CONTENTS
Review
- Stare i Nowe Sposoby Leczenia Chorób Zakaźnych: Wirusy Wszędzie i Nigdzie
Old and New Prescriptions for Infectious Diseases and the Newest Recipes for Biomedical Products in Plants (in Polish)
Hilary Koprowski, (Biotechnology Foundation Laboratories at Thomas Jefferson University, Philadelphia, Pennsylvania, USA)Abstract. The three antiviral vaccines discovered in the 18th century (smallpox), 19th century (rabies), and 20th century (polio) share a common feature: none would ever be licensed today for human vaccination. Yet Jenner’s smallpox vaccine led to the eradication of smallpox, Pasteur’s rabies vaccine represented the first successful post-exposure treatment of people bitten by rabid animals, and polio vaccine administered since its discovery in 1950 is leading to the eradication of polio (in the years 2004-2005) from the earth. However, in the case of rabies, efforts at complete eradication are unrealistic, despite the availability of a very effective vaccine, since rabies, unlike smallpox and polio, is not limited to humans and can infect all domestic and wild mammalian species. Rabies is probably the oldest known infectious disease, yet knowledge of the virus and the disease is far from complete. For instance, the appearance of 24 cases of „cryptic” rabies in the USA, i.e. cases not associated with any bite or scratch, with an incubation period in humans extending 6-8 years, is a puzzling phenomenon that cannot be readily explained. On the other hand, rabies is one of the few strictly neuronal infections and, as such, is an excellent model for the study of neurotropic virus distribution in the brain. Apoptosis induced by a rabies strain expressing high levels of glycoprotein spreads much more slowly through brain tissue than that induced by strains producing lower glycoprotein levels. Attenuated rabies virus constructed to express twice the normal glycoprotein levels is also an excellent antigen for induction of immune responses in the host. Foreign antigens using this vector may also produce highly immunogenic vaccines.
Global Approach to Immunization. Those monitoring the spread of AIDS in many parts of the world know that cost of treatment is one of the major problems in combating the disease. Vaccines against HIV face the same problem. In general, the price of vaccines and sera is exorbitant for the afflicted population in developing countries. In addition, the dearth of syringes, the unavailability of nurses and doctors to administer multiple vaccine injections, and other factors in these countries require a drastic change in current vaccine production approaches. About 12 years ago, plants became vehicles to produce biomedical reagents. Plants can be exposed directly to a construct containing a foreign gene and Agrobacterium to create a transgenic plant that, over several generations, produces the desired product. Alternatively, plants infected with a plant virus (e.g. alfalfa mosaic virus) fused with a foreign gene can propagate the foreign antigen as the virus multiplies. Extraction of the plant virus followed by purification provides the desired biomedical product. Our use of either of these systems has led to the creation of plants producing vaccines, sera, hormones, and other biological reagents. In two clinical trials at the Institute of Bioorganic Chemistry of the Polish Academy of Sciences in Poznan, volunteers who ingested lettuce expressing hepatitis B vaccine showed hepatitis B antibodies in their sera. In another trial carried out at the Biotechnology Foundation Laboratories in Philadelphia, volunteers ingesting a spinach-rabies vaccine showed an immunological priming effect, since only one injection of commercially available rabies vaccine significantly raised the level of rabies-specific antibodies. Vaccines against HIV gp120 and Tat have been produced in spinach, and a construct of gp120 with the CD4 receptor is now being adapted to this plant. Two types of antibodies against rabies and against colorectal cancer are being produced in tobacco and in lettuce. The suboptimal quality of the currently available anthrax vaccine prompted our efforts to produce the anthrax Protective Antigen (PA)in tobacco and lettuce. Quite clearly, plants will play a prominent role in producing a variety of biomedical reagents in the future.Keywords: vaccines; viruses; infectious diseases; bioproducts in plants.
- Predicting Outcome in Haematological Stem Cell Transplantation
Anne M. Dickinson1, James Cavet2, Hannah Cullup1, Xiao N. Wang1, Mark Jarvis1, Libset Sviland2 and Peter G. Middleton3 (1University Departmentt Haematology, Tyneside Leukaemia Research Laboratory, Royal Victoria Infirmary, Queen Victoria Road, Newcastle upon Tyne, UK, 2Avdeling for Patologi, Haukeland Sykehus, Bergen, Norway, 3Leukaemia Research Fund Laboratories, The Medical School, University of Newcastle, Newcastle-upon-Tyne, UK)Abstract. This present review summarises recent results investigating the role of certain cytokine gene polymorphisms, including TNFa, IFNg, IL-6, IL-10 and IL1 Ra, in allogeneic stem cell transplantation. The review discusses their role in predicting outcome and the development of a genetic risk index for Graft versus Host Disease in HLA matched sibling transplants. By the comparative use of an in vitro human skin explant model initial results suggest that certain cytokine gene polymorphisms may be associated with more severe disease.
Keywords: cytokine gene polymorphisms; predicting GvHD; an in vitro human skin explant model.
- Structures and Serology of the O-Specific Polysaccharides of Bacteria of the Genus Citrobacter
Yuriy A. Knirel1, Nina A. Kocharova1, Olga V. Bystrova1, Ewa Katzenellenbogen2 and Andrzej Gamian2 (1N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospekt 47, Moscow, 119991, Russia, 2Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland)Abstract. The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.
Keywords: Citrobacter; O-antigen; lipopolysaccharide; structure; immunospecificity
- Streptomyces and Escherichia coli, Model Organisms for the Analysis of the Initiation of Bacterial Chromosome Replication
Walter Messer1 and Jolanta Zakrzewska-Czerwińska2 (1Max-Planck-Institut für molekulare Genetik, D-14195 Berlin, Germany, 2Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland)Abstract. Streptomyces coelicolor A3(2) and Escherichia coli are quite different in their life-style and the structures of their genomes. Streptomyces exhibit complex multicellular development including formation of multigenomic hyphae during growth. These organisms possess a large linear (8.7 Mb) and GC-rich (~72%) chromosome. The genome sequence of S. coelicolor has just been completed. The difference between E. coli and Streptomyces making them an excellent model organisms for a comparison of their replication modes. In this review, we compare initiation of chromosome replication in both organisms. Their replication origins are different in size, but both have DnaA boxes – a binding motifs for initiator DnaA protein. The two DnaA proteins have practically the same biochemical properties. Many aspects of the control of initiation seem to be similar. A comparison of the two systems thus allows us to define those aspects of replication initiation that are universally used in the eubacterial kingdom.
Keywords: DnaA; Escherichia coli; oriC; Streptomyces coelicolor.
- Lactoferrin and Immunologic Dissonance: Clinical Implications
Marian L. Kruzel1 and Michał Zimecki2 (1Department of Integrative Biology and Pharmacology, University of Texas Health Science Center, Houston, TX 77030, USA, 2Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland)Abstract. Homeostasis is the maintenance of equilibrium in a biological system by means of positive and negative feedback control mechanisms that counteract influences tending toward physiological dissonance. At the molecular level, homeostasis is controlled by the network of the neuro-endocrine-immune system, in which lactoferrin plays a central role. The purpose of this review is to provide a comprehensive summary of a collaborative study established between the Hirszfeld Institute of Immunology and Experimental Therapy (Wrocław, Poland) and the University of Texas Health Science Center (Huston, USA) regarding lactoferrin and its role in homeostasis. In our studies we focused on the immunoregulatory functions of lactoferrin, both in vitro and in vivo,. We investigated the immune status of individuals subjected to different insults, including experimental endotoxemia in mice and surgery in humans. We also studied a lactoferrin-dependent delayed type hypersensitivity (DTH) response to evaluate some of the mechanisms by which lactoferrin can effectively substitute an adjuvant in vaccine.
Keywords: lactoferrin; inflammation; immunomodulation; oxidative stress; anti-aging.
- Experimental Therapies for Psoriasis
Khusru Asadullah1, Hans-Dieter Volk2, Markus Friedrich3 and Wolfram Sterry3 (1Corporate Research Business Area Dermatology, Schering AG, D-13342 Berlin, Germany, 2Department of Immunology, 3Department of Dermatology and Allergology, University Hospital Charité, Berlin Humboldt University, D-10098 Berlin, Germany)Abstract. There is a high medical need for better therapies for psoriasis. Based on new insight into the pathophysiology of this frequent immune disease, a number of novel systemic immunomodulatory therapies are currently in clinical development. These include approaches targeting antigen presentation and costimulation, T cell activation and leukocyte adhesion, action of proinflammatory mediators, and modulating the cytokine balance. Although mainly only preliminary data are available so far, these trials contribute to a further understanding of the disease and will eventually lead to new therapeutic options for psoriasis. Moreover, since psoriasis can be considered as a visible model disease for T cell-mediated disorders characterized by a type 1 cytokine pattern in general, such approaches may have impact for other immune disorders as well. Here we review the rationale and the initial clinical data of these important recent experimental therapies.
Keywords: psoriasis; immunotherapy; cytokines; costimulation; biologicals.
Original Paper
- Diminished Expression of the Type II Receptor for TGFb (TGFbRII) in T Lymphocytes from Patients with Sezary Syndrome Is Not Due to Mutations in the Receptor’s Poly-A Tract: Limitations of the Standard RT-PCR in cDNA Sequence Analysis of Homopolymeric Base Stretches
Qian Zhang1, Renold J. Capocasale1, Floyd E. Fox2, Vahe Bedian3, Eric C. Vonderheid4, Alain Rook2, Jonni S. Moore1, Peter C. Nowell1, Dale S. Haines5 and Mariusz A. Wasik1* (1Department of Pathology and Laboratory Medicine, 2Dermatology, and 3Department of Genetics, University of Pennsylvania Medical Center, Philadelphia, PA19104, USA, 4Department of Dermatology and 5Division of Hematology/Oncology, Hahnemann University)
Abstract. Peripheral blood lymphocytes from patients with Sezary syndrome (SzS) frequently demonstrate decreased surface expression of transforming growth factor b receptor II (TGFbRII). The mechanism of this low TGFbRII expression remains unknown. Because mutations within the poly-A tract of the TGFbRII sequence (nucleotides 709-718) were shown to result in diminished TGFbRII expression in other types of malignant tumors, we examined the sequence of the TGFbRII poly-A tract in two SzS-derived cell lines and in peripheral blood SzS cells from 17 SzS patients and 4 control, healthy individuals using DNA sequencing and single-stranded conformation polymorphism (SSCP) analysis. A standard bidirectional, automated sequence analysis of the RT-PCR-generated cDNA TGFbRII fragment showed a heterogenous population of the normal length, 10-, with admixed, shortened, 9-base poly-A stretches. Surprisingly, this mixture was present not only in the cells from 5 SzS patients and 2 SzS cell lines, but also in cells from 2 healthy control individuals. Importantly, the proportion of the shortened, 9-base fragments was markedly reduced or practically eliminated when the procedure was modified by usage of high-fidelity DNA polymerase, labeled primers and/or cloned RT-PCR products, which indicates that the presence of the shortened, 9-base fragments represented a procedural phenomenon rather than a true deletional mutation within an allele of the TGFbRII gene. Accordingly, SSCP analysis of genomic DNA did not reveal any mutations within the poly-A tract-containing region. These results indicate that a mechanism different from mutations in the polyadenine tract underlies the diminished TGFbRII expression in SzS cells and that the results of an unmodified, direct sequence analysis of homopolymeric base streaches in RT-PCR-derived cDNA should be interpreted with caution.
Keywords: type II receptor for TGFb;cutaneous T cell lymphoma; Sezary syndrome; poly-A tract sequence analysis.