Evaluation of sensitization to the crude extract of Dermatophagoides farinae and its derived allergens, Der f 2 and Zen 1, in dogs with atopic dermatitis in Southern Brazil.

BACKGROUND: Atopic dermatitis is associated with the production of IgE antibodies against environmental allergens and allergens of the house dust miteDermatophagoides farinae are frequently implicated in the disease. OBJECTIVES: We aimed to observe the allergen-specific IgE against crudeD. farinae, Der f 2 and Zen 1 in dogs with atopic dermatitis and report if these dogs are in contact with material that could shelter mite allergens. METHODS: 100 dogs with clinical diagnosis of atopic dermatitis were included after exclusion of other forms of pruritic skin disease and dogs that already received specific or non-specific immunotherapy. These dogs were of different breeds and ages and they were presented at a veterinary teaching hospital and a private service of veterinary dermatology, both located in Curitiba, Southern Brazil. At the time of anamnesis, some questions were applied to know the possibility of these dogs having had contact with furniture and textile material which could shelter house dust mites. Sera samples were obtained and further analyzed by ELISA assay to measure serum IgE levels against these allergens with an established cut-off of 0.200 IgE optical density. RESULTS: The allergen-specific IgE positivity against crudeD. farinae (92 %) and Zen 1 (77 %) was higher than Der f 2 (56 %). There was a correlation in sensitization to crude D. farinae and Zen 1 that was not observed between crude D. farinae and Der f 2 and Der f 2 and Zen 1. The sensitization to D. farinae and its allergens was associated with an unrestricted exposition to furniture and textile material. CONCLUSION & CLINICAL RELEVANCE: dogs with atopic dermatitis are frequently sensitized to D. farinae and its allergens, Der f 2 and Zen 1, may be considered major allergens in these dogs. Zen 1 may be the main allergen responsible for the sensitization to crude D. farinae.

Methods for Experimental Allergen Immunotherapy: Subcutaneous and Sublingual Desensitization in Mouse Models of Allergic Asthma.

Allergic asthma is characterized by airway hyperresponsiveness, remodeling, and reversible airway obstruction. This is associated with an eosinophilic inflammation of the airways, caused by inhaled allergens such as house dust mite or grass pollen. The inhaled allergens trigger a type-2 inflammatory response with the involvement of innate lymphoid cells (ILC2) and Th2 cells, resulting in high immunoglobulin E (IgE) antibody production by B cells and mucus production by airway epithelial cells. As a consequence of the IgE production, subsequent allergen reexposure results in a classic allergic response with distinct early and late phases, both resulting in bronchoconstriction and shortness of breath. Allergen-specific immunotherapy (AIT) is the only treatment that is capable of modifying the immunological process underlying allergic responses including allergic asthma. Both subcutaneous AIT (SCIT) as well as sublingual AIT (SLIT) have shown clinical efficacy in long-term suppression of the allergic response. Although AIT treatments are very successful for rhinitis, application in asthma is hampered by variable efficacy, long duration of treatment, and risk of severe side effects. A more profound understanding of the mechanisms by which AIT induces tolerance to allergens in sensitized individuals is needed to be able to improve its efficacy. Mouse models have been very valuable in preclinical research for characterizing the mechanisms of desensitization in AIT and evaluating novel approaches to improve its efficacy. Here, we present a rapid and reproducible mouse model for allergen-specific immunotherapy. In this model, mice are sensitized with two injections of allergen adsorbed to aluminum hydroxide, followed by subcutaneous injections (SCIT) or sublingual administrations (SLIT) of allergen extracts as an immunotherapy treatment. Finally, mice are challenged by intranasal allergen administrations. We will also describe the protocols as well as the most important readout parameters for the measurements of invasive lung function, serum immunoglobulin levels, isolation of bronchoalveolar lavage fluid (BALF), and preparation of cytospin slides. Moreover, we describe how to perform ex vivo restimulation of lung single-cell suspensions with allergens, flow cytometry for identification of relevant immune cell populations, and ELISAs and Luminex assays for assessment of the cytokine concentrations in BALF and lung tissue.

Imported fire ant immunotherapy prescribing patterns in a large health care system during an 11-year period.

BACKGROUND: The first large-scale evaluation of prescribing patterns for imported fire ant (IFA) in a large US health care system was published by Haymore et al in 2009. In this first evaluation of prescriptions from 1990 to 2007, the most often prescribed maintenance IFA prescription was 0.5 mL of 1:200 wt/vol. OBJECTIVE: To provide an updated description of IFA prescribing patterns over the ensuing 11 years from same large health care system. METHODS: We reviewed 1349 new IFA prescriptions written from 2007 to 2018, from a large nationwide health care system, with primary end points being maintenance prescription strength and prescribing patterns. RESULTS: In comparison to the data published by Haymore et al in 2009, which reported that 17% of the prescriptions were written for 0.5 mL of 1:100 wt/vol maintenance, we found that 69% (95% CI: 66.4%-71.4%) of IFA prescriptions written in the past 11 years were for the maintenance concentration of 0.5 mL of 1:100 wt/vol. We further studied the linear trend over time of percentage of prescriptions written for individual concentrations and observed that the percentage of 1:100 wt/vol prescriptions increased 3.5% yearly (R2 = 0.68, P < .001) from 2007 (40.0%, 95% CI: 24.6%-57.7%) to 2018 (84.4%, 95% CI: 77.4%-89.5%). CONCLUSION: Our study shows significant improvement in the accuracy and precision of IFA immunotherapy dosing for patients with IFA hypersensitivity, with ascendancy of 0.5 mL 1:100 wt/vol as the predominant treatment dose. A total of 87% of patients within our study were treated within the parameter recommendations, a stark improvement from findings in the 2009 Haymore study.

Similarities and differences in the effects of sensitisation and challenge with Dermatophagoides farinae and Dermatophagoides pteronyssinus extracts in a murine asthma surrogate.

Patients with atopic asthma may become sensitised to the grain storage mite Dermatophagoides farinae (Der f), the house dust mite Dermatophagoides pteronyssinus (Der p) or both, but thus far little attention has been paid to date to possible variation in their pathophysiological effects. Here we present a side by side comparison of the effects of extracts of these two dust mites in a murine surrogate of atopic asthma. Compared with the Der p-challenged mice, however, the mice-challenged with Der f had favour changes in lung tissue elasticity and expression in matrix metalloproteinases in lung tissue, while the mice challenged with Der p showed more neutrophils infiltrating around the airway and stronger expression of steroid-resistant related cytokines in the lung tissue. Our data suggest that different dust mite crude extracts might lead different pathological characteristics, at least in murine models of asthma.

Subcutaneous Immunotherapy With High-Dose Cat and Dog Extracts: A Real-life Study.

BACKGROUND AND OBJECTIVE: Background: Data on the efficacy of immunotherapy administered to patients with cat or dog allergy are scarce. Objective: We aimed to evaluate the safety and efficacy of subcutaneous immunotherapy (SCIT) in patients with allergy to cat and dog dander. METHODS: Consecutive patients with rhinitis and/or asthma related to sensitization to cat or dog dander were included in a pragmatic, real-life, prospective, observational study. All patients had specific IgE to cat, dog, or both. SCIT was administered using an infusion pump over 3 sessions as part of a rush protocol, followed by monthly administration over 12 months. We recorded adverse events, clinical outcomes, pulmonary function, FeNO, symptoms of rhinitis and asthma, quality of life (QoL), Asthma Control Test (ACT) score, and visual analog scale (VAS) score at baseline, 6 months, and 12 months. RESULTS: The study population comprised 66 patients (38 females, 46 allergic to cat and 20 to dog), with ages ranging from 9 to 59 years. During the up-dosing phase, in which the infusion pump was used, 8.1% of doses elicited a systemic reaction and 5.4% caused a local reaction, while 9.3% of doses administered during the maintenance phase (ie, without an infusion pump) induced a systemic reaction. No local reactions were recorded. A significant improvement in FEV1, symptoms of rhinitis and asthma, QoL, use of medication, VAS score, and ACT score was observed at 6 months and continued at 12 months. Clinical improvement with cat extract was significantly better than with dog extract. CONCLUSIONS: High-dose SCIT has substantial clinical value in many cat- and dog-allergic patients.

Optimal conditions for the storage of German cockroach extract.

Allergen extracts are commonly utilized for diagnosis and immunotherapy; however, the stability of protease­rich extracts is important for a precise diagnosis and treatment efficacy. The present study determines the optimal conditions for the storage of German cockroach allergen extract. Cockroach extracts were reconstituted in four buffers: normal saline (NS), 50% glycerol in NS, 0.3% phenol in NS, or 0.3% phenol and 50% glycerol in NS. The extracts in different buffers were stored either at room temperature (18­26˚C, RT) or refrigerated (2­8˚C). Subsequently, the protein concentration and allergen content (Bla g 1 and Bla g 2) in the extracts were examined for the course of one year. Extract potency was estimated by inhibition ELISA. At least 90.5% protein, 94.4% Bla g 1, 65.2% Bla g 2, and 91.4% potency remained after one year when 50% glycerol NS was added to the extract with refrigeration. However, less than 13.7% protein, 17.1% Bla g 1, 0% Bla g 2 and 32.5% potency were maintained after one year when 50% glycerol NS was not added to the extract and was maintained at RT. The addition of 0.3% phenol NS did not show significant effects on extract stability. The addition of 50% glycerol NS and refrigerated storage temperature were found to be important factors for increasing the shelf life of protease­rich cockroach extract.

Production of Inflammatory Cytokines and Nitric Oxide by Human Mast Cells Incubated with Toxoplasma gondii Lysate.

The roles of mast cells in allergic diseases and helminth infections are well known. However, the roles of mast cells in T. gondii infection is poorly understood. This study was focused on the production of pro-inflammatory cytokines (TNF-α, IL-4), chemokines (CXCL8, MCP-1) and nitric oxide (NO) by mast cells in response to soluble lysate of T. gondii tachyzoites. Production of CXCL8 (IL-8), MCP-1, TNF-α and IL-4 were measured by RT-PCR and ELISA. Western blot were used for detection of CXCR-1 and CXCR2. Our results showed that T. gondii lysates triggered mast cells to release CXCL8, MCP-1, TNF-α, IL-4 and to produce NO. This suggests that mast cells play an important role in inflammatory responses to T. gondii.

Necroptosis directly induces the release of full-length biologically active IL-33 in vitro and in an inflammatory disease model.

Interleukin-33 (IL-33) is a pro-inflammatory cytokine that plays a significant role in inflammatory diseases by activating immune cells to induce type 2 immune responses upon its release. Although IL-33 is known to be released during tissue damage, its exact release mechanism is not yet fully understood. Previously, we have shown that cleaved IL-33 can be detected in the plasma and epithelium of Ripk1-/- neonates, which succumb to systemic inflammation driven by spontaneous receptor-interacting protein kinase-3 (RIPK3)-dependent necroptotic cell death, shortly after birth. Thus, we hypothesized that necroptosis, a RIPK3/mixed lineage kinase-like protein (MLKL)-dependent, caspase-independent cell death pathway controls IL-33 release. Here, we show that necroptosis directly induces the release of nuclear IL-33 in its full-length form. Unlike the necroptosis executioner protein, MLKL, which was released in its active phosphorylated form in extracellular vesicles, IL-33 was released directly into the supernatant. Importantly, full-length IL-33 released in response to necroptosis was found to be bioactive, as it was able to activate basophils and eosinophils. Finally, the human and murine necroptosis inhibitor, GW806742X, blocked necroptosis and IL-33 release in vitro and reduced eosinophilia in Aspergillus fumigatus extract-induced asthma in vivo, an allergic inflammation model that is highly dependent on IL-33. Collectively, these data establish for the first time, necroptosis as a direct mechanism for IL-33 release, a finding that may have major implications in type 2 immune responses.

Lepista sordida Water Extract Enhances the Maturation of Mouse Dendritic Cells in Vitro and in Vivo.

BACKGROUND: Lepista sordida (LS) extract has been shown to possess anti-oxidant, anti-aging, and anti-tumor activities. However, the immunostimulatory effect of LS extract has not been elucidated. OBJECTIVE: To characterize the impact of a water extract of LS (WE-LS) on the maturation and function of mouse dendritic cell (DC) in vitro and in vivo. METHODS: Mouse bone marrow-derived DCs (BMDCs) were generated. Next, DC maturation was determined by flow cytometry, and cytokine production was measured by ELISA after WE-LS treatment. In addition, DC-induced OVA-specific T cell activation was assayed by [3H]-thymidine incorporation assay. Furthermore, the in vivo effects of WE-LS on DC maturation and Th1 responses in the spleens of mice were assessed by flow cytometry. RESULTS: WE-LS treatment up-regulated co-stimulatory (CD40 and CD80) and MHC class II molecules, increased the production of tumor necrosis factor-alpha (TNF-α), IL-6 and IL-12, and enhanced both the proliferation and IFN-γ secretion of allogenic T cells in BMDCs, partially mediated by the TLR2 and TLR4 signaling pathways. Moreover, the in vivo administration of WE-LS to mice enhanced the up-regulation of CD40, CD80 and MHC class II molecules in spleen DCs. WE-LS also increased the generation of T helper type 1 (Th1) cells in vivo. CONCLUSION: These results suggest that WE-LS might have the potential to promote immunity against infection and cancer or to serve as an adjuvant in vaccines and immunotherapies.

A Bioassay for Optimization of Macrophage-Conditioned Medium as a Culture Supplement to Promote Hybridoma Cell Survival and Growth.

Macrophage-conditioned medium (MCM) is an important cell culture supplement used to support the survival and growth of newly fused hybridoma cells. The use of macrophage cells, as a part of hybridoma technology, has proven to be an effective and inexpensive source of growth factors that promote the early survival and growth of hybridoma cells. Despite the widespread use of MCM as a hybridoma culture supplement, there is limited guidance and standardization for MCM production to achieve optimal hybridoma support. As an undefined supplement, significant variations in production of MCM may negatively impact hybridoma cell survival and growth. The lack of an available method for standardization of MCM bioactivity has limited validation, optimization, and commercial production. Consequently, variations in batch production of MCM may result in low-quality MCM that limits hybridoma viability and negatively impacts monoclonal antibody production. In this report, we describe a novel bioassay based on the newly generated, MCM-dependent RMH359 hybridoma cell line that can be used to validate MCM bioactivity and standardize production. We demonstrate the utility of the RMH359 bioassay (1) for evaluating MCM hybridoma bioactivity, (2) to define optimal conditions for production of MCM, and (3) as a method for MCM validation and standardization. In conclusion, the RMH359 cell bioassay provides a specific and sensitive assessment of MCM bioactivity in support of hybridoma cell survival and growth.

Treating cat allergy with monoclonal IgG antibodies that bind allergen and prevent IgE engagement.

Acute allergic symptoms are caused by allergen-induced crosslinking of allergen-specific immunoglobulin E (IgE) bound to Fc-epsilon receptors on effector cells. Desensitization with allergen-specific immunotherapy (SIT) has been used for over a century, but the dominant protective mechanism remains unclear. One consistent observation is increased allergen-specific IgG, thought to competitively block allergen binding to IgE. Here we show that the blocking potency of the IgG response to Cat-SIT is heterogeneous. Next, using two potent, pre-selected allergen-blocking monoclonal IgG antibodies against the immunodominant cat allergen Fel d 1, we demonstrate that increasing the IgG/IgE ratio reduces the allergic response in mice and in cat-allergic patients: a single dose of blocking IgG reduces clinical symptoms in response to nasal provocation (ANCOVA, p = 0.0003), with a magnitude observed at day 8 similar to that reported with years of conventional SIT. This study suggests that simply augmenting the blocking IgG/IgE ratio may reverse allergy.

Immunoreactivity of the Cold Water Extract of Royal Sun Culinary-Medicinal Mushroom, Agaricus brasiliensis Strain KA21 (Agaricomycetes), Assessed by Immunoglobulin Preparations for Intravenous Injection.

The royal sun medicinal mushroom, Agaricus brasiliensis, is a health food material that helps to improve quality of life. A. brasiliensis has long been used as a tea by extraction with cold and hot water. Our group has been investigating the immunopharmacological activities of the A. brasiliensis KA21 strain, which is cultivated outdoors. We prepared cold water (AgCWE) and hot water (AgHWE) extracts of this strain. AgCWE contained a larger proportion of proteins, including enzymes, and showed a brownish color during the extraction process. By contrast, chemical and immunochemical analyses revealed that AgHWE contained large amounts of ß-1,3-/1,6-glucans. In an attempt to elucidate the immunochemical characteristics of AgCWE, reactivities to immunoglobulin (Ig) preparations for intravenous injection were analyzed and compared with standard materials. To characterize brownish high-molecular weight components, standard phenol compounds such as caffeic acid (CA), trans-ferulic acid (FA), and coumaric acid (CouA) were polymerized to brownish polymerized polyphenols (PPPs) (i.e., polymerized CA, polymerized FA, and polymerized CouA) by laccase or peroxidase. The results obtained revealed that intravenous Ig reacted with all PPPs and PPPs cross-reacted with AgCWE and AgHWE. The isotype of the anti-PPP antibody was found to be IgG1, in contrast to that of the ß-glucan antibody, which was mainly IgG2. These results strongly suggest that A. brasiliensis extracts contain immunoreactive components against various classes of Igs.

Protective Effects of Ophiocordyceps lanpingensis on Glycerol-Induced Acute Renal Failure in Mice.

OBJECTIVE: Oxidative stress and immune response are associated with acute renal failure (ARF). Ophiocordyceps lanpingensis (OL) might be an antioxidant and immunopotentiator. In this study, we explored the protective effects of OL on glycerol-induced ARF. METHODS: Male mice were randomly divided into four groups, specifically, glycerol-induced ARF model group, low-dose OL-treated group (1.0 g/kg/d), high-dose OL-treated group (2.0 g/kg/d), and control group. Renal conditions were evaluated using kidney index, serum creatinine (Cr), blood urea nitrogen (BUN), and histological analysis. Rhabdomyolysis was monitored using creatine kinase (CK) level. Oxidative stress was determined using kidney tissue glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) levels. Immune status was evaluated using immune organ indices and immunoglobulin G (IgG) level. RESULTS: OL could relieve renal pathological injury and decrease the abnormal levels of kidney index, serum Cr, CK, BUN, and MDA, as well as increase the immune organ indices and the levels of IgG, GSH, and SOD. Treatment with a high dose of OL had more positive therapeutic effects on ARF than using a low dose of OL. CONCLUSION: OL could ameliorate renal dysfunction in glycerol-induced ARF in mice by inhibiting oxidative stress and enhancing immune response.

Accurate quantification of 5 German cockroach (GCr) allergens in complex extracts using multiple reaction monitoring mass spectrometry (MRM MS).

BACKGROUND: German cockroach (GCr) allergen extracts are complex and heterogeneous products, and methods to better assess their potency and composition are needed for adequate studies of their safety and efficacy. OBJECTIVE AND METHODS: The objective of this study was to develop an assay based on liquid chromatography and multiple reaction monitoring mass spectrometry (LC-MRM MS) for rapid, accurate, and reproducible quantification of 5 allergens (Bla g 1, Bla g 2, Bla g 3, Bla g 4, and Bla g 5) in crude GCr allergen extracts. RESULTS: We first established a comprehensive peptide library of allergens from various commercial extracts as well as recombinant allergens. Peptide mapping was performed using high-resolution MS, and the peptide library was then used to identify prototypic and quantotypic peptides to proceed with MRM method development. Assay development included a systematic optimization of digestion conditions (buffer, digestion time, and trypsin concentration), chromatographic separation, and MS parameters. Robustness and suitability were assessed following ICH (Q2 [R1]) guidelines. The method is precise (RSD < 10%), linear over a wide range (r > 0.99, 0.01-1384 fmol/µL), and sensitive (LLOD and LLOQ <1 fmol/µL). Having established the parameters for LC-MRM MS, we quantified allergens from various commercial GCr extracts and showed considerable variability that may impact clinical efficacy. CONCLUSIONS AND CLINICAL RELEVANCE: Our data demonstrate that the LC-MRM MS method is valuable for absolute quantification of allergens in GCr extracts and likely has broader applicability to other complex allergen extracts. Definitive quantification provides a new standard for labelling of allergen extracts, which will inform patient care, enable personalized therapy, and enhance the efficacy of immunotherapy for environmental and food allergies.

Transferon™, a peptide mixture with immunomodulatory properties is not immunogenic when administered with various adjuvants.

Transferon, a human dialyzable leukocyte extract (hDLE), is a biotherapeutic that comprises a complex mixture of low-molecular-weight peptides (< 10 kDa) and is used to treat diseases with an inflammatory component. Some biotherapeutics, including those composed of peptides, can induce anti-drug antibodies (ADA) that block or diminish their therapeutic effect. Nevertheless, few studies have evaluated peptide-derived drug immunogenicity. In this study, the immunogenicity of Transferon was examined in a murine model during an immunization scheme using the following adjuvants: Al(OH)3, incomplete Freund's adjuvant (IFA), or Titermax Gold. The inoculation scheme entailed three routes of administration (intraperitoneal, Day 1; subcutaneous, Day 7; and intramuscular, Day 14) using 200 µg Transferon/inoculation. Serum samples were collected on Day 21. Total IgG levels were quantitated by affinity chromatography, and specific antibodies against components of Transferon were analyzed by dot-blot and ELISA. Ovalbumin (OVA, 44 kDa) and peptides from hydrolyzed collagen (PFHC, < 17 kDa) were used as positive and negative controls, respectively, in the same inoculation scheme and analyses for Transferon. OVA, PFHC, and Transferon increased total IgG concentrations in mice. However, only IgG antibodies against OVA were detected. Based on the results, it is concluded that Transferon does not induce generation of specific antibodies against its components in this model, regardless of adjuvant and route of administration. These results support the safety of Transferon by confirming its inability to induce ADA in this animal model.

Fungal raw materials used to produce allergen extracts.

OBJECTIVE: To review the topic of fungal raw materials used for the production of allergen extracts and the associated challenges and highlight candidate areas for development before standardized fungal allergen extracts can be commercially produced. DATA SOURCES: A PubMed search was performed using focused keywords and combined with a review of regulatory documents and industry guidelines. Several books on mycology also were consulted. STUDY SELECTIONS: The information obtained through the literature, books, and industry was scrutinized and combined with personal experience and expertise to write this article. RESULTS: Fungi are complex ubiquitous organisms on Earth. They are beneficial and detrimental for humans. Fungi can cause hypersensitivity reactions, including types I, III, and IV. The procurement of fungal raw materials to prepare allergen extracts for diagnosis and possible allergen immunotherapy is complex owing to the intrinsic nature of fungi and their complex genome. Allergen manufacturers produce allergen extracts with variable qualitative and quantitative compositions, which can lead to unpredictable clinical outcomes. CONCLUSION: The clinician should be aware of the factors responsible for the qualitative and quantitative compositions of fungal allergen extracts and the reasons that currently preclude their standardization. Scientific advances and collaboration and cooperation between allergen manufacturing companies and regulatory agencies are necessary to improve the quality and consistency of fungal extracts. Moreover, clinicians should understand the limitations of currently available fungal extracts.

Efficacy of Sublingual Immunotherapy with Dermatophagoides Pteronyssinus: A Real-life Study.

Sublingual allergen immunotherapy (SLIT) is considered to be safer and more convenient than subcutaneus immunotherapy. SLIT trials with house dust mites involving patients with allergic rhinitis (AR) and asthma reported discordant results. The aim of the study was to investigate the clinical efficacy and safety of SLIT with Dermatophagoides pteronyssinus (D.pt) extract produced in Serbia and patient's satisfaction through open-label trial. Adult patients with allergic rhinitis were randomized into two groups: one received drugs and SLIT, while other received only drugs. Symptom score (SS), medication score (MS) and cumulative score (CS), skin prick tests (SPT) and serum level of D. pt specific IgE were assessed. One year after, the patients were re-evaluated. In total, 61 patients were enrolled in the study, but 52 of them were analyzed at the end of the year. CS (29.3%, p<0.001) and MS (54.3%, p<0.05) reduced significantly in the SLIT group. There was a significant improvement of MS and CS in the SLIT compared to control group (p<0.001 and p<0.05 respectively). There was no significant improvement of SS as well as specific slgE. Patients in the SLIT group were more satisfied with treatment (p<0.001). The incidence of mild adverse reaction was 38.4%. Specific lgG was not done. One year SLIT with D.pt extract was clinically efficient treatment in AR patients.

Improved Diagnosis of the Polysensitized Allergic Rhinitis Patients Using Component Resolved Diagnosis Method.

Allergy diagnosis needs to be improved in polysensitized patients due to the existence of possible confounding factors in this type of patients. Component resolved diagnosis (CRD) is a new concept in the investigation of polysensitized patients. The aim of this study was to evaluate if the utilization of ImmunoCAP ISAC improve the diagnosis of the polysensitized allergic rhinitis patients. Skin prick test (SPT) to 58 crude allergen extracts and CRD (ImmunoCAP ISAC) were carried out for 5 polysensitized allergic rhinitis patients. Two patients had a shellfish allergy and avoidance of shellfish was the only way to prevent an allergic reaction. In contrast, although the remaining three patients had low risk for shellfish allergy, but they were the best candidates for immunotherapy using mite extracts. CRD and particularly ImmunoCAP ISAC have proven to be a valuable diagnostic tool in polysensitized patients. ImmunoCAP ISAC helps refine the individual patient's sensitization profile and predict the potential risk of allergic reactions and improve the selection of patients for immunotherapy.

[Experimental modeling of nucleoprotein disposal disorders in systemic lupus erythematosus].

The objective of this research was to adapt the experimental model simulating the nucleoprotein disposal disorders in systemic lupus erythematosus (SLE) for further study of its extracorporeal correction, as well as to assess validity of the model by short-term experiment. Twenty to female Wistar rats were intraperitoneally injected with the chromatin-containing extract from bovine liver followed by intravenous administration of anti-DNA antibodies derived from SLE patients. After these procedures plasma concentrations of anti-dsDNA, circulating immune complexes and DNA became sharply increased, together with distinct elevation of leukocytes. On the contrary, changes in erythrocytes, platelets, total protein concentration, creatinine, asparagine and alanine aminotransferase activities, as well as blood coagulation time were changed insignificantly. Using direct immunofluorescence of cryosections, we detected human IgG deposition in rat kidneys treated in accordance with the simulation protocol. Thus, our model reproduces essential DNA disposal disorders in SLE without any animal death or the life-threatening changes in examined markers during short-term experiment.