. However, differentiation into mature cells is inhibited by negative signals originating from accessory or abnormal hemopoietic precursors in the non-adherent marrow fraction.

Unresponsiveness to human leukocytes in immunosuppressed mice by combined donor-derived human transferrin and antigens.

Previous work on the facilitation of xenogeneic and allogeneic bone marrow engraftment in irradiated mice and dogs with transferrins allowed the development of a model for induction of an apparently durable state of immunological unresponsiveness or 'tolerance' in chemically immunosuppressed mice. The system is based on the simultaneous and combined administration of donor-derived cell antigens, namely human leukocytes, and specific donor-derived or plasma pool human transferrin into BALB/c or C57BL/6 mice previously treated with prednisolone and cyclophosphamide on day 0 and day 1 of the experiment. A properly timed presentation of both donor-specific or plasma pool transferrin and leukocyte antigens into the mice on day 3 and day 16 of the experiment, in the course of initial restoration of their lymphohaemopoietic tissues and cells after severe immunosuppression, results 1-3 months later, in their inability to 'recognize' human donor lymphocytes and to mount an immediate or a delayed-type immune response against human antigens. This durable state of unresponsiveness was evaluated by a complement-mediated cytotoxicity assay, with a mixed lymphocyte culture method and confirmed by the abrogation of the humoral (antibody response to human erythrocytes) and of the cell-mediated (popliteal lymph node test) immune responses in vivo. Our findings demonstrate the capacity of human plasma-derived transferrins to induce a state of durable unresponsiveness (xenogeneic tolerance?) in mice when administered with human antigens in the course of regeneration of stem cells in the bone marrow and lymphatic organs.

Donor-derived plasma transferrin facilitates the engraftment of xenogeneic (rat) bone marrow in irradiated mice.

Endogenous factors originally found in the bone marrow (BM) and facilitating the engraftment of xenogeneic (rat) BM in lethally irradiated mice have been recently identified as transferrins (Tf). Tf have been separated and purified from plasma pools of inbred Rii/2 rats and injected in lethally irradiated BALB/c and C57BL/6 mice 1 h before the infusion of BM and for several days after BM transplantation. Other groups of irradiated mice have been similarly treated with human Tf, Tf from other strains of rats different from the BM donors and with human or rat serum albumin. A remarkable facilitation of BM engraftment and a durable graft-versus-host disease (GVHD)-free hemopoietic chimerism have been achieved in the irradiated mice when a combination of BM and Tf from the same donor rat (Rii/2) strain was used for transplantation. Durable survival and persistent chimerism were not observed in the control groups. It seems that donor Tf profoundly affects the outcome of BM transplantation when combined with donor BM. These results indicate that the mechanism by which Tf promotes engraftment of xenogeneic BM deserves investigation in order to improve this novel procedure and to extend it to other species and possibly to man.

"In vitro" and in animal model studies on a double virus-inactivated factor VIII concentrate.

To improve the safety of plasma derived factor VIII (FVIII) concentrate, we introduced a final super heat treatment (100 degrees C for 30 min) as additional virus inactivation step applied to a lyophilized, highly purified FVIII concentrate (100 IU/mg of proteins) already virus inactivated using the solvent/detergent (S/D) method during the manufacturing process. The efficiency of the super heat treatment was demonstrated in inactivating two non-lipid enveloped viruses (Hepatitis A virus and Poliovirus 1). The loss of FVIII procoagulant activity during the super heat treatment was of about 15%, estimated both by clotting and chromogenic assays. No substantial changes were observed in physical, biochemical and immunological characteristics of the heat treated FVIII concentrate in comparison with those of the FVIII before heat treatment.

[The action of defensins on the corticosterone level of the blood and on the immune response in stress]. / Vliianie defensinov na uroven' kortikosterona v krovi i immunnyi otvet pri stresse.

The influence of defensins on the level of corticosterone in the blood and immune response in stress was studied. Defensins were shown to have corticostatic activity in vivo in the models of stress- and ACTG-induced elevation of corticosterone. Also defensins abolished the immunosuppressive action of combining stress in animals. These results suggest that defensins may be endogenic peptides with stress-protective properties.

[The neuroendocrine system and specific factors of immunity in pesticide poisoning]. / Neiroendokrinnaia sistema i spetsificheskie faktory immuniteta pri otravlenii pestitsidami.

Experiments performed effects of chronic administration of organophosphate pesticide "Antio" (dose 1/100 LD50) on specific factors of immunity and corticosterone (CS) levels in the blood. Adult male Wistar rats were used in all experiments. The immunological indexes were increased, CS levels were decreased.

[Effect of intraventricular administration of naloxone on stress-induced hormonal reactions]. / Vliianie intraventrikuliarnogo vvedeniia naloksona na vyzvannye stressom gormonal'nye reaktsii.

The purpose of the work is to bring to light the role of naloxone-blocked CNS opioid receptors in the implementation of hormonal reactions caused by stress agents in male Wistar rats. Naloxone was administered at a dose of 10 micrograms/100 g body mass into the lateral cerebral ventricles via preliminarily implanted polyethylene cannulas. Stress stimuli (cooling at -20 degrees C for 10 min. and administration of a foreign protein/sheep erythrocytes at a dose of 10(9) cells) followed 30 min. after naloxone administration. Stress-induced changes in the concentrations of aldosterone and testosterone were shown to be significantly disturbed in animals receiving naloxone whereas changes in the concentrations of corticosterone and ACTH remained the same as those in animals not receiving the agent. Changes in hormonal functions and a type of stress showed correlation. It has been assumed that naloxone-blocked CNS receptors are involved in the implementation of stress-induced reactions of mineralocorticoid function of the hypothalamo-hypophyseo-adrenal system and androgenic function of the hypothalamo-hypophyseo-gonadal system.

[The mechanism of the effect of interleukin-1 on the glucocorticoid level of the blood: the absence of a direct action of interleukin-1 on adrenal cortex cells]. / K mekhanizmu vliianiia interleikina-1 na uroven' gliukokortikoidov v krovi: otsutstvie priamogo deistviia interleikina-1 na kletki kory nadpochechnikov.

The investigation is devoted to the problem of interaction of the body neuroendocrine and immune systems. A study was made of possible mechanisms of stimulating action of IL-1, a cellular mediator of the immune system, on glucocorticoid function of the hypothalamic-pituitary-adrenocortical system. RIA and fluorescence methods as well as methods for obtaining IL-1 from activated rabbit leukocytes were employed. IL-1 was shown to increase the plasma level of corticosterone in rats in ip administration. It also produced a direct effect on the secretion of adrenal cells without enhancing ACTH stimulating action on them. The results of the investigation and literature data show that IL-1 action on the hypothalamic-pituitary-adrenocortical system is implemented through its influence on neurons of the cerebral hypothalamic areas secreting the corticotropin releasing factor.

[The combined action of glucocorticoid hormones and interleukin-1 on the development of an immune response]. / Sochetannoe deistvie gliukokortikoidnykh gormonov i interleikina-1 na razvitie immunnogo otveta.

The effect of interleukin-I (IL-I) on glucocorticoid-induced inhibition of the immune response and reactions of cyclic nucleotides to antigen was studied on a model with the use of purified IL-I preparation. Hydrocortisone (HC) in the applied dose caused suppression of the immune response and sharp inhibition of antigen-induced reactions--cAMP in 10 minutes and cGMP on the 6th day after immunization. IL-I administered 10 minutes prior to HC and before immunization prevented the immunosuppressive effect of HC completely. Control administration of heated IL-I and in inactive fraction of the preparation failed to produce a similar effect. IL-I injected 10 minutes after HC also inhibited immunosuppression.