Vol. 53, No. 1, 2005
Immunology of viral co-infections with HIV
John W. Northfield, Gillian Harcourt, Michaela Lucas and Paul Klenerman
Abstract. Increasing clinical evidence is emerging that other persistent viral infections can act as important co-factors affecting the progression of human immunodeficiency virus-1 (HIV-1). It appears that hepatitis C (HCV) and cytomegalovirus (CMV) have a deleterious effect on HIV progression, whereas hepatitis G (GBV-C) benefits HIV-1 progression. At the same time, the aggressive nature of HCV infection in HIV is clearly recognized. Here we discuss this clinical evidence and go on to review scientific work pertaining to these interactions in the context of the known and theoretical immunological effects of these viruses. This is discussed at the level of the generation of adaptive immune responses and their effector functions. It is clear that co-infection with persistent viral infections may pose special problems for the human immune system, as pathogenic effects may not be specific to the actual eliciting virus and can therefore multiply the difficulties faced by host defenses. We also highlight the need for further therapies for HIV/HCV co-infected persons, as this is currently a complex and severe syndrome.
Keywords: HIV; GBV-C; CMV;hepatitis C;T cell; dendritic cell
Full-textPDF downloadStaphylococcal superantigens: do they play a role in sepsis?
Silva Holtfreter and Barbara M. Bröker
Abstract. In Staphylococcus aureus, 19 different superantigens (SAgs) have been described. Their genes are all located on mobile genetic elements, such as pathogenicity islands, plasmids, and phages. SAgs bypass conventional antigen recognition by directly cross-linking major histocompatibility complex class II (MHCII) molecules on antigen-presenting cells with T cell receptors. This leads to massive T cell proliferation and cytokine release, which may end in toxic shock syndrome. The role of SAgs in other forms of sepsis is less well defined. In animal models, SAgs and lipopolysaccharide (LPS) very efficiently synergize in the induction of lethal shock, and on the basis of these observations a two-hit model of sepsis has been proposed: LPS or another monocyte stimulus hits first, then SAg or another T cell stimulus hits. In clinical studies, however, evidence for an involvement of SAgs in sepsis has been difficult to obtain. This may have a number of reasons: differences between humans and rodents in their response to LPS and SAg, heterogeneity of SAg combinations in S. aureus clinical isolates, lack of tools to analyze SAg effects in patients, blocking anti-SAg serum antibodies, and MHCII polymorphisms.
Keywords: superantigen; two-hit model sepsis; Staphylococcus aureus; LPS; T cells
Full-textPDF downloadDimerization, ROS formation, and biological activity of o-methoxyphenols
Seiichiro Fujisawa, Toshiko Atsumi, Yukio Murakami and Yoshinori Kadoma
Abstract. o-Methoxyphenols are antioxidants widely used in the cosmetic and food industries. Dimers from 1, 2, or 3 were synthesized and their radical-scavenging and biological activities were compared with those of the original or other phenols. Radical-scavenging was evaluated from a kinetic induction period method (IPM). To simulate biomimetic thiolcooxidation with antioxidants, the behavior of mixtures of 1, 2, 4, or catechin with mercaptomethylimidazole (MMI), a thiol was investigated using IPM. Polyphenols 4 and catechin was accompanied by extensive oxygen uptake, suggesting the formation of thiyl radicals from MMI and their reaction with molecular oxygen. In contrast, 1 markedly enhanced radical-scavenging without oxygen uptake, probably because of the formation of EUGQM/MMI-conjugates. 2 showed relatively small oxygen uptake, probably resulting from the predominant formation of benzyl radicals. Intracellular reactive oxygen species (ROS) in cancer cells by 4, but not by compounds 1, 2, 6, 7, 8, 9, and 10 was found, suggesting a possible link between physicochemical oxygen-uptake and intracellular ROS. The induction of apoptosis by 4 in HL-60 cells was accompanied by intracellular ROS. Dimers 6 and 7 inhibited nuclear factor (NF)-?B activation stimulated by lipopolysaccharide (LPS) in RAW 264.7 cells. Also, 6, 7, and 9 inhibited LPS-induced cyclooxygenase-2 expression in RAW 264.7 cells in a dose-dependent manner, whereas 1, 2 and 3 did not. Dimerization of o-methoxyphenols may be a useful tool for the design of drugs to act as potent chemopreventive and anticancer agents.
Keywords: o-methoxyphenols; dimerization; stoichiometric factors; ROS formation; cooxidation of thiol; apoptosis; NF-?B; Cox-2
Full-textPDF downloadRNA interference – significance and applications
Justyna Stanisławska and Waldemar L. Olszewski
Abstract. RNA interference (RNAi) is a post-transcriptional, highly conserved process in eukaryotes that leads to specific gene silencing through degradation of the target mRNA. This mechanism is mediated by double-stranded RNA (dsRNA) that is homologous in sequence to the silenced gene. The dsRNA is processed into small interfering RNA (siRNA) by an enzyme called Dicer, and the siRNAs are then incorporated into a multi-component RNA- -induced silencing complex, which finds and cleaves the target mRNA. In plants and worms, amplification of the silencing signal and cell-to-cell RNAi spreading is observed. The proposed biological roles of RNAi include resistance to viruses, transposons (mainly in plants), and the silencing and regulation of gene expression, particularly during development. In developmental gene control, specific small RNAs (micro RNA and small temporal RNA) are involved, which are processed in the same way as dsRNAs but act at the level of translation. RNAi technology has become a powerful tool in functional genomic analyses and may prove to be a useful method to develop highly specific gene-silencing therapeutics against viral infections and cancer in the future.
Keywords: RNA interference; gene silencing; dsRNA; siRNA
Full-textPDF downloadAnti-tumor chemotherapy utilizing peptide-based approaches apoptotic pathways, kinases, and proteasome as targets
Francisco J. Mendoza, Paula S. Espino, Kendra L. Cann, Nicolle Bristow, Kristin McCrea and Marek Los
Abstract. The pharmacological sciences are taking advantage of recent discoveries that havedefined the molecular pathways governing apoptosis. These signaling cascades are frequentlyinactivated or distorted by mutations in cancer cells. Peptides derived from criticalinteraction, phosphorylation, or cleavage sites are the preferred leads (starting points)for the development of new drugs. In this review we summarize recent peptide-basedapproaches that target MDM2, p53, NF-?B, ErbB2, MAPK, as well as Smac/DIABLO, IAPBIR domains, and Bcl-2 interaction domains, with a specific focus on the BH3 domain.Separate parts of the review deal with proteasome inhibitors, integrin-derived peptides,and molecules that are being tested for tumor-selective delivery of anticancer drugs(“magic bullet” approach). The proteasome inhibitors and integrin-derived peptides showa variety of effects, targeting not only tumor growth, but also angiogenesis, metastasizingpotential, and other cancer cell functions. The last part of this review describes approachesthat use specific properties (surface receptors, increased enzymatic activities) of cancercells in order to target them specifically. These new generations of anticancer drugs provide the foundations for therapies with fewer side effects and higher efficacy.
Keywords: angiostatin; anti-angiogenic; Bortezomib; Velcade; EGFR; Endostatin; HMR1826 integrins; MDM2; p53
Full-textPDF downloadSEB-induced T cell apoptosis in atopic patients – correlation to clinical status and skin colonization by Staphylococcus aureus
Anna Kędzierska, Jolanta Kaszuba-Zwoińska, Zofia Słodowska-Hajduk, Monika Kapińska-Mrowiecka, Marzena Czubak, Piotr Thor, Kinga Wójcik and Juliusz Pryjma
Abstract. Introduction: We asked whether in atopic dermatitis (AD) increased T cell apoptosis in staphylococcal enterotoxin B (SEB)-activated cultures of peripheral blood mononuclear cells (PBMCs) is characteristic of the exacerbation of the disease or connected with skin colonization by Staphylococcus aureus.
Materials and Methods: The clinical status of the patients was evaluated using the SCORAD index. The number of bacteria colonizing patients’ skin lesions was determined by the cfu method. Mononuclear cells isolated from peripheral blood were stimulated by SEB and the apoptosis of CD3+ cells in culture was determined by flow cytometry using the monoclonal antibody APO2.7. The cytokine production in the culture supernatants was determined by ELISA and Cytometric Bead Array kits.
Results: T cell apoptosis was increased, while the production of interferon (IFN)-g was reduced in cultures of PBMCs of AD patients during exacerbation. The proportion of CD3+APO2.7+ cells positively correlated with the density of S. aureus recovered from skin lesions, but not with SCORAD index. By contrast, SCORAD index, but not S. aureus density, negatively correlated with IFN-g production. Furthermore it was found that the presence of S. aureus on uninvolved skin distinguishes a group of severe cases with high serum IgE level, increased T cell apoptosis, and reduced production of tumor necrosis factora in SEB- -stimulated cultures.
Conclusions: Among AD patients the increased activation-induced T cell apoptosis observed in SEB- -stimulated cultures is related to skin colonization by S. aureus. The presence of bacteria on uninvolved skin is a feature of a distinct group of AD patients.
Keywords: atopic dermatitis; apoptosis; T cells; Staphylococcus aureus colonization
Full-textPDF downloadAdvanced glycation end-products prepared in solution under high pressure contain epitopes distinct from those formed in the dry reaction at high temperature
Magdalena Staniszewska, Sławomir Jarosz, Marek Jon and Andrzej Gamian
Abstract. Introduction: Advanced glycation end-products play an important role in diseases related to diabetes and aging processes. Model compounds are synthesized in order to prepare the diagnostic and experimental tools for studying the mechanisms of pathogenesis. The objective of the present study was to accelerate glycation and upgrade its efficiency under high-pressure conditions.
Materials and Methods: Aqueous solutions of proteins were kept with carbohydrates under a pressure of up to 850 MPa for several hours. Then the high-pressure glycation (HPG) products were fractionated on a Sephadex G-200 column and characterized with SDS-PAGE and MALDI-TOF mass spectrometry.
Results:The low-molecular-mass fraction of glycated proteins was separated from the two fractions containing high- and intermediate-molecular-mass cross-linked products of glycation. The products were then compared with those obtained with the high-temperature glycation (HTG) procedure carried out in dry conditions with a lyophilized mixture of substrates. The fractionated products were used to prepare rabbit sera.
Conclusions: The immunoblotting experiments showed that the epitopes on the cross-linked glycation products formed in solution under high pressure differed from those originating in dry conditions at high temperature. Sera against the HPG products were specific to homologous material and did not interact with the fractions obtained by HTG. The antibodies against HTG products recognized HTG but not HPG products.
Keywords: glycation; Maillard reaction; high-pressure reaction; high-temperature reaction; antibodies anti-AGE; epitope
Full-textPDF downloadDifferent pro-inflammatory and immunogenic potentials of Propionibacterium acnes and Staphylococcus epidermidis: implications for chronic inflammatory acne
Anna Białecka, Monika Mak, Rafał Biedroń, Małgorzata Bobek, Andrzej Kasprowicz and Janusz Marcinkiewicz
Abstract. Introduction: Propionibacterium acnes (PA) and Staphyloccocus epidermidis (SE) are two major bacterial strains isolated from acne lesions. Nevertheless, only PA seems to be implicated in the pathogenesis of inflammatory acne vulgaris. Evidence for this, however, remains indirect and the precise role of PA in inflammatory acne is still a matter for conjecture. The aim of this study was to compare some pro-inflammatory and adjuvant properties of PA and SE.
Materials and Methods:To determine some of the pathogenic, immunostimulatory, and pro-inflammatory properties of PA and SE, two experimental models of inflammation were used. In vivo; chronic inflammation was induced by intradermal injection of living bacteria into the ear. In vitro; peritoneal macrophages elicited by the bacteria were examined for their ability to generate reactive oxygen species (ROS), nitric oxide (NO), and cytokines.
Results: PA, but not SE, evoked mild local inflammation of infected ears. Macrophages elicited with PA produced more tumor necrosis factor a and interleukin IL-12 than those inducedwith SE, while SE was a stronger inducer of IL-10 production. Both bacteria equally induced the generation of NO and ROS. In contrast, only PA showed adjuvant properties.
Conclusions: The results of these studies indicate that SE, in contrast to PA, does not exert pro-inflammatory properties. Thus it is unlikely that SE may be implicated in the pathogenesis of inflammatory acne vulgaris.
Keywords: acne vulgaris; Propionibacterium acnes; Staphyloccocus epidermidis; inflammation; TNF-a; IL-10; ROS
Full-textPDF downloadVol. 53, No. 2, 2005
Modulation of auxiliary signals to T cells as a mechanism to induce transplant tolerance
Reginald M. Gorczynski (Toronto Hospital and University Health Network, Toronto, Ontario, Canada)
Abstract. Advances in the treatment of transplant rejection, autoimmune disease, allergy, and other conditions of altered immunoregulation have come from our improved knowledge of the multi-faceted nature of lymphocyte activation, incorporating not merely antigen triggering of specific receptors, but a myriad of other accessory signals, all operating within a defined environmental (cytokine) milieu. The review below focuses on just one aspect of this, the ability to manipulate costimulatory signals, or regulatory signals, as a means to induce long-standing immune suppression. Emphasis is placed on the dominant suppression mediated following activation of any one of a number of regulatory signals as a potentially more rational approach to clinical therapy, as the redundancy in costimulatory signals suggests that blockade of any one of these may be unlikely to produce permanent unresponsiveness. The role of regulatory T ceillis, induced following antigen presentation in the presence of immunoregulatory signals, is also discussed.
Keywords: tolerance; immunoregulation; costimulation; CD200
Full-textPDF downloadMolecular basis of Trypanosoma cruzi and Leishmania interaction with their host(s): exploitation of immune and defense mechanisms by the parasite leading to persistence and chronicity, features reminiscent of immune system evasion strategies in cancer diseases
Ali Ouaissi and Mehdi Ouaissi
Abstract. A number of features occurring during host-parasite interactions in Chagas disease caused by the protozoan parasite, Trypanosoma cruzi, and Leishmaniasis, caused by a group of kinetoplastid protozoan parasites are reminiscent of those observed in cancer diseases. In fact, although the cancer is not a single disease, and that T. cruzi and Leishmania are sophisticated eukaryotic parasites presenting a high level of genotypic variability, the growth of the parasites in their host and that of cancer cells share at least one common feature, that is their mutual capacity for rapid cell division. Surprisingly, the parasitic diseases and cancers share some immune evasion strategies. Consideration of these immunological alterations must be added to the evaluation of the pathogenic processes. The molecular and functional characterization of virulence factors and the study of their effect on the arms of the immune system have greatly improved understanding of the regulation of immune effectors functions. The purpose of this review is to analyze some of the current data related to the regulatory components or processes originating from the parasite that control or interfere with host cell physiology. Attempts are also made to delineate some similarities between the immune evasion strategies that parasites and tumors employ. The elucidation of the mode of action of parasite virulence factors toward the host cell allow not only provide us with a more comprehensive view of the host-parasite relationships but may also represent a step forward in efforts aimed to identify new target molecules for therapeutic intervention.
Keywords: trypanosomatids; virulence factors; cancer cells; immunoregulation
Full-textPDF downloadzIL-15 and IL-15Ra in CD4+ T cell immunity
Tom Van Belle and Johan Grooten
Abstract. The cytokine IL-15 performs numerous functions, such as promotion of growth and survival, on a plethora of cell types from both the lymphoid and non-lymphoid compartments. Therefore, mice genetically engineered to either lack or overexpress functional IL-15 display reduced immunological responses and leukemia, respectively. Surprisingly, IL-15 protein is hardly found in serum or body fluids. Due to the lack of a clear demonstration of its presence as protein, IL-15 was often referred to as a “ghost cytokine”. Recently, however, membrane-bound IL-15 was detected in both a membrane-anchored form and an IL-15Ra-bound form on monocytes. Interestingly, the latter complex can be transpresented to cells expressing the intermediate-affinity IL-2/15Rß-gC receptor and thereby support the survival and proliferation of T cells. Moreover, overlapping promoter elements indicate a model of co-regulation of IL-15 and IL-15Ra by which IL-15 activities are controlled in a cell-contact-dependent manner. In this review, recent reports on IL-15 are combined with previous observations and discussed in terms of their functional consequences for CD4+ T cell responses.
Keywords: cytokines; T lymphocytes; IL-15; transpresentation; apoptosis
Full-textPDF downloadIon channels in T cells: from molecular pharmacology to therapy
Zoltán Krasznai
Abstract. Ion channels of a variety of cell types, such as cardiac and smooth muscle cells and neurons, serve as targets for many drugs used in therapy. T cells also express an assortment of ion channels that are in the focus of intensive research, as they may provide efficient ways to specifically manipulate T cell function and, consequently, immune responses. T cell activation relies on the operation of voltage-gated and Ca2+-activated potassium channels and Ca2+ release-activated Ca2+ channels. Many peptide toxin and small molecule blockers of these channels are known, but inhibitors of even higher affinity and selectivity would be needed for safe and effective clinical use. The recent discovery that the expression pattern of potassium channels in T cells is subset specific emphasizes the potential that these proteins have in immunomodulation. Compounds that could suppress T cells involved in autoimmunity without affecting T cells in normal immune responses would be of enormous value. In this paper the basic properties of these channels and compounds known to influence their operation are reviewed.
Keywords: T cell; activation; potassium channels; CRAC channels; channel blockers
Full-textPDF downloadInfluence of genetic factors on the susceptibility to HBV infection, its clinical pictures, and responsiveness to HBV vaccination
Irma Kacprzak-Bergman and Beata Nowakowska
Abstract. The association of genetic factors with hepatitis B virus (HBV) infection susceptibility, its different manifestations, and the different responses to hepatitis B antigen vaccination have been described by several authors. With regard to HLA class I molecules, association with HLA-B was especially observed. HLA-B35 and -B8 correlated with chronic active hepatitis (CAH) and with hepatitis B carriers. Correlation between HBV infection and HLA class II (loci DR and DQ) was also indicated, but results are not clear regarding the clinical pictures of the disease nor vaccination response. HLA class III (fourth complement component – C4, third complement component – C3, and properdin factor – BF) are associated with various manifestations of this disease. The gammaglobulin phenotype Gm(1, 2, 3, 10, 21) was more frequent in CAH. However, in only three publications was the impact of HLA on the efficacy of interferon therapy taken into account.
Keywords: HBV; genetics; HLA; C3; C4; BF; Gm; Inv
Full-textPDF downloadAnti-apoptosis function of TNF-ain chronic lymphocytic leukemia: lessons from Crohn’s disease and the therapeutic potential of bupropion to lower TNF-a
Irma Kacprzak-Bergman and Beata Nowakowska
Abstract. The association of genetic factors with hepatitis B virus (HBV) infection susceptibility, its different manifestations, and the different responses to hepatitis B antigen vaccination have been described by several authors. With regard to HLA class I molecules, association with HLA-B was especially observed. HLA-B35 and -B8 correlated with chronic active hepatitis (CAH) and with hepatitis B carriers. Correlation between HBV infection and HLA class II (loci DR and DQ) was also indicated, but results are not clear regarding the clinical pictures of the disease nor vaccination response. HLA class III (fourth complement component – C4, third complement component – C3, and properdin factor – BF) are associated with various manifestations of this disease. The gammaglobulin phenotype Gm(1, 2, 3, 10, 21) was more frequent in CAH. However, in only three publications was the impact of HLA on the efficacy of interferon therapy taken into account.
Keywords: HBV; genetics; HLA; C3; C4; BF; Gm; Inv
Full-textPDF downloadExpression profile analysis of human peripheral blood mononuclear cells in response to aspirin
Sunghee Choi, Hae-Sim Park, Myeong Sook Cheon and Kyunglim Lee
Abstract. Introduction: Aspirin is a popular nonsteroidal anti-inflammatory drug, but some patients suffer from hypersensitivity to it. This prompted us to identify the factors or molecules related to these responses.
Materials and Methods: A commercially available DNA microarray was used to study changes in gene expression in human peripheral blood mononuclear cells (PBMCs) after aspirin treatment. The PBMCs were collected from a patient with aspirin-intolerant asthma and one normal healthy control.
Results: We identified 61 and 107 genes respectively induced and repressed by aspirin treatment in the PBMCs derived from the normal control. In the patient showing aspirin-induced asthma responses, 31 genes were up-regulated and 6 were down-regulated after aspirin treatment. Among these, 1 gene was expressed with the same pattern in the control and the patient. In contrast, 19 genes showed different expression patterns, and it turned out that most of them were involved in immune responses, cell growth/proliferation, transcription/ translation, and signaling pathways.
Conclusions:These results show the molecules involved in hypersensitivity to aspirin and may lead to a better understanding of adverse responses to aspirin. Furthermore, they can provide clues for identifying novel therapeutic and/or preventive molecular targets of the adverse effects of aspirin.
Keywords: aspirin; asthma; human peripheral blood mononuclear cells; microarray
Full-textPDF downloadCartilage formed by syngeneic rat chondrocytes in joint surface defectsis rejected in animals sensitized with allogeneic chondrocytes: involvement of the synovial lining
Stanisław Moskalewski, Anna Osiecka-Iwan, Anna Hyc and Justyna Niderla
Abstract. Introduction: The aim of the study was to discover the mechanism of rejection of chondrocyte transplants introduced into articular cartilage defects.
Materials and Methods: Chondrocytes from 3–5-day-old Lewis or WAG rats were liberated by enzymatic digesandtion from articular-epiphyseal cartilage complexes and implanted into defects made in the subpatellar region of the femur condyle of naive Lewis rats. Syngeneic transplants were also done aftersensitization of the recipients with allogeneic chondrocytes injected intramuscularly. The transplants and synovial membrane were studied in periodate-lysineparaformaldehyde- fixed material with antibodies against B lymphocytes, CD4+ and CD8+ cells, NK cells, and macrophages. For detection of humoral response, chondrocyte lysates were subjected to protein electrophoresis and Western blotting with sera from the transplant recipients.
Results: Cartilage produced in intracartilaginous transplants of syngeneic chondrocytes did not show any signs of rejection. CD8+ lymphocytes and macrophages accumulated in the vicinity of cartilage produced by similar transplants in animals sensitized with intramuscular transplants of allogeneic WAG chondrocytes or bearing transplants of allogeneic WAG chondrocytes. CD8+ cells penetrated into the peripheral part of the cartilage, while macrophages advanced much more deeply. No specific anti-chondrocyte antibody was detected. The synovium from rats bearing intracartilaginous transplants of allogeneic chondrocytes or syngeneic chondrocytes after sensitization contained macrophages and CD8+ cells.
Conclusions: The rejection of cartilage formed by syngeneic chondrocyte transplants in sensitized animals argues in favor of a chondrocyte-specific antigen expression. The involvement of the synovial membrane during transplant rejection suggests that it should be included in observations of the behavior of chondrocyte transplants introduced into articular cartilage.
Keywords: chondrocyte transplants rejection; synovium; CD8+ cells; macrophage
Full-textPDF downloadQuantification of airborne birch (Betula sp.) pollen grains and allergens in Krakow
Jacek Madeja, Ewa Wypasek, Barbara Plytycz,Krzysztof Sarapata and Krystyna Harmata
Abstract. Introduction: Birch (Betula sp.) pollen grains are the main cause of seasonal allergies in northern andcentral Europe. The allergen particles released from the grains are often well distributed in the air. Due to their size, airborne protein particles can easily penetrate into the lower parts of the respiratory airways and may lead to symptoms of asthma. The purpose of this paper was to quantify both Betula sp. pollen grains and allergens in the air.
Materials and Methods: Materials for the investigation were collected in the spring of 2003 with two Hirst-type pollen volumetric traps. Tapes from one trap served for routine birch pollen grain counts, while those from the second for the immunodetection of birch allergens. As birch pollen allergen concentration is seen as dark spots on X-ray films densitometric measurements of the spots were used to quantify birch-pollen antigen concentrations in the air.
Results: In most instances, birch pollen counts corresponded with birch pollen allergen levels. However, on several occasions outside the pollen season, only grains or only allergens were detected. Apart from sampling variability, this could be due to faulty/dead pollen grains or submicronic airborne allergen particles.
Conclusions: Counting intact pollen grains and antibody-based detection of allergen molecules are efficient tools in controlled allergen avoidance.
Keywords: pollen allergy; pollen allergen release; immunodetection; pollen traps; birch allergens; Betula
Full-textPDF downloadINOS expression and NO production by neutrophils in cancer patients
Ewa Jabłońska, Wioletta Pużewska, Magdalena Marcińczyk, Zyta Grabowska and Jakub Jabłoński
Abstract. Introduction: The tumor-polymorphonuclear neutrophil (PMN) relationship can be altered by the release of toxic molecules, such as nitric oxide (NO). The aim of the present study was to examine the expression of the inducible synthase of NO (iNOS) and NO production by human neutrophils of patients with oral cavity cancer. For comparison we performed similar examinations in autologous peripheral blood mononuclear cells (PBMCs).
Materials and Methods: PMNs and PBMCs were isolated from the whole blood of 27 patients with squamous cell carcinoma of the oral cavity. iNOS protein expression in these cells was detected by Western blot. Total nitrite as an indicator of NO concentrations in the culture supernatants and the serum of patients was measured using a colorimetric assay.
Results: The PMNs of oral cavity cancer patients showed a significantly lower intensity of iNOS expression than those of healthy controls. The PBMCs of patients showed a more intensive expression of iNOS than the PMNs, but a lower intensity than the PBMCs of the controls. The expression of iNOS in rhIL-6 and rhIL-15-stimulated PMNs and PBMCs of patients increased in comparison with unstimulated cells. We observed lower productions of NO by PMNs and PBMCs of patients than those of the control group.
Conclusions: The results revealed that altered iNOS expression and NO production are more characteristic of PMNs than of PBMCs of patients with oral cavity cancer. Additionally, this study provided new information about IL-6 and IL-15 activity in a tumor-bearing host.
Keywords: neutrophils; peripheral blood mononuclear cells; squamous oral cavity cancer; nitric oxide; inducible synthase of nitric oxide
Full-textPDF downloadThe influence of immune system stimulation on encapsulated islet graft survival
Tadeusz Orłowski, Ewa Godlewska, Magda Tarchalska, Joanna Kinasiewicz, Magda Antosiak1 and Marek Sabat
Abstract. Introduction: The aim of this study was to determine the influence activating of the recipient immune system on the function of microencapsulated islet xenografts.
Materials and Methods: The skin of WAG or Fisher rats and WAG free or encapsulated (APA) Langerhans islets were transplanted to healthy or to streptozotocin diabetic BALB/c mice. Skin grafts were performed following the method of Billingham and Medawar. Rat islets were isolated from pancreas by the Lacy and Kostianovsy method and encapsulated with calcium alginate- poly-L-lysine-alginate according to the 3-step coating method of Sun.
Results: The transplantation of encapsulated WAG islets, despite activation of the host immune system, restored euglycemia for over 180±100 days. A subsequent skin graft taken from the same donor was rejected in the second set mode, but euglycemia persisted. In diabetic recipients, impaired immune response was corrected by successful encapsulated islet transplantation. In diabetic mice, strong stimulation with 2-fold skin transplantation induced primary non-function of grafted islets despite their encapsulation.
Conclusions: The survival of an islet xenograft depends on the level of activation of the recipient immune system. The immune response of diabetic mice was impaired, but increased after post-transplant restitution of euglycemia. Microencapsulation sufficiently protected grafted islets, and remission of diabetes was preserved. However, after strong specific or non- -specific stimulation of the host immune system, non-function of xenografted islets developed despite their encapsulation. Therefore, islet graft recipients should avoid procedures which could stimulate their immune systems. If absolutely necessary, the graft should be protected by exogenous insulin therapy at that time
Keywords: islet xenograft; islet encapsulation; type l diabetes mellitus
Full-textPDF download [/su_spoiler]Vol. 53, No. 3, 2005
Pneumolysin as a vaccine and drug target in the prevention and treatment of invasive pneumococcal disease
Riana Cockeran, Ronald Anderson and Charles Feldman
Abstract. Streptococcus pneumoniae (the pneumococcus) remains one of the major human pathogens and one of the most common causes of community-acquired pneumonia, otitis media, sinusitis, and meningitis. Aside from the threats posed by emerging antibiotic resistance and infection with the human immunodeficiency virus, the mortality rate among those patients with severe pneumococcal disease who receive seemingly appropriate antimicrobial chemotherapy remains unacceptably high. Because of its involvement in the pathogenesis of invasive disease, pneumolysin, one of the best-characterized virulence factors of the pneumococcus, represents not only a potential vaccine target, but also a target for adjunctive therapy to antibiotics in patients with acute pneumococcal disease. In this paper we review the cytolytic and pro-inflammatory properties of pneumolysin and their involvement in subversion of host defenses and extra-pulmonary dissemination of the pneumococcus, as well as strategies, both immunological and pharmacological, which may counter these harmful activities of the toxin.
Keywords: antibiotics • anti-inflammatory agents • community acquired pneumonia • conjugate vaccines • invasive pneumococcal disease • pneumolysin
Full-textPDF downloadPhagocyte NADPH oxidase: a multicomponent enzyme essential for host defenses
Jamel El-Benna, Pham My-Chan Dang, Marie-Anne Gougerot-Pocidalo and Carole Elbim Unité Inserm U479, Centre Hospitalo-Universitaire Xavier Bichat, Paris, France
Abstract. Phagocytes such as neutrophils and monocytes play an essential role in host defenses against microbial pathogens. Reactive oxygen species (ROS), such as superoxide anion, hydrogen peroxide, the hydroxyl radical, and hypochlorous acid, together with microbicidal peptides and proteases, constitute their antimicrobial arsenal. The enzyme responsible for superoxide anion production and, consequently, ROS generation, is called NADPH oxidase or respiratory burst oxidase. This multicomponent enzyme system is composed of cytosolic proteins (p47phox, p67phox, p40phox, and rac1/2) and membrane proteins (p22phox and gp91phox, which form cytochrome b558) which assemble at membrane sites upon cell activation. The importance of this enzyme in host defenses is illustrated by a life-threatening genetic disorder called chronic granulomatous disease in which the phagocyte enzyme is dysfunctional, leading to life-threatening bacterial and fungal infections. Also, because ROS can damage surrounding tissues, their production, and thus NADPH oxidase activation, must be tightly regulated. This review describes the structure and activation of the neutrophil NADPH enzyme complex.
Keywords: NADPH oxidase • neutrophils • phagocyte • CGD
Full-textPDF downloadSystemic lupus erythematosus – recent clues from congenic strains
Tamika Henry and Chandra Mohan
Abstract. Systemic lupus erythematosus is a polycongenic autoimmune disease characterized by the production of antinuclear antibodies that lead to subsequent end organ damage. The study of lupus is complicated by its polycongenic origin, contributions from hormones and the environment, epistasis among susceptibility loci, suppressive modifiers, and the fact that a single susceptibility locus may encompass multiple susceptibility genes. Murine models that develop lupus spontaneously have greatly contributed to our understanding of this disease. In particular, the advent of “congenic strains” has greatly simplified the study of this complex autoimmune disease. Thus, congenic strains bearing NZB/NZW/NZM2410, BXSB, and MRL lupus susceptibility loci are steadily replacing the traditionally studied murine lupus models as the models of choice for research. This review summarizes how researchers have used congenic strains over the past few years to dissect out and reconstruct the individual elements contributing to lupus pathogenesis.
Keywords: lupus • murine lupus • congenic strains • autoantibodies
Full-textPDF downloadThe biology and pathology of hypoxia-ischemia: an update
Andrew N. Clarkson, Brad A. Sutherland and Ian Appleton Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand
Abstract. After an hypoxic-ischemic (HI) insult, a multi-faceted complex cascade of events occurs that ultimately causes cell death and neurological damage to the central nervous system. The various cascades include, amongst others: immunological changes, such as the activation of the complement system and the generation of antibodies; increased inflammation through the actions of pro-inflammatory cytokines and chemokines; the production of reactive oxygen species leading to oxidative stress; and diminished mitochondrial function leading to the activation of apoptotic pathways and subsequent alteration in the function of neurons within the contralateral hemisphere. This review addresses the immunological aspects following HI, the role of various cytokines (both pro-inflammatory and anti-inflammatory) and chemokines after the induction of HI. In addition, the role of free radicals in producing HI-induced neurodegeneration and the contribution that mitochondrial dysfunction has in neuronal apoptotic cell death will be discussed. This review also covers the changes that the previously assumed “internal control”, the contralateral hemisphere, undergoes due to HI and describes the difficulties associated with therapy intended to prevent neuronal injury associated with HI.
Keywords: angiogenesis • immunity • inflammation • mitochondria
Full-textPDF downloadNew biodefense strategies by neutrophils
Fumio Ishikawa and Shuichi Miyazaki
Abstract. Chemokines and other chemotactic factors induce neutrophils, macrophages, and dendritic cells to migrate to an inflammatory site and efficiently ingest and destroy infective microorganisms. Moreover, antigen-presenting cells, such as macrophages and dendritic cells, present the microbial antigens via major histocompatibility complex class II molecules, resulting in the activation of specific CD4 T cells. Since neutrophils have a short life–span and are highly susceptible to apoptosis, their role in antigen presentation has been questioned. However, various pro-inflammatory cytokines, such as interleukin (IL)-1, IL-6, tumor necrosis factor α, and interferon γ, produced at the site of inflammation activate neutrophils and suppress apoptotic death. These cytokine-activated neutrophils show enhanced expression of cell surface molecules and become as competent as dendritic cells and macrophages in their ability of antigen presentation. Traditionally, neutrophils are known to be responsible for innate immunity, and recently they are also considered to be intimately associated with the establishment of acquired immunity. In the present review on the role of neutrophils we describe both classic innate and acquired immunity.
Keywords: neutrophils • chemotaxis • chemokines • antigen presentation
Full-textPDF downloadCellular responses to attaching and effacing bacteria: activation and implication of the innate immune system
Alain P. Gobert, Keith T. Wilson and Christine Martin
Abstract. During the last decade, research on attaching-effacing (A/E) bacteria/host cell interactions has revealed much of the molecular basis of colonization and lesion formation. The colonic mucosa represents the first line of defense against these pathogens, and its integrity is required to avoid translocation of bacteria or bacterial soluble factors into the infected host. Therefore, the cellular immune response to A/E pathogens plays an important role in bacterial pathogenesis since it can clear the bacteria or modulate the inflammatory processes. Data obtained from infected patients demonstrate a correlation between the production of pro-inflammatory cytokines and the severity of the disease. In vitro studies of infected epithelial cells have clearly elucidated A/E bacteria-induced host signal transduction events. However, the identification of the bacterial factors responsible for cellular activation remains a subject of controversy. Experimental studies with knock-out mice infected with Citrobacter rodentium, a rodent A/E pathogen, indicate that innate immunity is an essential component of pathogenesis. This review summarizes in vivo and in vitro evidence for the induction and potential role of the innate immune system during infection with A/E bacteria.
Keywords: EHEC • EPEC • inflammation • epithelial cells • macrophage • cytokine • nitric oxide
Full-textPDF downloadInnate immunity: cells, receptors, and signaling pathways
Zofia Błach-Olszewska
Abstract. Essential differences between the innate and acquired branches of immunity are described. These differences concern the detection system (receptors and pathogen structures) and the cells engaged in both systems as well as the effectory mechanisms. In contrast to those of the acquired system, receptors of the innate system, which developed during evolution, recognize unchanged structures on large groups of pathogens (e.g. lipopolysaccharide in Gram-negative bacteria). Two lineages, natural killer (NK) and dendritic cells (DCs), play important roles in the innate system. Phenotypic and functional differentiation is observed among NKs and DCs, so each of their sublineages plays a different role in the innate system. Every lineage of cells of the innate immune system express different stimulatory and sometimes also inhibitory receptors on their surfaces (e.g. NK cells). Among the stimulatory are Toll-like receptors (TLRs), mannose and scavenger receptors, and the stimulatory receptors of NK cells. All TLRs show similarity in structure and in the kind of molecules involved in intracellular signaling. The immune reactions of the innate system involve cytokine-dependent resistance of cells against infection with pathogen, production of cytokines (tumor necrosis factor, interferons, interleukins, chemokines) and MHC-independent killing. Although these reactions protect the host from invasion by microorganisms, they can also be responsible for significant tissue damage or may stimulate the development of autoimmunity. Therefore innate immunity must be under rigorous control. The possible regulatory mechanisms of innate immunity are discussed.
Keywords: innate immunity • receptors • intracellular signaling • immune reactions • regulation
Full-textPDF downloadEvaluation of interleukin-1 and -6 in the etiopathogenesis of idiopathic osteoporosis and osteopenia in children
Agnieszka Rusińska and Danuta Chlebna-Sokół
Abstract. Introduction: The aim of the study is to determine whether serum concentrations of interleukin (IL)-1 and IL-6 correlate with indices of bone mineral metabolism in children with idiopathic osteoporosis and osteopenia.
Materials and Methods: The study comprised 62 patients aged 6–18 years (20 with idiopathic osteoporosis, 22 with idiopathic osteopenia, and 20 controls). In 10 children, investigations were repeated after one year of treatment. Serum concentrations of IL-1(α, IL-1β, IL-1 receptor antagonist (IL-1ra), as well as IL-6 and its soluble receptor (IL-6sR) were determined by the ELISA method. In patients with decreased bone mass, selected calcium-phosphorus metabolism indices and bone turnover markers were assessed.
Results: Higher values of IL-6 were recorded in those with idiopathic osteoporosis than in controls (2.79 vs. 1.43; p<0.05). In these patients there was also a tendency towards higher values of IL-6sR (p=0.05). IL-1(α and IL-1β were not markedly elevated in any of the patients. No significant differences between groups regarding IL-1ra were observed. Negative correlation between IL-6, IL-1(α, cytokine/receptor indices, and spinal bone mineral density was determined. Positive correlation was found between IL-(α, IL-1/IL-1ra, and parathormon as well as between IL-1(α, IL-6sR, and bone formation markers. Increase in bone mass after treatment was accompanied by a decrease in IL-6sR.
Conclusions: The higher serum levels of IL-6 in children with idiopathic osteoporosis/osteopenia and the decrease in IL-6sR after treatment reveal an involvement of IL-6 in the etiopathogenesis of these disturbances. The results suggest that IL-1 may also participate in the primary decrease of bone mass in children.
Keywords: cytokine · bone mineralization · children
Full-textPDF downloadExpressions of selected adhesion molecules on peripheral blood leukocytes in patients with aggressive periodontitis
Malgorzata Pietruska, Janusz Żak, Jan Pietruski and Jolanta Wysocka
Abstract. Introduction: Aggressive forms of periodontitis lead to rapid bone destruction resulting in extensive losses in children’s and young adults’ dentition. Adhesion molecule deficiency syndrome and abnormalities in the expression of various adhesion molecules on peripheral blood leukocytes can be observed in prepubertal and aggressive periodontitis (AP) patients. The aim of the study was thus to assess the expression of selected cell adhesion molecules (CAMs; CD11a, CD11b, CD11c, CD54, and CD62L) on monocytes, neutrophils, and lymphocytes of the peripheral blood in patients with AP.
Materials Methods:The study involved 16 patients with AP and a control group of 13 generally healthy suband jects with healthy periodontium. CAM expressions were determined by flow cytometry and presented as mean fluorescence intensity (MFI) and percentage of cells showing expression of the assessed adhesion molecules.
Results: Neutrophil CAM expressions in AP patients were comparable with those of the control group. MFI of CD62L on monocytes in AP patients was significantly lower than that of the controls. Lymphocytes showed increased CD11b expression compared with the control group. The percentage of leukocytes showing CAM expression in both groups was similar. Only the percentage of lymphocytes with CD11b in AP patients was significantly higher than in healthy controls.
Conclusions: Because of the evident lack of differences between patients and controls and the great amount of individual dispersion of the results, the above CAMs on peripheral blood leukocytes in generally healthy patients with AP do not seem to be characteristic markers of this disease.
Keywords: adhesion molecules · peripheral blood leukocytes · aggressive periodontitis
Full-textPDF downloadEffect of cardiopulmonary bypass on neutrophil activity in pediatric open-heart surgery
Jarosław Paśnik, Krzysztof Siniewicz, Jadwiga Anna Moll, Jacek Moll, Zbigniew Baj, Andrzej Sysa and Krzysztof Zeman
Abstract. Introduction: The nature of the participation of neutrophils in the post-cardiopulmonary bypass (CPB) inflammatory response is not very clear. The aim of our study was to investigate alterations in neutrophil phagocytic activity and adhesion molecule expression on these cells in children during after CPB.
Materials and Methods: Twenty-one children aged 6–33 months with congenital heart disease, scheduled for priand mary corrective surgery, were enrolled. The expressions of CD11b adhesion molecules and Fcγ receptor on neutrophils and their phagocytic activity were evaluated. The studied markers were sequentially measured before, at the initiation of, and after CPB.
Results: During the course of the operation, CD11b molecule expression on neutrophils showed a slight elevation at the start of CPB (876.5±104.8 mean fluorescence intensity, MFI, vs. 768.1±178.2; p=0.0047), followed by a significant decrease to 689.01±166.7 MFI after completion of the procedure. The expression of CD11b molecule on neutrophils measured at the end of CPB inversely correlated with the duration of CPB (r= –0.68, p=0.00059). The expression of CD16 antigen dropped significantly at the start of CPB (1164.6±307.3 MFI vs. 1327.4±345.3 MFI; p=0.0007) and remained decreased until the end of CPB (814.0±198.1 MFI).
Conclusions: These findings suggest that the characteristics of the neutrophil response to cardiac surgery appear to depend on many factors. We demonstrated a link between the duration of CPB and adhesion molecule expression on neutrophils.
Keywords: cardiopulmonary bypass · systemic inflammatory response syndrome · neutrophil · phagocytic activity
Full-textPDF downloadVol. 53, No. 4, 2005
Human T cell leukemia virus type 1: the role of Tax in leukemogenesis
Cynthia A. Pise-Masison, Soo-Jin Jeong and John N. Brady
Abstract. Human T cell leukemia virus type 1 (HTLV-1) is a complex human retrovirus which is the causative agent of adult T cell leukemia (ATL). ATL occurs in about 4% of carriers and develops after a long latent period. Although the precise mechanism of HTLV-1 oncogenesis remains unclear, the pathogenesis has been linked to the pleiotropic activity of the viral transcriptional activator protein Tax. Tax has been shown to regulate viral and cellular gene expression and to functionally interfere with proteins involved in cell-cycle progression and DNA repair. This review will focus on the role of Tax in p53 inhibition.
Keywords: T cell; leukemia; Tax protein.
Full-textPDF downloadHemopoietic cell transplantation for the myelodysplastic syndromes
Bart L. Scott and H. Joachim Deeg
Abstract. Myelodysplastic syndromes (MDS) are hemopoietic stem cell disorders, and hemopoietic stem cell transplantation is currently the only therapeutic modality with curative potential. Among patients with less advanced/low-risk MDS (<5% marrow blasts), 3-year survivals of 65–70% are achievable with HLA-identical related and unrelated donors. The overall probability of disease recurrence in these patients is <5%. Among patients with more advanced disease (>=5% marrow blasts), the relapse probability is higher, ranging from 10–40%, and relapse-free survival is correspondingly lower. The criteria proposed by the International Prognostic Scoring System, derived from non-transplanted patients, also predict survival following transplantation. The development of reduced-intensity conditioning regimens and modification of conventional regimens, all aimed at optimizing the transplant approach, have permitted successful hemopoietic stem cell transplants even in patients 60–70 years of age. Improved survival with transplants from unrelated volunteer donors reflects to a large extent selection of donors on the basis of high resolution (allele-level) HLA typing. Graft-versus-host disease and associated problems remain major challenges after allogeneic transplantation. Autologous stem cell transplantation may be beneficial for selected patients who have obtained complete remissions with conventional chemotherapy.
Keywords: MDS; prognostic scoring systems; hemopoietic cell transplantation; conditioning regimens.
Full-textPDF downloadPeptide-based approaches to treat asthma, arthritis, other autoimmune diseases and pathologiesof the central nervous system
Kristin Hauff, Christina Zamzow, Warren J. Law, Jimmy De Melo, Kieron Kennedy1 and Marek Los
Abstract. In this review we focus on peptide- and peptidomimetic-based approaches that target autoimmune diseases and some pathologies of the central nervous system. Special attention is given to asthma, allergic rhinitis, osteoarthritis, and Alzheimer’s disease, but other related pathologies are also reviewed, although to a lesser degree. Among others, drugs like Diacerhein and its active form Rhein, Pralnacasan, Anakinra (Kineret), Omalizumab, an antibody “BION-1”, directed against the common b-chain of cytokine receptors, are described below as well as attempts to target b-amyloid peptide aggregation. Parts of the review are also dedicated to targeting of pathologic conditions in the brain and in other tissues with peptides as well as methods to deliver larger molecules through the “blood-brain barrier” by exploring receptor-mediated transport, or elsewhere in the body by using peptides as carriers through cellular membranes. In addition to highlighting current developments in the field, we also propose, for future drug targets, the components of the inflammasome protein complex, which is believed to initiate the activation of caspase-1 dependent signaling events, as well as other pathways that signal inflammation. Thus we discuss the possibility of targeting inflammasome components for negative or positive modulation of an inflammatory response.
Keywords: Anakinra; BION-1; b-amyloid; Diacerhein; Kineret; Omalizumab; osteoarthritis; Pralnacasan; Rhein; secretase; Zafirlukast.
Full-textPDF downloadMonocyte-related immunopathologies in trauma patients
Krzysztof Laudanski and Dorota Wyczechowska
Abstract. Mechanical trauma is one of the most important causes of morbidity in the developed world. The response of the immune system to mechanical insult is of paramount importance for the patient’s recovery. Shortly after trauma, the indiscriminate saystemic inflammatory response syndrome (SIRS) is mediated by circulating monocytes (MΦs) and other innate immunity components. Then acquired immunity, limited to the offending pathogen and the site of injury, gradually preponderates. SIRS is followed by the compensatory anti-inflammatory response syndrome (CARS), where the initial inflammatory response is quenched by anti-inflammatory mediators. This precisely regulated process of immune system activation in response to trauma can be easily deviated, resulting in multiorgan failure (MOF) and increased mortality. Excessive activation of inflammatory MΦs in the SIRS phase, premature or exorbitant CARS, a predominance of macrophages (Macs) in the blood stream and peripheral tissues, as well as a depletion of dendritic cells are often seen in trauma patients and contribute to the development of MOF. Here we explore several mechanisms of pathological MΦ activation in patients with severe mechanical traumatic injury without accompanying sepsis.
Keywords: mechanical trauma; monocyte; dendritic cell; macrophage; inflammatory monocyte.
Full-textPDF downloadNanovesicular vaccines: exosomes
Xiaobo Li, Zhiren Zhang, Thomas Beiter and Hermann J. Schluesener
Abstract. Exosomes are small membrane vesicles derived from late endosome. They are about 30–100 nm in diameter. The secretion of exosomes is a process in which multivesicular bodies fuse with the cell membrane, and all cells that contain multivesicular endocytic compartments could theoretically secrete exosomes. The surprising biological functions of exosomes are only slowly being unveiled, but it is already clear that they serve to remove obsolete membrane proteins and act as messages of inter-cellular communication. Exosomes derived from tumor or antigen-presenting cells have been extensively investigated. They are released into the extracellular environment and fuse with the membranes of neighboring cells, delivering membrane and cytoplasmic proteins from one cell to another. Exosomes carry immunorelevant structures which play important roles in immune response, such as MHC molecules, costimulatory molecules, heat shock proteins, and naive tumor antigens. Therefore they have been suggested as potential vaccines. Consequently, exosomes have shown considerable anti-tumor effect in several studies and are in phase I clinical trials.
Keywords: exosomes; immunotherapy; biogenesis; tumor; vaccine.
Full-textPDF downloadThe soluble CTLA-4 receptor: a new marker in autoimmune diseases
Edyta Pawlak, Iwona Ewa Kochanowska, Irena Frydecka, Marek Kiełbiński, Stanisław Potoczek and Małgorzata Bilińska
Abstract. A soluble form of cytotoxic T lymphocyte-associated antigen-4 (sCTLA-4) was recently found and shown to possess B7 binding activity. sCTLA-4 is generated by alternatively spliced mRNA. The mRNA encoding sCTLA-4 consists of 3 exons: exon 1 encodes a leader peptide, exon 2 the ligand binding domain, and exon 4 the cytoplasmic tail, but it lacks the transmembrane domain encoded by exon 3. The altered transcript is detected in resting CD4 and CD8 T cells and its expression is inhibited after 24–48 h of activation and returns to the prestimulation level after 72–120 h of activation. Low levels of sCTLA-4 have been detected in normal human serum and increased serum levels have been observed in several autoimmune diseases (e.g. Graves’ disease, myasthenia gravis, systemic lupus erythematosus, and systemic sclerosis). The biological significance of increased sCTLA-4 serum level has not been clarified. On one hand, sCTLA-4 may bind B7 expressed on antigen-presenting cells and is thus able to interfere with the B7:CD28-mediated costimulation of T cell responses. On the other hand, sCTLA-4 may also be capable of interfering with B7:CTLA-4 interactions, thereby blocking the negative signal imparted via the full-length form of CTLA-4. This double-edged nature of B7 blocking by sCTLA-4 may result in different outcomes of the clinical course of disease.
Keywords: sCTLA-4; alternative splicing; autoimmune diseases.
Full-textPDF downloadA mouse monoclonal antibody to the TSHR
Hanna Stankowiak-Kulpa, Jane Sanders, Hilde Depraetere, Jennifer Jeffreys, Michele Evans, Tonya Richards, Jadwiga Furmaniak and Bernard Rees Smith
Abstract. Introduction:
Mouse monoclonal antibodies (mAbs) with the ability to inhibit thyrotropin (TSH) binding to the TSH receptor (TSHR) are useful tools to study TSH-TSHR interaction. The 3C3 mAb we produced was found to inhibit binding of TSH to human (h)TSHR but not to porcine (p)TSHR.
Materials and Methods:
Purified 3C3 immunoglobulin G (IgG) and its antibody-binding fragment were prepared using standard methods and their ability to inhibit TSH binding to hTSHR or pTSHR was analyzed using a coated tube assay. The TSHR epitope reactive with 3C3 IgG was determined using Western blotting, ELISA based on peptides corresponding to the TSHR sequence, and the SPOT synthesis technique. RNA was isolated from 3C3 hybridoma cells and the mAb variable (V) region genes were sequenced and analyzed.
Results:
3C3 mAb had a 1×108l/mol binding affinity to the hTSHR as assessed by Scatchard analysis. 3C3 reacted with the hTSHR region between amino acids (aa) 212-230, and two aa differences were found between the corresponding regions in the hTSHR and pTSHR. The light chain (LC) genes of 3C3 were derived from the Vk21 germ-line (97.6% homology) and Jk2 genes. The heavy chain (HC) genes were from the V130 germ-line (94.6% homology) combined with a D gene (not identified) and JH3 gene. The replacement/silent mutation ratios of 6.0 and 6.5 for the LC and the HC V regions, respectively, indicated that 3C3 underwent antigen-driven maturation.
Conclusions:
Mouse mAbs of this type should be useful in studying the interactions between the TSHR, TSH, and mAbs in more detail.
Keywords: thyrotropin receptor; monoclonal antibodies; thyroid; Graves’s disease.
Full-textPDF downloadPrevalence of the intron 22 inversion of the factor VIII gene and inhibitor development in Polish patients with severe hemophilia A
Jadwiga Sawecka, Joanna Skulimowska, Jerzy Windyga, Stanisław Łopaciuk and Jerzy Kościelak
Abstract. Introduction:
Patients with severe hemophilia A often develop inhibitors (antibodies) against transfused factor VIII.
Materials and Methods:
One hundred thirteen Polish patients with severe hemophilia A, who had been treated on and Methods: demand with cryoprecipitate until 1992 and exclusively with factor VIII concentrates after 1995, were examined for intron 22 inversion by Southern blotting and the presence and magnitude of inhibitor activity in blood as determined by the Bethesda assay. The patients’ ages ranged 4–67 years (mean: 33.7±12.4 years, median: 32 years).
Results:
The number of patients with the inversion amounted to 57, while in 56 patients the mutation types were unknown; 47 patients had a distal and 10 patients a proximal type of inversion. Thirteen patients with inversions (22.8%) were found to have inhibitor in their blood. Most patients (14 out of 15) who developed inhibitors in the course of cryoprecipitate therapy were high responders. Conversely, 4 of 5 patients treated between 1992 and 1995 with both cryoprecipitate and intermediate-purity factor VIII concentrates were low responders. One multitransfused patient who had remained inhibitor-free on cryoprecipitate therapy developed inhibitor after receiving a large dose of factor VIII concentrate during surgery. None of these 5 patients developed inhibitors during their 12–40 years of treatment with cryoprecipitate, suggesting that it was less immunogenic than factor VIII concentrates.
Conclusions:
The prevalence of the intron 22 inversion mutation of the factor VIII gene in Polish hemophiliacs is similar to that in other European countries. Treatment regimens with either cryoprecipitate or virus-inactivated plasma-derived factor VIII concentrates may affect inhibitor formation in hemophilia A patients.
Keywords: hemophilia A; cryoprecipitate; factor VIII concentrates; intron 22 inversions; inhibitor in hemophilia.
Full-textPDF downloadAssessment of selected co-stimulatory, adhesion and activatory molecules and cytokines of Th1/Th2 balance in acute lymphoblastic leukemia in children
Włodzimierz Łuczyński, Anna Stasiak-Barmuta, Maryna Krawczuk-Rybak and Iwona Malinowska
Abstract. Introduction:
Recent years have seen a rise in the importance of cytokine production and co-stimulatory/activatory molecule expression in the immune response in leukemia. The aim of our study was to assess the function of T lymphocytes in children with acute lymphoblastic leukemia (ALL) during remission induction based on selected cytokine and co-stimulatory/activatory molecule expression.
Materials and Methods:
The study group consisted of 50 children with ALL (B cell precursor). Peripheral blood samples were taken before treatment (day 0), after the prednisone prophase (day 8), and during (day 15) and after (day 33) remission induction. The percentages of T cells with interferon (IFN)-γ (Th1), interleukin (IL)-4 (Th2) and IL-2 receptor (IL-2R), CD28, CTLA-4, CD38, ICAM-1, and HLA-DR expression were assessed by tricolor flow cytometry.
Results:
At the time of diagnosis we noted higher percentages of T cells with adhesion molecule ICAM-1, activation molecule CD38 expression, and an increased population of Th2 cells (IL-4) compared with the control group. During and after remission induction we observed a decreased population of CD38+ T cells, elevated percentages of helper T lymphocytes with IL-2R expression, and a rise in helper T lymphocytes producing IFN-γ (Th1). During fever/infection, higher levels of activated T lymphocytes (CD4+HLA-DR+, CD8+HLA-DR+), a rise in Th1, and no change in Th2 populations were observed.
Conclusions:
The results suggest T cell activation and Th2 predominance at the time of diagnosis and during remission induction in ALL in children. These results confirm the involvement of cellular immunity in the leukemic process and can be used in immune therapy in leukemia.
Keywords: immunosuppression; cancer; IFN-γ IL-4; co-stimulatory molecules.
Full-textPDF downloadSimultaneous transplantation of two allogeneic units of cord blood in an adult patient with acute myeloblastic leukemia. A case report
Wiesław Wiktor-Jędrzejczak, Małgorzata Rokicka, Elżbieta Urbanowska, Tigran Torosian, Elżbieta Graczyk-Pol, Agnieszka Tomaszewska, Małgorzata Król, Monika Paluszewska, Anna Gronkowska, Bogna Ziarkiewicz-Wróblewska, Justyna Jółkowska and Michał Witt
Abstract. Introduction:
The major obstacle to the therapeutic use of hematopoietic transplantation is the unavailability of matched, unrelated marrow donors for the large number of potential patients, although all of them have the chance to find sufficiently matched, unrelated cord blood units. However, the use of cord blood as a source of cells for transplantation is limited by its cell number, usually below 1 billion, which allows for routine transplantation only in children weighting less than 30 kg, while most potential recipients possess a higher body mass. This led to the idea of the simultaneous use of several units of cord blood which, combined, would fulfill the requirements for the necessary cell number for an adult recipient.
Materials and Methods:
We attempted to simultaneously transplant an adult patient with refractory acute myeloblastic leukemia utilizing two different cord blood units, one fully matched and one mismatched at one locus.
Results:
The patient became reconstituted with only one unit, the mismatched, as determined using microsatellite markers, and had no signs of relapse of leukemia. Unfortunately, he died of persistent fungal (brain aspergilloma) infection on day +103.
Conclusions:
The successful engraftment may suggest that a method based on the principle of using more than one cord blood unit for transplantation is feasible in large adult patients and may reach routine application.
Keywords: placental blood; bone marrow transplantation; hematopoiesis.
Full-textPDF downloadMeeting Reports
The 3rd Annual International Umbilical Cord Blood Transplantation SymposiumPDF download The 10th Congress of the European Hematology AssociationPDF downloadVol. 53, No. 5, 2005
Signal transduction in human pancreatic cancer: roles of transforming growth factor β, somatostatin receptors, and other signal intermediates
Min Li, Lauren S. Becnel, Wei Li, William E. Fisher, Changyi Chen and Qizhi Yao
Abstract. Pancreatic cancer is a devastating disease because of the lack of early detection markers and effective treatments. It is the fourth leading cause of cancer-related death in western countries, including the United States. The mechanisms of pancreatic cancer progression remain unknown. Transforming growth factor β (TGF-β), a multifunctional cytokine, regulates cell growth and differentiation in healthy tissues, yet fails to do so in pancreatic cancer. Alterations of the TGF-β and TGF-β receptor/Smad signal transduction pathway have been implicated in pancreatic cancer. Furthermore, both the TGF-β receptor and Smad proteins interact with a variety of cellular signal pathways, such as the somatostatin receptors (SSTRs), ERK1/2, and Wnt signal transduction cascades. This suggests that pancreatic cancer is a multi-gene-controlled malignancy and that effective treatments for pancreatic cancer should be aimed at multiple targets. In this review, we summarized the major signal intermediates involved in pancreatic cancer signal transduction pathways and specifically discussed how alterations in the regulatory functions of TGF-β and Smad proteins allow for pancreatic carcinogenesis.
Keywords: pancreatic cancer; TGF-β; SSTR.
Full-textPDF downloadInflammation in periodontal tissues in response to mechanical forces
Masaru Yamaguchi and Kazutaka Kasai
Abstract. Orthodontic forces are known to produce mechanical damage and inflammatory reactions in the periodontium and dental pulp, as well as inflammatory mediators, e.g. prostaglandins, interleukin (IL)-1, IL-6, tumor necrosis factor α, and receptor activator of nuclear factor κB ligand, in the periodontal ligament (PDL) and dental pulp. We have studied the effects of aging on the production of inflammatory mediators in the PDL using in vitro and in vivo methods and found that aging of PDL tissues may be an important factor in the severity of periodontal disease through a higher production of inflammatory mediators in response to mechanical forces. Further, the levels of inflammatory mediators in gingival crevicular fluid, an osmotically mediated inflammatory exudate found in the gingival sulcus, have been shown to be significantly elevated during orthodontic treatment. In order to reduce inflammation, low-level laser therapy has been recently studied in vivo and in vitro by many investigators as a substitute for anti-inflammatory drugs. Clinical and experimental studies have shown that low-level laser irradiation reduces orthodontic post-adjustment inflammation. We believe that orthodontic forces (mechanical forces) may play an important role in periodontal inflammation and that low-level laser therapy may be useful for its inhibition.
Keywords: inflammation; mechanical forces; cytokines; orthodontics; periodontal disease.
Full-textPDF downloadSurfactant proteins SP-A and SP-D in human health and disease
Uday Kishore, Andrés López Bernal, Mohammed F. Kamran, Shweta Saxena, Mamta Singh, P. Usha Sarma, Taruna Madan and Trinad Chakraborty
Abstract. Surfactant proteins A (SP-A) and D (SP-D) are lung surfactant-associated hydrophilic proteins that have been implicated in surfactant homeostasis and pulmonary innate immunity. They are collagen-containing C-type (calcium-dependent) lectins, called collectins, and are structurally similar to mannose-binding protein of the lectin pathway of the complement system. Being carbohydrate pattern-recognition molecules, they recognize a broad spectrum of pathogens and allergens via the lectin domain, with subsequent activation of immune cells via the collagen region, thus offering protection against infection and allergenic challenge. SP-A and SP-D have been shown to be involved in viral neutralization, clearance of bacteria, fungi, and apoptotic and necrotic cells, down-regulation of allergic reaction, and resolution of inflammation. Studies on single-nucleotide polymorphism, protein levels in broncho-alveolar lavage, and gene knock-out mice have clearly indicated an association between SP-A and SP-D and a range of pulmonary diseases. In addition, recent studies using murine models of allergy and infection have raised the possibility that the recombinant forms of SP-A and SP-D may have therapeutic potential in controlling pulmonary infection, inflammation, and allergies in humans.
Keywords: lung; surfactant; innate immunity; pathogen; allergy; disease; pregnancy.
Full-textPDF downloadToll-like receptor expression and function in airway epithelial cells
Catherine M. Greene and Noel G. McElvaney
Abstract. Toll-like receptors (TLRs) belong to a family of transmembrane proteins that can recognize and discriminate a diverse array of microbial antigens. Following their activation by specific ligands, TLRs initiate intracellular signaling cascades that culminate in the activation of transcription factors and ultimately lead to changes in pro-inflammatory gene expression. The TLR family constitutes an important component of the innate immune system and, although most commonly considered to be associated with immune cell responses, TLRs are also known to be functionally expressed on a variety of other cell types. Epithelial cells represent a significant component of the cellular content of the airways. These cells provide both a barrier to infection and an active defense mechanism against invading microbes. The expression and function of TLRs on airway epithelial cells has been an area of increasing interest in the recent past. This review will summarize advances in our understanding of the role of TLRs in airway epithelial cells.
Keywords: Toll-like receptors; airway epithelial cells; inflammatory lung disease.
Full-textPDF downloadHuman leukocyte antigens as psoriasis inheritance and susceptibility markers
Aneta Szczerkowska-Dobosz
Abstract. Psoriasis is a multifactoral and heterogenetically inherited disease. The role of hereditary transmission is supported by familial association, twin studies, and correlation with human leukocyte antigens (HLA). Numerous studies have proved that B13, B17, Cw6, and DR7 antigens are positively associated with psoriasis. Cw6 antigen has been repeatedly indicated to be the most significant marker for the risk prediction of the disease. On the basis of epidemiological studies and HLA analysis, a concept of two distinct disease patterns of psoriasis vulgaris was proposed. In type I psoriasis the disease has an early onset, strong correlation with Cw6, B13, B17, and DR7 antigens, and familiar inheritance. Type II psoriasis has a late onset, weak correlation with HLA antigens, and sporadic familiar occurrence. Both types seem to differ clinically. Moreover, some extended haplotypes were shown to be correlated with the disease, especially with the type I psoriasis. Although a psoriasis susceptibility gene(s) has not been yet identified, a number of candidate genes were studied, with evidence for a major locus located within the major histocompatibility complex (PSORS 1). Cw6 allele is the most extensively investigated candidate gene, but present evidence suggests that it is rather in strong linkage disequilibrium with the PSORS 1 gene than the susceptibility allele itself. This article reviews past and current data on the genetic background of psoriasis with special attention to its correlation with HLA antigens.
Keywords: Cw6; epidemiology; haplotypes; HLA; MHC; psoriasis.
Full-textPDF downloadMannose-binding lectin enhances the attachment and phagocytosis of mycobacteria in vitro
Agnieszka Bonar , Magdalena Chmiela , Wiesława Rudnicka and Barbara Różalska
Abstract. Introduction:
Phagocytosis is the critical first step in the Mycobacterium (M.) tuberculosis-phagocyte interaction. The process involves microbial ligands and phagocyte surface receptors. It is known that serum mannose-binding lectin (MBL), an innate immune system component, may enhance the uptake of microbes by phagocytic cells and activate the complement system. Since phagocytes are the replicative environment for mycobacteria and, as we described earlier, tuberculosis patients differ from controls in serum MBL level, we asked whether MBL plays a role in promoting M. tuberculosis access to phagocytic cells.
Materials and Methods:
To estimate the influence of MBL on the phagocytic process, FITC-labeled Mycobacterium bovis BCG was used as a model bacterium. Neutrophils from healthy individuals were used as phagocytes. Phagocytosis was performed in the presence or absence of recombinant MBL (rMBL; 2 or 20 μg/ml). The activation of complement was determined by dot-blot immune assay with monoclonal antibodies against C5b-C9.
Results:
We showed that phagocytosis of the bacteria was more intensive in the presence of human rMBL. Both attachment and ingestion of mycobacteria were enhanced when MBL and active complement components (fresh serum) were present in the medium. The dot-blot method showed that the bacteria slightly activated complement by themselves. This effect was enhanced in the phagocyte-bacteria co-cultures containing rMBL.
Conclusions:
It is possible that MBL may serve in vivo as one of the factors facilitating the entry of mycobacteria into phagocytes, pathogen spread, and the establishment of infection.
Keywords: tuberculosis; mycobacteria; phagocytes; collectins; mannose-binding lectin.
Full-textPDF downloadCharacterization of human hepatocytes isolated from non-transplantable livers
Anna Łaba, Alina Ostrowska, Dariusz Patrzałek, Leszek Paradowski and Andrzej Lange
Abstract. Introduction:
The successful use of hepatocytes depends on a reliable demonstration of the functional and morphological integrity of isolated cells. Herein we investigated whether the isolation and cryopreservation of primary human hepatocytes can compromise cell viability and liver-specific characteristics.
Materials and Methods:
Hepatocytes were isolated from encapsulated human liver segments by a modified 2-step perfusion technique. Isolated cells were Percoll-purified, cryopreserved, and stored in liquid nitrogen for 1–12 months. For rapid assessment of fresh and cryopreserve/thawed hepatocyte yield and viability, the cells were stained with trypan blue or labeled with fluorochromes. For immunocytochemical analysis, the cells were labeled with monoclonal antibodies for the presence of the following antigens and chemokines: CD3, CD45Ro, CD45Ra, CD34, CD68, CD90, CD95, CD20, HLA-DR, Ki67, PCNA, Bcl-2, p53, CXCR3, CXCR4, and SDF-1. The cells were tested for several specific functions, such as ureagenesis, energy status, MTT activity, lactate dehydrogenase leakage, and total CYP450 content.
Results:
Assessment of both freshly isolated (Percoll-purified) and cryopreserved/thawed hepatocytes revealed a low constitutive level of contamination by non-parenchymal cells compared with crude (unpurified) preparations and tissue sections. All viable hepatocytes showed intact morphology and retained CYP450 protein, energy status, and urea synthesis.
Conclusions:
Modifications in hepatocyte preparations, such as depletion of dead, damaged, and nonparenchymal cells, improves cell purity, which can be adapted to further evaluation of hepatocyte immunogenicity. These data illustrate the importance and feasibility of human hepatocyte banking.
Keywords: hepatocyte isolation; human; cryopreservation; non-parenchymal cells.
Full-textPDF downloadCorrelation of osteoprotegerin and sRANKL concentrations in serum and bone marrow of multiple myeloma patients
Maria Kraj, Katarzyna Owczarska, Urszula Sokołowska, Piotr Centkowski, Ryszard Pogłód and Barbara Kruk
Abstract. Introduction:
Recent studies suggest that multiple myeloma (MM) triggers osteoclastogenesis by disrupting the balance between the receptor activator of NF-;kappa&B ligand (RANKL) and osteoprotegerin (OPG), its natural antagonist.
Materials and Methods:
Determinations of bone marrow (BM) and serum OPG and sRANKL concentrations were performed in 133 MM patients and 42 healthy subjects by the ELISA method using Osteoprotegerin ELISA and sRANKL ELISA kits.
Results:
MM patients had elevated serum levels of OPG compared with controls (p<0.0001) and OPG levels were higher in patients with renal failure and patients with hipercalcemia (p<0.001 and p=0.04, respectively). Serum OPG levels correlated with age, serum β2-microglobulin, and BM OPG concentrations and did not correlate with the presence of osteolysis or with stage of disease. sRANKL serum levels in MM patients and in controls were not statistically different (p=0.42). In MM patients, serum OPG and sRANKL levels were similar at diagnosis and in the plateau phase of disease. There was a correlation between BM and serum sRANKL concentrations (p<0.001). Median values of the sRANKL/OPG ratio for BM and serum of MM patients were 0.14 and 0.11, respectively. The median value of the sRANKL/OPG ratio for the serum of controls was 0.11.
Conclusions:
In 20% of MM patients, serum OPG levels are elevated, and this may be a compensative reaction related to increased bone destruction. There is not statistically significant relationship between sRANKL serum and BM levels and the main clinical and laboratory parameters of the disease. Determination of BM and the serum sRANKL/OPG ratio seems to have no clinical value.
Keywords: multiple myeloma; osteoprotegerin; RANKL; osteolysis.
Full-textPDF downloadVol. 53, No. 6, 2005
Murine models of susceptibility to tuberculosis
Gillian L Beamer and Joanne Turner
Abstract: Approximately one third of the world’s population is infected with Mycobacterum tuberculosis, yet each year a small proportion of those individuals progress to an active disease state. Early identification and treatment of such individuals is essential to reduce transmission; however, genetic and immunological correlates of disease progression have not been well established in man. The murine model has been a central tool for the elucidation of protective immune mechanisms that are essential for controlling M. tuberculosis infection. Additionally, the study of inbred mice has revealed significant divergence in the susceptibility and disease progression of individual mouse strains to an infection with M. tuberculosis. The continued study of genetically disparate mouse strains has the potential to identify immune mechanisms that correlate with increasing susceptibility to tuberculosis. These mechanisms will be highly applicable to studies in man and assist in the early detection of individuals that are more vulnerable to the development of reactivation tuberculosis.
Keywords: tuberculosis; reactivation; murine; lung.
Full-textPDF downloadImmunotherapeutic approaches in ocular inflammatory diseases
Shree K. Kurup and Chi-Chao Chan
Abstract: This comprehensive review discusses immunotherapeutic approaches to ocular inflammatory diseases, updates information provided in the literature, and presents clinical experiences with an emphasis on autoimmune uveitis at the National Eye Institute, United States. Current medical and surgical therapeutic approaches, including medications such as corticosteroids, anti-metabolites, alkylating agents, calcineurin and purine synthesis inhibitors, biologics as well as some anti-infectious agents, are reviewed along with new modalities and experimental approaches. Most immunosuppressive therapies have significant adverse effects. Physicians must be familiar with the pharmacology of the available drugs and aware of the philosophies behind the treatment.
Keywords: inflammation; uveitis; autoimmune diseases; immunosuppressive medications; adverse effect; therapy.
Full-textPDF downloadRoles of galectin-3 in immune responses
Huan Yuan Chen, Fu-Tong Liu and Ri-Yao Yang
Abstract: Galectins are a family of animal lectins with conserved carbohydrate-recognition domains for β-galactoside. Galectin-3 is the only family member that is composed of a glycine/proline-rich N-terminal repeated sequence and a C-terminal carbohydrate-binding domain. Multiple functions of galectin-3 have been reported, depending on its location. Extracelluar galectin-3 can bind to cell surface through glycosylated proteins and thereby trigger or modulate cellular responses such as mediator release or apoptosis. Intracellular galectin-3 has been reported to inhibit apoptosis, regulate the cell cycle, and participate in the nuclear splicing of pre-mRNA. Recent studies have revealed that galectin-3 is expressed in a variety of cell types in the immune system, constitutively or in response to microbial invasion. These studies implicate galectin-3 in both innate and adaptive immune responses, where it participates in the activation or differentiation of immune cells. This review summarizes the roles of galectin-3 in the immune system and discusses the possible underlying mechanisms.
Keywords: galectin; galectin-3; immunity; immune response.
Full-textPDF downloadNeutrophils in the innate immune response
Scott D. Kobayashi, Jovanka M. Voyich, Christopher Burlak and Frank R. DeLeo
Abstract: Polymorphonuclear leukocytes (PMNs or neutrophils) are an essential component of the human innate immune system. Circulating neutrophils are rapidly recruited to sites of infection by host- and/or pathogen-derived components, which also prime these host cells for enhanced microbicidal activity. PMNs bind and ingest microorganisms by a process known as phagocytosis, which typically triggers production of reactive oxygen species and the fusion of cytoplasmic granules with pathogen-containing vacuoles. The combination of neutrophil reactive oxygen species and granule components is highly effective in killing most bacteria and fungi. Inasmuch as PMNs are the most abundant type of leukocyte in humans and contain an arsenal of cytotoxic compounds that are non-specific, neutrophil homeostasis must be highly regulated. To that end, constitutive PMN turnover is regulated by apoptosis, a process whereby these cells shut down and are removed safely by macrophages. Notably, apoptosis is accelerated following phagocytosis of bacteria, a process that appears important for the resolution of infection and inflammation. This review provides a general overview of the role of human neutrophils in the innate host response to infection and summarizes some of the recent advances in neutrophil biology.
Keywords: neutrophil; phagocytosis; inflammation; infection; apoptosis.
Full-textPDF downloadThe biological role and potential therapeutic application of interleukin 7
Agnieszka Krawczenko, Claudine Kieda and Danuta Duś
Abstract: Interleukin (IL)-7 is a pleiotropic, non-redundant cytokine necessary for the development of B and T lymphocytes, in particular γδ T cell receptor-positive cell differentiation. The cytokine can function as a cofactor during myelopoiesis and the generation of cytotoxic T cells and natural killer cells, can activate monocytes/macrophages, and support the survival of mature T cells. A role for IL-7 in promoting the formation of Peyer’s patch anlage has also been demonstrated. IL-7 is constitutively expressed in the thymus, bone marrow stromal cells, epithelial and dendritic cells, keratinocytes, as well as in fetal and adult liver. IL-7 acts on various cells through its receptor (IL-7R), a heterodimer consisting of an α chain (CD127) that specifically binds IL-7 and a common γc chain (CD132) shared by other cytokine receptors. The receptor is expressed on bone marrow progenitor cells, lymphoid T and B precursors, and mature T cells. IL-7 activity towards murine endothelial cells has been recently described. The presence of IL-7R on human endothelial cells has also been demonstrated. Several therapeutic applications of recombinant IL-7 have been proposed. These have focused on the enhancement of lymphopoiesis, promotion of stem cell engraftment, and the anti-tumor activity of the cytokine.
Keywords: interleukin 7; interleukin-7 receptor; B cell development; T cell development; endothelium; cytokine therapy.
Full-textPDF downloadReactive oxygen intermediates and serum antioxidative system in patients with chronic C hepatitis treated with IFN-α and thymus factor X
Elżbieta Jabłonowska, Henryk Tchórzewski, Przemysław Lewkowicz and Jan Kuydowicz
Abstract. Introduction:
In this study, the chemiluminescence (CL) of peripheral blood polymorphonuclear leukocytes (PMNLs) and the serum total antioxidative system (TAS) were assessed in patients with chronic C hepatitis (CCH) before and after 3 and 6 months of treatment with interferon (IFN)-α and thymus factor X (TFX).
Materials and Methods:
The study included 26 patients with CCH aged between 25–63 years (mean: 42.67). Combined therapy with IFN-α 2a and a TFX preparation was applied. PMNL metabolic activity was assessed applying the whole-blood CL method. We measured CL response of neutrophils unstimulated and stimulated by opsonized zymosan, N-formyl-methionylleucyl-phenylalanine (N-fMLP), and phorbol-myristate-acetate (PMA) without and after priming with tumor necrosis factor α (10 ng/ml). The assessment of serum TAS was performed directly before the beginning of therapy with IFN-α and TFX and after 3 and 6 months of the treatment. A colorimetric method based on the reduction of the cationic radical ABTS•+ (cation 2, 2’-azido-bis-[3-ethylobenzothiazolino-6-sulfonate]) in the presence of serum antioxidants was used.
Results:
As a result of the treatment with IFN-α and TFX, the formation of free oxygen radicals by resting (unprimed) neutrophils increased statistically significantly both without stimulation and following stimulation by fMLP and PMA. A statistically significant increase in the serum antioxidant capacity was observed, which suggests the induction of compensatory processes.
Conclusions:
Increased in vitro reactive oxygen species production by both stimulated and unstimulated peripheral blood neutrophils of patients with CCH was observed. Treatment with IFN-α and TFX resulted in a compensatory increase in serum antioxidative capacity.
Keywords: chronic hepatitis; HCV; chemiluminescence; neutrophils; free radicals.
Full-textPDF downloadSelective indices on non-specific cell-mediated immunity in rabbits immunized with Chlamydophila psittaci
Małgorzata Pawlikowska and Wiesław Deptuła
Abstract. Introduction:
Studies of non-specific immunity in infection or immunization with Chlamydophila (Chl.) psittaci in static experimental models showed changes in parameters of this immunity. The aim of this study was to evaluate selected parameters of non-specific immunity in rabbits immunized with Chl. psittaci.
Materials and Methods:
The study was performed on rabbits, divided into two groups of 10 animals each. The rabbits of the first group were immunized with Chl. psittaci strain 6BC and those of the second group were control animals. Blood samples were examined 9 times every 7 days. Adherence capacity and ingestion capacity of polymorphonuclear (PMN) cells (index of ingestion, percentage of ingestive cells) were determined in the blood. The cidal capacity of PMN cells was determined by evaluating the reduction of nitroblue tetrazolium test by cytochemical (spontaneous and stimulated tests) and spectrophotometric techniques. The presence of specific antibodies in the sera was estimated by the complement-fixation technique.
Results:
Analysis of the results of all the studied parameters of non-specific immunity showed tendencies to decrease or increase. These differences appeared on days 7–14 and lasted to days 42–56 of the experiment. Positive titers of specific antibodies appeared on day 42 after immunization, i.e. 5 weeks after the first changes in the parameters of non-specific immunity.
Conclusions:
The alterations noted in the parameters of non-specific immunity may prove useful in determining the condition of a macroorganism in contact with Chl. psittaci.
Keywords: Chlamydophila psittaci; rabbit; immune indices.
Full-textPDF downloadSerological classification and epitope specificity of Proteus penneri S29 lipopolysaccharide
Krystyna Zych , Małgorzata Siwińska and Zygmunt Sidorczyk
Abstract. Introduction:
Gram-negative bacteria of genus Proteus are common human intestinal and urinary tract pathogens. In the genus Proteus there are four clinically important named species: P. mirabilis, P. vulgaris, P. penneri, and P. hauseri, and three unnamed Proteus genomospecies: 4, 5, and 6. The clinical significance of P. penneri, described in 1982 as a new species, is poorly documented. The aim of this work is serological characterization and classification of a ceftriaxone-susceptible P. penneri S29 strain isolated from a 34-year-old patient with postneurosurgical meningitis. In this characterization we will also include a ceftriaxonresistant strain, P. penneri R15, isolated from the same patient after 12 days’ treatment with ceftriaxon and other antibiotics.
Materials and Methods:
Rabbit polyclonal O-antisera were obtained against these two strains and purified lipopolysaccharides (LPS) were extracted from the bacterial mass of the P. penneri S29 and R15 strains. In the serological investigations the following tests were used: enzyme immunosorbent assay (EIA), passive immunohemolysis (PIH), inhibition of these tests, absorption of rabbit O-antisera with the respective LPS, and repeated PIH, SDS/PAGE, and Western blot techniques.
Results:
The serological studies of the LPS extracted from both P. penneri strains showed the identity of both preparations of O-polysaccharides from LPS. In P. penneri S29 O-antiserum, four different types of antibodies were described and characterized.
Conclusions:
Both investigated P. penneri S29 and R15 strains were classified to the Proteus O31ab serogroup.
Keywords: Proteus penneri; lipopolysaccharide; O-serogroup; epitope; serological classification.
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