[Show abstract][Hide abstract] ABSTRACT: Traditional culture-based methods have incompletely defined the microbial landscape of common recalcitrant human fungal skin diseases, including athlete's foot and toenail infections. Skin protects humans from invasion by pathogenic microorganisms and provides a home for diverse commensal microbiota. Bacterial genomic sequence data have generated novel hypotheses about species and community structures underlying human disorders. However, microbial diversity is not limited to bacteria; microorganisms such as fungi also have major roles in microbial community stability, human health and disease. Genomic methodologies to identify fungal species and communities have been limited compared with those that are available for bacteria. Fungal evolution can be reconstructed with phylogenetic markers, including ribosomal RNA gene regions and other highly conserved genes. Here we sequenced and analysed fungal communities of 14 skin sites in 10 healthy adults. Eleven core-body and arm sites were dominated by fungi of the genus Malassezia, with only species-level classifications revealing fungal-community composition differences between sites. By contrast, three foot sites-plantar heel, toenail and toe web-showed high fungal diversity. Concurrent analysis of bacterial and fungal communities demonstrated that physiologic attributes and topography of skin differentially shape these two microbial communities. These results provide a framework for future investigation of the contribution of interactions between pathogenic and commensal fungal and bacterial communities to the maintainenace of human health and to disease pathogenesis.
[Show abstract][Hide abstract] ABSTRACT: Atopic dermatitis (AD) has long been associated with Staphylococcus aureus skin colonization or infection and is typically managed with regimens that include antimicrobial therapies. However, the role of microbial communities in the pathogenesis of AD is incompletely characterized. To assess the relationship between skin microbiota and disease progression, 16S ribosomal RNA bacterial gene sequencing was performed on DNA obtained directly from serial skin sampling of children with AD. The composition of bacterial communities was analyzed during AD disease states to identify characteristics associated with AD flares and improvement post-treatment. We found that microbial community structures at sites of disease predilection were dramatically different in AD patients compared with controls. Microbial diversity during AD flares was dependent on the presence or absence of recent AD treatments, with even intermittent treatment linked to greater bacterial diversity than no recent treatment. Treatment-associated changes in skin bacterial diversity suggest that AD treatments diversify skin bacteria preceding improvements in disease activity. In AD, the proportion of Staphylococcus sequences, particularly S. aureus, was greater during disease flares than at baseline or post-treatment, and correlated with worsened disease severity. Representation of the skin commensal S. epidermidis also significantly increased during flares. Increases in Streptococcus, Propionibacterium, and Corynebacterium species were observed following therapy. These findings reveal linkages between microbial communities and inflammatory diseases such as AD, and demonstrate that as compared with culture-based studies, higher resolution examination of microbiota associated with human disease provides novel insights into global shifts of bacteria relevant to disease progression and treatment.
Genome Research 03/2012; 22(5):850-9. · 13.85 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Retroviral gene therapy can restore immunity to infants with X-linked severe combined immunodeficiency (XSCID) caused by mutations in the IL2RG gene encoding the common gamma chain (gammac) of receptors for interleukins 2 (IL-2), -4, -7, -9, -15, and -21. We investigated the safety and efficacy of gene therapy as salvage treatment for older XSCID children with inadequate immune reconstitution despite prior bone marrow transplant from a parent. Subjects received retrovirus-transduced autologous peripherally mobilized CD34(+) hematopoietic cells. T-cell function significantly improved in the youngest subject (age 10 years), and multilineage retroviral marking occurred in all 3 children.
[Show abstract][Hide abstract] ABSTRACT: XSCID results from mutations in the IL2RG gene encoding the common gamma chain (γc) shared by several cytokine receptors. Gene therapy as primary treatment for XSCID infants can restore immunity, but 2 teenagers with XSCID did not benefit from gene transfer to autologous bone marrow CD34+ cells. We used retrovirally transduced autologous peripheral mobilized CD34+ cells to treat 3 XSCID preadolescents 10-14 years old with immunodeficiency despite haploidentical bone marrow transplants (BMT). After 6-24 m we found selective gene marking of T cells in all 3 patients, with variable evidence of immunologic improvement. Prior to gene therapy, they had growth failure, frequent infections, diarrhea, T lymphocytopenia, and poor T, NK and B cell function. Each received 30-40 million cells/kg transduced ex vivo with GALV-MFGS-γc. Patient 1 (P1) had a poly-A addition signal mutation allowing residual γc function and had rejected 4 BMTs. At 24 m post gene therapy P1 had provirus detected by quantitative PCR in 4.3% of T cells and 0.1% of both B and myeloid cells. In P2 proviral marking at 18 m was 110% (or average copy number of 1.1), 5% and 0.1% in T, B and myeloid cells, respectively. P3 at 6 m had 22% marking in T cells, with low levels in other leukocyte lineages. CD4+ T cell counts in all patients have increased, but remain below the normal range. P1 has experienced improvement in well-being and resolution of lifelong diarrhea, abdominal distention and rashes; he has not had infections except for an otitis externa 12 m after treatment. A 0.5 cm submandibular nodule consistent with a lymph node arose 6 m post gene therapy and has not changed. After gene therapy P2 has had less frequent diarrhea and respiratory infections. At 6 m post treatment his T cells developed normal proliferative responses to mitogens and Candida antigen. At 12 m, P2 acquired detectable CD4+CD45RA+ T cells and T cell receptor excision circles, indicating new thymic output; donor chimerism assay showed autologous T cell expansion from 49% before gene therapy to 87% at 12 m and beyond. P3 has also reported improved well-being, exercise tolerance and appetite at 6 m; he has had increases in T cell number and proliferative responses to mitogens. Analysis of retroviral insertion sites in P1 and P3 consistently showed polyclonality. T cells of P2 had an oligoclonal pattern by LAM-PCR, but linker-mediated PCR yielded a wide variety of insertions. Furthermore, both Vβ TCR usage and spectratyping showed increasing diversity after gene therapy. Proviral insertions near LMO2, implicated in European XSCID leukemia cases, have not been identified. Our preliminary results suggest that gene therapy may benefit some XSCID patients who have failed previous BMT treatment. Prior production of allogeneic T cells in P2 may have helped to preserve his thymic function. Further follow-up will establish long-term safety, efficacy and indications for XSCID gene therapy in preadolescent subjects with XSCID.
[Show abstract][Hide abstract] ABSTRACT: Chronic graft-versus-host disease (cGVHD) remains a problematic complication of allogeneic hematopoietic stem cell transplantation. Laboratory parameters correlated with cGVHD have not been fully defined, although changes in CD4/CD8 ratios occur and a decrease in CD4(+) central memory T cells has been noted. Extracorporeal photopheresis (ECP) is an effective therapy for steroid-refractory cGVHD. We have noted changes in lymphocyte subsets after ECP. CD4(+) and CD8(+) T-cell central and effector memory populations were enumerated by flow cytometry in a cohort of 37 patients postallogeneic transplantation with symptomatic cGVHD. Of the patients with symptomatic cGVHD, 7 were treated with ECP over 6 months and prospectively assessed for changes in lymphocyte subsets. There was a highly significant correlation of an increase in CD8(+) central memory cells and a concomitant decrease in CD4(+) central memory cells in patients with symptomatic cGVHD. These changes were not detected in patients without cGVHD posttransplantation. In all, 7 patients with cGVHD followed up prospectively during ECP treatment showed a statistically significant normalization of the pattern of CD4(+) and a trend toward normalization of CD8(+) central memory T cells coincident with improvement of cGVHD. These data indicate a high correlation between disturbances in the balance of central and effector memory populations and cGVHD suggesting use in following up responses to therapy. The normalization of central and effector memory populations in response to ECP coincident with clinical improvement of cGVHD support a correlation between these laboratory parameters and cGVHD. Further studies are needed to demonstrate whether laboratory measurements of the magnitude of changes in central and effector memory populations are useful prognostically or can be used to guide response to therapy. The contrasting change in central memory cells (CD8(+) increased versus CD4(+) decreased) in cGVHD provide support for recent reports suggesting unique differences in the differentiation pathways for CD8(+) versus CD4(+) T cells.
Biology of Blood and Marrow Transplantation 02/2006; 12(1 Suppl 2):22-30. · 3.35 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Since antiquity, the genus Commiphora is composed of more than 200 species, and has been exploited as a natural drug to treat pain, skin infections, inflammatory conditions, diarrhea, and periodontal diseases. In more recent history, products derived from Commiphora myrrha and various other species of Commiphora are becoming recognized to possess significant antiseptic, anesthetic, and antitumor properties. Traditional practice and evidence-based research have supported that these properties are directly attributable to terpenoids (especially furanosesquiterpenes), the active compounds present in myrrh essential oil. More recently, current studies have focused on applying clinical trial methodologies to validate its use as an antineoplastic, an antiparasitic agent, and as an adjunct in healing wounds.
Holistic nursing practice 21(6):308-23. · 0.52 Impact Factor