CD4+ T Cell Effects on CD8+ T Cell Location Defined Using Bioluminescence

T lymphocytes of the CD8+ class are critical in delivering cytotoxic function and in controlling viral and intracellular infections. These cells are “helped” by T lymphocytes of the CD4+ class, which facilitate their activation, clonal expansion, full differentiation and the persistence of memory. In this study we investigated the impact of CD4+ T cells on the location of CD8+ T cells, using antibody-mediated CD4+ T cell depletion and imaging the antigen-driven redistribution of bioluminescent CD8+ T cells in living mice. We documented that CD4+ T cells influence the biodistribution of CD8+ T cells, favoring their localization to abdominal lymph nodes. Flow cytometric analysis revealed that this was associated with an increase in the expression of specific integrins. The presence of CD4+ T cells at the time of initial CD8+ T cell activation also influences their biodistribution in the memory phase. Based on these results, we propose the model that one of the functions of CD4+ T cell “help” is to program the homing potential of CD8+ T cells

Identification and Visualization of CD8+ T Cell Mediated IFN-γ Signaling in Target Cells during an Antiviral Immune Response in the Brain

CD8+ T cells infiltrate the brain during an anti-viral immune response. Within the brain CD8+ T cells recognize cells expressing target antigens, become activated, and secrete IFNγ. However, there are no methods to recognize individual cells that respond to IFNγ. Using a model that studies the effects of the systemic anti-adenoviral immune response upon brain cells infected with an adenoviral vector in mice, we describe a method that identifies individual cells that respond to IFNγ. To identify individual mouse brain cells that respond to IFNγ we constructed a series of adenoviral vectors that contain a transcriptional response element that is selectively activated by IFNγ signaling, the gamma-activated site (GAS) promoter element; the GAS element drives expression of a transgene, Cre recombinase (Ad-GAS-Cre). Upon binding of IFNγ to its receptor, the intracellular signaling cascade activates the GAS promoter, which drives expression of the transgene Cre recombinase. We demonstrate that upon activation of a systemic immune response against adenovirus, CD8+ T cells infiltrate the brain, interact with target cells, and cause an increase in the number of cells expressing Cre recombinase. This method can be used to identify, study, and eventually determine the long term fate of infected brain cells that are specifically targeted by IFNγ. The significance of this method is that it will allow to characterize the networks in the brain that respond to the specific secretion of IFNγ by anti-viral CD8+ T cells that infiltrate the brain. This will allow novel insights into the cellular and molecular responses underlying brain immune responses

Genotoxicity assessment of a pharmaceutical effluent using four bioassays

Pharmaceutical industries are among the major contributors to industrial waste. Their effluents when wrongly handled and disposed of endanger both human and environmental health. In this study, we investigated the potential genotoxicity of a pharmaceutical effluent, by using the Allium cepa, mouse- sperm morphology, bone marrow chromosome aberration (CA) and micronucleus (MN) assays. Some of the physico-chemical properties of the effluent were also determined. The A. cepa and the animal assays were respectively carried out at concentrations of 0.5, 1, 2.5, 5 and 10%; and 1, 5, 10, 25 and 50% of the effluent. There was a statistically different (p < 0.05), concentration-dependent inhibition of onion root growth and mitotic index, and induction of chromosomal aberrations in the onion and mouse CA test. Assessment of sperm shape showed that the fraction of the sperm that was abnormal in shape was significantly (p < 0.05) greater than the negative control value. MN analysis showed a dose-dependent induction of micronucleated polychromatic erythrocytes across the treatment groups. These observations were provoked by the toxic and genotoxic constituents present in test samples. The tested pharmaceutical effluent is a potentially genotoxic agent and germ cell mutagen, and may induce adverse health effects in exposed individuals

The primary headaches: genetics, epigenetics and a behavioural genetic model

The primary headaches, migraine with (MA) and without aura (MO) and cluster headache, all carry a substantial genetic liability. Familial hemiplegic migraine (FHM), an autosomal dominant mendelian disorder classified as a subtype of MA, is due to mutations in genes encoding neural channel subunits. MA/MO are considered multifactorial genetic disorders, and FHM has been proposed as a model for migraine aetiology. However, a review of the genetic studies suggests that the FHM genes are not involved in the typical migraines and that FHM should be considered as a syndromic migraine rather than a subtype of MA. Adopting the concept of syndromic migraine could be useful in understanding migraine pathogenesis. We hypothesise that epigenetic mechanisms play an important role in headache pathogenesis. A behavioural model is proposed, whereby the primary headaches are construed as behaviours, not symptoms, evolutionarily conserved for their adaptive value and engendered out of a genetic repertoire by a network of pattern generators present in the brain and signalling homeostatic imbalance. This behavioural model could be incorporated into migraine genetic research

Molecular mechanics potential functions for drug design: DNA minor-groove binders

Therapeutic agents that interfere with replication and transcription functions by binding to double-stranded DNA afford the possibility of effective cancer treatment in which toxic side-effects are minimised. We report on the parameterization of a molecular mechanics potential function for minor groove binders to DNA. The parameters were adjusted to reproduce the results of ab initio quantum chemical calculations of electrostatic potentials and torsional barriers. Non-planar minimum- energy geometries were found for several fragments, and may play a role in the binding modes to DNA. The derived force field should be of use in understanding fundamental aspects of DNA-targeted drug design

Brazil pre-salt, santos basin: Feasibility study for the application ofborehole gravity to improve reservoir monitoring

Brazilian Pre-Salt reservoir characterization and monitoring is a challenge for surface geophysical methodsdue to the inherently minute observable signals from the reservoirs, located below the massive salt, in depthranges > 5000 m. The status of a program to develop a wireline deployed 3-axis gravity sensor with a target sensitivity of~ 5 μGal is firstly introduced. This is followed by a feasibility study for the potential deployment as a time-lapse gravity survey (4D gravity monitoring) within the pre-salt. A conceptual pre-salt reservoir model forthe Libra field, Santos Basin, the offshore of Brazil, is developed, built on available pre-salt knowledgeprovided from seismic imaging and well log data, including reservoir and production characteristics. Themodel is based on water substituting oil over 6 months within and through the reservoir, with adoption ofan oil production rate for the field published in 2017 by the ANP. These data are used to forward modelgravity to understand the potential for a detectable signal and thereby establish a baseline for a time-lapsegravity survey (4D gravity) that could be used to monitor the Libra Field. A clear gravity response >80 μGal is observed over a six month period in the reservoir, due to waterreplacing oil, at the defined oil production rate. In a three-axis measurement the vertical axis is directlyrelated to the magnitude of the fluid substitution and the two horizontal axes are sensitive to the fluidmovement directions. Together, these suggest that an annual survey with a limited well stock could beeffective in monitoring this type of reservoir and that a wireline deployed 3-axis gravity tool is likelyto provide significant additional surveillance to constrain a reservoir production strategy through betterappreciation of the direction of water movement. The follow-on step would be to model further field dataand run a baseline survey to develop a novel reservoir surveillance method within the Pre-Salt