Publications

Among pollen allergens, grass pollen allergens are some of the most frequent contributors to allergic symptoms. Substantial progress has been made since the 1960s in the identification and characterization of the grass allergens. Members of this group belong to the Poaceae family, and have been classified into 13 distinct groups based on their structure, and their biological and immunologic properties. The major contributors to allergy and, hence, most studied among the grass allergens, are those belonging to groups 1 and 5. This review is focused on the structure and immunobiology of the grass allergens and highlights how recent advances in the field have contributed to superior diagnosis and immunotherapy for allergy to grass pollens.

BACKGROUND: The natriuretic hormone peptide (NHP)(99-126), a C-terminal peptide of pro-atrial natriuretic factor (proANF), induces bronchodilatory effects in people with asthma. Recently, another plasmid-encoded C-terminal peptide, pNHP(73-102), was shown to induce a long-lasting bronchoprotective effect in a mouse model of allergic asthma.

OBJECTIVE: This study was carried out to determine the role of lung epithelial cells in the bronchoprotective and anti-inflammatory activity of these peptides.

METHODS: Human type II alveolar epithelial cells (A549) and normal human bronchial epithelial (NHBE) cells were transfected with pNHP(73-102) to test the effect of this peptide on activation of these cells. After transfection, cells were analyzed for changes in Ca(++) and nitric oxide (NO) levels. Also, activation of NFkappaB and the extracellularly regulated kinase (ERK) 1, 2 signaling pathway was examined by luciferase reporter assay and phosphorylation studies respectively.

RESULTS: Analysis of intracellular Ca(++) levels in pNHP(73-102) -transfected A549 or NHBE showed that the peptide increases release. This Ca(++) release was accompanied by an increase in the production of NO. Also, overexpression of pNHP(73-102), but not pVAX control, in phorbol myristate acetate-activated A549 cells resulted in a significant decrease in expression of a cotransfected nuclear factorkappaB (NFkappaB)-luciferase reporter. Similarly, pNHP(73-102) decreased TNF-alpha-induced NFkappaB activation in NHBE cells. Furthermore, NHP(73-102) but not atrial natriuretic peptide decreased phosphorylation of Erk-1, 2 in A549 cells.

CONCLUSIONS: Overexpression of pNHP(73-102) in epithelial cells causes increased production of intracellular Ca(++) and NO, with a concomitant decrease in activation of NFkappaB and ERK1, 2. These results suggest a bronchodilatory and anti-inflammatory activity of this peptide.

BACKGROUND: The need for safe and effective treatment of dengue virus (DEN), a class A agent that causes dengue hemorrhagic fever/dengue shock syndrome, has been a critical global priority. An effective vaccine for DEN is not yet available. In this study the possibility of attenuating DEN infection using adeno-associated virus (AAV)-encoded short interfering RNAs (siRNA) was examined in Vero cells and human dendritic cells (DCs). METHODS: A cassette encoding siRNA targeted to a 3' untranslated sequence common to all DEN serotypes was designed and tested for its ability to attenuate DEN infection by use of AAV delivery. RESULTS: Vero cells or DCs infected with AAV-siRNA showed a significant, dose-dependent reduction in DEN infection. Treatment of DCs with AAV-siRNA also decreased the DEN-induced apoptosis of DCs and did not induce significant inflammation. CONCLUSION: These results demonstrate that AAV-mediated siRNA delivery is capable of reducing DEN infection in cells and may be useful in decreasing DEN replication in humans.

With the development of genomic and proteomic technologies, the prospect for gene therapy has progressed rapidly. This has been partly possible due to the emergence of a diverse array of polymeric and non-polymeric nanoparticles that are being investigated for their ability to deliver genes and drugs. In this review, particles have been pragmatically divided as chitosan-related and chitosan-unrelated nanomaterials. The state of the art in terms of the development, characterisation and evaluation of their in vitro and/or in vivo potential is discussed for each of these various particles. Although substantial progress has been made, the potential of these particles in the clinical arena and human responses remain to be evaluated. It is hoped that this review will provide an impetus for further studies of these particles, with the ultimate intent that one or more of these diverse nanoparticle-based non-viral approaches for gene transfer will translate from 'bench to bedside' in the future.

Respiratory syncytial virus (RSV) infection up-regulates the expression of genes encoding proinflammatory mediators in bronchial epithelial cells. However, the specific signaling events immediately following RSV exposure are poorly understood. Herein, we report that RSV attachment to A549 cells activates both ERK-1 and ERK-2 pathways within 5 min. Inhibition of ERK pathways significantly decreases RSV infection of these cells compared to controls. These results demonstrate that the activation of the ERK-1/2 is required in RSV-induced early gene expression.

Respiratory syncytial virus (RSV) infection activates protein kinase C (PKC), but the precise PKC isoform(s) involved and its role(s) remain to be elucidated. On the basis of the activation kinetics of different signaling pathways and the effect of various PKC inhibitors, it was reasoned that PKC activation is important in the early stages of RSV infection, especially RSV fusion and/or replication. Herein, the role of PKC-alpha during the early stages of RSV infection in normal human bronchial epithelial cells is determined. The results show that the blocking of PKC-alpha activation by classical inhibitors, pseudosubstrate peptides, or the overexpression of dominant-negative mutants of PKC-alpha in these cells leads to significantly decreased RSV infection. RSV induces phosphorylation, activation, and cytoplasm-to-membrane translocation of PKC-alpha. Also, PKC-alpha colocalizes with virus particles and is required for RSV fusion to the cell membrane. Thus, PKC-alpha could provide a new pharmacological target for controlling RSV infection.

Serum IgE (total and five specific) and eosinophil cationic protein (ECP) levels were compared in elderly physician-diagnosed patients with asthma with non-asthmatic controls matched by age and gender to ascertain whether elevated levels are indicators of asthma in the elderly. All subjects and controls were non-smokers. The subjects were participants in the Florida Geriatric Research Program (FGRP), a longitudinal aging study that tracks the health status of people 65 years and older. Frozen sera from 33 randomly selected asthmatic patients and 21 controls, none of whom had any other chronic respiratory disease, such as chronic obstructive pulmonary disease (COPD), all between the ages of 65 and 90, were assessed for total IgE; five specific IgE concentrations (for cat, ragweed, German cockroach, Dermatophagoides pteronyssinus (Dp) and live oak); and ECP levels using the Pharmacia Unicap System. The odds of an elderly asthmatic patient having a total IgE of > 100 KU/L were higher than that for a non-asthmatic patient (odds ratio (OR) = 13.0; Mantel-Haenszel (MH) p = 0.005). The odds of elderly asthmatic patients having at least one positive serum specific IgE compared to elderly age-matched non-asthmatic patients were higher (OR = 21.2; MH p = 0.001). Among the five specific IgE concentrations, only IgE for Dp was higher in asthmatic than in non-asthmatic patients (OR = 13.00; MH p = 0.005). The ECP level was not significantly different between elderly asthmatic and non-asthmatic patients (asthmatic mean = 20.7 microg/L, SE = 0.48; control mean = 19.5 microg/L. SE = 0.76) (mean for younger adults 4.4 microg/L, Pharmacia Diagnostics). The serum of elderly asthmatic patients is more likely to have elevated total IgE and a positive specific IgE to Dp. ECP is elevated in elderly subjects but is not an indicator of asthma.

Exposure to allergens and infections contribute to early immune development. However, knowledge of the role of cellular metabolic, physiologic, and endocrinologic factors in controlling immune development and asthma is limited. Immune cells, including macrophages, dendritic cells, and T lymphocytes, express receptors for atrial natriuretic peptide (ANP) both in the fetal and neonatal lymphoid organs. ANP has garnered much attention for its cardiovascular effects, but its apparently significant role in the physiology and immunity of the lung has been underappreciated. Studies indicate that ANP also plays a significant role in shaping the early immune responses to environmental antigens. The C-terminal prohormone natriuretic peptide ANP (or NP(99-126)), which possesses bronchodilatory properties, is involved in polarizing dendritic cells to produce a T(H)2 response. Also, de novo overexpression of another pro-ANP peptide, NP(73-102), provides persistent bronchoprotection and induces significant anti-inflammatory activities in the lung epithelial cells. Thus natriuretic peptides appear to play a pivotal role in the genesis and control of asthma, and they might provide an important target to modulate allergen-induced immune responses in allergic patients.