A Rac1 inhibitory peptide suppresses antibody production and paw swelling in the murine collagen-induced arthritis model of rheumatoid arthritis

Introduction

The Rho family GTPase Rac1 regulates cytoskeletal rearrangements crucial for the recruitment, extravasation and activation of leukocytes at sites of inflammation. Rac1 signaling also promotes the activation and survival of lymphocytes and osteoclasts. Therefore, we assessed the ability of a cell-permeable Rac1 carboxy-terminal inhibitory peptide to modulate disease in mice with collagen-induced arthritis (CIA).

Methods

CIA was induced in DBA/1 mice, and in either early or chronic disease, mice were treated three times per week by intraperitoneal injection with control peptide or Rac1 inhibitory peptide. Effects on disease progression were assessed by measurement of paw swelling. Inflammation and joint destruction were examined by histology and radiology. Serum levels of anti-collagen type II antibodies were measured by enzyme-linked immunosorbent assay. T-cell phenotypes and activation were assessed by fluorescence-activated cell sorting analysis. Results were analyzed using Mann-Whitney U and unpaired Student t tests.

Results

Treatment of mice with Rac1 inhibitory peptide resulted in a decrease in paw swelling in early disease and to a lesser extent in more chronic arthritis. Of interest, while joint destruction was unaffected by Rac1 inhibitory peptide, anti-collagen type II antibody production was significantly diminished in treated mice, in both early and chronic arthritis. Ex vivo, Rac1 inhibitory peptide suppressed T-cell receptor/CD28-dependent production of tumor necrosis factor α, interferon γ and interleukin-17 by T cells from collagen-primed mice, and reduced induction of ICOS and CD154, T-cell costimulatory proteins important for B-cell help.

Conclusions

The data suggest that targeting of Rac1 with the Rac1 carboxy-terminal inhibitory peptide may suppress T-cell activation and autoantibody production in autoimmune disease. Whether this could translate into clinically meaningful improvement remains to be shown.     

The seed banks of English lowland calcareous grasslands along a restoration chronosequence

We investigated whether the seed banks of ex-arable lowland calcareous grasslands underwent restoration similar to that of the above-ground restoration, and whether this was influenced by seed-sowing or environmental conditions. We compared 40 sites, where some form of restoration work had been implemented between 2 and 60 years previously, with 40 paired reference sites of good quality calcareous grassland with no history of ploughing or agricultural improvement. We analysed differences between sites and between above- and below-ground vegetation using both a multivariate approach and proportions of selected plant attributes. Seed banks of reference sites were more characteristic of late successional communities, with attributes such as stress tolerance, perenniality and a reliance on fruit as the germinule form more abundant than in restoration sites. In restoration sites, these tended to decrease with restoration site isolation and increase with restoration site age and where soil nutrient conditions were more similar to reference sites (i.e. with relatively low phosphorus and high nitrogen). Seed bank communities of all sites differed considerably from above-ground communities, however, and no overall significant responses to site age, isolation or soil nutrients were detected by multivariate analyses of similarity of species between pairs of sites. Responses to different seeding methods were also barely detectable. While there is some indication from the plant attribute data that the regeneration potential contained in the seed banks of restored sites increasingly resembles that of references sites over time, even seed banks of good quality calcareous grassland are dominated by ruderal species. It is likely, therefore, that permanent seed banks do not facilitate the restoration of ex-arable grasslands.     

Major morphological effects of a regulatory gene: Pgm1-t in rainbow trout

We have investigated the morphological effects of a genetic locus, Pgm1- t, that affects the expression of a phosphoglucomutase locus (Pgm1) in liver of rainbow trout (Salmo gairdneri). We have previously shown that embryos with liver Pgm1 expression hatch earlier than those without liver Pgm1 expression. We predicted that this difference in developmental rate should cause a reduction in meristic counts in the more rapidly developing fish with liver Pgm1 expression. Eight meristic (countable) characters in nine full-sib groups segregating for the presence or absence of liver Pgm1 expression are in agreement with this prediction. In eight of the nine families, there is a significant difference in the multivariate distribution of the eight meristic counts between full sibs with and without liver Pgm1 expression. This separation in multivariate space is based on a tendency for lower meristic counts in fish with liver Pgm1 expression. The magnitude of these morphological differences is similar to that between two subspecies of cutthroat trout (Salmo clarki) that show substantial genetic divergence at structural loci encoding enzymes (Nei's D = 0.34). These data support the view that small changes in the developmental process caused by genetic differences at regulatory genes can have large effects on morphology.      

Two common invertebrate predators show varying predation responses to different types of sentinel prey

Sentinel prey (an artificially manipulated patch of prey) are widely used to assess the level of predation provided by natural enemies in agricultural systems. Whilst a number of different methodologies are currently in use, little is known about how arthropod predators respond to artificially manipulated sentinel prey in comparison with predation on free‐living prey populations. We assessed how attack rates on immobilized (aphids stuck to cards) and artificial (plasticine lepidopteran larvae mimics) sentinel prey differed to predation on free‐moving live prey (aphids). Predation was assessed in response to density of the common invertebrate predators, a foliar‐active ladybird Harmonia axyridis (Coleoptera: Coccinellidae), and a ground‐active beetle Pterostichus madidus (Coleoptera: Carabidae). Significant increases in attack rates were found for the immobilized and artificial prey between the low and high predator density treatments. However, an increased predator density did not significantly reduce numbers of free‐living live aphids included in the mesocosms in addition to the alternate prey. We also found no signs of predation on the artificial prey by the predator H. axyridis. These findings suggest that if our assessment of predation had been based solely on the foliar artificial prey, then no increase in predation would have been found in response to increased predator density. Our results demonstrate that predators differentially respond to sentinel prey items which could affect the level of predation recorded where target pest species are not being used.     

Long-term change in calcareous grassland vegetation and drivers over three time periods between 1970 and 2016

Plant Ecology - Analysis of long-term vegetation change is limited. Furthermore most studies evaluating change only examine two snapshots in time, which makes it difficult to define rates of change...     

Effects of vegetation structure and floristic diversity on detritivore,herbivore and predatory invertebrates within calcareous grasslands

Calcareous grasslands in Europe have shown wide scale declines in their extent and quality as a result of modern agricultural practices, increased atmospheric eutrophication and lack of management. In addition to being a key habitat for specialist plants, calcareous grasslands are also important for many threatened invertebrates. In this UK based study, we investigated the impact of military vehicle activity, floral species richness and vegetation structure on assemblages of detritivore, herbivore and predatory invertebrates. We also consider the impact that disturbance by military vehicle activity on the proportion of invertebrate species capable of flight, a surrogate for dispersal ability. Sward height was negatively correlated with detritivore, herbivore and predator species richness. Herbivores species richness was positively correlated with both forb and grass species richness. Spatial variation in the number of plant species was negatively correlated with herbivore species richness. Those sites most heavily disturbed by military vehicle activity supported the lowest proportions of flightless invertebrates. Successful management for calcareous grassland invertebrates should aim to maintain short swards with high floristic diversity, in terms of both the forbs and grasses. It should be noted, however, that these findings refer to principally surface rather than sward active invertebrates. While disturbance associated with military vehicle activity was not found to affect invertebrate species richness, it has negative consequences for the structure of invertebrate assemblages by selecting against invertebrates with low dispersal ability. To support invertebrate diversity in calcareous grasslands we emphasise the need for variety in the timing and type of management applied to promote heterogeneity in sward structure.