Molecular characterization of interleukin 12.

Interleukin 12 (IL-12), formerly known as cytotoxic lymphocyte maturation factor and natural killer cell stimulatory factor, is a cytokine secreted by a human B lymphoblastoid (NC-37) cell line when induced in culture with phorbol ester and calcium ionophore. This factor has been purified to homogeneity and shown to synergize with low concentrations of interleukin 2 in causing the induction of lymphokine-activated killer cells. In addition, purified IL-12 stimulated the proliferation of human phytohemagglutinin-activated lymphoblasts by itself and exerted additive effects when used in combination with suboptimal amounts of interleukin 2. The protein is a heterodimer composed of a 40- and a 35-kDa subunit. Amino acid sequence analysis confirmed predicted sequences from the cloned cDNAs of each subunit. Chemical and enzymatic deglycosylation of the heterodimer demonstrated that the 40- and 35-kDa subunits contain 10 and 20% carbohydrate, respectively. Structural analysis of IL-12 using site-specific chemical modification revealed that intact disulfide bonds are essential for bioactivity. The 40-kDa subunit of IL-12 was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by immunoblotting as being present in NC-37 cell supernatant solutions in relatively large amounts uncomplexed to the 35-kDa subunit. Previously it had been shown that the 40-kDa subunit alone does not cause the proliferation of activated human T lymphocytes or enhance the cytolytic activity of human natural killer cells. However, results obtained by site-specific chemical modification suggesting that a tryptophan residue is at or near the active site of IL-12 may imply a direct role of the subunit in interacting with the IL-12 receptor. These data may support the recent proposal (D.P. Gearing and D. Cosman (1991) Cell 66, 9-10) that IL-12 consists of a complex of cytokine and soluble receptor.     

Molecular epidemiology of infectious diseases

Data on molecular epidemiology of bacterial infections is summarized. The term definitions of "molecular epidemiology", "taxonomic species" are given, the limits and species structure of prokaryotes are described. The basic mechanisms of the prokaryotes variability in the epidemic process, as well as the possibilities and limitations of microbiological, molecular-biological and population--genetic typing methods, are characterized. The tactics of molecular-biological studies in analyzing the population structure on the global, regional and local levels is presented. The economic effectiveness of measures taken with due consideration of information on the clonal structure of causative agents of hospital infections is shown.     

Molecular and cellular basis of human olfaction

The human olfactory systems recognize and discriminate a large number of different odorant molecules. The detection of chemically distinct odorants begins with the binding of an odorant ligand to a specific receptor protein in the ciliary membrane of olfactory neurons. To address the problem of olfactory perception at a molecular level, we have cloned, functionally expressed, and characterized some of the human olfactory receptors from chromosome 17. Our results show that a receptor protein is capable of recognizing the particular chemical substructure of an odor molecule and, therefore, is able to respond only to odorants that have a defined molecular structure. These findings represent the beginning of the molecular understanding of odorant recognition in humans. In the future, this knowledge could be used for the design of synthetic ideal receptors for specific odors (biosensors), or the perfect odor molecule for a given receptor.     

Recombinant viruses as tools to induce protective cellular immunity against infectious diseases.

Infections by intracellular pathogens such as viruses, some bacteria and many parasites, are cleared in most cases after activation of specific T cellular immune responses that recognize foreign antigens and eliminate infected cells. Vaccines against those infectious organisms have been traditionally developed by administration of whole live attenuated or inactivated microorganisms. Nowadays, research is focused on the development of subunit vaccines, containing the most immunogenic antigens from the particular pathogen. However, when purified subunit vaccines are administered using traditional immunization protocols, the levels of cellular immunity induced are mostly low and not capable of eliciting complete protection against diseases caused by intracellular microbes. In this review, we present a promising alternative to those traditional protocols, which is the use of recombinant viruses encoding subunit vaccines as immunization tools. Recombinant viruses have several interesting features that make them extremely efficient at inducing immune responses mediated by T-lymphocytes. This cellular immunity has recently been demonstrated to be of key importance for protection against malaria and AIDS, both of which are major targets of the World Health Organization for vaccine development. Thus, this review will focus in particular on the development of new vaccination protocols against these diseases.     

Outside-host phage therapy as a biological control against environmental infectious diseases

Background

Environmentally growing pathogens present an increasing threat for human health, wildlife and food production. Treating the hosts with antibiotics or parasitic bacteriophages fail to eliminate diseases that grow also in the outside-host environment. However, bacteriophages could be utilized to suppress the pathogen population sizes in the outside-host environment in order to prevent disease outbreaks. Here, we introduce a novel epidemiological model to assess how the phage infections of the bacterial pathogens affect epidemiological dynamics of the environmentally growing pathogens. We assess whether the phage therapy in the outside-host environment could be utilized as a biological control method against these diseases. We also consider how phage-resistant competitors affect the outcome, a common problem in phage therapy. The models give predictions for the scenarios where the outside-host phage therapy will work and where it will fail to control the disease. Parameterization of the model is based on the fish columnaris disease that causes significant economic losses to aquaculture worldwide. However, the model is also suitable for other environmentally growing bacterial diseases.

Results

Transmission rates of the phage determine the success of infectious disease control, with high-transmission phage enabling the recovery of the host population that would in the absence of the phage go asymptotically extinct due to the disease. In the presence of outside-host bacterial competition between the pathogen and phage-resistant strain, the trade-off between the pathogen infectivity and the phage resistance determines phage therapy outcome from stable coexistence to local host extinction.

Conclusions

We propose that the success of phage therapy strongly depends on the underlying biology, such as the strength of trade-off between the pathogen infectivity and the phage-resistance, as well as on the rate that the phages infect the bacteria. Our results indicate that phage therapy can fail if there are phage-resistant bacteria and the trade-off between pathogen infectivity and phage resistance does not completely inhibit the pathogen infectivity. Also, the rate that the phages infect the bacteria should be sufficiently high for phage-therapy to succeed.

     

Immunizations against infectious diseases and childhood cancers

Summary A study based upon an unusually large series of childhood cancers and matched controls found a significant deficit of case/control pairs in which the cancer case had fewer immunizations against infectious diseases than the matched control. All types of immunizations and cancers were affected but the case/control differences were more pronounced for older cases with late immunizations than for younger cases with early immunizations, and more pronounced for solid tumours than leukaemia. Therefore there may be immune system responses to immunizations (or simulated infections) which make it difficult for small clones of cancer cells to enlarge and are more successful in preventing localised tumours in adolescents than childhood leukaemias.     

Molecular basis of gestational trophoblastic diseases

Gestational trophoblastic disease (GTD) encompasses a diverse group of lesions with specific pathogenesis, morphological characteristics and clinical features. The modified World Health Organization-classification of GTD includes complete and partial hydatidiform mole, invasive mole, choriocarcinoma, placental site trophoblastic tumor, epithelioid trophoblastic tumor, exaggerated placental site, and placental site nodule. The various forms of gestational trophoblastic disease can be defined and related to discrete pathologic aberrations occurring at different stages of trophoblastic differentiation. Some of these lesions are true neoplasms, whereas others represent abnormally formed placentas with a predisposition for neoplastic transformation of the trophoblast. Except hydatidiform moles in which the cytogenetic studies have been extensively reported, the pathogenesis of other trophoblastic lesions is poorly understood. Recent studies have shed light on the molecular mechanisms of trophoblastic function, especially as it relates to trophoblastic disease. This review will focus on these advances with special emphasis on the pathogenesis of each specific form of GTD. In addition, the morphology and clinical behavior of each of these entities will be briefly discussed.     

Molecular basis of cerebral neurodegeneration in prion diseases

The biochemical nature and the replication of infectious prions have been intensively studied in recent years. Much less is known about the cellular events underlying neuronal dysfunction and cell death. As the cellular function of the normal cellular isoform of prion protein is not exactly known, the impact of gain of toxic function or loss of function, or a combination of both, in prion pathology is still controversial. There is increasing evidence that the normal cellular isoform of the prion protein is a key mediator in prion pathology. Transgenic models were instrumental in dissecting propagation of prions, disease-associated isoforms of prion protein and amyloid production, and induction of neurodegeneration. Four experimental avenues will be discussed here which address scenarios of inappropriate trafficking, folding, or targeting of the prion protein.     

感染性疾病单克隆抗体的研究进展

近年来,感染性疾病(infectious disease)已成为一种严重威胁人类健康的疾病。有研究显示,预计感染性疾病引起的死亡人数在2050年将达到1 000万。由于超级细菌及新型病毒的出现,推动了现有感染性疾病治疗方法的发展。其中,单克隆抗体(monoclonal antibody, McAb)以其具有特异性强、副作用小、灵敏性高等优势,成为对抗各种感染源持续攻击的治疗方法之一。随着抗体技术的发展,有很多针对感染性疾病的McAb进入研发或临床试验阶段,希望用于预防与治疗感染性疾病。目前,抗生素价格低廉,McAb治疗成本高、诊断学并行发展的需求是其主要的发展障碍。因此,McAb在治疗感染性疾病中找到较合适的定位非常重要,很多国内外公司通过分子工程设计等方式改善抗体的治疗潜力,成为McAb药物的发展策略。现就McAb治疗感染性疾病的研究现状与药物研发策略作一综述。     

Utilizing bacterial flagellins against infectious diseases and cancers

The flagellum is the organelle providing motility to bacterial cells and its activity is coupled to the cellular chemotaxis machinery. The flagellar filament is the largest portion of the flagellum, which consists of repeating subunits of the protein flagellin. Receptors of the innate immune system including Toll like receptor 5, ICE protease activating factor, and neuronal apoptosis inhibitory protein 5 signal in response to bacterial flagellins. In addition to inducing innate immune responses, bacterial flagellins mediate the development of adaptive immune responses to both flagellins and coadministered antigens. Therefore, these proteins have intensively been investigated for the vaccine development and the immunotherapy. This review describes the utilization of bacterial flagellins for the construction of vaccines against infectious diseases and cancer immunotherapy. Furthermore, the key factors affecting the performance of these systems are highlighted.     

Heat shock protein-peptide interaction as the basis for a new generation of vaccines against cancers and infectious diseases

Heat shock proteins (HSPs) are associatedin vivo with the entire repertoire of peptides (antigenic and otherwise) generated within that cell. Immunization with such HSP-peptide complexes is unusually efficient in eliciting cellular immune responses against the antigenic peptides associated with the HSPs. This broad and general principle is the basis for a new generation of vaccines against cancers and infectious diseases and circumvents the need for identification of the T cell epitopes for any given cancer or infectious agent.     

Molecular basis of bacteriocins production and activity

Bacteriocins--bacterial proteins or peptides--are envisaged as candidates for the next generation of effective antimicrobials. Analysis of characteristics of natural and genetically engineered bacteriocins with regard to the molecular basis of their production and activity has been performed. Most bacteriocins have narrow spectrum of the inhibitory activity. Some of the broad-spectrum bacteriocins have circular molecular structure (C- and N-terminals of the aminoacid chain are joined by a peptide sequence). Fixed position of molecules' ends possibly accounts for the ability of the proteins to bind with various receptors on the surface of the target cells. Genes encoding bacteriocins and functionally associated proteins can be expressed in heterologous cells including eukaryotic cells. Also there were reports of changing bacteriocin characteristics by the use of site-specific mutagenesis.