Prevalence of immunoglobulin E against cross-reactive carbohydrate determinants and the impact of a blocker in canine sera.

OBJECTIVE: Cross-reactive carbohydrate determinants (CCD) cause multiple positive results in seasonal in vitro allergy tests. False positive/clinically irrelevant results have been identified due to the binding of immunoglobulin E against CCD (anti-CCD IgE) when testing for pollen allergens. The aim of this study was to investigate the prevalence of polysensitised serum samples and evaluate the impact of a CCD inhibitor/blocker in seasonal allergy test results. MATERIAL AND METHODS: A total of 4614 canine serum samples, submitted from July 2017 to June 2018 for seasonal in vitro allergy test via ELISA Fc-Ε receptor technology, were studied. Samples were grouped into polysensitised (group A) and non-polysensitised (group B). Polysensitised samples were retested after adding a modified glycoprotein plant extract (blocker). To determine the impact of the blocker for each allergen, results prior and post blocking were investigated in 96 randomly selected samples. RESULTS: Polysensitisation to seasonal allergens was diagnosed in 818 (17.7 %) serum samples. The blocker eliminated the binding of anti-CCD IgE to allergen extracts (49 %) or suppressed the reaction classes (40 %) which are indicative of the presence of clinically relevant IgE. Negative reactions after blocking were less common when testing for antibodies against a mixture of 6-grass mix (29 %), rye (22 %), and sheep sorrel (20 %) in comparison to nettle (82 %), willow (70 %), birch-hazel (65 %), mugwort-ragweed (63 %) and English plantain (57 %). CONCLUSION AND CLINICAL RELEVANCE: Blocking should be used in the case of polysensitized results to improve the quality of seasonal in vitro allergy tests and avoid the use of allergen-specific immunotherapy (ASIT) for clinically irrelevant allergens.

[Prevalance of immunoglobulin E against cross-reactive carbohydrate determinants (CCD) and impact of a blocker in seasonal allergy tests]. / Prävalenz von Immunglobulin E gegen kreuzreagierende Kohlenhydrat-Seitenketten (CCD) und die Wirkung eines Blockers im saisonalen In-vitro-Allergietest.

OBJECTIVES: Cross-reactive carbohydrate determinants (CCD) cause false positive/clinically irrelevant results in seasonal in vitro allergy tests due to the binding of immunoglobulin IgE against CCD (anti-CCD IgE).There is no study regarding the presence of this phenomen in cats. The aim of this study was to investigate the prevalence of polysensitization in serum samples and evaluate the impact of a CCD inhibitor/blocker in multi-positive seasonal allergy test results in cats. MATERIALS AND METHODS: A total of 472 feline sera, submitted from July 2017 to June 2018 for seasonal in vitro allergy test via ELISA Fc-Ε receptor technology, were studied. Samples were grouped into polysensitized (group A) and non-polysensitized (group B). Polysensitized samples (A) were retested after adding a modified glycoprotein plants extract (blocker). To determine the impact of the blocking to each allergen, the results in 48 randomly selected samples in cats prior- and post-blocking were investigated. RESULTS: Polysensitization to seasonal allergens was diagnosed in 137 (29 %) samples. No discrepancy in presence of polysensitization was seen in different seasons. Blocking eliminated the binding of anti-CCD IgE and produced either negative test results (49 %) or a decrease of 1-4 reaction classes (41 %) which is indicative of the simultaneous presence of clinically relevant allergen specific IgE. Total negative reactions after blocking were less common in 6-grass mix (31 %), rye (23 %) and sheep sorrel (25 %) in comparison to willow und birch-hazel (67 %), mugwort-ragweed und nettle (65 %), as well as English plantain (54 %). CONCLUSION AND CLINICAL RELEVANCE: In order to improve the quality of seasonal in vitro allergy test, blocking should be employed in cases of polysensitized results resulting in an avoidance of the administration of non-offending allergens during allergen-specific immunotherapy (ASIT).

Carriage of meticillin-resistant staphylococci between humans and animals on a small farm.

BACKGROUND: Meticillin-resistant staphylococci (MRS) are pathogens of increasing importance to human and animal health worldwide. Transmission of meticillin-resistant Staphylococcus aureus (MRSA) between animals and humans has been well documented. By contrast, information about transmission of other Staphylococcus spp. is limited. HYPOTHESIS/OBJECTIVES: The aim of this study was to screen animals and humans on a small farm for nasal carriage of MRS and to assess interspecies exchange. METHODS: After detection of MRSA in a lung sample of a deceased cat, which lived on a small mixed farm, nasal swabs were taken within two weeks, four and 16 months from other animals of various species and humans living on the farm. Swabs were cultured for MRS which were then characterized molecularly. RESULTS: MRSA and meticillin-resistant coagulase negative staphylococci (MRCoNS), including Staphylococcus haemolyticus, S. epidermidis and S. fleurettii, were isolated from humans and different animal species. Typing of the MRS revealed isolates with the same characteristics in different human and animal hosts. CONCLUSIONS AND CLINICAL IMPORTANCE: To the best of the authors' knowledge, this is the first report of carriage of both MRSA and MRCoNS among humans and various animals within a shared environment. The detection of strains with indistinguishable molecular characteristics strongly suggested transmission of these MRS between the various animal species and humans.

Lactoferrin, a glycoprotein with immunomodulatory and mast cell stabilising properties, in skin of horses suffering from Culicoides hypersensitivity.

Lactoferrin (LF), a glycogen of the transferrin family with anti-bacterial and immunomodulatory properties, is expressed in various secretions and tissues. Cutaneous LF serves as a mast cell stabilising compound, modulates T cell activity and is found during IgE-mediated late phase reactions at allergen challenged sites. Culicoides hypersensitivity (CHS) in horses is a common IgE-mediated allergic dermatitis, characterised by an early and late phase cutaneous reaction upon allergen challenge. The aim of the study presented here was to examine whether LF mRNA expression in skin biopsies from horses affected by CHS prior to and 4h following intradermal challenge with a commercial C. nubeculosus extract is modified in comparison to skin biopsies from non-affected horses. In order to obtain reliable data, real time PCR was performed and genes of interest were normalized using three different housekeeping genes, beta-actin, GAPDH, beta-2-microglobulin. In comparison to non-affected horses, higher variation in LF mRNA levels both prior to and post-intradermal challenge with C. nubeculosus extract was seen in horses affected by CHS. However, the statistical analysis demonstrated that LF mRNA expression was not significantly different between CHS affected and non-affected horses prior to intradermal challenge with C. nubeculosus extract. Intradermal injection of C. nubeculosus extract did not result in local upregulation of LF mRNA at 4h post-injection. LF mRNA expression was therefore not significantly different pre- or post-intradermal challenge with C. nubeculosus extract in either group. Our data indicate that clinically normal skin of horses affected by CHS is not characterized by modified maintenance levels of LF mRNA. In contrast to human skin allergen challenged sites, LF mRNA levels in horses affected by CHS are not significantly different to that of control sites at 4h post-injection of C. nubeculosus extract.

Increased interleukin-1beta mRNA expression in skin biopsies of horses with Culicoides hypersensitivity following challenge with Culicoides nubeculosus extract.

Interleukin-1beta (IL-1beta) is a primary cytokine of the skin that has a pivotal role in keratinocyte differentiation, epidermal wound healing and host defense. Pathological increase of cutaneous IL-1beta is associated with edema formation, epidermal hyperproliferation and atopic dermatitis in humans. However, in horses the role of cutaneous IL-1beta in edema formation and allergic skin disease has not been characterised so far. Particularly in Culicoides hypersensitivity (CHS), intradermal injection of Culicoides extract may be associated with enhanced transcription of local IL-1beta. To examine the mRNA expression of IL-1beta and its receptor antagonist IL-1RA in the skin of horses, biopsy specimens of horses affected and non-affected by CHS prior and following intradermal challenge with a commercial C. nubeculosus extract were examined. Our hypothesis was that cutaneous IL-1beta mRNA was significantly upregulated in horses with CHS in response to Culicoides allergen. Biopsies were taken from sites prior to and 4 h following intradermal challenge with C. nubeculosus extract. In order to obtain reliable data, real time PCR was performed and genes of interest were normalized using three different housekeeping genes, beta-actin, GAPDH, beta-2-microglobulin. No significant difference was detected in non-challenged cutaneous IL-1beta mRNA and IL-1RA mRNA levels between CHS affected and non-affected horses. Intradermal injection of C. nubeculosus extract resulted in local upregulation of IL-1beta mRNA both in horses with typical history, characteristic clinical signs for CHS and a positive intradermal skin test (IDT), and non-affected horses with a negative IDT. However, the difference in prior and post challenged site IL-1beta mRNA levels only reached statistical significance in the affected horses (p=0.01 versus 0.7). In contrast, IL-1RA mRNA levels did not demonstrate any modification following intradermal injection with C. nubeculosus in either group. In contrast to human atopic dermatitis, clinically normal skin of horses affected by CHS is not characterized by increased maintenance levels of IL-1beta mRNA. C. nubeculosus stimulates local IL-1beta transcription in all horses independent from disease, but the extent of upregulation from basal levels only reaches statistical significance in horses affected by CHS and active stage of disease.