Infectious bronchitis virus infection in chicken: viral load and immune responses in Harderian gland, choanal cleft and turbinate tissues compared to trachea.

1. The role of the Harderian gland (HG), choanal cleft (CC) and turbinate in terms of IBV M41 viral load compared to the trachea, and immune (innate, cellular and mucosal) responses were studied in 21-day-old commercial broiler chickens.2. After virulent IBV M41 challenge, the antigen concentration detected either by quantitative RT-PCR or immunohistochemistry peaked at 2-3 days post challenge (dpc) in all tissues. Significant increases of lachrymal IBV-specific IgA and IgY levels were found at 4-5 dpc.3. Gene transcription showed a significant up-regulation of TLR3, MDA5, IL-6, IFN-α and IFN-ß, where patterns and magnitude fold-change of mRNA transcription were dependent on the gene and tissue type.4. The results demonstrated active IBV M41 replication in the HG, CC and turbinate, comparable to levels of replication found in the trachea. Data on immune-related genes in head-associated tissues provide further understanding on the immunobiology of IBV and offer opportunities to identify their use as quantitative biomarkers in pathogenicity and vaccination-challenge studies.

An innate defense peptide BPIFA1/SPLUNC1 restricts influenza A virus infection.

This corrects the article DOI: 10.1038/mi.2017.45.

An innate defense peptide BPIFA1/SPLUNC1 restricts influenza A virus infection.

The airway epithelium secretes proteins that function in innate defense against infection. Bactericidal/permeability-increasing fold-containing family member A1 (BPIFA1) is secreted into airways and has a protective role during bacterial infections, but it is not known whether it also has an antiviral role. To determine a role in host defense against influenza A virus (IAV) infection and to find the underlying defense mechanism, we developed transgenic mouse models that are deficient in BPIFA1 and used these, in combination with in vitro three-dimensional mouse tracheal epithelial cell (mTEC) cultures, to investigate its antiviral properties. We show that BPIFA1 has a significant role in mucosal defense against IAV infection. BPIFA1 secretion was highly modulated after IAV infection. Mice deficient in BPIFA1 lost more weight after infection, supported a higher viral load and virus reached the peripheral lung earlier, indicative of a defect in the control of infection. Further analysis using mTEC cultures showed that BPIFA1-deficient cells bound more virus particles, displayed increased nuclear import of IAV ribonucleoprotein complexes, and supported higher levels of viral replication. Our results identify a critical role of BPIFA1 in the initial phase of infection by inhibiting the binding and entry of IAV into airway epithelial cells.

Alveolar macrophages are the main target cells in feline calicivirus-associated pneumonia.

Feline calicivirus (FCV) is a pathogen of felids and one of the most common causative agents of feline upper respiratory disease (URD). Reports of natural FCV pneumonia in the course of respiratory tract infections are sparse. Therefore, knowledge on the pathogenesis of FCV-induced lung lesions comes only from experimental studies. The aim of the present study was to assess the type and extent of pulmonary involvement in natural respiratory FCV infections of domestic cats and to identify the viral target cells in the lung. For this purpose, histology, immunohistochemistry and RNA-in situ hybridisation for FCV and relevant cell markers were performed on diagnostic post-mortem specimens collected after fatal URD, virulent systemic FCV or other conditions. All groups of cats exhibited similar acute pathological changes, dominated by multifocal desquamation of activated alveolar macrophages (AM) and occasional type II pneumocytes with fibrin exudation, consistent with diffuse alveolar damage (DAD). In fatal cases, this was generally seen without evidence of epithelial regeneration. In cats without clinical respiratory signs, type II pneumocyte hyperplasia was present alongside the other changes, consistent with the post-damage proliferative phase of DAD. FCV infected and replicated in AM and, to a lesser extent, type II pneumocytes. This study shows that lung involvement is an infrequent but important feature of FCV-induced URD. AM are the main viral target cell and pulmonary replication site, and their infection is associated with desquamation and activation, as well as death via apoptosis.

Surface heterogeneity of rat sperm during maturation.

Rat sperm isolated from the caput and caudal epididymis and the vas deferens were subjected to multiple partition in aqueous two-phase systems. The technique was used to reveal heterogeneity of a sperm population with respect to particular surface properties. Sperm from all three regions gave broad distributions indicative of heterogeneous cell populations. Greatest heterogeneity was observed for cauda sperm with caput and vas sperm producing similar distributions. Following multiple partition sperm from different regions of the distribution profiles were immunostained with three antibodies known to recognise maturation antigens. The results show that some antigens are acquired during epididymal transit whilst others are present throughout. The partition (surface heterogeneity) seen cannot therefore be explained solely by the distribution of the antigens recognized by 2D6, 6B2 and 3D5.

The homeobox gene Hox 7.1 has specific regional and temporal expression patterns during early murine craniofacial embryogenesis, especially tooth development in vivo and in vitro.

Hox 7.1 is a murine homeobox-containing gene expressed in a range of neural-crest-derived tissues and areas of putative epithelial-mesenchymal interactions during embryogenesis. We have examined the expression of Hox 7.1 during craniofacial development in the mouse embryo between days 8 and 16 of development. Whereas facial expression at day 10 of gestation is broadly localised in the neural-crest-derived mesenchyme of the medial nasal, lateral nasal, maxillary and mandibular processes, by day 12 expression is restricted to the mesenchyme immediately surrounding the developing tooth germs in the maxillary and mandibular processes. Hox 7.1 expression in the mesenchyme of the dental papilla and follicle is maximal at the cap stage of development and progressively declines in the bell stage prior to differentiation of odontoblasts and ameloblasts. Hox 7.1 expression in tooth germs is independent of overall embryonic stage of development but is dependent on stage of development of the individual tooth. Similar patterns of transient Hox 7.1 expression can also be detected in tooth germs in vitro in organ cultures of day 11 first branchial arch explants cultured for up to 7 days. Hox 7.1 is also expressed early in development (days 10/11) in the epithelium of the developing anterior pituitary (Rathke's pouch), the connective tissue capsule and meninges of the developing brain, and specific regions of neuroepithelium in the developing brain.

Changes in the rat lymphoid tissues after Leydig cell destruction by ethane-1,2-dimethanesulphonate.

Since ethane-1,2-dimethanesulphonate (EDS) causes Leydig cell destruction which may elicit an immune response the lymphoid tissues of sexually mature male rats were examined after an intraperitoneal injection of EDS. The changes observed in the testicular regional lymph nodes confirm an immune response while those observed in the thymus may indicate involvement in the immunological response or be a toxic reaction to EDS.

Cellular proliferation in the lymphoid tissues of an inbred (RT1a) rat strain during the oestrous cycle.

Changes in body weight, uterine weight and tissue weight, cell content and cellular proliferation of the thymus and uterine regional and popliteal lymph nodes were examined at daily intervals during the oestrous cycle of the DA (RT1a) inbred rat strain. Thirty-nine sexually mature virgin animals, aged between 13 and 15 weeks were used in this investigation. During dioestrus body weight and uterine weight fell significantly, while intrathymic and intranodal cellular proliferation increased significantly. These findings are discussed in relation to ovarian hormone secretion and it is suggested that increased thymocyte and lymphocyte proliferation occurs in response to rising oestrogen levels. This proliferative response prepares the female for the immunological challenge with allogeneic spermatozoa should mating occur during the subsequent oestrous phase.

A dioestrous increase in thymocyte proliferation during the oestrous cycle.

Coitus, which precedes internal fertilisation, is a unique physiological event which allows motile allogeneic spermatozoa to enter the female host and invade her tissues. The cyclic cellular proliferation observed in the thymus of the female rat may be an important preparation of her immune system for this event.

The cell content and proliferative response of the rat thymus during first syngeneic and allogeneic pregnancy, and the effects of strain difference.

Two hundred female rats, 91-101 days of age, were used in this investigation which provided data on cell content and proliferative response of the thymus. The observations began in oestrous phase virgin females from two highly inbred strains AO(RT1u/AgB2) and DA(RT1a/AgB4), continued at five day intervals throughout syngeneic and allogeneic pregnancy in both strains and were completed in the immediate post-partum period. This experimental protocol permitted sequential observations throughout gestation and allowed syngeneic: allogeneic and interstrain comparisons to be made. The syngeneic: allogeneic comparisons demonstrated significant differences in the thymic proliferative responses while the AO:DA comparisons demonstrated significant differences in the thymic cell contents.

Thymic and body weight during first syngeneic and allogeneic pregnancy in the rat, and the effects of strain difference.

Two hundred female rats, 91-101 days of age, were used in this investigation which provided data on thymic weight, body weight and where appropriate the number of progeny. The observations began in oestrus phase virgin females from the two highly inbred strains AO (RT1u/AgB2) and DA (RT1a/AgB4), continued at five day intervals throughout syngeneic and allogeneic pregnancy in both strains and were completed in the immediate post-partum period. This experimental protocol permitted sequential observations throughout gestation and allowed syngeneic:allogeneic and interstrain comparisons to be made. The syngeneic:allogeneic comparisons demonstrated significant differences in the body weight response and the number of progeny while the AO:DA comparisons demonstrated significant differences in the thymic weight and body weight responses.

A specific cytolytic T cell response induced by subcutaneous challenge with allogeneic rat epididymal spermatozoa.

Groups of virgin female rats were killed on days 2-5 after footpad injection with either thoracic duct lymphocytes or epididymal spermatozoa from allogeneic male rats and the cellular response in the regional lymph nodes determined by the specific cytoadhesion technique. After challenge with both thoracic duct lymphocytes and epididymal spermatozoa a specific cytolytic T cell response to the male strain histocompatibility antigens was evident on the second day and increased until the fifth day. The weight and total lymphocyte content of the nodes were significantly greater than the unimmunized control values throughout the observation period. These results provide further evidence for the expression of histocompatibility antigens by epididymal spermatozoa.