[Reactivation of persistent herpes virus infection as a factor of endogenous uveitis in children].

Two hundred and fifty-five children with endogenous uveitis, aged 2 months to 15 years, were examined. 73-93% of children were chronically infected by different viruses of the human herpes group; mycoplasmal past-infection was detected in only 13% of them and Chlamydia past-infection _ in 3.7% of them. Herpes simplex virus of type 1 reactivated reliably more often versus other types of Herpesviridae. Reactivation of cytomegalovirus infection prevailed in mothers who gave birth to children with intrauterine uveitis. A prolonged active replication of herpes virus was primarily observed in children with a Suppressed cell antiviral immunity component. Uveitis in such children was notable for a severe clinical course and trend towards often relapses.

[Diagnosis and importance of the opportunistic herpes-virus infection in etiology and pathogenesis of different eye diseases]. / Gerpesvirusnaia infektsiia v étiopatogeneze razlichnykh zabolevanii glaz.

A total of 439 patients (adults--184, children--255) with herpetic keratitis as well as with uveitis and choriorenitis of different geneses and localizations were examined. An aggravation of HSV-infection was determined by detecting, in the blood), anti-bodies to the unstructured early viral antigens by using the immune enzyme analysis (IEA). The efficiency of the test was demonstrated during the examination of patients with different ophthalmic pathologies. An active infection was detected in 16-39% of patients with various ocular diseases of non-herpetic etiology. The pathogenetic value of reactivated HSV infection is diverse: it can function as an etiological or trigger factor and it can aggravate the main disease through causing, postoperatively, complications.

[Prevalence and clinical significance of active cytomegalovirus infection in patients with inflammatory ophthalmopathy]. / Rasprostranennost' i klinicheskoe znachenie aktivnoi tsitomegalovirusnoi infektsii u bol'nykh s oftal'mopatologiei vospalitel'nogo kharaktera.

A total of 374 patients with different inflammatory diseases of the eyes were examined (227 children and 147 adults). For detecting active cytomegalovirus (CMV) infection, sera were tested for antibodies to nonstructural intact CMV proteins. Antibodies were detected in 26 of 147 adults (17.7%) and 15 of 227 children (6.6%). Difference in the frequency of detection of active CMV infection in adults and children is statistically significant (p < 0.01). Three groups of patients with active CMV were distinguished: children with congenital diseases, immunocompromised patients, and the most numerous group of patients with isolated ocular diseases without manifest signs of immunodepression. In this latter group active CMV was detected in patients with different clinical diagnoses, including ophthalmic herpes. Russian commercial enzyme immunoassay kit CMV-Control proved to be a highly sensitive, simple, and cheap test for the diagnosis of exacerbations of chronic CMV infection.

[Cytomegalovirus infection in children with endogenous uveitis]. / Tsitomegalovirusnaia infektsiia u detei s éndogennymi uveitami.

A total of 405 children aged 3 months to 15 years with uveitis of different origin and localization and 50 mothers of children with intrauterine uveitis were tested for cytomegaloviruses (CMV). Chronic CMV infection was detected in 79% children and 88% mothers. Active CMV infection was diagnosed in 7.1% children with various clinical forms of uveitis; it was somewhat more frequent in cases with grave posterior uveitis and panuveitis complicated by detachment of the retina and vitreous fibrosis. Anti-CMV IgG antibodies indicate an infection, but their detection is insufficient for identifying the etiology of uveitis. Active CMV infection can be a cause of uveitis or aggravate uveitis of another etiology and favor the development of postoperative complications. In many cases active CMV infection was detected in children during remission without clinical signs of uveitis activity. Individual analysis of clinical laboratory data is needed for each patient in order to evaluate the etiological and pathogenetic role of active CMV infection.

[Diagnosis of the active form of cytomegalovirus infection based on determination of IgG antibodies to the immediate-early cytomegalovirus protein by lanthanide immunofluorescence analysis]. / Diagnostika aktivnoi formy tsitomegalovirusnoi infektsii na osnove opredeleniia IgG-antitel k predrannemu belku tsitomegalovirusa metodom lantanidnogo immunofliuorestsentnogo analiza.

A system for time-resolved fluoroimmunoassay (TR-FIA) is developed for detecting early IgG during the active stage of cytomegalovirus infection on the basis of CMV-control enzyme immunoassay system. The antigen is cloned and expressed in E. coli recombinant main immediate early protein of human CMV. The active form of CMV infection was detected additionally in 5 out of 25 women who gave birth to sick babies by means of the new test system in complex with other laboratory diagnostic methods. The test is recommended for prenatal diagnosis of active CMV infection in pregnant women and for predicting the risk of neonatal disease.

[Cloning the human cytomegalovirus immediate-early gene and expression of it in Escherichia coli]. / Klonirovanie gena osnovnogo predrannego belka tsitomegalovirusa cheloveka i ékspressiia ego v kletkakh Escherichia coli.

The major immediate early gene of human cytomegalovirus has been cloned. The fragment of the immediate early protein containing 351 amino acids has been fused with cro-beta-galactosidase and expressed in Escherichia coli cells. The obtained recombinant protein reacts in immunoblotting with the antibodies in human sera from patients suffering from acute cytomegalovirus infection.

[The production of recombinant polypeptides from the herpes simplex viruses types 1 and 2]. / Poluchenie rekombinantnykh polipeptidov virusa gerpesa prostogo tipov 1 i 2.

The genes IE175, IE63, IE68, IE12, UL29 and UL19 of herpes simplex virus types 1 and 2 were cloned. Fragments of these genes were cloned into open reading frame expression vector pEX1-3. Recombinant proteins containing amino acid sequences of ICP4, ICP27, ICP22, ICP47, major DNA-binding protein and major capsid protein were expressed in E. coli cells by fusion with cro-beta-galactosidase proteins.

[Cloning and expression of certain herpes simplex virus type 1 genes in Escherichia coli]. / Klonirovanie i ékspressiia v kletkakh Escherichia coli nekotorykh genov virusa gerpesa prostogo tipa 1.

Complete genes IE12, IE63, IE68, IE175, UL19, UL29 of herpes simplex virus type I or their fragments have been cloned in Escherichia coli cells. The peptides expressed were shown to be fused with cro-beta-galactosidase proteins. The recombinant proteins containing amino acid sequences of ICP4, ICP27 and ICP47, major capsid protein, and major DNA-binding protein react in immunoblotting with the anti-HSVI serum from hyperimmune rabbit. The recombinant proteins can be used for creation of diagnosticums and other scientific and practical purposes. The immunological properties of the recombinant proteins are being investigated.

[The determination of the antigenic activity of recombinant virus-specific polypeptides from the herpes simplex virus types 1 and 2 and their use in immunoenzyme analysis]. / Opredelenie antigennoi aktivnosti rekombinantnykh virusspetsificheskikh polipeptidov virusa gerpesa prostogo tipov 1 i 2 i ispol'zovanie ikh v immunofermentnom analize.

The structural and nonstructural recombinant proteins of HSV type 1 and type 2 were expressed in E. coli by fusion with cro-beta-galactosidase proteins. These proteins containing amino acid sequences of ICP4, ICP27, ICP47, ICP22, major capsid protein and major DNA-binding proteins of HSV types 1 and 2 reacted in immunoblotting with the corresponding anti-HSV sera from hyperimmune rabbits. The nonstructural recombinant proteins can be used for enzyme immunoassay detection of HSV1 or HSV2 type-specific antibodies in sera from patients with acute HSV infection.

[Isolation and properties of hybridomas producing monoclonal antibodies to the Leningrad-16 measles virus]. / Poluchenie i svoistva gibridom, produtsiruiushchikh monoklonal'nye antitela k virusu kori Leningrad-16.

A panel of 18 hybridomas producing MCA to measles virus (the L-16 strain) was produced by somatic hybridization method. Thirteen hybridomas produce immunoglobulins of the G class represented by all known subclasses of mouse immunoglobulins; the subclass G 2a is predominant. Five hybridomas produce immunoglobulins of the M class. The resulting preparations of MCA are highly reactive in several serological tests: IFA, NIF, HI, NT. MCA have been divided into 3 groups according to their capacity to combine with three strains of morbilli viruses: L-16, LEC, and carnivora distemper virus (strain EPM).

[A simple and rapid method of determining the titer of herpes simplex virus antibodies in a study by immunoenzyme analysis of a single serum dilution]. / Prostoi i bystryi metod opredeleniia titra antitel k virusu gerpesa prostogo pri issledovanii v immunofermentnom analize odnogo razvedeniia syvorotki.

The result obtained in the study of the possibility of using the method for the determination of the titer of antibodies to herpes simplex virus by EIA techniques in a single dilution of the serum under test are presented. This method is based on the determination of the optical density of the serum titer (rcut) in different groups of sera with the use of the assay system, permitting the evaluation of the positive results obtained in the determination of their final dilution. The results obtained with the use of this method showed that error was 50% for high-titer sera, 60% for medium-titer sera and 30% for low-titer sera.

[Dynamics of the formation of antibodies to surface and internal proteins of the herpes simplex virus in a model experiment on rabbits]. / Dinamika obrazovaniia antitel k poverkhnosti i vnutrennim belkam viru sa gerpesa prostogo v model'nom ékpserimente na krolikakh.

It has been shown by immunoblotting that antibodies to HSV-1 nucleocapsid proteins (39-45K) predominate at the early stages of infection (up to day 21 after infection) in rabbits. At later stages (up to day 75) the intensity of bands corresponding to virus-specific glycoproteins (presumably gD, gE, gC) increases. At the same time the level of antibodies to nucleocapsid proteins diminishes. In ELISA the same pattern was obtained using envelope and nucleo-capsid proteins denatured by SDS and 2-mercaptoethanol. The possibility of using individual HSV antigens for the development of highly specific diagnostic ELISA test-systems is discussed.

[Isolation and use in virological research of a peroxidase conjugate with protein A]. / Poluchenie i ispol'zovanie v virusologicheskikh issledovaniiakh peroksidaznogo kon''iugata s belkom A.

On the basis of proteins A produced at the Mechnikov Central Research Institute for Vaccines and Sera (Moscow) and obtained from Pharmacia AB (Sweden), several batches of peroxidase conjugate have been prepared by a modified method of periodate oxidation. The preparations thus obtained have been evaluated by means of the corresponding enzyme immunoassay systems for the detection of antibodies to herpes simplex and measles viruses. The results of this investigation indicate that serum antibody titers, determined in the assays with antispecific conjugates and with protein A-based based conjugates, coincide. The comparative study of conjugates prepared on the basis of Soviet and imported proteins A has demonstrated their similar activities and specificities.

[Preparation and investigation of some properties of influenza virus nucleoid]. / Poluchenie i izuchenie nekotorykh svoistv nukleoida virusa grippa.

Influenza virus nucleoid ("core") was obtained by the treatment of virions with hydrolytic enzymes without the use of fixing agents. The "core" buoyant density in sucrose density gradient was 1.24 g/cm3. The subvirus particles are completely devoid of surface proteins, glycoproteins, and their polypeptide composition is represented by group P proteins; NP and M proteins. RNA in the core is insensitive to the effect of RNase. Morphological analysis by electron microscopy revealed the presence of spherical compact particles without spikes on their surface.

[Characteristics of the structural organization and transcriptase activity of the influenza virus nucleoid]. / Osobennosti strukturnoi organizatsii i transkriptaznoi aktivnosti nukleoida virusa grippa.

The state of the secondary structure of RNA and proteins comprising nucleoids (cores) and RNP of influenza virus was evaluated comparatively. The identity of RNA conformation in these particles and differences from free RNA conformation due to less marked secondary structure were found. Core proteins were predominantly represented by the beta-framework, and RNP as an alpha-helix. The specific transcriptase activity of the core is significantly lower than RNP activity of influenza virus.

[Secondary structure of RNA of influenza virus in free form and in ribonucleoprotein]. / Izuchenie vtorichnoi struktury RNK virusa grippa v svobodnom sostoianii i v sostave ribonukleoproteidov.

Large amounts of RNA and RNP isolated from influenza virus were obtained. This has allowed us to undertake detailed physical studies of the secondary structure of RNA of influenza virus in free form and in RNP. Analysis of CD spectrum and the hypochromic effect after thermal denaturation of RNA indicated that RNA in free form contains 58--62% double-stranded regions. By comparative studies of the secondary structure of RNA in RNP, it was estimated that 12--14% of the RNA exists in double-stranded form.