Serological Cross-Reactions between Expressed VP2 Proteins from Different Bluetongue Virus Serotypes.

Bluetongue (BT) is a severe and economically important disease of ruminants that is widely distributed around the world, caused by the bluetongue virus (BTV). More than 28 different BTV serotypes have been identified in serum neutralisation tests (SNT), which, along with geographic variants (topotypes) within each serotype, reflect differences in BTV outer-capsid protein VP2. VP2 is the primary target for neutralising antibodies, although the basis for cross-reactions and serological variations between and within BTV serotypes is poorly understood. Recombinant BTV VP2 proteins (rVP2) were expressed in Nicotiana benthamiana, based on sequence data for isolates of thirteen BTV serotypes (primarily from Europe), including three 'novel' serotypes (BTV-25, -26 and -27) and alternative topotypes of four serotypes. Cross-reactions within and between these viruses were explored using rabbit anti-rVP2 sera and post BTV-infection sheep reference-antisera, in I-ELISA (with rVP2 target antigens) and SNT (with reference strains of BTV-1 to -24, -26 and -27). Strong reactions were generally detected with homologous rVP2 proteins or virus strains/serotypes. The sheep antisera were largely serotype-specific in SNT, but more cross-reactive by ELISA. Rabbit antisera were more cross-reactive in SNT, and showed widespread, high titre cross-reactions against homologous and heterologous rVP2 proteins in ELISA. Results were analysed and visualised by antigenic cartography, showing closer relationships in some, but not all cases, between VP2 topotypes within the same serotype, and between serotypes belonging to the same 'VP2 nucleotype'.

Serological Detection of SARS-CoV-2 Antibodies in Naturally-Infected Mink and Other Experimentally-Infected Animals.

The recent emergence of SARS-CoV-2 in humans from a yet unidentified animal reservoir and the capacity of the virus to naturally infect pets, farmed animals and potentially wild animals has highlighted the need for serological surveillance tools. In this study, the luciferase immunoprecipitation systems (LIPS), employing the spike (S) and nucleocapsid proteins (N) of SARS-CoV-2, was used to examine the suitability of the assay for antibody detection in different animal species. Sera from SARS-CoV-2 naturally-infected mink (n = 77), SARS-CoV-2 experimentally-infected ferrets, fruit bats and hamsters and a rabbit vaccinated with a purified spike protein were examined for antibodies using the SARS-CoV-2 N and/or S proteins. From comparison with the known neutralization status of the serum samples, statistical analyses including calculation of the Spearman rank-order-correlation coefficient and Cohen's kappa agreement were used to interpret the antibody results and diagnostic performance. The LIPS immunoassay robustly detected the presence of viral antibodies in naturally infected SARS-CoV-2 mink, experimentally infected ferrets, fruit bats and hamsters as well as in an immunized rabbit. For the SARS-CoV-2-LIPS-S assay, there was a good level of discrimination between the positive and negative samples for each of the five species tested with 100% agreement with the virus neutralization results. In contrast, the SARS-CoV-2-LIPS-N assay did not consistently differentiate between SARS-CoV-2 positive and negative sera. This study demonstrates the suitability of the SARS-CoV-2-LIPS-S assay for the sero-surveillance of SARS-CoV-2 infection in a range of animal species.

Selection for environmental variance of litter size in rabbits involves genes in pathways controlling animal resilience.

BACKGROUND: Environmental variance (VE) is partially under genetic control, which means that the VE of individuals that share the same environment can differ because they have different genotypes. Previously, a divergent selection experiment for VE of litter size (LS) during 13 generations in rabbit yielded a successful response and revealed differences in resilience between the divergent lines. The aim of the current study was to identify signatures of selection in these divergent lines to better understand the molecular mechanisms and pathways that control VE of LS and animal resilience. Three methods (FST, ROH and varLD) were used to identify signatures of selection in a set of 473 genotypes from these rabbit lines (377) and a base population (96). A whole-genome sequencing (WGS) analysis was performed on 54 animals to detect genes with functional mutations. RESULTS: By combining signatures of selection and WGS data, we detected 373 genes with functional mutations in their transcription units, among which 111 had functions related to the immune system, stress response, reproduction and embryo development, and/or carbohydrate and lipid metabolism. The genes TTC23L, FBXL20, GHDC, ENSOCUG00000031631, SLC18A1, CD300LG, MC2R, and ENSOCUG00000006264 were particularly relevant, since each one carried a functional mutation that was fixed in one of the rabbit lines and absent in the other line. In the 3'UTR region of the MC2R and ENSOCUG00000006264 genes, we detected a novel insertion/deletion (INDEL) variant. CONCLUSIONS: Our findings provide further evidence in favour of VE as a measure of animal resilience. Signatures of selection were identified for VE of LS in genes that have a functional mutation in their transcription units and are mostly implicated in the immune response and stress response pathways. However, the real implications of these genes for VE and animal resilience will need to be assessed through functional analyses.

Evaluation on two types of paramyosin vaccines for the control of Haemaphysalis longicornis infestations in rabbits.

BACKGROUND: Haemaphysalis longicornis is an obligate hematophagous ectoparasite that transmits a variety of pathogens causing life-threatening diseases in humans and animals. Paramyosin (Pmy) is not only an invertebrate-specific myofibrillar protein but also an important immunomodulatory protein. Therefore, it is one of the ideal candidate antigens for vaccines. METHODS: We conducted two vaccine trials to evaluate the protective efficacy of Pmy recombinant protein (rPmy) and peptide vaccine (KLH-LEE). Each rabbit was immunized with three doses of rPmy or KLH-LEE adjuvanted with Freund's complete/incomplete at 500 µg/dose at 2-week intervals before challenge with 40 female H. longicornis/rabbit. PBS plus adjuvant, Trx or KLH was used as control group. The antibodies of rabbits were detected by ELISA. Then, female ticks were fed on the rabbits until detachment. RESULTS: ELISA results showed that both vaccines induced rabbits to produce antibodies. Compared with the Trx group, the engorgement weight, oviposition and hatchability of the rPmy group decreased by 8.87%, 26.83% and 38.86%, respectively. On the other hand, engorgement weight, oviposition and hatchability of female ticks in the KLH-LEE group correspondingly resulted in 27.03%, 53.15% and 38.40% reduction compared with that of the KLH group. Considering the cumulative effect of vaccination on the evaluated parameters, results showed 60.37% efficacy of the rPmy vaccine formulation and 70.86% efficacy in the KLH-LEE group. CONCLUSIONS: Pmy and particularly epitope LEE have potential for further development of an effective candidate vaccine to protect the host against tick infection. GRAPHIC ABSTARCT.

Rabbit Elastase Aneurysm: Imaging and Histology Correlates for Inflammation and Healing.

OBJECTIVE: Aneurysmal subarachnoid hemorrhage remains a devastating event with poorly understood pathophysiology. Previous studies have suggested that aneurysm wall inflammation may play a part in the development and potential rupture of aneurysms. The rabbit elastase aneurysm model is a well-established model, which produces aneurysms closely mimicking human cerebral aneurysms in flow dynamics and histopathology. The primary aim of this study was to correlate inflammatory changes after aneurysm formation using sequential vessel wall imaging with histopathologic analysis. A secondary aim was to evaluate the potential effect of gender and anti-inflammatory treatment with aspirin on this inflammatory response. METHODS: Twenty-seven New Zealand rabbits underwent surgery to create an aneurysm using elastase infusion at the right common carotid artery origin. Vessel wall imaging and histopathologic analysis was obtained at different time points after aneurysm creation. The rabbits were also randomized by gender and to treatment groups with or without aspirin. RESULTS: Histopathologic analysis revealed 3 distinct phases after aneurysm formation. These phases were an initial inflammatory phase, followed by a regeneration phase, and finally a connective tissue deposition phase. Vessel wall imaging demonstrated 2 distinct imaging patterns. No appreciable differences were seen in histology or imaging when comparing gender or treatment with aspirin. CONCLUSIONS: Inflammatory changes induced by the rabbit elastase aneurysm model can be correlated with histopathologic findings and observed on noninvasive vessel wall imaging. This may provide a method to study the inflammatory pathway as it pertains to aneurysmal development and subsequent rupture.

Molecular stability of the rabbit and chicken egg yolk immunoglobulins.

Antibodies play a vital role in a variety of applications from diagnostics, imaging, and therapeutics. The stability of antibodies is one of the most important key attributes for its prolonged activity and functionality. Here, we demonstrate a detailed comparative study of the molecular stability of the rabbit immunoglobulin G (IgG) and chicken egg yolk immunoglobulin Y (IgY) at different pH, temperatures, and time points. The molecular stability of IgG and IgY is compared on the basis of its binding activity and conformational changes. The optimum temperature and time were found to be 30 °C, and 37 °C, up to 8 h for both IgY and IgG antibodies. With increasing temperature and time, IgG antibodies were found to be significantly less stable in comparison to IgY antibodies. IgY is almost twenty-fold more stable than IgG at 60° C for up to 8 hours. The extra domain present in the heavy chain of IgY plays a significant role in providing more molecular stability as compared to IgG under the above-mentioned experimental conditions. The results show that, as compared with IgG, the IgY are more stable, are less expensive to make in high yield and exhibit minimal conformational changes and hence are cost effective for use in a diverse array of purposes.

Aspergillus awamori positively impacts the growth performance, nutrient digestibility, antioxidative activity and immune responses of growing rabbits.

In this study, we explored the effects of dietary administration of Aspergillus awamori on the growth, intestinal histomorphology, immune parameters and antioxidant enzyme activity of growing rabbits. The rabbits of 5 weeks of age (body weight, 855 ± 5.53 g) were allotted into four groups (12 rabbits per group) and fed four experimental diets enriched with A. awamori at 0, 50, 100 and 150 mg per kg diet for 8 weeks. After the feeding trial, an equal number of male APRI rabbits (3 months old; 4 groups, 5 rabbits per group) were housed in metabolism cages and fed the test diets for 14 days for the digestibility trial. All A. awamori-enriched diets induced a significant increase in the average body weight and weight gain of rabbits and significantly decreased the feed conversion ratio. In rabbits fed with A. awamori at 100 or 150 mg per kg diet, protein, lipid and fibber digestibility coefficients significantly increased compared with the control group. Intestinal villi measurements (length and thickness) were also enhanced in all rabbits fed with A. awamori. Adding A. awamori as a food supplement generally did not affect rabbit haematology and blood biochemistry values; however, at 150 mg per kg diet, it significantly increased the levels of haemoglobin and total protein, as well as red blood cell count. Furthermore, all enriched diets significantly increased rabbits' phagocytic activity and their phagocytic index. Rabbits fed with A. awamori also showed decreased malondialdehyde but increased catalase activity. In conclusion, A. awamori administered as feed supplement at 100-150 mg per kg of growing rabbits' diet enhances their growth, intestinal health and nutrient digestibility, and it raises the levels of their immune and antioxidative responses.

In vivo Induction of Functional Inhibitory IgG Antibodies by a Hypoallergenic Bet v 1 Variant.

Allergic sensitization to the major allergen Bet v 1 represents the dominating factor inducing a vast variety of allergic symptoms in birch pollen allergic patients worldwide, including the pollen food allergy syndrome. In order to overcome the huge socio-economic burden associated with allergic diseases, allergen-specific immunotherapy (AIT) as a curative strategy to manage the disease was introduced. Still, many hurdles related to this treatment exist making AIT not the patients' first choice. To improve the current situation, the development of hypoallergen-based drug products has raised attention in the last decade. Herein, we investigated the efficacy of the novel AIT candidate BM4, a hypoallergenic variant of Bet v 1, to induce treatment-relevant cross-reactive Bet v 1-specific IgG antibodies in two different mammals, Wistar rats and New Zealand White rabbits. We further analyzed the cross-reactivity of BM4-induced Wistar rat antibodies with the birch pollen-associated food allergens Mal d 1 and Cor a 1, and the functional capability of the induced antibodies to act as IgE-blocking IgG antibodies. Enzyme-linked immunosorbent assay (ELISA) was used to determine the titers of rat IgG1, IgG2a, IgG2b, and IgE, as well as rabbit IgG and IgE antibodies. To address the functional relevance of the induced IgG antibodies, the capacity of rat sera to suppress binding of human IgE to Bet v 1 was investigated by using an inhibition ELISA and an IgE-facilitated allergen-binding inhibition assay. We found that the treatment with BM4 induced elevated Bet v 1-specific IgG antibody titers in both mammalian species. In Wistar rats, high BM4-specific IgG1, IgG2a, and IgG2b titers (104 to 106) were induced, which cross-reacted with wild-type Bet v 1, and the homologous allergens Mal d 1 and Cor a 1. Rat allergen-specific IgG antibodies sustained upon treatment discontinuation. Sera of rats immunized with BM4 were able to significantly suppress binding of human IgE to the wild-type allergens and CD23-mediated human IgE-facilitated Bet v 1 binding on B cells. By contrast, treatment-induced IgE antibody levels were low or undetectable. In summary, BM4 induced a robust IgG immune response that efficiently blocked human IgE-binding to wild-type allergens, underscoring its potential therapeutic value in AIT.

B and T lymphocytes in rabbits change according to the sex and throughout the year.

The European rabbit (Oryctolagus cuniculus) is a good model in biomedicine used in research on several human diseases. The reference values of B and T cells and their subpopu- lations are very important to understand how the adaptive immune system is responding to infectious agents. The aim of this study was to determine values of B and T cells and their subpopulations in Polish mixed-breed rabbits, considering seasons of the year and sex. The study was performed on 200 Polish mixed-breed rabbits and the percentage of B and T lymphocytes was measured cytometrically using mouse anti-rabbit antibodies. The study revealed that the season of the year and sex of the animals affected the percentage of B- and T-cells and their subpopulations in peripheral blood. Statistically significant values of CD19+ B-cells in spring and autumn, of T CD5+ cells in spring and winter, of T CD4+ in spring, summer, autumn and winter, of T CD8+ in winter and of T CD25+ in spring were noted. Generally the highest values were found mainly in warm part of the year, while the lowest in colder months. A statistical significance was also observed between males and females - changes were found in T CD4+ and T CD25+ lymphocytes in spring, T CD8+ cells in winter and higher percentage was generally obtained in females than in males. The only exception was the T CD5+ subpopulation in which no differences were observed between the sexes and throughout the year. This is the first paper on adaptive immune system cell values in the European rabbit of domestic breeds.

The Anti-Neuron-Specific Enolase Antibody Induced Neuronal Cell Death in a Novel Fashion.

Suppression of ubiquitin proteasome pathway (UPP) and stimulation of caspase-3 are involved in neurodegeneration. Can UPP activators and caspase-3 inhibitors ameliorate neurodegeneration? Here, we found a novel neuronal cell death accompanied with UPP activation and caspase-3 inhibition. Recently, plasmalemmal neuron-specific enolase (NSE) has been identified as one of membrane targets of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2). 15d-PGJ2 induces neuronal apoptosis via activating caspase-3 and inactivating UPP, whereas the anti-NSE antibody inactivated caspase-3, activated UPP, and caused neuronal cell death. The anti-NSE antibody activated caspase-1 (pyroptosis marker), but not condense chromatin (apoptosis marker). The anti-NSE antibody declined intracellular level of ATP, which is not altered in pyroptosis. The intracellular level of calcium is elevated in necrosis and pyroptosis, but its chelator did not ameliorate the neurotoxicity of anti-NSE. Thiol antioxidants such as N-acetyl cysteine and glutathione reduced the neurotoxicity of 15d-PGJ2 but enhanced that of the anti-NSE antibody. The anti-NSE antibody incorporated propidium iodide into neurons through the disrupted plasma membrane, which are not observed in ferroptosis and autophagic cell death. Thus, the anti-NSE antibody induced neuronal cell death in a novel fashion distinguished from necrosis, necroptosis, apoptosis, pyroptosis, ferroptosis, and autophagic cell death.

Influence of allicin administration on reproductive efficiency, immunity and lipid peroxidation of rabbit does under high ambient temperature.

This study investigated the effect of daily oral administration with allicin levels (0, 5 and 10 mg/kg of female body weight), 30 days pre-insemination, on reproductive performance in vivo and in vitro, immunity, and oxidative stress of rabbit does under high ambient temperature. Niliparous NZW does (n = 105) were randomly divided into three groups (35 in each) treated with 0, 5 and 10 mg allicin dissolved in 2 ml distilled water, respectively, for 30 days pre-insemination. At the end of treatment (30 days), does were artificially inseminated with fresh diluted semen of 20 fertile NZW bucks. Reproductive performance and ovulatory response parameters were determined. Serum biochemicals, enzyme activity, immunoglobulins (IgG and IgM) and antioxidant status were determined on day 30 of treatment. Serum progesterone and prolactin were determined pre-insemination (30 days of treatment), on 15 days of pregnancy and 7 days post-partum. Results showed that both allicin levels increased live litter size, and bunny viability rat and litter size at birth and weaning. Allicin levels increased ovulation rate and improved embryo quality. Number of total follicles decreased only with 10 mg allicin. Progesterone increased pre-insemination, 15 days of pregnancy and 7 days post-partum progesterone by allicin levels. Prolactin pre-insemination and on day 7 post-partum increased with 10 mg allicin. Serum total proteins, albumin, globulin, IgG and IgM increased, while glucose, aspartate and alanine aminotransaminases, and thiobarbituric acid reaction decreased by both allicin levels. In conclusion, the mechanism by which allicin administration 30 days pre-insemination to improve the reproductive performance of rabbit does is based on that allicin can play an important role, as a natural exogenous antioxidant, increasing immune response and reducing lipid peroxidation.

Functional characterization of NLRX1 in rabbit during enterohemorrhagic Escherichia coli infection.

Nucleotide oligomerization domain (NOD) like receptor X1 (NLRX1) is a member of pattern recognition receptor, which has been linked to viral response, cancer, and inflammatory diseases. In this study, rabbit NLRX1 (rNLRX1) was firstly cloned from RK-13 cells, which protein contained a NACHT domain and seven LRRs. rNLRX1 was widely expressed in tissues of rabbits, and highly increased in liver, spleen, kidney, and colon after infected with enterohemorrhagic Escherichia coli (EHEC). Overexpression of rNLRX1 negatively regulated NF-κB signaling, and impaired the expression of pro-inflammatory cytokines and defensins. Moreover, deficient of rNLRX1 in RK-13 cells was performed to investigate the possible roles of rNLRX1. Upon EHEC stimulation, knockdown of rNLRX1 markedly enhanced NF-κB activation and downstream responsive cytokines (IL1ß and TNFα) and ß-defensins (DEFB114, DEFB124, and DEFB125). Furthermore, overexpression of rNLRX1 promoted the proliferation of EHEC, whereas knockdown of rNLRX1 inhibited its growth. Our study identified that rNLRX1 acts as a negative regulatory in anti-microbial responses after EHEC infection.

Early deviations in performance, metabolic and immunological indicators affect stayability in rabbit females.

The main purpose of this study was to find several early factors affecting stayability in rabbit females. To reach this goal, 203 females were used from their first artificial insemination to their sixth parturition. Throughout that period, 48 traits were recorded, considered to be performance, metabolic and immunological indicators. These traits were initially recorded in females' first reproductive cycle. Later, removed females due to death or culling and those that were non-removed were identified. A first analysis was used to explore whether it was possible to classify females between those reaching and those not reaching up to the mean lifespan of a rabbit female (the fifth reproductive) cycle using information from the first reproductive cycle. The analysis results showed that 97% of the non-removed females were classified correctly, whereas only 60% of the removed females were classified as animals to be removed. The reason for this difference lies in the model's characteristics, which was designed using early traits and was able to classify only the cases in which females would be removed due to performance, metabolic or immunologic imbalances in their early lives. Our results suggest that the model defines the necessary conditions, but not the sufficient ones, for females to remain alive in the herd. The aim of a second analysis was to find out the main early differences between the non-removed and removed females. The live weights records taken in the first cycle indicated that the females removed in their first cycle were lighter, while those removed in their second cycle were heavier with longer stayability (-203 and +202 g on average, respectively; P < 0.05). Non-removed females showed higher glucose and lower beta-hydroxybutyrate concentrations in the first cycle than the removed females (+4.8 and -10.7%, respectively; P < 0.05). The average lymphocytes B counts in the first cycle were 22.7% higher in the non-removed females group (P < 0.05). The females removed in the first reproductive cycle presented a higher granulocytes/lymphocytes ratio in this cycle than those that at least reached the second cycle (4.81 v. 1.66; P < 0.001). Consequently, non-removed females at sixth parturition offered adequate body development and energy levels, less immunological stress and a more mature immune function in the first reproductive cycle. The females that deviated from this pattern were at higher risk of being removed from the herd.

Effect of colibacillosis on the immune response to a rabbit viral haemorrhagic disease vaccine.

Viral haemorrhagic disease (VHD) and colibacillosis are common diseases in rabbits that cause economic losses worldwide. The effect of colibacillosis on the immune response of vaccinated rabbits against rabbit haemorrhagic disease virus (RHDV) was studied. Four groups (G1-G4) were included. G1 was the negative control group; G2 was the RHDV vaccine group; G3 was the E. coli-infected group; and G4 was the E. coli-infected + RHDV vaccine group. The E. coli infection and RHDV vaccination were simultaneously performed, with another previous infection, 3 days before vaccination. At 28 days post-vaccination (PV), the rabbits (G2-G4) were challenged intramuscularly with 0.5 ml of RHDV at a dose of 103 50% median lethal dose (LD50)/rabbit. The rabbits were observed for clinical signs, body weight gain and mortality rates. Tissue, blood, serum, and faecal samples and rectal swabs were collected at 3, 5, 7, 14, 21 and 28 days PV. Significant clinical signs and mortality and a decrease in BW were observed in the infected + RHDV vaccine group. On the 3rd day post-infection (PI), compared with all the other groups, the vaccinated group (G2) had significantly upregulated hepatic tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels; however, the infected + RHDV vaccine group had significantly higher intestinal levels of TNF-α and IL-6 than the other groups. Furthermore, E. coli infection in vaccinated rabbits led to immunosuppression, as shown by significant decreases (P < 0.05) in heterophil phagocytic activity, the CD4+/CD8+ ratio, and HI antibody responses to RHDV and a significant increase in the heterophil to lymphocyte (H/L) ratio. In conclusion, colibacillosis leads to immunosuppression involving a shift in the equilibrium of cytokines and reduced weight gain and mortality in vaccinated rabbits and could be a contributing factor in RHDV vaccination failure in rabbit farming.

Dietary addition of garlic straw improved the intestinal barrier in rabbits1.

Weanling rabbits frequently exhibit diarrhea or flatulence. Our experiment was conducted to investigate the effect of garlic straw on the performance and intestinal barrier of rabbits. Hyla rabbits (60 d, n = 160) with similar body weight were divided into 4 groups (4 replicates per group and 10 rabbits per replicate): fed a basal diet (control) or fed an experimental diet with 5%, 10%, or 15% garlic straw powder supplement. The results showed that the dietary addition of garlic straw increased significantly the average daily gain and average daily feed intake. Compared with the control, dietary addition of 10% and 15% garlic straw decreased significantly the death rate of rabbit. Rabbits in 10% garlic straw group had a higher secretory immunoglobulins A and immunoglobulins G concentration in jejunum and ileum than control while lower tumor necrosis factor α (TNFα) concentration in jejunum. Compared with the control, dietary addition of 10% garlic straw increased significantly genes expression of zonula occluden protein 2 (ZO2) in jejunum and ileum and mucin4 in ileum while did not alter the genes expression of junctional adhesion molecule 2 (JAM2), JAM3, ZO1, occluding, claudin1, mucin1, mucin6, and toll-like receptor 4 in jejunum and ileum and mucin4 in jejunum. In conclusion, dietary supplement of garlic straw modulates immune responses and enhances intestinal barrier, meanwhile inhibits the synthesis of pro-inflammatory cytokine of TNFα. Besides, our experiment offers positive evidence in improving rabbit health of garlic instead of antibiotics.

Epstein-Barr Virus and Monoclonal Gammopathy of Clinical Significance in Autologous Stem Cell Transplantation for Multiple Sclerosis.

INTRODUCTION: Autologous hematopoietic stem cell transplantation (AHSCT) with anti-thymocyte globulin (ATG) conditioning as treatment of active multiple sclerosis (MS) is rapidly increasing across Europe (EBMT registry data 2017). Clinically significant Epstein-Barr virus reactivation (EBV-R) following AHSCT with ATG for severe autoimmune conditions is an underrecognized complication relative to T-cell deplete transplants performed for hematological diseases. This retrospective study reports EBV-R associated significant clinical sequelae in MS patients undergoing AHSCT with rabbit ATG. METHODS: Retrospective data were analyzed for 36 consecutive MS-AHSCT patients at Kings College Hospital, London. All patients routinely underwent weekly EBV DNA polymerase chain reaction monitoring and serum electrophoresis for monoclonal gammopathy (MG or M-protein). EBV-R with rising Epstein-Barr viral load, M-protein, and associated clinical sequelae were captured from clinical records. RESULTS: All patients had evidence of rising EBV DNA-emia, including 7 who were lost to long-term follow-up, with a number of them developing high EBV viral load and associated lymphoproliferative disorder (LPD). Nearly 72% (n = 18/29) developed de novo MG, some with significant neurological consequences with high M-protein and EBV-R. Six patients required anti-CD20 therapy (rituximab) with complete resolution of EBV related symptoms. Receiver operating characteristics estimated a peak EBV viremia of >500 000 DNA copies/mL correlated with high sensitivity (85.5%) and specificity (82.5%) (area under the curve: 0.87; P = .004) in predicting EBV-R related significant clinical events. CONCLUSION: Symptomatic EBV reactivation increases risk of neurological sequelae and LPD in MS-AHSCT. We recommend regular monitoring for EBV and serum electrophoresis for MG in MS patients in the first 3 months post-AHSCT.

A review of complement sources used in serum bactericidal assays for evaluating immune responses to meningococcal ACWY conjugate vaccines.

Invasive meningococcal disease is rare and potentially devastating but often vaccine-preventable. Evaluation of meningococcal vaccine effectiveness is impractical owing to relatively low disease incidence; protection is therefore estimated using serum bactericidal antibody (SBA) assays. Original experiments on natural immunity established a titer of ≥4 as the correlate of protection for SBA assays using human complement (hSBA), but human complement is relatively difficult to obtain and standardize. Use of baby rabbit complement (rSBA assays), per standard guidelines for serogroups A and C, generally results in comparatively higher titers. Postlicensure effectiveness data for serogroup C conjugate vaccines support acceptance of rSBA titers ≥8 as the correlate of protection for this serogroup, but no thresholds have been formally established for serogroups A, W, and Y. Studies evaluating MenACWY-TT (Nimenrix®; Pfizer Inc, Sandwich, UK) immunogenicity have used both hSBA and rSBA assays, and ultimately suggest that rSBA may be more appropriate for these measurements.

Dietary addition of Artemisia argyi reduces diarrhea and modulates the gut immune function without affecting growth performances of rabbits after weaning1.

The present study was conducted to investigate the effect of Artemisia argyi on the production performance and intestinal barrier of rabbits. Weaned Hyla rabbits (30 d, n = 160) of similar body weight were divided into 4 groups (40 rabbits per treatment), and they were fed a control diet or fed an experimental diet supplemented with 3%, 6%, or 9% A. argyi. The results showed that the dietary supplementation with A. argyi did not affect the rabbits' food intake and body weight gain regardless of the inclusion level but decreased the diarrhea rate and diarrhea index (P < 0.05). Dietary addition of A. argyi increased the small intestine length and villus height/crypt depth, regardless of the inclusion level (P < 0.05). Compared with the control, the A. argyi supplementation increased the gene expression of zonula occludens 1 (ZO-1) and claudin 1 in all segments of the small intestine and regardless of the level of A. argyi (P < 0.05). In the duodenum, a dietary supplementation with 6% and 9% A. argyi increased the immunoglobulins A (IgA) content (P < 0.05). In the jejunum, the A. argyi supplementation decreased interleukin 2 (IL2) and IL6 content regardless of the inclusion level (P < 0.05). In the ileum, a 3% A. argyi addition decreased IL2 content, whereas a 6% A. argyi addition decreased IL6 content (P < 0.05). Furthermore, 6%-9% A. argyi supplementation increased the IgA content in the ileum (P < 0.05). In conclusion, dietary addition of A. argyi reduces diarrhea and modulates the gut immune function without affecting growth performances of rabbits.

Parallel adaptation of rabbit populations to myxoma virus.

In the 1950s the myxoma virus was released into European rabbit populations in Australia and Europe, decimating populations and resulting in the rapid evolution of resistance. We investigated the genetic basis of resistance by comparing the exomes of rabbits collected before and after the pandemic. We found a strong pattern of parallel evolution, with selection on standing genetic variation favoring the same alleles in Australia, France, and the United Kingdom. Many of these changes occurred in immunity-related genes, supporting a polygenic basis of resistance. We experimentally validated the role of several genes in viral replication and showed that selection acting on an interferon protein has increased the protein's antiviral effect.

Probiotic Clostridium butyricum Improves the Growth Performance, Immune Function, and Gut Microbiota of Weaning Rex Rabbits.

Probiotics could promote animal growth and enhance immune function. This study investigated the effects of Clostridium butyricum (CB) on the growth performance, intestinal immune, and gut microbiota of weaning rex rabbits. A total of 60 healthy female rabbits (5-month-old) were divided equally into four groups and mated on the same day: control group (CTRL, fed with basal feed), low-dose group (LDG, fed with basal feed + 1.0 × 103 CFU/g CB), middle-dose group (MDG, fed with basal feed + 1.0 × 104 CFU/g CB), and high-dose group (HDG, fed with basal feed + 1.0 × 105 CFU/g CB). Then, 30 weaning rex rabbits (35-day-old) were collected from each group for this experiment, and they were offered the same feeds as their mother. The results demonstrated that high-dose CB treatment significantly increased average daily weight gain of weaning rex rabbits. Further studies suggested that CB enhanced small intestinal digestive enzyme activity and improved mucosal morphology and antioxidant status. Supplemented with CB, small intestinal barrier function was maintained with the upregulation of mRNA levels of ZO-1, claudin, and occludin as well as the increase of sIgA production. Moreover, the relative expressions of MyD88, TLR2, and TLR4 were elevated in HDG; simultaneously, pro-inflammatory cytokines including IL-6, INF-γ, and TNF-α were decreased after CB administration. In addition, CB showed beneficial effects in improving weaning rex rabbit intestinal microflora via increasing the abundance of beneficial bacteria. Therefore, our results indicated CB can promote rex rabbit growth, which is likely to the enhancement of immune function and the improvement of intestinal microbiota.