Immunoglobulin classes synthesised by the chicken Harderian gland after local immunisation.

The immunoglobulin (Ig) levels in tears and sera were compared after antigen administration (salmonella O antigen) by eyedrop and injection into the nictitating membrane, to determine the Ig classes synthesised by the plasma cells in the chicken Harderian gland. Samples of tears and sera were collected from immunised and control birds between 24 hours and 24 days after the antigen or sterile saline was administered. Samples were assayed for IgA, IgG and IgM concentrations using radial immunodiffusion. It is suggested that most of the IgG found in tears after local immunisation has an extraglandular origin.     

切除结膜结合淋巴组织鸡对新城疫疫苗点眼免疫反应 Ⅱ.组织学与免疫组织化学研究

60只 1日龄健康雏鸡随机分为 A、B、C三组。A组于 1日龄手术切除下眼睑结膜。A、B两组于 12日龄时结扎鼻泪管 ,用新城疫克隆 30苗点眼。C组为对照组。 2 3日龄分别采取其 CAL T和哈德氏腺 ,比较观察其组织细胞。结果 ,1日龄手术切除下眼睑结膜后造成鸡 CAL T缺失 ,点眼免疫后 ,哈氏腺的免疫细胞也较正常免疫鸡少 ,而正常接种 ND克隆 30 (B组 )后 ,CAL T发生早 ,生长快 ,淋巴细胞数量多 ,故本实验证实 CAL T对哈氏腺免疫细胞的数量与成分有一定的调控作用。 CAL T缺失鸡是研究黏膜免疫系统比较理想的模型     

切除结膜结合淋巴组织鸡对新城疫疫苗的免疫反应Ⅰ.局部与循环抗体检测

实验将 60只 1日龄健康雏鸡随机分为A、B、C 3组。A组为CALT缺失组 ,B组为免疫组 ,C组为非免疫对照组。A组于 1日龄手术切除下眼睑结膜 ,A、B两组于 1 2日龄结扎鼻泪管后进行点眼免疫接种新城疫克隆 30疫苗。2 3日龄时 ,收集A、B、C各实验组鸡的血清和泪液 ,做红细胞凝集抑制试验 (HI试验 )、酶联免疫吸附试验(ELISA)。比较观察各实验组雏鸡血清和泪液抗体HI效价与IgA、IgG、IgM含量的变化特点。结果 ,1日龄手术切除下眼睑结膜后造成鸡CALT缺失 ,CALT缺失鸡点眼免疫后 ,泪液抗体HI效价与IgA、IgG、IgM含量比免疫组雏鸡低。但对雏鸡的抗体生成的规律并不产影响。故实验证实了CALT对眼区局部抗体的产生及其种类有重要的调控作用 ,协同哈氏腺完成局部免疫的功能     

网状内皮增殖病病毒(REV)感染SPF雏鸡后局部免疫组织抗体生成细胞的变化

对 SPF雏鸡人工感染 REV后局部免疫组织盲肠扁桃体和哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量动态变化进行了检测。结果发现 ,REV感染 SPF雏鸡后 7～ 4 9天消化道局部免疫组织盲肠扁桃体和呼吸道相关局部免疫组织哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量明显低于未感染对照雏鸡。表明雏鸡感染 REV后消化道局部组织盲肠扁桃体和呼吸道相关免疫组织哈德氏腺体液免疫机能均显著下降。     

Local immunity in the respiratory tract of the chicken. II. The secretory immune response to newcastle disease virus and the role of IgA

The nature, specificity and characteristics of the secretory immune response in the respiratory tract of the chicken were investigated in young specific-pathogen-free chickens after vaccination with live lentogenic and inactivated Newcastle disease virus (NDV). Virus-neutralizing (VN) activity considerably exceeding transudation levels from serum were detected in lachrymal fluid, saliva and tracheal washes following infection by both ocular and oral routes. Heat inactivated virus inoculated into the trachea evoked neither serum nor secretory VN activity, whereas a commercial inactivated virus vaccine in mineral oil adjuvant stimulated high titres of serum antibody and some VN activity in tracheal fluids.Antibody in secretions limited, but did not prevent, reinfection of the trachea when birds were challenged 2 weeks later. In contrast to an elevation of circulating antibody titre, challenge induced only a repeated primary response of secreted antibody.All secretions contained IgA which, at least in saliva, accounted for 85% of its activity, the remainder being due to IgG. Fluorescent localization of immunoglobulin producing cells (IPC) demonstrated large numbers containing IgA in association with the upper respiratory tract, particularly in the Harderian gland which contained dense aggregations of plasma cells, many of which were producing IgA.It is concluded that the respiratory tract of the chicken possesses an antibody mediated secretory immune system analogous to that of mammalian species.     

Expression of different classes of immunoglobulin in intraepithelial plasma cells of the Harderian gland of domestic ducks Anas platyrhynchos

The Harderian gland of chickens contains numerous plasma cells and is considered as a peripheral lymphoid organ. Data about this gland in other avian species are scarce or inexistent. Considering that ducks show some unique characteristics regarding the immune system, which are important in evolutionary context, and that unusual location of plasma cells into the epithelium was recently described in primitive avian species, here we investigated the occurrence and characterized intraepithelial plasma cells in the Harderian gland of ducks, according to the immunoglobulin produced. Numerous intraepithelial plasma cells were found confined to the Harderian gland ducts. Plasma cells were also found in the ducts lamina propria. IgM-positive cells were the most abundant into the epithelium. In contrast, IgY- or IgA-positive cells were predominant in the lamina propria. The constancy of intraepithelial plasma cells in all specimens examined indicates that they may be essential mediator for an effective immunesurvaillance of the ocular mucosa.     

Failure to stimulate a local immune response in the mammary gland of the sow using intraperitoneal "priming"

An experiment was carried out to determine whether a local immune response could be produced in the mammary gland of sows by "priming" the animals with an intraperitoneal injection of antigen without adjuvant. Adjuvant was not used in the immunization regimen because it had been shown to induce acute peritonitis and multiple abscess formation when injected intraperitoneally in pigs. Four pregnant sows were "primed" by intraperitoneal injection of ferritin; a second dose of the antigen was given two weeks later into the lumen of the jejunum. Another group of pregnant sows was given two intra-muscular injections of ferritin two weeks apart and a third group of sows served as non-immunized controls. Antibody titres and concentrations of IgG, IgA and IgM in colostral and milk whey suggested that a local immune response had not been stimulated in the mammary glands of these sows. Mammary tissue was examined for the presence of immunoglobulin-containing and specific antibody-containing plasma cells. Results of these studies confirmed that intraperitoneal/intrajejunal injection of ferritin without adjuvant was not successful in inducing a local immune response in the mammary gland.     

Local and systemic immune responses following infection of broiler-type chickens with avian Metapneumovirus subtypes A and B

Infections with avian Metapneumovirus (aMPV) are often associated with swollen head syndrome in meat type chickens. Previous studies in turkeys have demonstrated that local humoral and cell-mediated immunity plays a role in aMPV-infection. Previous experimental and field observations indicated that the susceptibility of broilers and their immune reactions to aMPV may differ from turkeys. In the presented study local and systemic immune reactions of broilers were investigated after experimental infections with subtypes A and B aMPV of turkey origin. Both virus subtypes induced a mild respiratory disease. The recovery from respiratory signs correlated with the induction of local and systemic aMPV virus-neutralizing antibodies, which began to rise at 6 days post infection (dpi), when the peak of clinical signs was observed. In a different manner to the virus neutralizing (VN) and IgG-ELISA serum antibody titres, which showed high levels until the end of the experiments between 24 and 28 dpi, the specific IgA-ELISA and VN-antibody levels in tracheal washes decreased by 10 and 14 dpi, respectively, which may explain the recurring aMPV-infections in the field. Ex vivo cultured spleen cells from aMPV-infected broilers released at 3 and 6 dpi higher levels of IFN-γ after stimulation with Concanavalin A as compared to virus-free birds. In agreement with studies in turkeys, aMPV-infected broilers showed a clear CD4+ T cell accumulation in the Harderian gland (HG) at 6 dpi (P<0.05). In contrast to other investigations in turkeys aMPV-infected broilers showed an increase in the number of CD8alpha+ cells at 6 dpi compared to virus-free birds (P<0.05). The numbers of local B cells in the Harderian gland were not affected by the infection. Both aMPV A and B induced up-regulation of interferon (IFN)-γ mRNA-expression in the nasal turbinates, while in the Harderian gland only aMPV-A induced enhanced IFN-γ expression at 3 dpi. The differences in systemic and local T cell and possibly natural killer cell activity in the HG between turkeys and chickens may explain the differences in aMPV-pathogenesis between these two species.     

Immune response of chickens to various routes of administration of Australian infectious bronchitis vaccine

Groups of 100 two-week-old cockerels were vaccinated with the A₃ strain of IB vaccine by conjunctival, intranasal, in-contact, drinking water, or aerosol routes, or were left as unvaccinated controls. Three weeks after vaccination, each group of chickens was challenged with the Appin strain of IBV.
Vaccination by the conjunctiva!, intranasal and in-contact routes induced a good resistance to challenge, concurring with an obvious stimulation of the Harderian gland, while the drinking water route led to a low resistance to challenge, with minor changes in the gland. The results of no immune response and no resistance to challenge in the birds vaccinated by aerosol route was due to unsuccessful vaccination in the group. Application of the vaccine by the conjunctival route would appear to be a most effective route for the application of Australian IB vaccines, while the in-contact method appears worthy of further study.     

Evaluation of a soluble sustained-release ophthalmic delivery unit in the dog.

A 5-mg hydroxypropyl cellulose soluble ocular insert was evaluated by placement in the upper and the lower conjunctival and bulbar nictitating membrane fornices in normal dogs. The insert was easily placed in the conjunctival fornices, but was difficult to place in the bulbar nictitating membrane fornix. The conjunctival inserts did not produce irritation of the cornea and the conjunctiva. Placement of the inserts beneath the nictitating membrane produced local irritation and chemosis. The sites of highest to lowest retention for the insert were upper conjunctival fornix, bulbar nictitating membrane fornix, and lastly, lower conjunctival fornix. Dissolution was nearly complete in 8 hours.     

Everted third eyelid cartilage in a cat: a case report and literature review

An 8-year-old neutered female British Blue cat was presented with a presumed diagnosis of a prolapsed nictitans gland and associated ocular irritation and epiphora. However, during surgery, the apparent nictitans gland protrusion was determined to be an everted cartilage of the nictitating membrane. The scrolled portion of the cartilage was removed through an incision through the conjunctiva on the bulbar aspect of the third eyelid, as previously described in the dog. This operation resolved the ocular irritation occurring, and the third eyelid returned to its anatomically correct position.     

A study of the distribution patterns and levels of Salmonella enteritidis in the immune organs of ducklings after oral challenge by serovar-specific real-time PCR

The objective of this study was to understand the distribution patterns and levels of Salmonella Enteritidis (SE) in the immune organs of ducklings after oral challenge. We conducted serovar-specific real-time polymerase chain reaction (PCR) for SE to detect the genomic DNA of SE in the blood and immune organs, including the bursa of Fabricius, thymus, spleen, and Harderian gland, from ducklings after oral challenge at different time points. The results showed that SE was consistently detected in all the samples. The Harderian gland and spleen tested positive at 8 hr postinoculation (PI). The organism was detected in the blood, bursa of Fabricius, and thymus at 10 hr PI. The copy number of SE DNA in each tissue reached a peak at 24-36 hr PI. The spleen, blood, and Harderian gland contained high concentrations of SE, whereas the thymus and bursa of Fabricius had low concentrations. SE populations began to decrease and were not detectable at 2 days PI, but they were still present up to 9 days PI in the spleen, without producing any apparent symptoms. To validate these results, the indirect immunofluorescent antibody (IFA) technique was used, and the IFA results were similar to those of the fluorescent quantitative-PCR. In conclusion, the results provided insight into the SE life cycle in the immune organs; furthermore, the Harderian gland and spleen were determined to be the primary sites of invasion among the immune organs of normal ducklings after oral SE challenge. This study will help in understanding the pathogenesis of SE infection in vivo and may help in the development of a live Salmonella vaccine in the future.     

IgA and secretory component (SC) in the third eyelid of domestic animals: a comparative study

Objective The third eyelid of domestic animals is important for the production and distribution of tears, in removing ocular debris and in protection of the globe, and has significant immunologic functions. Although it is known that tears contain antibodies of the immunoglobulin A (IgA) isotype which are produced mainly by plasma cells of the lacrimal gland, very little is known about the antibody repertoires in the third eyelid of domestic animals. To assess whether IgA is derived from local synthesis, we analyzed the location of IgA‐producing cells and the cellular distribution of secretory component (SC) in the third eyelid of domestic animals in a comparative study. Animal studied A total of 83 third eyelids of dogs, cats, pigs, cows, sheep, goats and horses were investigated in the course of this study. Procedures Third eyelids were obtained immediately after death, cut length‐wise, fixed overnight and processed for immunohistochemical detection of IgA and SC by the ABC technique. Results The results show that IgA‐producing plasma cells are densely populated in subepithelial spaces of the surface epithelium as well as in the nictitating gland in a species‐specific manner. In contrast, the SC could be demonstrated exclusively in glandular acinar and ductal epithelial cells and in different cell types of the surface epithelium, preferentially located on the bulbar side of the nictitating membrane. Conclusion It is suggested that most of the SC is locally produced by resident plasma cells and subsequently transferred through the surface epithelium and glandular duct cells by transcytosis. This indicates that the third eyelid is an important member of the secretory immune system in domestic animals.     

Investigations on the conjunctival goblet cells and the characteristics of the glands associated with the eye in chinchillas (Chinchilla Laniger)

Objective To investigate the density and distribution of conjunctival goblet cells (GC) and study the anatomy and microscopic characteristics of glands associated with the eye in chinchillas (Chinchilla Laniger). Procedure 12 chinchillas were included in the study. Conjunctiva (divided into four regions), eyelids, and glands were embedded in paraffin wax, sectioned, stained, and analyzed. Results Highest GC densities were found in the palpebral region of the nasal and temporal conjunctiva of both eyelids (GC index: 25.1-18.2%), and lowest densities, in the bulbar and marginal region of the nasal and temporal conjunctiva of both eyelids (GC index: 1.5-0.0%). Meibomian glands extend along the entire length of both eyelids, and the whole glandular complex broadens toward the temporal canthus. This is macroscopically visible through the conjunctiva. The openings of the Meibomian glands are macroscopically not discernible. The light pink, smooth, and crescent-shaped lacrimal gland lies next to the aforementioned broadened part of the Meibomian glands in the temporal canthus. The whitish, 0.9-cm-long, smooth Harderian gland is firmly attached to the posterior part of the globe and extends nasally from the optic nerve to the equator. Furthermore, chinchillas possess two lacrimal puncta, situated on the inner conjunctival surface of both eyelids near the medial canthus. A pigmented lacrimal canaliculus originates from each punctum. The vestigial nictitating membrane is supported by a hyaline cartilage and is pigmented at its free margin. Conclusions Chinchillas possess a Harderian gland, a lacrimal gland, and Meibomian glands. The GC density in the nasal and temporal palpebral conjunctiva is higher than in guinea pigs.     

Normal anatomical and histochemical characteristics of the lacrimal glands in the American bison and cattle

Dorsal lacrimal glands, superior glands of the third eyelid and Harderian glands (deep gland of the third eyelid) from 19 bison and 18 cattle free of apparent ocular disease were examined to compare the normal anatomical properties of these glands. All glands were characterized and measured (length and width). The gross anatomy of the dorsal lacrimal glands was similar, with the exception of a bipartite gland in cattle. The bison's superior gland of the third eyelid and Harderian gland was longer as compared with cattle. A subset of the bison and cattle samples (five bison and five cattle) was sectioned for histological and histochemical analysis. The histology of the dorsal lacrimal and superior gland of the third eyelid revealed tubuloalveolar cells with basophilic vacuolated cytoplasm in bison and eosinophilic granular cytoplasm in cattle. The Harderian glands consisted of a tubuloalveolar anterior part combined with large lumens acini lined with cuboidal epithelium in the posterior part; the posterior part of the bison Harderian gland was more predominant than in cattle samples. Mucosubstance histochemistry revealed acidic and neutral glycoproteins with similar staining patterns in all glands of both species.     

鸡贫血因子病局部粘膜免疫功能变化的研究

用鸡贫血病毒感染１日龄ＡＡ雏鸡,以未感染同龄ＡＡ雏鸡的对照,在感染后７,１４,２１,２８,３５,４２,４９日龄检测其哈德尔腺及盲肠扁桃体Ｔ细胞和ＩｇＧ＾＋、ＩｇＭ＾＋、ＩｇＡ＾＋抗体生成细胞数量,泪液,气管液、肠液中ＩｇＭ、ＩｇＭ、ＩｇＡ含量的动态变化。     

马立克氏病疫苗免疫后鸡免疫器官组织抗体生成细胞的变化

本实验分别用马立克氏病（ＭＤ）三价苗和ＨＶＴ疫苗肌肉注射免疫１日龄雏鸡，在１０、２０、４０、６０和９０日龄以兔抗鸡ＩｇＧ、ＩｇＭ和ＩｇＡ重链抗血清为一抗，用彩色免疫金银染色法检测法氏囊、脾脏、盲肠扁桃体和哈德尔腺的ＩｇＧ、ＩｇＭ和ＩｇＡ抗体生成细胞的动态变化。结果发现：雏鸡ＭＤ疫苗免疫后，法氏囊和脾脏的ＩｇＧ、ＩｇＭ和ＩｇＡ抗体生成细胞较对照鸡显著增多，盲肠扁桃体以ＩｇＡ抗体生成细胞为主、哈德尔腺以ＩｇＧ抗体生成细胞居多的三种抗体生成细胞数量均明显升高；三价苗免疫鸡的抗体生成细胞显著多于ＨＶＴ疫苗免疫鸡。说明ＭＤ疫苗免疫鸡全身免疫器官、呼吸道和消化道相关局部免疫组织的体液免疫反应显著增强，三价苗免疫鸡的体液免疫应答水平明显高于ＨＶＴ疫苗免疫鸡。     

Histological and immunological studies on the fowl lacrimal gland following surgical excision of Harder's gland.

Surgical removal of the Harderian gland of the domestic fowl resulted in increased secretory activity in the lacrimal gland and also in an increase in goblet cell numbers along the length of the lacrimal gland duct. Plasma cells were more numerous in the lacrimal glands of operated birds and they were capable of antibody responses to both systemically and topically applied bovine serum albumin (BSA). Gel diffusion studies showed the presence of anti-BSA activity in the tears and serum of fowls stimulated after surgical removal of the Harderian gland.     

Effect of Harderian adenectomy on the statistical analyses of mouse brain imaging using positron emission tomography

Positron emission tomography (PET) using 2-deoxy-2-[¹⁸F] fluoro-D-glucose (FDG) as a radioactive tracer is a useful technique for in vivo brain imaging. However, the anatomical and physiological features of the Harderian gland limit the use of FDG-PET imaging in the mouse brain. The gland shows strong FDG uptake, which in turn results in distorted PET images of the frontal brain region. The purpose of this study was to determine if a simple surgical procedure to remove the Harderian gland prior to PET imaging of mouse brains could reduce or eliminate FDG uptake. Measurement of FDG uptake in unilaterally adenectomized mice showed that the radioactive signal emitted from the intact Harderian gland distorts frontal brain region images. Spatial parametric measurement analysis demonstrated that the presence of the Harderian gland could prevent accurate assessment of brain PET imaging. Bilateral Harderian adenectomy efficiently eliminated unwanted radioactive signal spillover into the frontal brain region beginning on postoperative Day 10. Harderian adenectomy did not cause any post-operative complications during the experimental period. These findings demonstrate the benefits of performing a Harderian adenectomy prior to PET imaging of mouse brains.     

Symblepharon with aberrant protrusion of the nictitating membrane in the snowy owl (Nyctea scandiaca)

Two young snowy owl chicks were presented with aberrant protrusion of the nictitating membranes. This was caused by conjunctival adhesions causing symblepharon secondary to a previous septicemia episode. While symblepharon has been noted in birds before, this unusual presentation of the nictitating membrane has not been reported. Surgical intervention ameliorated the clinical signs, allowing vision in one bird by removal of the nictitating membranes, a technique which appeared to have no deleterious effects on the ocular surface.