Immunogenicity and safety of the adult tbe vaccine «tick-e-vac¼.

About 3,000 cases of TBE are registered annually in the Russian Federation. Vaccination is the main way to prevent the tick-borne encephalitis disease. Comparative study of the reactogenicity and immunogenicity of a new vaccine «Tick-E-Vac¼ was held. Volunteers aged from 16 years old were twice immunized with the vaccines «Tick-E-Vac¼ or «Encevir¼ derived from strains of Far East subtype of TBE virus, according to standard and emergency schemes. The clinical study was randomized, comparative, blind, and controlled. The frequency, intensity, time of occurrence, and duration of local and general reactions had been recorded. The titers of antiviral antibodies in ELISA had been determined to assess the immunological efficacy of vaccination. According to the results of the clinical study, the severity of local and general reactions in initial seronegative recipients was weak or moderate. The symptoms were usually manifested within 1-2 days after injection and persisted for not more than 4 days, after which time the symptoms disappeared. There was no statistically significant difference in the reactogenicity of the vaccines after the first and after the second injection. The reactogenicity also did not depend on the gender of recipients. After the first immunization, the level of seroprotection was not less than 43%; the average geometric titer of antibodies (GTA), not less than 1:200. After the second injection, the level of seroprotection reached 90-100%; GTA, not less than 1:500. The data on the reactogenicity and immunogenicity to the original seropositive recipients is not significantly different from the data for the initial seronegative recipients. The data indicate weak reactogenicity of the vaccines «Tick-E-Vac¼ and «Encevir¼. Double vaccination with an interval of 14 or 30 days leads to the formation of expressed immune response. Thus, differences in the level of seroprotection and in antiviral titers in the cases of the standard and emergency vaccination schedules are not statistically significant. The correlation between the development in recipients of local and general symptoms and the immunological efficacy of the vaccines has not been identified.

[The epizootological and virological characteristics of a natural hantavirus infection focus in the subtropic zone of the Krasnodarsk Territory].

A natural focus of hantavirus infection was detected and examined during the studies conducted in 2000-2002 around the Sochi (the western spurs of the Great Caucasus Ridge, which descended to the Black Sea (the Krasnodar Territory of Russia). At least 4 rodent species, such as Microtus majori, A. (S.) ponticus, A. agrarius, A. (S.) ciscaucasicus, were shown to participate in the circulation of hantaviruses. A comparative analysis of the nucleotide sequences of genomic S- and M-segments of hantaviruses has provided evidence that 13 viral RNA isolates from the A. (S.) ciscaucasicus belong to the Dobrava/Belgrade virus clade; however the RNA isolate from the Microtus majori belong to the Tula virus clade.

Evaluation of the new control methods for oral poliomyelitis vaccine.

In the draft recommendations for production and control of OPV the WHO proposed new control methods: (i) mutant analysis with PCR and restriction enzyme cleavage (MAPREC) assay that allows evaluation of poliovirus population heterogeneity at the molecular level; (ii) neurovirulence (NV) test using transgenic mice susceptible to polioviruses and (iii) control of the seed lots for the presence of the simian virus 40 (SV40) DNA sequence. This paper is focused on our experience in the practical implementation of the new methods at the Institute of Poliomyelitis and Viral Encephalitides (IPVE). Using methods based on PCR we have demonstrated that working seed viruses used by IPVE for OPV production are free from SV40 DNA sequences. Our experience on the conduction of the OPV type 3 control using TgPVR21 mice NV test (seven vaccine lots) and the MAPREC assay (more than 150 samples of single harvests and monovalent bulks) showed that these methods may be used instead of the monkey NV test, because they could not pass the vaccine failed monkey NV test. The necessity for single harvests control is discussed.

Poliomyelitis in Russia in 1998-1999.

After introducing surveillance for poliomyelitis and AFP cases in the Russian Federation in 1998, 740 AFP cases have been registered in 1998-1999, and 18 of that number were considered as vaccine-associated paralytic poliomyelitis (VAPP). Of 18 cases 11 were classified as VAPP of vaccine recipients and confirmed by virus isolation; from two of the vaccine recipients virus was not isolated, and five were poliomyelitis cases in contact non-vaccinated children. In all the cases the disease was characterised with the typical clinical picture with residual pareses and paralyses. One case was fatal. Vaccine virus type 3 has been isolated from all the vaccine recipients. The MAPREC test has shown that the quality of monovaccine type 3 bulks used for vaccinating these children did not differ from the quality of other bulk vaccines produced by the Chumakov Institute of Poliomyelitis. Patients surveyed for gammaglobulin were positive. Polioviruses type 1 isolated from two of the contact cases had changed antigenic properties and were recombinants of types 1 and 2.

Outbreak of paralytic poliomyelitis in the Chechen Republic in 1995.

An outbreak of poliomyelitis with 146 cases among children of whom six died occurred in the Chechen Republic in 1995. Sporadic cases of poliomyelitis have been reported in the neighbouring Ingush Republic. The outbreak lasted for five months (from May to September) and the maximum number of cases was registered in July. The age of the patients did not exceed 11 years, and more than 90% of the patients were children aged from one month to four years. The overwhelming majority of the patients had not been vaccinated in the routine OPV immunization programme. The outbreak was due to wild poliovirus type 1 belonging to genotype T previously known to circulate in the territory of the former Soviet Union (FSU). Chechen and Ingush isolates were very closely related to each other and to isolates from Central Asia, Tajikistan, 1994. Only a very distant relatedness of the Chechen and Ingush isolates was found with the strains isolated at about the same time outside the FSU (China 1994, Pakistan 1995). The presence of high numbers of non-vaccinated/poorly vaccinated persons and the poor sanitary and hygienic conditions for civilians due to the military conflict were factors that had a role in the outbreak.

[Geotyping of wild poliovirus strains on the territory of Russia and CIS countries in 1987-1995]. / Geotipirovanie dikikh shtammov virusa poliomielita, izolirovannykh na territorii Rossii i SNG v 1987-1995 gg.

33 wild poliovirus strains isolated on the territory of the former USSR were subjected to the comparative sequence analysis of the 150 bp genome fragment VP1/2A on the junction of the genome regions coding basic capsid protein VP1 and viral protease 2A. The dendrograms characterizing the similarity of nucleotide sequences revealed the existence of 3 geographical genotypes (geotypes) of wild poliovirus strains of type 1 (A, G, T) and broad geotype (C) of wild poliovirus strains of type 3. The comparison of the analyzed strains with strains circulating in the neighboring countries at the same period provided information on their genetic relationship. The data thus obtained made it possible to establish the pathways of the transmission of wild poliovirus strains in the common epidemic area of the Russia u CIS.

[Strain-specific antibodies to wild poliomyelitis viruses as an index of the circulation of these viruses in a surveyed territory]. / Shtammospetsficheskie antitela k dikim virusam poliomielita kak pokazatel' tsirkuliatsii étikh virusov na obsleduemoi territorii.

In response to vaccination or infection with wild poliovirus, some children can finely differentiate the virus structure and respond by forming strain-specific antibodies. The analysis of strain-specific antibodies indicates that in epidemically unfavorable regions strain-specific antibodies to wild poliovirus strains may prevail over those to vaccine poliovirus strains in children or occur with equal frequency. In epidemically favourable regions strain-specific antibodies to vaccine poliovirus strain significantly prevail in children or may be absent at all. The analysis of strain-specific antibodies makes it possible to retrospectively judge on the circulation of wild viruses in the surveyed region.

[Detection of poliomyelitis viral strains in natural isolates and identification of them by polymerase chain reaction]. / Obnaruzhenie shtammov virusa poliomielita v prirodnykh izoliatakh i ikh identifikatsiia s pomoshch'iu polimeraznoi tsepnoi reaktsii.

One hundred and fifty nucleotide-long VP1/2A junction regions were sequenced in the RNAs of 19 strains isolated in 1990-1991 from patients with paralytic poliomyelitis in different regions of the former USSR. On the basis of the alignments of these sequenced RNAs, four pairs of 19-25 base-long oligodeoxynucleotide PCR primers were designed capable of detecting polio RNAs in isolated strains and of discriminating between polio genotypes. PCR with 520 polio virus strains isolated from patients, normal subjects, and environmental objects showed 428 of these strains to be related to Sabin's vaccine strains, whereas the rest were referred to A (30), T (24), and G (1) genotypes of serotype 1 and to C-genotype (37) of serotype 3. The designed primers were highly specific and did not cross-react between themselves and with primers specific for Sabin's vaccine strains in PCR.

Geographical genotypes (geotypes) of poliovirus case isolates from the former Soviet Union: relatedness to other known poliovirus genotypes.

A 150 nucleotide long region corresponding to adjoining segments of the genes encoding polypeptides VP1 and 2A of 84 poliovirus strains recently isolated from patients with paralytic poliomyelitis over the territory of the former Soviet Union (FSU) were characterized by sequencing and/or PCR amplification using specially designed primers. Eighteen isolates were found to be very closely related to one or another of the three Sabin vaccine strains. Three distinct classes of geographical genotypes (geotypes) were discerned among 42 wild-type (non-Sabin) strains of serotype 1. One such geotype (called A) was widely circulating in 1990-91 in the Caucasian (Azerbaijan and Georgia) as well as Asian (Kyrgyzstan and Turkmenistan) Republics; this geotype exhibited only weak relatedness to known strains isolated outside the FSU. On the other hand, a subset of strains belonging to another geotype (T) of serotype 1, which circulated in 1991 in Tajikistan, demonstrated very close relatedness to contemporaneous strains isolated in Pakistan, India and Jordan. Strains that were somewhat different, but belonging to the same T-geotype, were found also in Moldova and Georgia. Strikingly, the primary structure of the VP1/2A junction of certain T-geotype isolates differed from the corresponding region of Sabin 1 only in 13-15% of positions, thereby not reaching the upper limit accepted for a geotype. This observation raises, though does not prove, the possibility that at least the relevant segment of the T-geotype RNA originated from the vaccine strain. The third geotype of serotype 1 was represented by a single, perhaps imported, isolate. Four distinct subsets of a common geotype (C) were discerned among 24 wild-type isolates belonging to serotype 3. These strains exhibited a broad geographical distribution being found, in particular, in Armenia, Azerbaijan, Georgia, Turkmenistan and Tajikistan; on the other hand, the C-geotype strains exhibited only a relatively distant relatedness to a strain isolated outside of the FSU (in Oman).

Electropherotypes and serotypes of human rotavirus in Estonia in 1989-1992.

The electropherotypes and serotypes of human rotaviruses circulating in Tallinn (Estonia) in 1989-1992 have been studied. Rotaviruses were found in 372 (25.8%) of 1,442 faecal specimens of pediatric patients with acute diarrhea. Polyacrylamide gel electrophoresis of RNA from 318 isolates has revealed 41 electropherotypes. Frequent genomic alterations, including a shift of predominant electropherotypes, were observed during the study period. The serotype of 158 (50.3%) of 314 tested rotavirus isolates was identified using ELISA with VP7-specific monoclonal antibodies against 4 established human rotavirus serotypes. Serotype G1 was found to be largely predominant and accounted for 70.9% of the typeable specimens, serotypes G4, G2, and G3 accounted for 12%, 9.5%, 7.6%, respectively. A major shift to serotype G4 took place in 1990-1991. Serotype G1 was represented by the largest number of electropherotypes. All G1, G3 and G4 isolates were of "long" and all G2 isolates were of "short" electropherotypes. According to our results the isolates of an identical electropherotype belong to the same serotype.

Russian contribution to OPV.

Field trials and the first mass immunization campaign using Oral Poliomyelitis Vaccine (OPV) from the strains derived by Albert Sabin were carried out in the former U.S.S.R. in 1959 under the guidance of Mikhail Chumakov. The results obtained during these trials contributed greatly to the general acceptance of OPV as a safe and efficient preparation. Mechanisms responsible for the attenuated phenotype of the Sabin strains have long been studied in Russia. For the first time, recombination analysis was used for mapping genetic determinants of attenuation, which were found to reside largely in the 5' half of the Sabin strains RNA. It has been established that the phenotype effect of a subset of the major attenuating mutations is due to the impairment of the translation initiation.

Postinfection treatment with antiviral serum results in survival of neural cells productively infected with virulent poliovirus.

The death of human neuroblastoma cells undergoing productive infection with virulent poliovirus was prevented by addition of antiserum against the virus a few hours after the onset of infection; this treatment, however, did not prevent reproduction of the virus. Despite the presence of the viral antigen, the cells retained the ability to divide. Upon further cultivation in the absence of antiserum, the cells developed specific postinfection immunity or resistance to superinfection with poliovirus.

[The biochemical indices of the blood serum in experimental hepatitis A in tamarins]. / Biokhimicheskie pokazateli syvorotki krovi pri éksperiemental'nom gepatite A u tamarinov.

Serum biochemical parameters were studied in 42 healthy wild-caught adult tamarins (S. mystax), males and females, to determine the normal values. Blood samples were drawn repeatedly, and the serum was tested for aspartate aminotransferase, alanine aminotransferase, isocitric dehydrogenase, serum glucose, serum urea, triglyceride, cholesterol, albumin, and total protein. The results indicated that serum chemistry values were similar to those reported as normal for both humans and other Callitrichidae species. The study of serum biochemical parameters in tamarins with experimental hepatitis A indicated that serum enzyme activities alone reflected the hepatic damage, while other biochemical parameters were of no real clinical importance. The experimental results showed the levels of serum urea to be indicative of the pathological involvement of the kidneys in experimental hepatitis A in some cases.

Frequent isolation of intertypic poliovirus recombinants with serotype 2 specificity from vaccine-associated polio cases.

Five representatives from a collection of 21 Sabin type 2-like poliovirus strains isolated from paralytic poliomyelitis cases in two regions of the USSR have been subjected to limited nucleotide sequencing. All proved to be intertypic recombinants having the genes encoding capsid proteins of Sabin 2 origin and a 3'-end portion of the genome derived from either type 1 (3 isolates) or type 3 (2 isolates) Sabin strains. The crossover points in all the 5 genomes have been mapped to different loci of the P3 region. At least 6 additional isolates from the same collection (and 2 isolates from healthy contacts), appeared to have a type 2/type 1 recombinant genome, as judged by oligonucleotide mapping. The biological significance of frequent occurrence of recombinants among field isolates of vaccine-related strains is discussed.

Immunogenicity trial of inactivated hepatitis A virus vaccine in human volunteers.

An inactivated hepatitis A virus (HAV) vaccine was tested on a group of human adult volunteers. The vaccine was administered subcutaneously, and a control group received a placebo (aluminium hydroxide). The vaccine was found to be relatively well tolerated and non-reactogenic, and levels of anti-HAV were comparable to those in other studies.

Analysis of rotaviral gastroenteritis in Tbilisi.

Electrophoretypes of 107 rotaviral isolates collected in Tbilisi for 18 months revealed seven patterns from which 4 were "long" and 3 "short". The "long" electrophoretypes represented 74.8% of total number of the isolates analysed. One of the "long" electrophoretypes dominated for the whole investigation period. Differences in the seasonal distribution of the isolates with various electrophoretypes were demonstrated and appearance of rotaviral isolates was registered with a "short" electrophoretype which had never been detected before.

Use of protamine sulphate for elimination of substrate DNA in poliovaccines produced on continuous cell lines.

Cell substrate DNA was shown to be an abundant contaminant in the clarified preparations of the Sabin type 1, 2 and 3 poliovaccines produced on a continuous cell line (4647). The size of the DNA, as evaluated for the Sabin type 1 poliovaccine, was highly heterogeneous, ranging from 100 to 20,000 base pairs. In view of potential oncogenicity of this DNA a simple and efficient procedure for its elimination is proposed. The method is based on use of protamine sulphate which at the concentration of 2.0 mg ml-1 precipitated cell DNA almost completely without affecting the virus titres.