BackgroundWe previously identified a MUC5B gene promoter-variant that is a risk allele for sporadic and familial Idiopathic Pulmonary Fibrosis/Usual Interstitial Pneumonia (IPF/UIP). This allele was strongly associated with increased MUC5B gene expression in lung tissue from unaffected subjects. Despite the strong association of this airway epithelial marker with disease, little is known of mucin expressing structures or of airway involvement in IPF/UIP.MethodsImmunofluorescence was used to subtype mucus cells according to MUC5B and MUC5AC expression and to identify ciliated, basal, and alveolar type II (ATII) cells in tissue sections from control and IPF/UIP subjects. Staining patterns were quantified for distal airways (Control and IPF/UIP) and in honeycomb cysts (HC).ResultsMUC5B-expressing cells (EC) were detected in the majority of control distal airways. MUC5AC-EC were identified in half of these airways and only in airways that contained MUC5B-EC. The frequency of MUC5B+ and MUC5AC+ distal airways was increased in IPF/UIP subjects. MUC5B-EC were the dominant mucus cell type in the HC epithelium. The distal airway epithelium from control and IPF/UIP subjects and HC was populated by basal and ciliated cells. Most honeycombing regions were distinct from ATII hyperplasic regions. ATII cells were undetectable in the overwhelming majority of HC.ConclusionsThe distal airway contains a pseudostratified mucocilary epithelium that is defined by basal epithelial cells and mucus cells that express MUC5B predominantly. These data suggest that the HC is derived from the distal airway.

Multipotential (MP) differentiation is one characteristic of a tissuespecific stem cell (TSC). Lineage tracing of tracheobronchial basal cells after naphthalene (NA) injury or in the postnatal period demonstrated that basal cells were MP progenitors for Clara-like and ciliated cells. These studies, as well as reports of spatially restricted, label-retaining basal cells, and MP differentiation by human bronchial cells support the hypothesis that a TSC maintained and repaired the tracheobronchial epithelium. However, differences in basal cell phenotype (keratin [K] 51 versus K141), age (postnatal versus adult), health status (normal versus injured), and injury type (acid, detergent, NA) limited comparisons among studies and thus diminished the strength of the TSC argument. The finding that K14 was up-regulated after NA injury was a caveat to our previous analysis of reparative (r)K14-expressing cells (EC). Thus, the present study lineage traced steady-state (s)K14EC and evaluated differentiation potential in the normal and repairing epithelium. We showed that sK14EC were unipotential in the normal epithelium and MP after NA, sK14EC-dervied clones were not restricted to putative TSC niches, sK14EC cells were a direct progenitor for Claralike and ciliated cells, MP-sK14EC clones accumulated over time, and sK14EC-derived Clara-like cells were progenitors for ciliated cells.

Objective. Because the magnetic resonance imaging (MRI) transverse relaxation time (T2) of cartilage is sensitive to organization of collagen fibers in the cartilage, it may be a noninvasive image marker for senescent changes in cartilage collagen and early cartilage degeneration. The purpose of this study was to determine age-dependent differences in cartilage T2 values in healthy asymptomatic women.Methods. Quantitative T2 maps of patellar cartilage from 30 asymptomatic women ages 22-86 years were obtained using a 3.0T MRI scanner. The study population was stratified by age into 4 cohorts: 18-30, 31-45, 46-65, and 66-86 years. Spatial differences in cartilage T2 were determined as a function of normalized distance from bone. Older groups were compared with the 18-30-year-old group to determine the effects of age on cartilage T2 values. Regions were considered statistically significantly different if the mean T2 values between groups differed at P < 0.05.Results. Mean cartilage T2 profiles were nearly identical for the 2 youngest cohorts. Compared with the 18-30-year-old group, T2 values were statistically significantly longer in the superficial 40% of cartilage in the 46-65-year-old group and over the entire cartilage thickness in the 66-86-year-old group.Conclusion. The location of T2 elevations in women over the age of 45 years is consistent with the theory that senescent changes of cartilage collagen begin near the articular surface and progress to the deeper cartilage with advancing age.Age is a recognized risk factor in the development of osteoarthritis (OA) (1). Epidemiology studies demonstrate a linear increase in OA in individuals younger than 45 years of age, with an exponential increase in those of older ages (1). Understanding of both OA and the basic aging of connective tissue would be of benefit if specific parameters in cartilage could be identified that reflect senescent modification of tissue. Because magnetic resonance imaging (MRI) can directly visualize articular cartilage, it is likely to be a useful modality in the study of cartilage aging and OA. Current clinical MRI techniques demonstrate joint anatomy and can be used to determine morphologic parameters such as cartilage volume, thickness, and presence of focal, superficial cartilage lesions (2). More recently, techniques for generating spatially localized quantitative maps of the MRI relaxation times of cartilage have been described (3). These MRI parametric mapping techniques have the potential to identify and localize specific biochemical and structural changes within the extracellular matrix of cartilage.The transverse relation time (T2) is a measurable MRI time constant that is sensitive to the slow molecular motion of mobile protons. In articular cartilage, the T2 relaxation time has a linear correlation with the water content of tissue (4) and is sensitive to loss of collagen content (5) and to the orientation of collagen fibers (6) in the extracellular matrix. There is a strong inverse correlation between spatially localized cart...

Tissue-specific stem cell (TSC) behavior is determined by the stem cell niche. However, delineation of the TSC-niche interaction requires purification of both entities. We reasoned that the niche could be defined by the location of the TSC. We demonstrate that a single CD49f bright /Sca1 1 /ALDH 1 basal cell generates rare label-retaining cells and abundant label-diluting cells. Label-retaining and labeldiluting cells were located in the rimmed domain of a unique clone type, the rimmed clone. The TSC property of self-renewal was tested by serial passage at clonal density and analysis of clone-forming cell frequency. A single clone could be passaged up to five times and formed only rimmed clones. Thus, rimmed clone formation was a cell-intrinsic property. Differentiation potential was evaluated in air-liquid interface cultures. Homogenous cultures of rimmed clones were highly mitotic but were refractory to standard differentiation signals. However, rimmed clones that were cocultured with unfractionated tracheal cells generated each of the cell types found in the tracheal epithelium. Thus, the default niche is promitotic: Multipotential differentiation requires adaptation of the niche. Because lung TSCs are typically evaluated after injury, the behavior of CD49f bright / Sca1 1 /ALDH 1 cells was tested in normal and naphthalene-treated mice. These cells were mitotically active in the normal and repaired epithelium, their proliferation rate increased in response to injury, and they retained label for 34 days. We conclude that the CD49f bright /Sca1 1 /ALDH 1 tracheal basal cell is a TSC, that it generates its own niche in vitro, and that it participates in tracheal epithelial homeostasis and repair.