Impact of bacterial autoregulatory molecules (homoserine lactones and alkylhydroxybenzenes) on the oxidative metabolism of the cell effectors of natural immunity

We investigated the impact of bacterial regulators homoserine lactones (HSLs) and alkylhydroxybenzenes (AHBs) (which are present in human fluids at pico- and nanomolar concentrations) on neutrophile oxidative metabolism. The HSL and AHB effects were determined using a test based on induced luminol-dependent chemoluminescence of neutrophiles in human peripheral blood. In this test, neutrophiles were preincubated with chemical analogs of bacterial autoregulators with different lengths of the hydrocarbon radical, such as HSL · HCl, C6- and C12-HSL, and C1-, C6-, and C12-AHB. We revealed that they suppressed the chemoluminescence and, accordingly, the oxidative metabolism of neutrophiles. This effect was more significant with HSLs than with AHBs. Within each of the two groups, the effect increased with an increase in the length of the hydrocarbon chain of the homologues. High concentrations of long-chain autoregulators of both types produce a cytotoxic effect that is associated with apoptosis in the case of C12-HSL and with cell membrane damage in the case of C12-AHB. The effects of low HSL and AHB concentrations involve their protein-modifying properties and result in changes in the activities of neutrophile oxidative enzymes. To a lesser extent, these effects are due to the pro- and antioxidant activities of HSLs and AHBs, respectively. In light of the results obtained, the HSL and AHB effects are to be considered as a novel mechanism of regulating the activities of cell effectors of natural innate immunity. In symbiotic and parasitic systems, the mechanism involves the bimodal pattern of the effects of HSLs and AHBs that vary depending on their structure and concentrations.     

小鼠中性粒细胞在抗体及补体参与下对日本血吸虫童虫杀伤作用的体外实验

用光镜及电镜观察小鼠中性粒细胞及中性粒细胞依赖抗体及补体对体外培养的日本血吸虫童虫 的作用。结果表明：单纯中性粒细胞很少粘附到童虫表面,仅个别十分疏松地粘附在童虫表面,被粘附 的童虫结构正常。提示：单纯中性粒细胞对童虫无明显作用,在抗体及补体协同下,中性粒细胞成群且 紧密地粘附在童虫体表,在细胞集聚的周围,虫体体被出现隧道样及火山口样变化,紧贴童虫的中性 粒细胞伸出伪足,虫体体棘紊乱,皮层变平,体被剥脱,虫体变形,说明中性粒细胞在抗体及补体协同 下,对童虫有杀伤作用、文中对杀伤机制进行了扼要的讨论。     

Effect of antibiotic therapy and vaccine therapy on the phagocytic reaction in mice infected with Staphylococcus

The time course of changes in the activity, intensity and completeness of phagocytosis with leukocytes of the peritoneal exudate was studied on mice with experimental staphylococcal infection treated with rifampicin, lincomycin and inactivated staphylococcal vaccine used alone or in combination. It was shown that immunization of the animals with inactivated staphylococcal vaccine promoted stimulation of the phagocytic defense. Rifampicin and lincomycin applied therapeutically induced a decrease in the activity, intensity and completeness of phagocytosis. It should be noted that rifampicin had a less pronounced inhibitory effect than lincomycin. The combined use of vaccine and antibiotics with therapeutic purposes promoted an increase in phagocytosis as compared to the use of the antibiotics alone. The combined therapy sometimes resulted in completeness of phagocytosis making it reach the control values (the 10th and 15th days, rifampicin and vaccine). It should be noted that a more pronounced stimulation of the activity, intensity and completeness of the phagocytosis was observed with the use of the combination of rifampicin and the vaccine.     

Ultrastructure of blood cells in children exposed to radiation in utero and resided in radionuclide contaminated zones

The electron-microscopic examinations of lymphocytes and neutrophiles in circulating blood of children irradiated during intrauterine development and on posterior stages of ontogenesis were performed in dynamics after accidental period. Some changes in lymphocyte ultrastructure were revealed; namely the increased undulation of nuclear contours with dilated perinuclear space and some reconstructions of their cellular organoids (the increased density of mitochondrial matrix, crist disorganization, dilation of clear space in cisterns of cytoplasmic net, disorders in the structure of electron-dense granules). The peculiarities of neutrophile ultrastructure that are characterized by centre dilated perinuclear space, decrease in glycosomes, disorders in granular structure, vacuole presence, degradation of cytoplasm and nuclear hypersegmentation were determined.     

Genetic divergence and phylogenetic independence of Far Eastern species in subfamily Leuciscinae (Pisces: Cyprinidae) inferred from mitochondrial DNA analyses

Classification of freshwater fish in the subfamily Leuciscinae, Cyprinidae is hampered by complexity or lack of morphological diversity. In this study, analyses based on mtDNA sequences were undertaken to clarify phylogenetic relationships among Far Eastern, North American and European species in the Leuciscinae. Evolutionary rate in cytochrome b gene (Cyt-b) and D-loop sequences appear to be almost constant in Leuciscinae. The topology of phylogenetic trees generated by neighbor-joining (NJ) and maximum likelihood (ML) methods based on Cyt-b gene and D-loop sequences was similar. Five major clades, designated clades 1-5, and a minor clade were discriminated. Most of the Far Eastern, North American and European species were included in the major clades. Clade 1, comprised almost entirely of Far Eastern phoxinins, is monophyletic and greatly diverged from the other species of Leuciscinae. From the present phylogenetic relationships and the previous studies, we present the following hypotheses with respect to the evolutionary history of the Far Eastern phoxinins. The Far Eastern species should be classified into Far Eastern-specific genera, although ichthyologists have still insisted that the species should be included in the European genera. The Far Eastern clade 1 consists of two subclades, including genera Pseudaspius-Tribolodon and Far Eastern Phoxinus species. According to our phylogenetic analyses, Pseudaspius leptocephalus and Tribolodon species should be reclassified into the same genus. On the basis of evolutionary rate in Cyt-b gene in Cyprinidae, it is estimated that the Far Eastern lineage diverged approximately 10-14 million years ago (mya) from the common ancestor of Leuciscinae. It is deduced that speciation of the Far Eastern species occurred until approximately 4 mya, in relation to the formation of the Sea of Japan and the Japanese Islands.     

Experimental study of cellular immunity after administration of inactivated and live virus vaccine]

Experiments were conducted on guinea pigs with the use of cell migration inhibition test of the peritoneal exudate; stimulation of a definite level of cell immunity in response to the administration of both live and of inactivated vaccine virus was shown. The results obtained are used for the interpretation of the action mechanism of the inactivated preparation in two-stage smallpox vaccination.     

Immunomodulatory effect of selenosemicarbazides and selenium inorganic compounds, distribution in organs after selenium supplementation

Antioxidant properties of selenium producing a protective barrier against free radicals play an important role in numerous metabolic and immunologic processes associated with oxidation- reduction reactions which take place during intracellular digestion of phagocyted bacteria. The aim of our study was to examine the properties of an organic compound of selenium, 4-(o-tolilo)- selenosemicarbazide of p-chlorobenzoic acid in terms of its retention in organs, effect on erythropoesis and phagocytic abilities of neutrophiles as well as antioxidant properties in neutrophiles tested with NBT test. This compound as well as inorganic sodium selenate was given to Swiss mice at the dose of 10^–3 g Se/kg for the period of 10 days. The concentrations of selenium in livers of mice treated with sodium selenate and selenosemicarbazide were found to be higher than in controls (18,7 g lg^–1 and 23.2 g lg^–1 vs. 12 g lg^–1, respectively). Analysis of blood cells count has shown a significant decrease in neutrophile levels in both groups treated with selenium. The influence of selenium compounds on phagocytosis and especially NBT test has been determined (3.8% of positive cells in the controls vs. 2.2% and 0.9% in the groups treated with sodium selenate and selenosemicarbazide, respectively). Our preliminary investigations suggest that selenosemicarbazides are biologically active compounds and can modify neutrophile functions.     

Specific prophylaxis of influenza with inactivated split vaccine Vaxigrip

Influenza remains a serious health problem, annually causing epidemics embracing up to 10% of the population of the world, and at the periods of pandemics this number may rise 4- to 6-fold. In the overwhelming majority of the countries the prophylaxis of influenza is carried out at present out with the use of inactivated vaccines. One of such vaccines is the highly purified split vaccine Vaxigrip (Aventis-Pasteur, France), permitted for use in Russia since 1992. The article contains the review of the data of literature, as well as the materials provided by the authors, which indicate that the preparation has low reactogenicity (also for children starting from the age of 6 months) and high reactogenic properties, leading to antibody formation at protective levels with respect to all three components of the vaccine. The vaccine has been found to ensure pronounced prophylactic efficacy for 70-90% of vaccinees and a decrease in influenza morbidity even in case of using the preparation a week before the onset of the epidemic. This shows the advantage of Vaxigrip in comparison with other inactivated vaccines.     

Assessment of newcastle disease vaccination of houbara bustard breeders (Chlamydotis undulata undulata)

The houbara bustard (Chlamydotis undulata undulata) is endangered in North Africa. Through a captive-breeding program established in Morocco by The Emirates Center for Wildlife Propagation, wild populations are being supplemented by the releasing of captive-reared birds. Newcastle disease, which is caused by Newcastle disease virus (NDV; Avian paramyxovirus type 1), can infect houbara bustards and is a significant threat through contact with backyard poultry and possibly wild birds. Three vaccination schedules for Newcastle disease were evaluated by serologic monitoring to assess the efficiency and safety of various types of vaccines (live vs. inactivated), vaccine strains (Hitchner B1 and Clone 30), and administration routes (intranasal vs. injection). We evaluated antibody titers in 211 adult houbara bustards for 10 mo. Antibody titers to NDV in both sera and egg yolks were monitored by hemagglutination inhibition test. The inactivated vaccine provided a high, homogeneous, and durable serologic response in breeders; titers were higher than log2 11 after 4 wk and remained higher than log2 7 after 10 mo. The response to the two live vaccines was similar, and antibody titers did not exceed log2 6 at sero-conversion. Maternally derived antibodies were efficiently transmitted in vitellus, further confirming that offspring of females hyperimmunized with the inactivated vaccine received high titers of maternal antibodies.     

Toxicities of influenza vaccine: peripheral leukocytic response to live and inactivated influenza viruses in mice.

Intraperitoneal or intravenous inoculation of live or inactivated influenza virus induced two characteristic responses of the peripheral leukocytes in mice, an early appearing leukopenic response and late appearing lymphopenia. The former response usually developed and subsided within several hours, though the change in leukocyte population was fairly complicated depending upon the activity of the inoculated material, while the latter began several hours after inoculation and reached its minimum level in 10 to 20 hr. The agent responsible for the former may be virus pyrogen, while the latter seems to be caused by some substance(s) other than that. The early appearing leukopenic response was similar to that due to bacterial endotoxin in respect to the characteristic pattern of the change in peripheral leukocyte population, though it was relatively easy to distinguish one from the other by the length of the latent period and by the heat stability of the causative agent. Live or inactivated influenza virus causing the early appearing leukopenic response was found also to have the mouse body weight-decreasing toxicity. The significance of these findings in the laboratory control test of influenza vaccine for untoward reactions often observed in human inoculated with some inactivated influenza vaccines was discussed. The possible roles of the two agents, virus pyrogen and endotoxin, in the febrile response were mentioned.     

Effect of antibiotic and vaccine therapies on the succinate dehydrogenase and phosphatase activity of liver cells of mice infected with Staphylococcus

Changes in the activity of succinate dehydrogenase (SDH), total and acid phosphatase (TP and AP) were studied in treatment of laboratory animals with rifampicin, lincomycin and with inactivated staphylococcal vaccine used alone or in combinations. It was shown that immunization of the animals with inactivated staphylococcal vaccine under conditions of experimental staphylococcal infection promoted stimulation of the enzyme activity. Rifampicin and lincomycin used for the treatment of such animals lowered the activity of the enzymes. The suppressing effect of the antibiotics increased with an increase in the period of their use. It should be noted that the inhibitory effect of rifampicin on the activity of SDH, TP and AP was less pronounced than that of lincomycin. The combined use of the vaccine and antibiotics for the treatment of the animals promoted an increase in the enzyme activity as compared to the use of the antibiotics alone. Sometimes the activity of SDH, TP and AP reached the control levels in such animals or the levels observed in the animals treated with the vaccine alone. Stimulation of the enzyme activity was more pronounced when the vaccine was used in combination with rifampicin.     

Genetic status of Asiatic black bear (Ursus thibetanus) reintroduced into South Korea based on mitochondrial DNA and microsatellite loci analysis

The Asiatic black bear is one of the most endangered mammals in South Korea owing to population declines resulting from human exploitation and habitat fragmentation. To restore the black bear population in South Korea, 27 bear cubs from North Korea and Russian Far East (Primorsky Krai) were imported and released into Jirisan National Park, a reservoir of the largest wild population in South Korea, in 2004. To monitor the success of this reintroduction, the genetic diversity and population structure of the reintroduced black bears were measured using both mitochondrial and nuclear DNA markers. Mitochondrial D-loop region DNA sequences (615 bp) of 43 Japanese black bears from previous study and 14 Southeast Asian black bears in this study were employed to obtain phylogenetic inference of the reintroduced black bears. The mitochondrial phylogeny indicated Asiatic black bear populations from Russian Far East and North Korea form a single evolutionary unit distinct from populations from Japan and Southeast Asia. Mean expected heterozygosity (H(E)) across 16 microsatellite loci was 0.648 for Russian and 0.676 for North Korean populations. There was a moderate but significant level of microsatellite differentiation (F(ST) = 0.063) between black bears from the 2 source areas. In addition, genetic evidences revealed that 2 populations are represented as diverging groups, with lingering genetic admixture among individuals of 2 source populations. Relatedness analysis based on genetic markers indicated several discrepancies with the pedigree records. Implication of the phylogenetic and genetic evidences on long-term management of Asiatic black bears in South Korea is discussed.     

Equine Abortion (Herpes) Virus: Strain Differences in Susceptibility to Inactivation by Dithiothreitol

The infectivity of equine abortion (herpes) virus (EAV) was inactivated by treatment with reduced dithiothreitol (DTT). According to their susceptibility to DTT, the EAV strains could be divided into three groups. The vaccine strain RAC-H (419) proved to be more resistant to DTT than all of the other 14 strains tested. The hemagglutinin of EAV was also inactivated by DTT; no strain differences were observed in this respect.     

Active synthesis of hemagglutinin-specific immunoglobulin A by lung cells of mice that were immunized intragastrically with inactivated influenza virus vaccine

Intragastric inoculation with whole-virion vaccine of inactivated influenza virus resulted in production of hemagglutinin (HA)-specific immunoglobulin A (IgA) and IgG both in lung lavage fluids and in serum samples of mice. HA-specific IgA was the predominant isotypic antibody secreted in the lung lavage fluids (average IgA/IgG ratio, 13:1), whereas HA-specific IgG was the major antibody class in serum (average IgA/IgG ratio, 0.3:1). These responses were similar to the antibody responses stimulated by intranasal infection with live influenza virus. In vitro cultures of lymphoid cells from lungs and Peyer's patches, but not from spleens, in the presence of homologous antigen, from mice vaccinated intragastrically synthesized mostly HA-specific IgA. Mice immunized parenterally with inactivated influenza virus produced only IgG in lung lavage fluids and sera. Cultures of lymphoid cells from their spleens, but not their lungs, synthesized HA-specific IgG upon antigenic stimulation in vitro; neither synthesized IgA. These in vitro cell culture results, as well as the inverse relationship of IgA/IgG ratios in lung lavage fluids and sera, demonstrated that the IgA antibody in lung lavage fluids was actively synthesized locally in the lungs of intragastrically immunized mice. This finding was consistent with the migratory distribution of antigen-primed lymphoid cells from Peyer's patches to distant lymphoid tissue such as lung. Intragastric vaccination conferred protection against intranasal challenge with a lethal dose of virulent virus.     

Transcutaneous immunisation with herpes simplex virus stimulates immunity in mice

Herpes simplex virus (HSV) is common throughout the world and is a target for vaccine development. Transcutaneous immunisation is a novel technique that uses the application of vaccine antigens in solution on the skin in the presence of cholera toxin (CT) as an adjuvant. This study investigated the potential of transcutaneous immunisation in C3H mice, using CT co-administered with whole inactivated HSV-1 (CT+HSVi) or HSV-1 antigens extracted from infected Vero cells (CT+HSVag) or a control protein (CT+BSA). The application of any of the three vaccines on to bare mouse skin resulted in the migration of Langerhans cells from the epidermis and in the production of serum antibodies to CT. Both HSV preparations generated serum and mucosal (faecal) antibodies to HSV, with the CT+HSVi vaccine being a more potent stimulator of humoral immunity. The CT+HSVag vaccine, however, was the more potent stimulator of cell-mediated immunity, giving rise to a strong delayed type hypersensitivity response and lymphocyte proliferation in vitro. When the mice were challenged by epidermal inoculation of HSV, the CT+HSVag vaccine induced a higher level of protection than the CT+HSVi vaccine, a result which may indicate that the efficacy of HSV vaccines depends on stimulation of cell-mediated rather than humoral responses. The success of topical vaccination suggests that the transcutaneous route may offer a promising potential for novel vaccine delivery which merits further investigation.     

Le Grand Nord et la religion

The Far North, considered as the northern limit of the human ecumene characterized by an arctic climate, extended to contemporaneous temperate zones during glacial periods. Yet humans, an animal originating in intertropical zones, spread further in the Far North over 2 million years, confronting, even during full glacials, exceptional climatic constraints in order to take advantage of the abundant herbivores. Soon after 30,000, humans deeply penetrated within the Arctic Circle, both in Europe and Siberia. It was during the Last Glacial Maxium, in the Far North and Eurasia, that the first archaeological evidence for religious representations as well as significant social and technological innovations appeared. It was also via the Far North that the North American continent was reached.     

Efficacy of inactivated whole-virus and subunit vaccines in preventing infection and disease caused by equine infectious anemia virus.

We report here on a series of vaccine trials to evaluate the effectiveness of an inactivated equine infectious anemia virus (EIAV) whole-virus vaccine and of a subunit vaccine enriched in EIAV envelope glycoproteins. The inactivated vaccine protected 14 of 15 immunized ponies from infection after challenge with at least 10(5) 50% tissue culture-infective doses of the homologous prototype strain of EIAV. In contrast, it failed to prevent infection in any of 15 immunized ponies that were challenged with the heterologous PV strain. Levels of PV virus replication and the development of disease, however, were significantly reduced in 12 of the 15 ponies so challenged. The subunit vaccine prevented infection from homologous challenge in four of four ponies tested but failed to prevent infection in all four challenged with the PV strain. Two of the four subunit vaccinates had more severe symptoms of equine infectious anemia than nonimmunized ponies infected in parallel. Both vaccines stimulated EIAV-specific cell-mediated immunity. The in vitro lymphoproliferative response was shown to be mediated by T lymphocytes and appeared to be indistinguishable from that induced by EIAV infection. Significant differences were observed in the in vivo lymphocyte responses following challenge with the two virus strains. While peripheral blood mononuclear cells from the inactivated virus vaccinates were equally stimulated by both the prototype and PV strains, the subunit vaccinates challenged with PV exhibited lower levels of spontaneous proliferation and serine esterase activity. This diminished cellular response to PV was correlated with more severe clinical disease in the same ponies. These studies demonstrate for the first time that both an EIAV inactivated whole-virus vaccine and a viral envelope glycoprotein-based subunit vaccine can provide protection against rigorous challenge levels of homologous virus but are unable to protect against similar challenge levels of a heterologous virus. Moreover, the data demonstrate that protection can be achieved in the absence of detectable levels of virus-specific neutralizing antibody in the vaccine recipients at the time of virus challenge. While vaccine-induced virus-specific cell-mediated immune responses were detected, their role in conferring protection was not obvious. Nevertheless, protection from disease appeared to be correlated with the induction of high levels of serine esterase activity following challenge. A significant observation is that while the whole-virus vaccine was usually capable of preventing or markedly moderating disease in the PV-infected ponies, the subunit vaccine appeared to have a high potential to enhance the disease induced by PV infection.(ABSTRACT TRUNCATED AT 400 WORDS)     

Novel dry powder preparations of whole inactivated influenza virus for nasal vaccination

The purpose of these studies was to enhance mucosal and systemic antibody production in response to increased local residence time of a whole inactivated influenza virus administered as a dry powder nasal vaccine formulation. Spray-freeze-drying (SFD) particles suitable for nasal delivery were characterized for physico-chemical properties and stability. Mucoadhesive compounds (MA) were characterized for their effects on nasal residence time of vaccine powders in rats compared with published in vitro data and elicited immune responses. SFD particles (D(50) = 26.9 microm) were spherical with a specific surface area of 1.25 m(2)/g. Thermal analysis indicated SFD powders were amorphous and demonstrated improved stability with respect to liquid formulations under various storage conditions. In vitro physico-chemical studies and in vivo scintigraphic imaging experiments indicated sodium alginate (SA) and carboxymethylcellulose-high molecular weight (CMC-HMW) powder formulations most significantly increased residence time in Brown Norway rats. Intramuscular delivery provided equivalent serum antibody titers to intranasal (IN) powder without MA, in the presence of CMC-HMW, SA, and hydroxypropyl methylcellulose (HPMC-HMW) after initial dosing and all formulations except IN powder with chitosan after boosting. IN liquid provided equivalent serum antibody titers to all IN powders after the initial vaccination and significantly greater serum antibody titers than IN powder with chitosan after boosting. Trends were consistent between residence time studies and immune response; however, no statistically significant differences between powder and liquid formulations were observed. It was concluded that enhanced serum and mucosal antibody responses were elicited by a dry powder nasal vaccine, specifically, administered in the presence of sodium alginate.     

新城疫壳聚糖微球疫苗免疫效果的研究

鸡新城疫是由新城疫病毒引起的鸡的一种急性、烈性、高度接触性传染病,是危害养禽业的最严重疫病之一。控制新城疫最根本的措施是进行有效的疫苗接种,目前常用的疫苗是弱毒活疫苗和灭活疫苗,但二者在实际应用中均存在一定的局限性。口服微球疫苗可以诱导较强的粘膜免疫;同时还能够诱导产生系统的体液免疫和细胞免疫,已成为ND疫苗研究的热点。以壳聚糖为囊材,新城疫La Sota抗原液为芯材,戊二醛为交联剂,制备出新城疫壳聚糖微球疫苗,通过了实验室安全检验和效力检验。将新城疫壳聚糖微球疫苗与LaSota活疫苗和新城疫油乳剂灭活苗分别免疫SPF鸡,利用MTT、血凝抑制法(HI)和ELISA等分别检测不同疫苗免疫后的细胞免疫、体液免疫和粘膜免疫抗体IgA,并在当免疫鸡HI抗体降到23的情况下进行了攻毒试验。结果表明,新城疫壳聚糖微球疫苗安全性好,免疫后可刺激机体产生较强的细胞免疫、体液免疫和局粘膜免疫,具有较好的保护作用。