Round Cells of the Epidermis: Clues from Studies on Neoplastic Lymphocytes of Cutaneous T Cell Lymphoma

Neoplastic cells of cutaneous T cell lymphoma (CTCL) appear to be of monoclonal origin and frequently are nonspecific helpers of normal B cell differentiation. A natural progression from epidermotropic (mycosis fungoides and Sézary syndrome) to nonepidermotropic, more widely disseminated T cell neoplasms generally occurs. Affinity of CTCL cells for the epidermis may result from their having membrane receptors for histocompatibility (Ia) antigens present in skin. Cultured human epidermal cells produce a thymopoietin-like molecule, an indication of a role for skin in T cell differentiation

Discovery of a ~5 day characteristic timescale in the Kepler power spectrum of Zw 229-15

We present time series analyses of the full Kepler dataset of Zw 229-15. This Kepler light curve --- with a baseline greater than three years, composed of virtually continuous, evenly sampled 30-minute measurements --- is unprecedented in its quality and precision. We utilize two methods of power spectral analysis to investigate the optical variability and search for evidence of a bend frequency associated with a characteristic optical variability timescale. Each method yields similar results. The first interpolates across data gaps to use the standard Fourier periodogram. The second, using the CARMA-based time-domain modeling technique of Kelly et al. (2014), does not need evenly-sampled data. Both methods find excess power at high frequencies that may be due to Kepler instrumental effects. More importantly both also show strong bends ({\Delta}{\alpha} ~ 2) at timescales of ~5 days, a feature similar to those seen in the X-ray PSDs of AGN but never before in the optical. This observed ~5 day timescale may be associated with one of several physical processes potentially responsible for the variability. A plausible association could be made with light-crossing, dynamical or thermal timescales, depending on the assumed value of the accretion disk size and on unobserved disk parameters such as {\alpha} and H/R. This timescale is not consistent with the viscous timescale, which would be years in a ~10^7 Solar mass AGN such as Zw 229-15. However there must be a second bend on long (>~1 year) timescales, and that feature could be associated with the viscous timescale.Comment: 10 pages, 5 figures, 1 table. To appear in the Astrophysical Journal, Part

Induction of an Immature T-Cell Phenotype in Malignant Helper T Cells by Cocultivation With Epidermal Cell Cultures

The possible inductive effect of epidermal cells on T-cell maturation has been examined employing an in vitro co-cultivation technique. Mononuclear cells from 6 patients with cutaneous T-cell lymphoma (CTCL) and from 12 healthy volunteers were studied. In the 6 CTCL patients, all showed an expansion of the helper T-cell subpopulation and in one patient with leukemic CTCL, there was almost complete replacement of peripheral blood mononuclear cells by malignant cells with a helper T-cell phenotype. Epidermal cells derived from normal human skin were cultured to confluent monolayers, and were cocultivated with the mononuclear cells from CTCL patients or normal controls for 48h at a density of 106/ml. Following cocultivation, the surface phenotype of the cells from the 12 healthy volunteers and 5 of the patients with CTCL showed no significant phentotypic change. In the patient with leukemic CTCL, however, the surface phenotype of the malignant T cells had changed, with the acquisition of the T6 antigen by the majority of the cells. Cells cocultivated in medium alone and with human fibroblast monolayers showed no change in surface phenotype. The malignant T cells from the leukemic CTCL patient failed to react in a mixed lymphocyte culture to lymphocytes from 2 different healthy donors, and showed no phenotypic change following culture with these lymphocytes, indicating that the phenotypic change seen was not due to allogeneic stimulation

Rapid construction of a dendritic cell vaccine through physical perturbation and apoptotic malignant T cell loading

We have demonstrated that adherence and release of monocytes from a plastic surface drives their differentiation into immature dendritic cells (DC,) that can mature further during overnight incubation in the presence of apoptotic malignant T cells. Based on these results, we sought to develop a clinically, practical, rapid means for producing DC loaded with malignant cells. A leukapheresis harvest containing the clonal, leukemic expansion of malignant CD4(+ )T cells was obtained from the blood of patients with cutaneous T cell lymphoma (CTCL). CTCL cells were purified with a CD3-magnetic bead column where CD3 engagement rendered the malignant T cells apoptotic. The monocyte fraction was simultaneously activated by column passage, re-added to the apoptotic CTCL cells and co-cultured overnight. CTCL cell apoptosis, DC differentiation and apoptotic malignant T cell ingestion were measured by immunostaining. The results demonstrate that as monocytes passed through the column matrix, they became activated and differentiated into semi-mature DC expressing significantly increased levels of class II, CD83 and CD86 (markers associated with maturing DC) and reduced expression of the monocyte markers CD14 and CD36. Apoptotic malignant T cells were avidly engulfed by the phagocytic transitioning DC. The addition of supportive cytokines further enhanced the number of DC that contained apoptotic malignant T cells. Functional studies confirmed that column passaged DC increased class II expression as shown by significantly enhanced stimulation in mixed leukocyte culture compared to control monocytes. In addition, DC loaded with apoptotic CTCL cells stimulated an increase in the percentage and absolute number of CD8 T cells compared to co-cultivation with non-loaded DC. After CD8 T cells were stimulated by DC loaded with malignant cells, they mediated increased apoptosis of residual CTCL cells and TNF-α secretion indicating development of enhanced cytolytic function. We report a simple one-step procedure where maturing DC containing apoptotic malignant T cells can be prepared rapidly for potential use in vaccine immunotherapy. Ready access to both the DC and apoptotic cells provided by this system will allow extension to other malignancies through the addition of a variety of apoptotic tumor cells and maturation stimuli

The Radio Properties of Seyfert Galaxies in the 12--Micron and CfA Samples

We report the results of 20, 6, and 2 cm VLA and 1.5 cm OVRO observations of the optically-selected CfA Seyfert galaxies and the bolometric-flux-limited 12-Micron active galaxy sample. Every object observed was detected at 6 cm. Only 6-8% of the 12um sample Seyferts (3-4 objects) are radio-loud (and none of the CfA sample), as compared to 15-20% for the BQS quasars. These radio-loud objects are compact and have flat spectra, distinguishing them from the more common radio-quiet objects. The 6-20 cm slopes of the Seyfert 1s and 2s are similar, with average values of about -0.7. Although several Seyfert 1s are significantly flatter than this in their 6-20 and/or 1.5-6 cm slopes, there is no systematic trend for either Seyfert type to display upward or downward spectral curvature. Excluding the radio-loud quasars, the integrated 6 cm radio luminosity is linearly proportional to the 60um luminosity over several orders of magnitude, with on average twice the radio power of normal spirals of the same far-infrared power. About half of the objects show extended 6 cm emission, contributing on average 33% of the total flux. Thus the luminosities of these extended components alone are comparable to normal spirals of similar infrared luminosities. The 12um sample radio luminosity function is slightly higher than that of the CfA sample. The integrated space density of Seyfert 2s is about 2 times that of Seyfert 1s over their common range in luminosity. In terms of the standard unified model, this ratio in space density corresponds to a typical half-angle of the torus of about 48 degress.Comment: AASTeX, 16 pages. Text and Tables 3-4 only. PostScript versions of this file, figures, and Tables 1-2, and a latex version of Tables 1-2 are available at ftp://ftp.astro.ucla.edu/pub/incoming/rush/, mget rme_*. Also get longtable.sty & fancyheadings.sty from there if you don't have them. Accepted by ApJ ~1996 July

KSwAGS: A Swift X-ray and UV Survey of the Kepler Field. I

We introduce the first phase of the Kepler-Swift Active Galaxies and Stars survey (KSwAGS), a simultaneous X-ray and UV survey of ~6 square degrees of the Kepler field using the Swift XRT and UVOT. We detect 93 unique X-ray sources with S/N>3 with the XRT, of which 60 have observed UV counterparts. We use the Kepler Input Catalog (KIC) to obtain the optical counterparts of these sources, and construct the X-ray to optical flux ratio as a first approximation of the classification of the source. The survey produces a mixture of stellar sources, extragalactic sources, and sources which we are not able to classify with certainty. We have obtained optical spectra for thirty of these targets, and are conducting an ongoing observing campaign to fully identify the sample. For sources classified as stellar or AGN with certainty, we construct SEDs using the 2MASS, UBV and GALEX data supplied for their optical counterparts by the KIC, and show that the SEDs differ qualitatively between the source types, and so can offer a method of classification in absence of a spectrum. Future papers in this series will analyze the timing properties of the stars and AGN in our sample separately. Our survey provides the first X-ray and UV data for a number of known variable stellar sources, as well as a large number of new X-ray detections in this well-studied portion of the sky. The KSwAGS survey is currently ongoing in the K2 ecliptic plane fields.Comment: Accepted for publication in the Astronomical Journal. 19 pages, 8 figures, 3 table

Regulation of Transgenic Class II Major Histocompatibility Genes in Murine Langerhans Cells

I-E is a class II major histocompatibility complex molecule normally expressed by Langerhans cells, A series of transgenic mice were developed previously that carry Eαd gene constructs with promoter-region deletions that cause expression of I-E by different cell types when maintained on a B6 (I-E[–]) genetic background. To study cis-acting gene sequences that regulate expression of class II proteins by Langerhans cells, we identified trans genie I-E expression by tissue immunoperoxidase staining and by epidermal cell suspension lmmunofluorescence cytometry. Mice with a transgene containing 1.4 kilobase pairs (kb) of flanking sequence 5' to the Eα initiation site expressed barely detectable levels of I-E on a tiny percentage of Langerhans cells, indicating that sequences promoting Langerhans cell expression of Eα exist between 2.0 and 1.4 kb 5' of the Eα initiation site. Removal of an additional 170 bp of 5' flanking sequence caused near-normal levels of expression by approximately one third of epidermal Langerhans cells, which contrasts with studies that showed minimal transgene expression by splenic dendritic cells in these animals, Thus, sequences between 1.4 and 1.23 kb 5' of the Eα initiation site decrease expression of I-E by epidermal Laugerhans cells, but enable I-E expression by splenic dendritic cells, These studies identify Langerhans cell-specific regulatory sequences and genetic regions controlling major histocompatibility complex class II gene expression in Langerhans cells and splenic dendritic cells. The genetic regions identified may be particularly important because differential regulation of class II major histocompatibility complex protein synthesis by Langerhans cells and dendritic cells may be crucial to immune functions of intact animals

Improved generation of anti-tumor immunity by antigen dose limitation

BACKGROUND: The malignant cells of cutaneous T cell lymphoma (CTCL) display immunogenic peptides derived from the clonal T cell receptor (TCR) providing an attractive model for refinement of anti-tumor immunization methodology. To produce a clinically meaningful anti-tumor response, induction of cytotoxic anti-CTCL cells must be maximized while suppressive T regulatory cells (Treg) should be minimized. We have demonstrated that engulfment of apoptotic CTCL cells by dendritic cells (DC) can lead to either CD8 anti-CTCL responses or immunosuppressive Treg induction. Treg generation is favored when the number of apoptotic cells available for ingestion is high. METHODS: In this study, we sought to determine whether the balance between immunity and immunosuppression could be shifted towards a CD8 anti-CTCL response by lowering the ratio of apoptotic CTCL cells available for DC ingestion. CTCL cell apoptosis was produced by engagement of the TCR by anti-CD3 antibody affixed to magnetic beads. RESULTS: The physical perturbation inherent in passage through a separation column induced monocytes to differentiate into DC, demonstrated by increased expression of class II and CD86 and decreased expression of the monocyte marker CD14. The immature DC internalized and processed apoptotic CTCL cells and could potentially present the tumor-derived peptides in the context of MHC class I and II. As the number of apoptotic cells increased, there was a dose-dependent increase in the expression of Treg markers CTLA-4, CD25, and FoxP3, with a ratio of apoptotic cell/DC loading of > 10:1 corresponding to the greatest Treg induction. These inducible phenotypic Treg also functionally inhibited CD8-mediated perforin expression in vitro. At lower levels of apoptotic cell/DC loading of < 5:1, there was an expansion of the CD8 T cell compartment with increased perforin expression and increased CTCL cell death, indicating anti-tumor activity. CONCLUSION: These findings demonstrate that the ratio of apoptotic cells supplied to DC is an important determinant of whether CD8 anti-tumor immunity or immunosuppression is generated

X-ray Observations of the Broad-Line Radio Galaxy 3C 390.3

We present the data and preliminary analysis for a series of 90 ROSAT HRI and two ASCA observations of the broad-line radio galaxy 3C 390.3. These data were obtained during the period 1995 January 2 to 1995 October 6 as part of an intensive multiwavelength monitoring campaign. The soft X-ray flux in the ROSAT band varied by nearly a factor of four during the campaign, and the well-resolved light-curve shows several distinct features. Several large amplitude flares were observed, including one in which the flux increased by a factor of about 3 in 12 days. Periods of reduced variability were also seen, including one nearly 30 days long. While the HRI hardness ratio decreased significantly, it is apparently consistent with that expected due to the detector during the monitoring period. The two ASCA observations were made on 1995 January 15 and 1995 May 5. The 0.5-10.0 keV spectra can be adequately described by an absorbed power-law. There is no evidence for a soft excess in the ASCA spectra, indicating that the ROSAT HRI is sampling variability of the X-ray power-law. A broad iron line was observed in a longer 1993 ASCA observation, and while there is statistical evidence that the line is present in the 1995 spectra, it could not be resolved clearly. There is evidence, significant at >90% confidence, that the photon index changed from 1.7 to 1.82 while the flux increased by 63%. The spectral change can be detected in the spectra below 5 keV, indicating that the origin cannot be a change in ratio of reflected to power-law flux. A compilation of results from ASCA and Ginga observations show that on long time scales the intrinsic photon index is correlated with the flux.Comment: 17 pages using (AASTeX) aaspp4.sty and 4 Postscript figures. Astrophysical Journal, in pres