IgG subclass antibodies to human and bacterial HSP60 are not associated with disease activity and progression over time in axial spondyloarthritis

Introduction

Spondyloarthritis (SpA), an interrelated group of rheumatic diseases, has been suggested to be triggered by bacterial infections prior to the development of an autoimmune response that causes inflammation of the spinal and peripheral joints. Because human heat shock protein 60 (HSP60), recently renamed HSPD1, and bacterial HSP60 are highly homologous, immunological cross-reactivity has been proposed as a mechanism of disease initiation. However, previous investigations of the humoral immune response to HSP60 in SpA patients have lacked determination of immunoglobulin G (IgG) subclasses and patient follow-up. In this study, we have focused on these parameters in a cohort of axial SpA patients with a well-established set of clinical characteristics, including MRI changes and human leukocyte antigen B27.

Methods

IgG subclass antibodies (IgG1, IgG2, IgG3 and IgG4) against recombinant HSP60 of three reactive arthritis-related bacteria; human HSP60; and the microorganisms Chlamydia trachomatis and C. pneumoniae were determined by ELISA. Serum samples collected from 2004 to 2006 and in 2010 and 2011 from 39 axial SpA patients were analyzed and compared with samples from 39 healthy controls. The Mann-Whitney U test and Wilcoxon matched pairs test were used to compare the antibody levels in different and paired groups, respectively. P < 0.01 was considered significant. The Spearman nonparametric correlation was used to determine correlation between antibody levels and between antibody levels and the disease parameters.

Results

Elevated levels of IgG1 and IgG3 to human HSP60 and IgG1 to HSP60 of Salmonella enterica Enteritidis were observed in SpA patients compared with healthy controls at both time points. The antibody levels were almost constant over time for IgG1, whereas high levels of IgG3 to human HSP60 tended to decrease over time. The antibody response to human HSP60 was predominantly of the IgG3 subclass, and patients with high levels of IgG3 to this antigen had low levels of IgG1, indicating an inverse association. Different IgG subclasses were produced against bacterial and human HSP60 in the same serum sample, IgG1 and IgG3, respectively, indicating that there was no cross-reaction.

Conclusions

A significant association was observed between axial SpA and the presence of IgG1/IgG3 antibodies to human HSP60 and of IgG1 to S. enterica Enteritidis and C. trachomatis. Generation of antibodies to human HSP60 was independent of the presence of antibodies to bacterial HSP60. No association was observed between clinical and MRI changes with antibodies over time. Altogether, such antibodies do not reflect the disease activity in these patients.This study has been approved by the Regional Research Ethics Committee of Central Jutland, Denmark. Trial registration numbers: 20050046 and 20100083     

The influence of crop rotation on the onset of early blight (Alternaria solani)

Field experiments were carried out in 2016 and 2017 to determine the effect of crop rotation on the onset of early blight in three potato varieties in Denmark. Six and seven fields with different histories with potato (rotation‐fields) were used for the experiments in 2016 and 2017, respectively. The results showed that variety and the interaction between variety and rotation‐field had no significant effect on the onset of early blight. Rotation‐field, on the other hand, had a significant effect on the onset of early blight in both years of the study. Early blight occurred earlier in fields with <2 years between subsequent potato crops than in fields where potatoes have not been grown for at least 2 years. The onset of early blight was not different between the fields where there had not been potatoes for at least 2 years. For fields that were preceded by potatoes, early blight occurred earlier in fields in which the severity of early blight in the previous years was higher than that in the fields in which the severity of early blight in the previous years was low or moderate. In conclusion, the results of this study suggest that at least a 2‐year interval between subsequent potatoes in a rotation cycle is necessary to delay the onset of early blight. Moreover, the severity of early blight in the previous years can affect the onset of early blight.     

Invasion of terrestrial enchytraeids into two postglacial tundras: North-eastern Greenland and the Arctic Archipelago of Canada (Enchytraeidae, Oligochaeta)

The aim was to study the invasion of terrestrial enchytraeids into the two postglacial tundras, North-eastern Greenland and the Arctic Archipelago of Canada. The majority of species recorded have a wide distribution in the Northern Hemisphere and the species diversity is considerably lower than that of potential source regions indicating either strong selection or varied dispersal ability. It appears that the Archipelago is influenced mainly from North America and North-eastern Greenland from Europe while the specialized fauna of the ancient Beringian tundra is of minor importance. The two alternative scenarios: (a) survival of a prepleistocene fauna in protected refugia within the area or (b) a postglacial re-invasion from outside are discussed, but the available data do not discriminate between these two possibilities. A total of 24 terrestrial enchytraeid taxa are recorded of which 22 are identified to the species level. Bryodrilus archipelagicus and Hemifridericia bivesiculata are described as new to science and Henlea groenlandica Černosvitov is revalidated. We hesitate to conclude that the new taxa described are endemic.     

Soil fauna communities and microbial respiration in high Arctic tundra soils at Zackenberg, Northeast Greenland

The soil fauna communities were described for three dominant vegetation types in a high arctic site at Zackenberg, Northeast Greenland. Soil samples were extracted to quantify the densities of mites, collembolans, enchytraeids, diptera larvae, nematodes and protozoa. Rates of microbial respiration were also assessed. Collembolans were found in highest densities in dry heath soil, about 130,000 individuals m⁻², more than twice as high as in mesic heath soils. Enchytraeids, diptera larvae and nematodes were also more abundant in the dry heath soil than in mesic heath soils, whereas protozoan densities (naked amoeba and heterotrophic flagellates) were equal. Respiration rate of unamended soil was similar in soil from the three plots. However, a higher respiration rate increase in carbon + nutrient amended soil and the higher densities of soil fauna (with the exception of mites and protozoa) in dry heath compared to the mesic heath soils indicated a higher decomposition rate here.     

MobiSeq: De novo SNP discovery in model and non‐model species through sequencing the flanking region of transposable elements

In recent years, the availability of reduced representation library (RRL) methods has catalysed an expansion of genome‐scale studies to characterize both model and non‐model organisms. Most of these methods rely on the use of restriction enzymes to obtain DNA sequences at a genome‐wide level. These approaches have been widely used to sequence thousands of markers across individuals for many organisms at a reasonable cost, revolutionizing the field of population genomics. However, there are still some limitations associated with these methods, in particular the high molecular weight DNA required as starting material, the reduced number of common loci among investigated samples, and the short length of the sequenced site‐associated DNA. Here, we present MobiSeq, a RRL protocol exploiting simple laboratory techniques, that generates genomic data based on PCR targeted enrichment of transposable elements and the sequencing of the associated flanking region. We validate its performance across 103 DNA extracts derived from three mammalian species: grey wolf (Canis lupus), red deer complex (Cervus sp.) and brown rat (Rattus norvegicus). MobiSeq enables the sequencing of hundreds of thousands loci across the genome and performs SNP discovery with relatively low rates of clonality. Given the ease and flexibility of MobiSeq protocol, the method has the potential to be implemented for marker discovery and population genomics across a wide range of organisms—enabling the exploration of diverse evolutionary and conservation questions.     

Mutations in FLS2 Ser-938 Dissect Signaling Activation in FLS2-Mediated Arabidopsis Immunity

FLAGELLIN-SENSING 2 (FLS2) is a leucine-rich repeat/transmembrane domain/protein kinase (LRR-RLK) that is the plant receptor for bacterial flagellin or the flagellin-derived flg22 peptide. Previous work has shown that after flg22 binding, FLS2 releases BIK1 kinase and homologs and associates with BAK1 kinase, and that FLS2 kinase activity is critical for FLS2 function. However, the detailed mechanisms for activation of FLS2 signaling remain unclear. The present study initially identified multiple FLS2 in vitro phosphorylation sites and found that Serine-938 is important for FLS2 function in vivo. FLS2-mediated immune responses are abolished in transgenic plants expressing FLS2_S938A, while the acidic phosphomimic mutants FLS2_S938D and FLS2_S938E conferred responses similar to wild-type FLS2. FLS2-BAK1 association and FLS2-BIK1 disassociation after flg22 exposure still occur with FLS2_S938A, demonstrating that flg22-induced BIK1 release and BAK1 binding are not sufficient for FLS2 activity, and that Ser-938 controls other aspects of FLS2 activity. Purified BIK1 still phosphorylated purified FLS2_S938A and FLS2_S938D mutant kinase domains in vitro. Phosphorylation of BIK1 and homologs after flg22 exposure was disrupted in transgenic Arabidopsis thaliana plants expressing FLS2_S938A or FLS2_D997A (a kinase catalytic site mutant), but was normally induced in FLS2_S938D plants. BIK1 association with FLS2 required a kinase-active FLS2, but FLS2-BAK1 association did not. Hence FLS2-BIK1 dissociation and FLS2-BAK1 association are not sufficient for FLS2-mediated defense activation, but the proposed FLS2 phosphorylation site Ser-938 and FLS2 kinase activity are needed both for overall defense activation and for appropriate flg22-stimulated phosphorylation of BIK1 and homologs.     

A comparative ecological investigation of two related mayfly nymphs

Zusammenfassung Heptagenia sulphurea wird auf Steinen, H. fuscogrisea in der Vegetation von Fließgewässern gefunden (Fig. 2).In Experimenten wählte H. sulphurea Steine und groben Kies. H. fuscogrisea wählte Batrachium als Substrat, nur ein geringer Anteil der Larven fand sich auf Callitriche (Fig. 3).Die Zeit, die zwischen Loslassen und erneutem Anhaften auf dem Substrat im strömenden Milieu verstreicht, war unterschiedlich bei den zwei Spezies. Die auf Steinen lebende Art reagierte schneller als die aus der Vegetation. H. sulphurea verhielt sich, wenn sie von der Strömung ergriffen worden war, ziemlich passiv, doch trug die turbulente Strömung das Tier dem Substrat zu, so hatte es gewöhnlich sicheren Halt.Im Strömungsaquarium bevorzugt H. sulphurea die strömungsexponierten Bereiche, aber viele waren auch im ruhigen Wasser (Fig. 5).Wurde H. fuscogrisea von der Strömung ergriffen, so führte sie Schwimmbewegungen aus. Erreicht sie eine Haltmöglichkeit an strömungsexponiertem Platz, so wird sie oft weitergetragen. Die Tiere lassen sie meist in ruhigem Wasser nieder.Im Strömungsaquarium bevorzugt H. fuscogrisea überwiegend die Regionen ruhigen Wassers (Fig. 5).Die Zeit zwischen Loslassen und Festsetzen war deutlich durch die Temperatur beeinflußt (Fig. 6).Der dichte Haarbesatz an der hinteren Kante der Femur von H. sulphurea wird als Anpassung an das Leben in der Strömung angesehen, weil er die Wirbelbildung herabsetzen kann. Der Haarbesatz entlang der Tibia bei H. fuscogrisea wird als Anpassung zum Schwimmen angesehen, zu dem these Spezies befähigt ist, wenn sie von der Strömung ergriffen wird (Fig. 8).Die Spezies von Steinen ist etwas mehr abgeflacht als die aus der Vegetation (Fig. 7).     

The role of cladocerans in tracking long-term change in shallow lake trophic status

Shallow lakes have been affected by a variety of human activities profoundly altering their ecological structure and function. Cladocerans have been used to track change resulting from a variety of drivers at a number of time scales. Aquatic macrophytes are well recognised as reflecting the ecological condition of a lake. Here, we compare the plant macrofossils with the sub-fossil cladoceran assemblages from 20 dated sediment cores. Co-correspondence analysis was used to determine the degree of commonality of change in community composition of the two biological groups through time. This analysis revealed very high levels of agreement in the nature and timing of change at all the sites examined with very high correlation coefficients between the axis 1 scores for macrofossils and cladocerans. Furthermore, at all sites a high proportion of the variance (min 20%, max 54%) in the macrofossil data was explained by the change in the cladoceran assemblage. Sub-fossil macrofossil and cladoceran assemblages, from at least from 1700 AD onwards, were examined in more detail at three sites: Ormesby Great Broad, Felbrigg Lake and Lake Søbygaard. There was very good accord in the main shifts of the cladoceran and macrofossil assemblages at all three sites. This may reflect the long-term shift in the principal focus of primary production from the benthic to the pelagic habitat. We suggest that the combination of their central position in the food-web and the presence of both pelagic and benthic taxa make cladocerans a strong candidate as the single best indicator of (palaeo) ecological condition related to changing trophic status and alteration in food-web structure in shallow lakes.