Virosomes for antigen and DNA delivery

Specific targeting and delivery as well as the display of antigens on the surface of professional antigen-presenting cells (APCs) are key issues in the design and development of new-generation vaccines aimed at the induction of both humoral and cell-mediated immunity. Prophylactic vaccination against infectious diseases in general aims at the induction of humoral immune responses to prevent infection. This humoral immune response is mediated by antibody-producing B cells. On the other hand, therapeutic immunisation against virally infected cells and tumour cells requires the induction of cytotoxic T lymphocytes (CTLs) that can specifically recognise and lyse infected cells or transformed tumour cells. The induction of Major Histocompatibility Complex (MHC) class I restricted CTL activity is optimally achieved by synthesis of antigens within APCs, for example, after immunisation with live attenuated virus. However, immunisation with live vaccines bears the risk of causing disease. Therefore, alternative vaccine delivery systems, which enable introduction of nonreplicating antigen into the MHC class I presentation pathway, are sought. Furthermore, for the induction of effective humoral and cellular responses, MHC class II restricted activation of T helper cells (Th cells) is required. Among other delivery systems, as described in this theme issue of Advanced Drug Delivery Reviews, virosomes seem ideally suited for delivery of antigens into both MHC pathways. In this review, we will focus on the use of virosomes as carrier vehicles for the intracellular delivery of protein antigens and DNA, and the induction of a cellular immune response against encapsulated protein antigens and proteins expressed by virosome-associated plasmids.     

Osteopontin distribution in the canine skeleton during growth and structural maturation

The distribution of osteopontin (OPN) was studied immunohistochemically in cells and extracellular matrix in the humerus, scapula, and lumbar vertebrae of growing (age: 6 weeks, 12 weeks, 4.5 months) and adult dogs. OPN was expressed in hypertrophic chondrocytes of epiphyseal cartilage and in chondrocytes of the deep zone of mature articular cartilage, where extracellular matrix was also stained. OPN expression was strongest in 4.5-month-old puppies in cells of the osteoblastic lineage. It also varied with microlocation and was pronounced in areas prone to resorption due to modelling and remodelling activities. Osteoclasts were always strongly labelled with OPN. OPN deposition in extracellular bone matrix was detected particularly as a delineation of cartilage cores within secondary trabeculae and as a lining of the trabecular surfaces in resorption microlocations. The OPN distribution pattern is discussed here for each cell population with regard to its functional implications.     

Hypoxic tolerance in air-breathing invertebrates

Terrestrial invertebrates experience hypoxia in many habitats and under a variety of physiological conditions. Some groups (at least insects) are much more capable of recovery from anoxia than most vertebrates, but there is still a tremendous unexplained variation in hypoxia/anoxia tolerance among terrestrial invertebrates. Crustaceans and arachnids may be less often confronted with hypoxic environments than insects and myriapods and also seem to be less hypoxia/anoxia tolerant. Tracheated groups, especially those that are able to ventilate their tracheal system like many insects, cope with lower critical PO2 than nontracheated groups. Modulation of oxygen carrier proteins is normally not important in hypoxia resistance. Recent application of genetic and cellular tools are revealing that many of the same pathways documented for mammals (e.g. HIF, nitric oxide) function to regulate morphological and biochemical responses to hypoxia/anoxia in insects.     

Ghrelin receptor gene: identification of several sequence variants in extremely obese children and adolescents, healthy normal-weight and underweight students, and children with short normal stature

GH secretagogue receptor (GHSR, ghrelin receptor) is involved in regulation of body weight and GH secretion. We initially analyzed two single-nucleotide polymorphisms of the GHSR in up to 184 extremely obese children and adolescents and up to 184 healthy underweight students. The frequency of the 171T allele of rs495225 was higher in our obese samples (75.0%) than in the underweight individuals (70.2%; nominal P = 0.14). This trend could not be substantiated in an additional association study in 270 obese and 145 underweight and normal weight individuals and in a transmission disequilibrium test based on 387 obesity trios (transmission rate of 171T, 51.8%; nominal P = 0.53). Additionally, the coding region of GHSR was systematically screened, and seven sequence variants were identified in 93 obese, 96 normal weight, and 94 underweight individuals and 43 children with short normal stature (SNS). Five silent single-nucleotide polymorphisms showed similar genotype frequencies in the different weight groups and SNS children (all nominal P > 0.3). Two novel missense variants were detected only in one obese carrier and one SNS child, respectively. In conclusion, we did not obtain conclusive evidence for an involvement of the ghrelin receptor gene in body weight regulation or SNS in our study groups.     

Autologous stem-cell transplantation restores the functional properties of CD14+CD16+ monocytes in patients with myeloma and lymphoma

The CD14+CD16+ monocytes appear to be important to immune defense against infection, as these cells are very potent with respect to tumor necrosis factor (TNF) production, phagocytosis, and antigen presentation. Myeloablative high-dose chemotherapy (HDT) and subsequent autologous stem-cell transplantation (ASCT) are being used increasingly for therapy of hematological malignancies, but the pronounced immunosuppression renders the patients prone to infection. To determine the functional properties of CD14+CD16+ monocytes under these conditions, 15 patients with lymphoma or myeloma were examined. Before HDT, the ratio of CD14+CD16+ cells to the population of the classical CD14++ monocytes was 0.28 +/- 0.12; this ratio changed during the course of HDT and ASCT in favor of the CD14+CD16+ monocytes to a maximum of 12.4 +/- 7.8 (P<0.001) on day 3.5 +/- 1.6 after transplanation (Tx) and returned to 0.11 +/- 0.07 (P<0.001) after engraftment on day 11.3 +/- 2.2. Although the absolute number of classical CD14++ monocytes declined to less than 1/microl at the nadir, the number of CD14+CD16+monocytes fell from 29.7 +/- 9.8/microl to 4.5 +/- 3.0/microl at the nadir and increased to 13.8 +/- 9.8/microl at the day of discharge from the hospital. Flow cytometric analysis of phagocytosis of fluorescein isothiocyanate (FITC)-labeled Escherichia coli showed that 30 +/- 10% CD14+CD16+ monocytes of patients were FITC-positive before Tx, and at engrafment, the percentage of FITC-positive cells had doubled to 60 +/- 6% (healthy controls, 41+/-7%). When determining generation of reactive oxygen species after E. coli ingestion, the CD14+CD16+ monocytes showed a decreased response before Tx (32+/-12% positve cells), which increased to 53 +/- 24% after ASCT. The median fluorescence intensity of human leukocyte antigen (HLA)-DR expression on the CD14+CD16+ monocytes increased from 11 +/- 6 before Tx to 17 +/- 11 after Tx, and the production of TNF after lipopolysaccharide showed no remarkable difference (46+/-13 vs. 49+/-14 channels). At the same time, expression of TNF and of HLA-DR showed a dramatic decrease in the CD14++ monocytes. Taken together after stem-cell Tx, the function of the CD14++ monocytes is impaired, and the functional properties of CD14+CD16+ monocytes recover, indicating that these cells may be important for defense against infections post-ASCT.     

Predicting long-term outcome in group treatment of atopic dermatitis

BACKGROUND: Psychological group treatments are effective adjuncts to dermatological treatments in improving dermatological symptoms and quality of life. However, no data are available on variables that may predict which patients benefit from such group treatments. METHODS: On the basis of data from the Marburg group treatment study, we explored which variables are associated with successful treatment. Treatment outcome was defined by a marked reduction of dermatological symptoms at 1-year follow-up. RESULTS: Regardless of the treatment condition, a high pretreatment level of scratching, a low serum IgE level, low scores on the internal Health Locus of Control Scale and a high frequency of coping-related cognitions regarding itching predicted good outcome at 1 year. Logistic regression analyses indicated that the predictive power of locus of control and itch-related cognitions differed between different treatment conditions. CONCLUSION: In patients with atopic dermatitis, indication for psychological treatment may be based on the assessment of psychological target problems including excessive scratching and dysfunctional cognitions.     

The beta-N-acetylhexosaminidase of Entamoeba histolytica is composed of two homologous chains and has been localized to cytoplasmic granules

We have purified a beta-N-acetylhexosaminidase from trophozoites of Entamoeba histolytica to homogeneity. In SDS-PAGE, the enzyme yielded a single protein band at an apparent M(r) of 64,000. The elution behaviour of the native enzyme upon molecular sieve chromatography corresponded to a molecular mass of approximately 132,000 suggesting that the enzyme is a dimer. Upon sedimentation velocity centrifugation, hexosaminidase activity sedimented at 12S, implying aggregation to a higher molecular mass complex with an apparent M(r) of approximately 400,000. Based on the N-terminal sequence of the purified enzyme and on data extracted from the E. histolytica genomic data base, we amplified and cloned two genes (EhHEXA and EhHEXB) coding for two presumptive, highly similar hexosaminidase chains which we designated as Ehhexalpha and Ehhexbeta. Northern blot analysis indicated that the two genes were expressed to a similar level, and Western blotting with chain-specific antisera showed that the trophozoites synthesize both proteins. By cell fractionation, the hexosaminidase was found to be a major component of cytoplasmic granules; these contain tissue-destructive factors and are released after collagen-induced exocytosis to the cell surface. In agreement with this observation, immunocytochemistry with an antiserum cross-reacting with both hexosaminidase chains revealed strong fluorescence in surface patches, which we interpret as released granules, and in vesicles throughout the cell. Its localization in cytoplasmic granules strengthens the notion that the hexosaminidase complex may contribute to amoebic pathogenicity.     

Serum lipids and hippocampal volume: The link to Alzheimer's disease?

The association between hippocampal volume (as a presumed index of Alzheimer's disease pathology) with serum total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol was studied in 86 elderly subjects with a range of cognitive functions. High-density lipoprotein cholesterol, but not low-density lipoprotein cholesterol or total cholesterol, was associated with hippocampal volume and dementia. This is compatible with protective effects of high-density lipoprotein cholesterol on hippocampal atrophy and Alzheimer's disease.