Revealing weak A and B antigens in patients with knee and hip joint arthroplasty

АНТИГЕНЫКРОВЬАНТИТЕЛА АНТИИДИОТИПИЧЕСКИЕАНТИ-АНТИТЕЛААНТИГАММА-ГЛОБУЛИНОВЫЕ АНТИТЕЛААНТИГЛОБУЛИНЫАНТИИДИОТИПНЫЕ АНТИТЕЛАКОМПЛЕМЕНТКОМПЛЕМЕНТБЕЛКИ КОМПЛЕМЕНТАРНЫЕКОМПЛЕМЕНТА БЕЛКИЭРИТРОЦИТЫЦель. Провести анализ методов выявления слабых А и В антигенов на эритроцитах в системе АВ0. Материал и методы. Проведено обследование пациентов с эндопротезированием коленного и тазобедренного суставов с целью определения группоспецифической принадлежности с выявлением слабых А и В антигенов на эритроцитах. Использованы методы абсорбции, реакция агглютинации, агглютинация в присутствии комплемента и антиглобулиновый тест. Результаты. Антиглобулиновый тест с использованием как поликлональной сыворотки, так и сыворотки, содержащей только IgG антитела, позволил выявить слабые A и B подгруппы на эритроцитах при 37{o}С. В реакции абсорбции с анти-А, анти-В поликлональными сыворотками у некоторых пациентов на эритроцитах выявлены А и В антигены, также проявившиеся в реакции агглютинации при 37{o}С, но не обнаруженные при инкубации при комнатной температуре. Агглютинационный тест с использованием комплемента и сыворотки с IgG антителами также способствовал проявлению слабых антигенов. Присутствие IgG антител было определено после обработки сыворотки унитиолом в антиглобулиновом тесте. Наличие в сыворотке только IgG антител, соответствующих антигенам, при участии комплемента приводило к более выраженным изменениям эритроцитов по сравнению с присутствием обоих классов антител – IgM и IgG. Появление гемолиза ассоциировалось с увеличением размеров эритроцитов и гипохромией. Присутствие слабых подгрупп в большинстве случаев было ассоциировано с гемолизирующими, а не агглютинирующими свойствами сыворотки пациента, а также наличием комплементсвязывающих IgG антител. Заключение. Применение абсорбции, агглютинация при 37{o}С, антиглобулиновый тест при 37{o}С с сывороткой, как подвергшейся, так и не подвергшейся обработке унитиолом, а также реакция агглютинации с использованием комплемента способствовали определению слабых антигенов в системе АВ0.Objective. To analyze the methods of revealing weak A and B antigens on the erythrocytes in AB0 system. Methods. Patients after knee and joint arthroplasty were examined on group-specific characteristics with revealing weak A and B antigens on the erythrocytes. Methods of absorption, agglutination, agglutination with complement and antiglobulin test were used. Results. Antiglobulin test with the use of polyclonal serum as well as serum containing the only IgG allowed revealing weak A and B subgroups on erythrocytes at 37{o}С. In some patients A and B antigens on erythrocytes were found while absorption with anti-A, anti-B polyclonal sera and also revealed in agglutination at 37{o}С, but were not revealed while incubation at the room temperature. Agglutination test with the use of complement and IgG was also helpful in determining the weak antigens. Presence of IgG antibodies was revealed by the treatment of the serum with unithiol in antiglobulin test. Presence of complement and only IgG antibodies corresponding to the antigens led to the more expressed changes of erythrocytes as compared to the presence of both types of antibodies – IgM and IgG. Appearance of hemolysis was associated with the increased sizes of erythrocytes and hypochromia. Presence of weak subgroups was mostly associated with hemolytic rather than agglutinating abilities of the patient’s serum, as well as with the presence of complement binding IgG antibodies. Conclusions. Absorption, agglutination at 37{o}С, antiglobulin test at 37{o}C with serum both treated and non-treated with unithiol and agglutination with use of the complement helped to define weak antigens in AB0 system

Evaluation of probiotic lactobacillus as adjuvants for nasal immunization with chimeric pneumococcal vaccine

Vaccine protection against photogenic gram-positive bacteria including different species ofstreptococci is an important problem of contemporary molecular biology. Streptococcal infections are mostcommon bacterial infections surpassing by the economic losses all the infections excluding influenza. The gatesof streptococcal infection, oral cavity or vagina, are covered with immune and non-immune mucosal cells thatare the first line of defenses. Subcutaneous immunization not always stimulate the local immunity on mucosalsurfaces. On the other hand, mucosal vaccination can provide an appropriate local immune response togetherwith systemic protection. However, mucosal immunization often requires usage of special and effectiveadjuvants especially in case of vaccines based on recombinant proteins.For protection against Streptococcus pneumoniae infection, two chimeric recombinant proteins (PSPF andPSP) have been tested as vaccines. Recombinant proteins PSPF and PSP carry immunogenic epitopes fromthe respiratory pathogen including PspA, Spr1875 and PsaA. PSPF structure also carries a fraction of flagellin-FliC molecule in comparison with PSP, which does not have this fragment. This portion of PSPF was includedas internal adjuvant intended for the stimulation of Toll-like receptor 5.In this work, the adjuvant capacity of two probiotic strains, Lactobacillus rhamnosus CRL1505 andL. rhamnosus L32 was evaluated. It was demonstrated that both lactic acid bacteria strains were able to provideadjuvant effects by enhancing the mucosal and systemic immune responses after their co-administration withthe recombinant chimeric protein PSPF. The adjuvant effect of both Lactobacillus strains was significantlydecreased after their thermal inactivation. However, the cell walls of bacteria showed a marked adjuvant activity.An improved protection against several S. pneumoniae serotypes after mucosal immunization of infant micewith PSPF vaccine with probiotic strains or their cell walls was also demonstrated here.The recombinant chimeric protein PSPF administered with immunomodulatory probiotic strains or theirbacterial components would be a promising vaccine for immunization of humans against S. pneumoniae,particularly in children.Одной из наиболее актуальных задач медико-биологической науки является создание вакцинных препаратов против патогенных стрептококков – самых распространенных бактериаль-ных возбудителей заболеваний человека, экономический ущерб от которых уступает лишь потерям от гриппозной инфекции. Входными воротами стрептококковой инфекции являются слизистые обо-лочки респираторного и мочеполового тракта. Парентеральный способ введения вакцин не всегда позволяет добиваться одинаково эффектив-ной стимуляции местного иммунитета на слизистых оболочках, а вакцины, вводимые через слизи-стые оболочки, способны эффективно стимулировать иммунную защиту в области введения, а также обеспечить развитие системного иммунного ответа. Введение через слизистые оболочки вакцинных препаратов белковой природы требует исполь-зования специальных эффективных и безопасных адъювантов, поскольку рекомбинантные белки обычно проявляют недостаточную ммуногенность при таком способе введения. В работе в качестве вакцинных адъювантов при мукозальной иммунизации лабораторных животных пневмококковыми химерными рекомбинантными белками PSPF и PSP были апробированы два штамма пробиотиков – Lactobacillus rhamnosus CRL1505 и L32. Рекомбинантные химерные белки PSPF и PSP несут в своей структуре несколько иммуногенных эпитопов PspA, Spr1875, PsaA и предназначены для вакцинации против инфекции Streptococcus pneumoniae. Белки, отличие которых связано с присутствием в струк-туре PSPF участка молекулы флагеллина – FliC, по-разному стимулировали иммунный ответ при совместном введении с двумя штаммами пробиотиков. Установлено, что оба исследованных штам-ма L. rhamnosus были способны оказывать адъювантный эффект при интраназальном введении вак-цинных белков, проявлявшийся в усилении секреторного и гуморального иммунного ответа на со-вместно введенный рекомбинантный химерный белок PSPF. Выраженной стимуляции продукции специфических IgA носовых смывов и IgG сыворотки крови на PSP под влиянием L. rhamnosus L32 не происходило. Адъювантный эффект от вводимых лактобациллярных препаратов существенно сни-жался после температурной инактивации бактерий, однако препарат клеточных стенок L. rhamnosus CRL1505 проявлял выраженную активность. Стимуляция иммунного ответа адъювантами приводила к усилению протективного эффекта вакцины в экспериментах на лабораторных животных, инфици-рованных S. pneumoniae. Установлено, что некоторые штаммы лактобацилл, в частности Lactobacillus rhamnosus CRL1505 и L32, могут быть использованы в качестве адъювантов в составе мукозальных вакцин, однако эта способность зависит от свойств вакцинного препарата и формы введения пробиотиков.Fil: Leontieva, G. F.. Institute of Experimental Medicine; RusiaFil: Kramskaya, T. A.. Institute of Experimental Medicine; RusiaFil: Grabovskaya, K. B.. Institute of Experimental Medicine; Rusia; RusiaFil: Filimonova, V. Yu.. Institute of Experimental Medicine; Rusia; RusiaFil: Laiño, Jonathan Emiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Danilenko, V. N.. Academy of Sciences. Institute of General Genetics. Head, Division of Fundamental Genetic Studies in Biotechnology; RusiaFil: Suvorov, A. N.. St. Peterburg State University; Rusia. Institute of Experimental Medicine. Head, Division of Molecular Microbiology; Rusi

INFLUENCE OF PROBIOTICS ON CYTOKINE PRODUCTION IN THE IN VITRO AND IN VIVO SYSTEMS

Modulatory effects of three probiotic bacterial strains (Lactobacillus rhamnosus K32 (L), Bifidobacterium longum GT15 (B, Enterococcus faecium L-3 (E) on expression level and contents of key cytokines were studied using PCR techniques with reverse transcription, and enzyme-linked immunosorbent assay. Both cell cultures and an experimental model of intestinal dysbiosis were used in this study.The genes encoding bacteriocins, surface membrane component, pili and exopolysaccharides involved in host immune system modulation were previously identified in the B and Ebacterial strains.Investigation of probiotic strains and effects of their supernatants expression of cytokines in cell cultures of promonocyte origin (HTP-1) showed increased expression of TNFα, due to E and L supernatants. Moreover, the Bl culture induced IL-8 and IL-10 expression.In a model of Wistar rats with ampicillinand metronidazole-induced intestinal dysbiosis corrected with probiotics we have shown that the dysbiosis was accompanied by sufficient alterations in microbiota composition (Klebsiella spp. overgrowth and low contents of Faecalobacterium prausnitzii) that were observed only in the animals untreated with probiotics (control), or after administration of L.In contrast to these results, the animals treated with E and B, the following changes were revealed: 1) low expression of proinflammatory cytokines IL-8, TNFα, MCP-1 inmesenteric lymph nodes and appropriate changes of their serum contents, 2) increased serum content of the anti-inflammatory TGFβ cytokine. Hence, the present study, having used two complementary models, has detected some individual features of immune modulation produced by the probiotictic strains of L. rhamnosus K32, B. longum GT15 и E. faecium L-3 which exert differential effects upon the intestinal microbiota

Sustained proliferation in cancer: mechanisms and novel therapeutic targets

Proliferation is an important part of cancer development and progression. This is manifest by altered expression and/or activity of cell cycle related proteins. Constitutive activation of many signal transduction pathways also stimulates cell growth. Early steps in tumor development are associated with a fibrogenic response and the development of a hypoxic environment which favors the survival and proliferation of cancer stem cells. Part of the survival strategy of cancer stem cells may manifested by alterations in cell metabolism. Once tumors appear, growth and metastasis may be supported by overproduction of appropriate hormones (in hormonally dependent cancers), by promoting angiogenesis, by undergoing epithelial to mesenchymal transition, by triggering autophagy, and by taking cues from surrounding stromal cells. A number of natural compounds (e.g., curcumin, resveratrol, indole-3-carbinol, brassinin, sulforaphane, epigallocatechin-3-gallate, genistein, ellagitannins, lycopene and quercetin) have been found to inhibit one or more pathways that contribute to proliferation (e.g., hypoxia inducible factor 1, nuclear factor kappa B, phosphoinositide 3 kinase/Akt, insulin-like growth factor receptor 1, Wnt, cell cycle associated proteins, as well as androgen and estrogen receptor signaling). These data, in combination with bioinformatics analyses, will be very important for identifying signaling pathways and molecular targets that may provide early diagnostic markers and/or critical targets for the development of new drugs or drug combinations that block tumor formation and progression

Comparing individual-based approaches to modelling the self-organization of multicellular tissues.

The coordinated behaviour of populations of cells plays a central role in tissue growth and renewal. Cells react to their microenvironment by modulating processes such as movement, growth and proliferation, and signalling. Alongside experimental studies, computational models offer a useful means by which to investigate these processes. To this end a variety of cell-based modelling approaches have been developed, ranging from lattice-based cellular automata to lattice-free models that treat cells as point-like particles or extended shapes. However, it remains unclear how these approaches compare when applied to the same biological problem, and what differences in behaviour are due to different model assumptions and abstractions. Here, we exploit the availability of an implementation of five popular cell-based modelling approaches within a consistent computational framework, Chaste (http://www.cs.ox.ac.uk/chaste). This framework allows one to easily change constitutive assumptions within these models. In each case we provide full details of all technical aspects of our model implementations. We compare model implementations using four case studies, chosen to reflect the key cellular processes of proliferation, adhesion, and short- and long-range signalling. These case studies demonstrate the applicability of each model and provide a guide for model usage

Androgen Receptor Drives Cellular Senescence

The accepted androgen receptor (AR) role is to promote proliferation and survival of prostate epithelium and thus prostate cancer progression. While growth-inhibitory, tumor-suppressive AR effects have also been documented, the underlying mechanisms are poorly understood. Here, we for the first time link AR anti-cancer action with cell senescence in vitro and in vivo. First, AR-driven senescence was p53-independent. Instead, AR induced p21, which subsequently reduced ΔN isoform of p63. Second, AR activation increased reactive oxygen species (ROS) and thereby suppressed Rb phosphorylation. Both pathways were critical for senescence as was proven by p21 and Rb knock-down and by quenching ROS with N-Acetyl cysteine and p63 silencing also mimicked AR-induced senescence. The two pathways engaged in a cross-talk, likely via PML tumor suppressor, whose localization to senescence-associated chromatin foci was increased by AR activation. All these pathways contributed to growth arrest, which resolved in senescence due to concomitant lack of p53 and high mTOR activity. This is the first demonstration of senescence response caused by a nuclear hormone receptor