Quantification of the humoral alloimmune response is generally achieved by measuring serum HLA antibodies, which provides no information about the cells involved in the humoral immune response. ...Therefore, we have developed an HLA‐specific B‐cell ELISPOT assay allowing for quantification of B cells producing HLA antibodies. We used recombinant HLA monomers as target in the ELISPOT assay. Validation was performed with human B‐cell hybridomas producing HLA antibodies. Subsequently, we quantified B cells producing HLA antibodies in HLA‐immunized individuals, non‐HLA‐immunized individuals and transplant patients with serum HLA antibodies. B‐cell hybridomas exclusively formed spots against HLA molecules of corresponding specificity with the sensitivity similar to that found in total IgG ELISPOT assays. HLA‐immunized healthy individuals showed up to 182 HLA‐specific B cells per million total B cells while nonimmunized individuals had none. Patients who were immunized by an HLA‐A2‐mismatched graft had up to 143 HLA‐A2‐specific B cells per million total B cells. In conclusion, we have developed and validated a highly specific and sensitive HLA‐specific B‐cell ELISPOT assay, which needs further validation in a larger series of transplant patients. This technique constitutes a new tool for quantifying humoral immune responses.
The authors show that peripheral blood B cells of defined HLA specificity can be enumerated using a novel, monomeric HLA molecule‐based assay.
•This retrospective records study investigated 116 dogs diagnosed with meningoencephalitis of unknown aetiology (MUA).•Thirty of 114 (26%) of dogs died within 1 week of diagnosis of MUA.•Negative ...prognostic factors were decreased mentation and seizures at presentation.•An increased neutrophil percentage in CSF was also a negative prognostic factor.
Although long-term outcomes of meningoencephalitis of unknown aetiology (MUA) in dogs have been evaluated, little is known about short-term survival and initial response to therapy. The aim of this study was to evaluate possible prognostic factors for 7-day survival after diagnosis of MUA in dogs. Medical records were reviewed for dogs diagnosed with MUA between 2006 and 2015. Previously described inclusion criteria were used, as well as 7-day survival data for all dogs. A poor outcome was defined as death within 1 week. Of 116 dogs that met inclusion criteria, 30 (26%) died within 7 days of diagnosis. Assessed variables included age, sex, bodyweight, duration of clinical signs and treatment prior to diagnosis, venous blood glucose and lactate levels, white blood cell count on complete blood count, total nucleated cell count/total protein concentration/white blood cell differentiation on cerebrospinal fluid (CSF) analysis, presence of seizures and cluster seizures, mentation at presentation, neuroanatomical localisation, imaging findings and treatment after diagnosis. Multivariate analysis identified three variables significantly associated with poor outcome; decreased mentation at presentation, presence of seizures, and increased percentage of neutrophils on CSF analysis. Despite initiation of appropriate treatment, more than a quarter of dogs died within 1 week of diagnosis of MUA, emphasising the need for evaluation of short-term prognostic factors. Information from this study could aid clinical staff to provide owners of affected dogs with prognostic information.
To ensure safety tolerance induction protocols are accompanied by conventional immunosuppressive drugs (IS). But IS such as calcineurin inhibitors (CNI), for example, cyclosporin A (CsA), can ...interfere with tolerance induction. We investigated the effect of an additional transient CsA treatment on anti‐CD4mAb‐induced tolerance induction upon rat kidney transplantation. Additional CsA treatment induced deteriorated graft function, resulting in chronic rejection characterized by glomerulosclerosis, interstitial fibrosis, tubular atrophy and vascular changes. Microarray analysis revealed enhanced intragraft expression of the B cell attracting chemokine CXCL13 early during CsA treatment. Increase in CXCL13 expression is accompanied by enhanced B cell infiltration with local and systemic IgG production and C3d deposition as early as 5 days upon CsA withdrawal. Adding different CNIs to cultures of primary mesangial cells isolated from glomeruli resulted in a concentration‐dependent increase in CXCL13 transcription. CsA in synergy with TNF‐α can enhance the B cell attracting and activating potential of mesangial cells. Transient B cell depletion or transfer of splenocytes from tolerant recipients 3 weeks after transplantation could rescue tolerance induction and did inhibit intragraft B cell accumulation, alloantibody production and ameliorate chronic rejection.
Transient coapplication of cyclosporine A abrogates CD4‐specific monoclonal antibody‐mediated transplant tolerance by inducing intragraft CXCL13 expression leading to B cell attraction, alloantibody production and complement‐mediated tissue destruction.
Micro-algae are seen as one of the major future fuel sources. Culture and growth of oil rich micro-algae and catalytic process for the conversion of their crude oils or biomass is reviewed here. ...While there is a significant literature on growth and extraction of oil from the resultant biomass the literature on the problems of refining these oils is diverse and needs collation. It is clear that previous work has been focused on the two green algae
Botryococcus braunii and
Chlorella protothecoides containing terpenoid hydrocarbons and glyceryl lipids as their major crude oils, respectively, both of which will need different refinery technology for upgrading. Studies show a number of conventional catalysts in the petroleum refining industry including transition metals, zeolites, acid and base catalysts can be used with variable effect. These have been employed for cracking, hydrocracking, liquefaction, pyrolysis and transesterification processes to produce diesel, jet fuel and petrol (gasoline). However there is strong evidence that new nano-scale materials containing a high number of active sites and high surface areas may offer more potential.
•Track potential behavioural changes throughout a patients’ epilepsy treatment.•Consider qualities of the chosen anti-epileptic drug (AED); seek a reduced behavioural adverse effect.•A behavioural ...modification plan should be trialled in the first instance.•For anxiety, consider starting fluoxetine at 0.125–0.167mg/kg/day.•Treatment of behavioural comorbidities may not effect seizure control and vice versa.
Psychiatric comorbidities affect a large percentage of people with epilepsy and have a detrimental impact on their quality of life. Recently, behavioural comorbidities, with similar characteristics to human psychiatric diseases, have been identified in dogs with idiopathic epilepsy. In particular, behaviours motivated by the fear–anxiety emotional system have been found to be associated with the occurrence of idiopathic epilepsy in both dogs receiving anti-epileptic drugs, and drug-naïve dogs. There has been little research into the relationship between epilepsy and behavioural signs, and even less into potential treatment protocols. The following article will review available literature from human medicine to describe the current state of knowledge about the bi-directional relationship between anxiety and epilepsy, draw parallels from reported anxiogenic and anxiolytic properties of anti-epileptic drugs and attempt to provide pharmaceutical and behavioural guidance for veterinary patients with epilepsy and comorbid anxiety.
Adoptive immunotherapy with regulatory T cells (Treg) is a new option to promote immune tolerance following solid organ transplantation (SOT). However, Treg from elderly patients awaiting ...transplantation are dominated by the CD45RA(-) CD62L(+) central memory type Treg subset (TregCM), and the yield of well-characterized and stable naïve Treg (TregN) is low. It is, therefore, important to determine whether these TregCM are derived from the thymus and express high stability, suppressive capacity and a broad antigen repertoire like TregN. In this study, we showed that TregCM use a different T cell receptor (TCR) repertoire from conventional T cells (Tconv), using next-generation sequencing of all 24 Vβ families, with an average depth of 534 677 sequences. This showed almost no contamination with induced Treg. Furthermore, TregCM showed enhanced suppressive activity on Tconv at early checkpoints of immune activation controlling activation markers expression and cytokine secretion, but comparable inhibition of proliferation. Following in vitro expansion under mTOR inhibition, TregCM expanded equally as well as TregN without losing their function. Despite relatively limited TCR repertoire, TregCM also showed specific alloresponse, although slightly reduced compared to TregN. These results support the therapeutic usefulness of manufacturing Treg products from CD45RA(-) CD62L(+) Treg-enriched starting material to be applied for adoptive Treg therapy.
In the brain, the efflux transporter P-glycoprotein (Pgp) is predominantly located on the luminal membrane of endothelial cells lining brain microvessels and forming the blood–brain barrier. Many ...lipophilic drugs, including antiepileptic drugs, are potential substrates for Pgp. Overexpression of Pgp in endothelial cells of the blood–brain barrier has been determined in patients with drug resistant forms of epilepsy such as temporal lobe epilepsy and rodent models of temporal lobe epilepsy and suggested to lead to reduced penetration of antiepileptic drugs into the brain. Expression of Pgp after seizures has also been described in astrocytes, whereas it is not clear whether neurons can express Pgp. In the present study, Pgp expression was studied by immunohistochemistry in rats 24 h after a status epilepticus induced by either pilocarpine or kainate, widely used models of temporal lobe epilepsy. Unexpectedly, in addition to endothelial Pgp staining, intense Pgp staining was found in neurons in the CA3c/CA4 sectors and hilus of the hippocampus formation, but not in other brain regions examined. The neuronal Pgp staining was confirmed by two different Pgp antibodies. Double immunolabeling and confocal microscopy showed that Pgp was colocalized with the neuronal marker neuronal nuclear antigen, but not with the glial marker glial fibrillary acidic protein. No neuronal Pgp staining was seen in control rats. The expression of Pgp in neurons after limbic seizures was substantiated by determining Pgp encoding genes (
mdr1a,
mdr1b) in neurons by real time quantitative RT-PCR. Increased Pgp expression in hippocampal neurons is likely to affect the action of drugs with intraneuronal targets and, in view of recent evidence from other cell types, could be associated with prevention of apoptosis which is involved in neuronal damage developing after seizures such as produced by pilocarpine.
Immunomodulatory therapies in sepsis Kox, W J; Volk, T; Kox, S N ...
Intensive care medicine,
01/2000, Letnik:
26 Suppl 1, Številka:
1
Journal Article
Recenzirano
Despite advances in critical care medicine, mortality from sepsis in ICU patients remains high. In response to several infectious and non-infectious stimuli, monocytes/ macrophages release a number ...of mediators, including cytokines, involved in the proinflammatory response that underlies sepsis. The excessive release of these mediators results in the development of whole body inflammation, and plays an important role in the pathogenesis of sepsis and septic shock. In addition, patients with sepsis also undergo an anti-inflammatory phase (the compensatory anti-inflammatory response syndrome) and at times, a mixed response with both pro-and anti-inflammatory components (the mixed antagonistic response syndrome). The initial systemic hyperinflammation is caused by production of inflammatory cytokines, especially tumour necrosis factor-a (TNF-alpha), and also interleukin-1 (IL-1), IL-6, and interferon gamma, which act synergistically with TNF-alpha in inducing shock in animal models. However, clinical trials aimed at downregulating these mediators using antibodies against endotoxin, TNF-alpha, antagonists of IL-1 or platelet activating factor have proved to be uniformly disappointing. Not only have these agents been found to have no effect, but they may also increase mortality. One of the reasons for such failure may be the lack of precise immunological monitoring during the course of sepsis. We have recently demonstrated that sepsis shows a biphasic immunological pattern during the initial and later phase: the early hyperinflammatory phase is counterbalanced by an anti-inflammatory response which may lead to a hypoinflammatory state. The latter is associated with immunodeficiency that is characterised by monocytic deactivation, so-called immunoparalysis. Interferon gamma-1 b has an immunoregulatory effect in patients with immunoparalysis during the compensatory anti-inflammatory response syndrome, not only restoring levels of HLA-DR expression but also reestablishing the ability of monocytes to secrete cytokines such as TNF-alpha. By monitoring immune status in septic patients, targeted intervention may lead to more success in immunomodulation of sepsis.
MAPKAP kinase 2 (MK2) is one of several kinases that are regulated through direct phosphorylation by p38 MAP kinase. By introducing a targeted mutation into the mouse MK2 gene, we have determined the ...physiological function of MK2 in vivo. Mice that lack MK2 show increased stress resistance and survive LPS-induced endotoxic shock. This is due to a reduction of approximately 90% in the production of tumor necrosis factor-alpha (TNF-alpha) and not to a change in signalling from the TNF receptor. The level and stability of TNF-alpha mRNA is not reduced and TNF-alpha secretion is not affected. We conclude that MK2 is an essential component in the inflammatory response which regulates biosynthesis of TNF-alpha at a post-transcriptional level.
Spinal disease in dogs is commonly encountered in veterinary practice. Numerous diseases may cause similar clinical signs and presenting histories. The study objective was to use statistical models ...to identify combinations of discrete parameters from the patient signalment, history and neurological examination that could suggest the most likely diagnoses with statistical significance. A retrospective study of 500 dogs referred to the Queen Mother Hospital for Animals before June 2012 for the investigation of spinal disease was performed. Details regarding signalment, history, physical and neurological examinations, neuroanatomical localisation and imaging data were obtained. Univariate analyses of variables (breed, age, weight, onset, deterioration, pain, asymmetry, neuroanatomical localisation) were performed, and variables were retained in a multivariate logistic regression model if P<0.05. Leading diagnoses were intervertebral disc extrusion (IVDE, n=149), intervertebral disc protrusion (n=149), ischaemic myelopathy (IM, n=48) and neoplasms (n=44). Multivariate logistic regression characterised IM and acute non-compressive nucleus pulposus extrusions as the only peracute onset, non-progressive, non-painful and asymmetrical T3-L3 myelopathies. IVDE was most commonly characterised as acute onset, often deteriorating, painful and largely symmetrical T3-L3 myelopathy. This study suggests that most spinal diseases cause distinctive combinations of presenting clinical parameters (signalment, onset, deterioration, pain, asymmetry, neuroanatomical localisation). Taking particular account of these parameters may aid decision making in a clinical setting.
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