Ocular infection of mice with an avirulent recombinant HSV-1 expressing IL-4 and an attenuated HSV-1 strain generates virulent recombinants in vivo

PurposeTo assess the relative impact of overexpression of interleukin 2 (IL-2), interleukin 4 (IL-4), and interferon gamma (IFN-γ) expressing recombinant herpes simplex virus type 1 (HSV-1) on altering immune responses in ocularly infected mice.MethodsBALB/c mice were co-infected ocularly with avirulent HSV-1 strain KOS and avirulent recombinant HSV-1 expressing murine IL-4 (HSV-IL-4). Controls mice were co-infected with KOS + HSV-IL-2 or KOS + HSV-IFNγ. Following ocular infection, virus replication in the eye, corneal scarring (CS), and survival were determined. We also isolated recombinant viruses from eye and trigeminal ganglia of KOS + HSV-IL-4 infected mice.ResultsIn this study we found that ocular infection of BALB/c mice with a mixture of HSV-IL-4 and KOS resulted in increased death and increased eye disease. In contrast, when mice were infected in one eye with KOS and the other eye with HSV-IL-4 no death or eye disease was seen. Intraperitoneal co-infection of mice with KOS and HSV-IL-4 also did not result in HSV-1 induced death. Interestingly, ocular infection of mice with a mixture of HSV-IL-2 and KOS did not have any effect on severity of the disease in infected mice. We isolated recombinant viruses from KOS + HSV-IL-4 infected mice eye and trigeminal ganglia. Some of the isolated viruses were more neurovirulent then either parental virus. Infection of macrophages with IL-4 expressing virus down-regulated IL-12 production by macrophages.ConclusionsThese results suggest a role for IL-4 in suppression of immune response and generation of virulent viruses in vivo

A role for the JAK-STAT1 pathway in blocking replication of HSV-1 in dendritic cells and macrophages

<p>Abstract</p> <p>Background</p> <p>Macrophages and dendritic cells (DCs) play key roles in host defense against HSV-1 infection. Although macrophages and DCs can be infected by herpes simplex virus type 1 (HSV-1), both cell types are resistant to HSV-1 replication. The aim of our study was to determine factor (s) that are involved in the resistance of DCs and macrophages to productive HSV-1 infection.</p> <p>Results</p> <p>We report here that, in contrast to bone marrow-derived DCs and macrophages from wild type mice, DCs and macrophages isolated from signal transducers and activators of transcription-1 deficient (STAT1^-/-) mice were susceptible to HSV-1 replication and the production of viral mRNAs and DNA. There were differences in expression of immediate early, early, and late gene transcripts between STAT1^+/+and STAT1^-/-infected APCs.</p> <p>Conclusion</p> <p>These results suggest for the first time that the JAK-STAT1 pathway is involved in blocking replication of HSV-1 in DCs and macrophages.</p

Therapeutic Periocular Vaccination with a Subunit Vaccine Induces Higher Levels of Herpes Simplex Virus-Specific Tear Secretory Immunoglobulin A Than Systemic Vaccination and Provides Protection against Recurrent Spontaneous Ocular Shedding of Virus in Latently Infected Rabbits

AbstractRabbits latently infected with herpes simplex virus type 1 (HSV-1) were vaccinated either periocularly or systemically with a subunit vaccine (gB2 + gD2) plus adjuvant or adjuvant alone. Tear films were collected daily to measure recurrent infectious HSV-1 shedding. After systemic vaccination, the latently infected rabbits were not protected against recurrent ocular viral shedding (HSV-1-positive tear film cultures/total cultures) compared with either the systemic or periocular adjuvant controls (systemic vaccination = 49 of 972, 5.0%; systemic control = 46 of 972, 4.7%; periocular control = 43 of 930, 4.6%;P> 0.8). In contrast, latently infected rabbits vaccinated periocularly with the same vaccine had significantly reduced recurrent shedding (20 of 1026, 2.0%) compared with controls (P< 0.001) or systemic vaccination (P= 0.0002). Thus, recurrent HSV-1 shedding was significantly reduced by therapeutic local periocular subunit vaccination but not by therapeutic systemic subunit vaccination. Neutralizing antibody titers in the serum of systemically and ocularly vaccinated rabbits was similar. In contrast, HSV-specific tear secretory immunoglobulin A was significantly higher in the ocularly vaccinated group (P< 0.01). These results strongly suggest that in the rabbit, and presumably in humans, the local ocular (mucosal) immune response is much more important than the systemic immune response for therapeutic protection against recurrent ocular HSV-1. Thus development of a therapeutic vaccine against recurrent ocular HSV-1 should be directed at enhancing the local ocular (mucosal) immune response

The Gene That Encodes the Herpes Simplex Virus Type 1 Latency-Associated Transcript Influences the Accumulation of Transcripts (Bcl-x\u3csub\u3eL\u3c/sub\u3e and Bcl-x\u3csub\u3es\u3c/sub\u3e) That Encode Apoptotic Regulatory Proteins

The herpes simplex virus type 1 latency-associated transcript (LAT) inhibits apoptosis. We demonstrate here that LAT influences the accumulation of the Bcl-xL transcript versus the Bcl-xS transcript in Neuro-2A cells. Bcl-xL encodes an antiapoptotic protein, whereas Bcl-xS encodes a proapoptotic protein. Promoting the accumulation of Bcl-xL in neurons may inhibit apoptosis, thus enhancing the latency-reactivation cycle

The Gene That Encodes the Herpes Simplex Virus Type 1 Latency-Associated Transcript Influences the Accumulation of Transcripts (Bcl-x\u3csub\u3eL\u3c/sub\u3e and Bcl-x\u3csub\u3es\u3c/sub\u3e) That Encode Apoptotic Regulatory Proteins

The herpes simplex virus type 1 latency-associated transcript (LAT) inhibits apoptosis. We demonstrate here that LAT influences the accumulation of the Bcl-xL transcript versus the Bcl-xS transcript in Neuro-2A cells. Bcl-xL encodes an antiapoptotic protein, whereas Bcl-xS encodes a proapoptotic protein. Promoting the accumulation of Bcl-xL in neurons may inhibit apoptosis, thus enhancing the latency-reactivation cycle

Identification of Herpes Simplex Virus Type 1 Latency-Associated Transcript Sequences That both Inhibit Apoptosis and Enhance the Spontaneous Reactivation Phenotype

The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) gene is essential for the high spontaneous and induced reactivation phenotype of HSV-1 in the rabbit ocular model and for the high induced reactivation phenotype in the mouse ocular model. Recently we showed that LAT has an antiapoptosis function, and we hypothesized that LAT’s ability to inhibit apoptosis played an important role in LAT’s ability to enhance the reactivation phenotype. Expression of just the first 1.5 kb of the 8.3-kb LAT gene is sufficient for both inhibition of apoptosis in an in vitro transient-transfection assay and the high spontaneous reactivation phenotype in vivo. Here we show the results of more complex mapping studies in which inhibition of apoptosis and the enhanced spontaneous reactivation phenotype also appear to be linked. The HSV-1 mutant virus dLAT371 has a high spontaneous reactivation phenotype in rabbits, suggesting that the LAT region deleted in this mutant (LAT nucleotides 76 to 447) is not required for this phenotype. The LAT3.3A viral mutant (which expresses LAT nucleotides 1 to 1499) also has a high spontaneous reactivation phenotype, suggesting that the region of LAT not expressed by this mutant (LAT nucleotide 1500 to the end of LAT) is also not required for this phenotype. Surprisingly, LAT2.9A, which is a combination of dLAT371 and LAT3.3A (i.e., it expresses LAT nucleotides 1 to 76 and 447 to 1499), has a low spontaneous reactivation phenotype indistinguishable from that of LAT null mutants. We report here that consistent with the low spontaneous reactivation phenotype of LAT2.9A, a plasmid expressing the identical LAT RNA did not inhibit caspase 9-induced apoptosis. In contrast, plasmids containing the same deletion but able to transcribe up to or past LAT nucleotide 2850 (rather than just up to LAT nucleotide 1499) inhibited caspase 9-induced apoptosis, consistent with the high spontaneous reactivation phenotype of dLAT371. Thus, LAT2.9A may have a low spontaneous reactivation phenotype because the LAT RNA that is made cannot block apoptosis, and dLAT371 apparently has a high spontaneous reactivation phenotype because the LAT RNA made has significant antiapoptosis activity. Furthermore, LAT appeared to have at least two regions capable of interfering with caspase 9-induced apoptosis. One region partially overlaps LAT nucleotides 76 to 447. The second region is partially (or completely) downstream of LAT nucleotide 1499

Inclusion of CD80 in HSV Targets the Recombinant Virus to PD-L1 on DCs and Allows Productive Infection and Robust Immune Responses

CD80 plays a critical role in stimulation of T cells and subsequent control of infection. To investigate the effect of CD80 on HSV-1 infection, we constructed a recombinant HSV-1 virus that expresses two copies of the CD80 gene in place of the latency associated transcript (LAT). This mutant virus (HSV-CD80) expressed high levels of CD80 and had similar virus replication kinetics as control viruses in rabbit skin cells. In contrast to parental virus, this CD80 expressing recombinant virus replicated efficiently in immature dendritic cells (DCs). Additionally, the susceptibility of immature DCs to HSV-CD80 infection was mediated by CD80 binding to PD-L1 on DCs. This interaction also contributed to a significant increase in T cell activation. Taken together, these results suggest that inclusion of CD80 as a vaccine adjuvant may promote increased vaccine efficacy by enhancing the immune response directly and also indirectly by targeting to DC

A Critical Assessment of Stellar Mass Measurement Methods

In this paper we perform a comprehensive study of the main sources of random and systematic errors in stellar mass measurement for galaxies using their Spectral Energy Distributions (SEDs). We use mock galaxy catalogs with simulated multi-waveband photometry (from U-band to mid-infrared) and known redshift, stellar mass, age and extinction for individual galaxies. Given different parameters affecting stellar mass measurement (photometric S/N ratios, SED fitting errors, systematic effects, the inherent degeneracies and correlated errors), we formulated different simulated galaxy catalogs to quantify these effects individually. We studied the sensitivity of stellar mass estimates to the codes/methods used, population synthesis models, star formation histories, nebular emission line contributions, photometric uncertainties, extinction and age. For each simulated galaxy, the difference between the input stellar masses and those estimated using different simulation catalogs, $\Delta\log(M)$, was calculated and used to identify the most fundamental parameters affecting stellar masses. We measured different components of the error budget, with the results listed as follows: (1). no significant bias was found among different codes/methods, with all having comparable scatter; (2). A source of error is found to be due to photometric uncertainties and low resolution in age and extinction grids; (3). The median of stellar masses among different methods provides a stable measure of the mass associated with any given galaxy; (4). The deviations in stellar mass strongly correlate with those in age, with a weaker correlation with extinction; (5). the scatter in the stellar masses due to free parameters are quantified, with the sensitivity of the stellar mass to both the population synthesis codes and inclusion of nebular emission lines studied.Comment: 33 pages, 20 Figures, Accepted for publication in Astrophysical Journa

The Relation Between SFR and Stellar Mass for Galaxies at 3.5 ≤z≤\le z\le 6.5 in CANDELS

Distant star-forming galaxies show a correlation between their star formation rates (SFR) and stellar masses, and this has deep implications for galaxy formation. Here, we present a study on the evolution of the slope and scatter of the SFR-stellar mass relation for galaxies at $3.5\leq z\leq 6.5$ using multi-wavelength photometry in GOODS-S from the Cosmic Assembly Near-infrared Deep Extragalactic Legacy Survey (CANDELS) and Spitzer Extended Deep Survey. We describe an updated, Bayesian spectral-energy distribution fitting method that incorporates effects of nebular line emission, star formation histories that are constant or rising with time, and different dust attenuation prescriptions (starburst and Small Magellanic Cloud). From $z$=6.5 to $z$=3.5 star-forming galaxies in CANDELS follow a nearly unevolving correlation between stellar mass and SFR that follows SFR $\sim$ $M_\star^a$ with $a = 0.54 \pm 0.16$ at $z\sim 6$ and $0.70 \pm 0.21$ at $z\sim 4$. This evolution requires a star formation history that increases with decreasing redshift (on average, the SFRs of individual galaxies rise with time). The observed scatter in the SFR-stellar mass relation is tight, $\sigma(\log \mathrm{SFR}/\mathrm{M}_\odot$ yr$^{-1})< 0.3\ -$ 0.4 dex, for galaxies with $\log M_\star/\mathrm{M}_\odot > 9$ dex. Assuming that the SFR is tied to the net gas inflow rate (SFR $\sim$ $\dot{M}_\mathrm{gas}$), then the scatter in the gas inflow rate is also smaller than 0.3$-$0.4 dex for star-forming galaxies in these stellar mass and redshift ranges, at least when averaged over the timescale of star formation. We further show that the implied star formation history of objects selected on the basis of their co-moving number densities is consistent with the evolution in the SFR-stellar mass relation.Comment: 31 pages, 24 figures, accepted for publication in Ap

CANDELS Observations of the Environmental Dependence of the Color-Mass-Morphology Relation at z = 1.6

We study the environmental dependence of color, stellar mass, and morphology by comparing galaxies in a forming cluster to those in the field at z = 1:6 with Hubble Space Telescope near-infrared imaging in the CANDELS/UDS field. We quantify the morphology of the galaxies using the effective radius, reff, and S\'ersic index, n. In both the cluster and field, approximately half of the bulge-dominated galaxies (n > 2) reside on the red sequence of the color-magnitude diagram, and most disk-dominated galaxies (n < 2) have colors expected for star-forming galaxies. There is weak evidence that cluster galaxies have redder rest-frame U - B colors and higher stellar masses compared to the field. Star-forming galaxies in both the cluster and field show no significant differences in their morphologies. In contrast, there is evidence that quiescent galaxies in the cluster have larger median effective radii and smaller S\'ersic indices compared to the field with a significance of 2?. These differences are most pronounced for galaxies at clustercentric distances 1 Mpc < Rproj < 1.5 Mpc, which have low S\'ersic indices and possibly larger effective radii, more consistent with star-forming galaxies at this epoch and in contrast to other quiescent galaxies. We argue that star-forming galaxies are processed under the influence of the cluster environment at distances greater than the cluster-halo virial radius. Our results are consistent with models where gas accretion onto these galaxies is suppressed from processes associated with the cluster environment.Comment: ApJ accepted, 19 pages, 10 figure