Particle uptake by conjunctiva-associated lymphoid tissue (CALT) in turkeys

Uptake of tracer particles was assessed in lower eyelid conjunctiva-associated lymphoid tissue (CALT) of 3-week-old turkeys. Tracer particle suspensions, including carbon, iron oxide, or three sizes of latex beads (0.81 micron, 1.7 microns, and 2.9 microns), were placed into the experimentally sealed conjunctival space. After 5, 15, or 30 minutes, eyelids were removed and CALT was examined by light microscopy. Uptake was confirmed for all tracers and occurred within the lymphoepithelium of CALT. The uptake of latex beads was not as frequent as for carbon and iron. Tracers were increasingly evident in lymphoepithelium and subepithelial macrophage clusters as contact time increased. These findings provide additional evidence that CALT is capable of antigen uptake and may play a role in paraocular and upper respiratory immunity in turkeys.     

Conjunctiva-associated lymphoid tissue (CALT) in normal and Bordetella avium-infected turkeys

Conjunctiva-associated lymphoid tissue (CALT) was characterized in normal and Bordetella avium-infected turkey poults during the first 5 weeks of life. At 1, 5, 12, 19, 25, and 33 days post-hatching (DPH), upper and lower eyelids were examined by gross, histologic, and electron microscopic techniques. CALT was confined to the proximal part of the lower eyelid near the conjunctival fornix; it appeared by 5 DPH as individual lymphoid nodules and as dense masses by 19 DPH. In the upper eyelid, CALT was present only as isolated nodules. Histologically, CALT was composed of dense lymphocyte infiltrates within subepithelial connective tissue, intraepithelial lymphocytes, and flattened lymphoid-associated epithelium that lacked goblet cells. Germinal centers were in CALT by 19 DPH. By scanning electron microscopy, epithelial cells over lymphoid areas were flat and had short, irregular microvilli; non-lymphoid areas were covered by cells with tall, regular microvilli. Transmission electron microscopy revealed that with increasing age of birds, the epithelium over conjunctival lymphoid infiltrates became progressively flattened and infiltrated by lymphocytes. Some blood vessels in CALT had high endothelial cells; lymphocytes were in the lumen and between or beneath endothelial cells. In B. avium-infected poults, CALT was increased, developed earlier, and contained more germinal centers than in normal poults. We conclude that CALT of turkeys closely resembles other mucosal lymphoid tissues and may serve as a site for local antigen uptake.(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantification of particle uptake by conjunctiva-associated lymphoid tissue (CALT) in chickens

Tracer particle uptake by conjunctiva-associated lymphoid tissue (CALT) was quantified in the lower eyelids of 1-, 3-, 5-, and 7-week-old broiler chickens. CALT was measured histologically by computerized image analysis in all birds and in additional 1-day-old and 16-week-old chickens not subjected to uptake assessment. Suspensions of carbon or iron oxide were placed in contact with CALT for 5, 15, or 30 minutes (contact time). After eyelid removal, tracer uptake was scored by light microscopy, CALT was measured, and a mathematically derived uptake index was evaluated statistically. At each age examined, computer-generated measurements showed a significant increase in the proportion of CALT lymphoepithelium within proximal eyelids. Within the conjunctival sites evaluated, tracer uptake was significantly greater in lymphoepithelium than in non-lymphoepithelium at all ages and at all contact times. Uptake increased significantly between 3 and 5 weeks of age and between 5 and 15 minutes of tracer contact. Based on these uptake data for CALT in chickens, a minimal age-specific maturity is suggested that may influence function in mucosal immunity.     

Characterization of membranous (M) cells in normal feline conjunctiva-associated lymphoid tissue (CALT)

Objective To characterize conjunctival lymphoid nodules obtained from the nictitans of healthy cats to determine if the follicle‐associated epithelium (FAE) of conjunctiva‐associated lymphoid tissue (CALT) in this species contains membranous (M)‐cells analogous to those described in other regions of mucosa‐associated lymphoid tissue (MALT). Methods Lymphoid follicles from nictitan bulbar surfaces of 10 healthy cats (20 eyes total) were examined. Nictitans from five cats were harvested immediately post‐mortem and a minimum of 12 lymphoid nodules from each third eyelid were isolated using a Zeiss operating microscope. At least three lymphoid follicles from each eye were examined using light microscopy (LM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) using standard fixation and embedding protocols. Nictitan‐lymphoid follicles from another five healthy cats were processed for immunohistochemistry to characterize the distribution of T‐ and B‐lymphocytes present beneath the FAE. Results The FAE overlying CALT from 10 healthy cats demonstrated morphology characteristic of M‐cells including attenuated apical cell surface with blunted microvilli and microfolds, invaginated basolateral membrane forming a cytoplasmic pocket, and diminished distance between the apical and pocket membrane. Immunohistochemistry of lymphoid tissue subtending the FAE demonstrated B‐cell dependent regions in the germinal centers surrounded by T‐cell dependent interfollicular zones. Conclusions Healthy feline CALT contains morphologic features analogous to those described in other regions of MALT. Documentation of feline conjunctival M‐cells is of clinical relevance in the study of primary infectious, allergic, and autoimmune ocular diseases, as well as a potential means of vaccination or drug delivery.     

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。23日龄分别采取其CALT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CALT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，CALT发生早，生长快，淋巴细胞数量多，故本实验证实CALT对哈氏腺免疫细胞的数量与成分有一定的调控作用。CALT缺失鸡是研究黏膜免疫系统比较理想的模型。     

Mucosa-associated lymphoid tissue lymphoma of the third eyelid conjunctiva in a dog

A 4-year-old, neutered female Cocker Spaniel was presented to the veterinary clinic for protrusion of the left third eyelid. When the third eyelids from both eyes were everted, lobulated masses were present on the bulbar surface. The left third eyelid had a larger protrusion. There was no apparent associated ocular or systemic involvement. The tumor of left third eyelid was removed and referred for histological examination. Histologically, there were proliferations of lymphoid follicles surrounded by lymphoid cells forming a marginal zone. Those lymphoid cells occasionally infiltrated into conjunctival epithelium. A few apoptotic bodies with karyopyknotic and karyorrhexic nuclei were observed in the germinal center of lymphoid follicles. Mitotic figures were rare. On immunohistochemistry, tumor cells expressed CD79a but not CD3. A diagnosis of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) of the third eyelid was established based on the histological and immunophenotypical features. At the 1-year follow-up, there was no evidence of recurrence of the mass at the area of excision of the left third eyelid and the remaining tumor of the right third eyelid was still a similar size. The dog still showed no significant findings, except those of the tumor, and no evidence of systemic involvement. To the authors' knowledge, this is the first reported case of MALT lymphoma of the third eyelid in a dog.     

Larynx-associated lymphoid tissue (LALT) in young cattle

This paper describes the presence of lymphoid tissue in the mucosa of the bovine larynx. A total of 15 bovine larynges were examined both macroscopically after tissue fixation in acetic acid and microscopically using histology. It was found that no paraepiglottic tonsil was present in cattle, although a few lymphoid follicles were present in the mucosa at the base of the epiglottis. This result is in accordance with previous reports. In contrast, numerous lymphoid follicles were seen in the mucosa of the epiglottis and the corniculate processes of the arytenoid cartilages. This suggests that larynx-associated lymphoid tissue is present in cattle. Our observation could be of clinical importance, e.g. in the framework of the development of aerosolized vaccines.     

NALT (nasal cavity-associated lymphoid tissue) in the rabbit

Due to its many advantages, interest in intranasal vaccination of domestic mammals and humans is currently increasing. Successful stimulation of the immune system by intranasal vaccines requires, however, the presence of lymphoid tissue in the nasal cavity. This nasal cavity-associated lymphoid tissue (NALT) has already been described in humans and many laboratory rodents, but data about rabbits are very scarce. For this purpose, histological sections of the nasal cavities of 10 female adult New Zealand White rabbits were examined for the presence of lymphoid tissue. Primary (I) and secondary (II) lymphoid follicles divided by interfollicular regions were mainly present at the bottom of the ventral nasal meatus and the nasopharyngeal meatus from 1 to 3.3 cm from the tip of the nose. In this region intraepithelial and lamina propria lymphocytes, and isolated lymphoid follicles (ILF's) were additionally seen at the dorsal and dorsolateral sides of the nasopharyngeal meatus and within the mucosae of the nasal conchae and the lateral nasal walls. Intraepithelial and lamina propria lymphocytes, and ILF's were, just like in humans, randomly distributed along the entire nasal mucosa. The rabbit NALT is more voluminous compared to rodents in which lymphoid tissue is only present at the bottom of the nasopharyngeal meatus. Since the relative volume of the rabbit nasal cavity is also similar to that of humans, the rabbit could be a valuable research model not only for animal but also for human intranasal vaccine development.     

Electron microscopic structure of mesangioproliferative glomerulonephritis (MesPGN) in animals

In a murine lupus model, the kidneys of 42 MRL mice of different age groups (substrains MRL/Mp-lpr/lp and -+/+) were studied by light and transmission electron microscopy. The results demonstrate the ultrastructural feature of the mesangial-proliferative glomerulonephritis which is one of the observed forms of glomerulonephritis. The most important alterations are a severe proliferation of mesangial cells (with an increase in rough endoplasmic reticulum and mitochondria) and electron dense deposits in different sites of the glomerular basement membrane (subepithelial, subendothelial, intramembranous). These deposits are proposed to be immune complexes. Osmiophilic intracytoplasmic inclusions of the mesangial cells are indications of an altered renin production. Alterations of the glomerular epithelial cells are characterized by fusions of the epithelial pedicles, an increase of microvilli and intracytoplasmic concentrations of electron dense material which are also proposed to be immune complex deposits. The morphological feature of the mesangial-proliferative glomerulonephritis is completed by an activation and focal edema of endothelial cells. The described alterations are discussed and compared with findings in other species.     

Investigation of bovine gut associated lymphoid tissue (GALT) using monoclonal antibodies against bovine lymphocytes

Gut associated lymphoid tissue of the small and large intestine of calves and cows has been compared morphologically and quantitatively using monoclonal antibodies to bovine lymphocytes. B cells were significantly decreased in the ileum of the cow compared to the calf. Significantly increased numbers of T cells were present in cell suspensions of all lymphoid areas of the cow compared to the calf. T lymphocyte subsets were quantified into cryostat sections of lymphoid tissues expressing BoT4, and BoT8 antigens demonstrated increased numbers in follicular and dome areas of the discrete Peyer's patches of the small and large intestine of the cow. BoT4+, BoT8+, and the non-BoT4/BoT8+ T cell subsets were increased in the mucosa of the cow as compared to the calf. Similarities in structure and lymphocyte composition of the discrete Peyer's patches of the small intestine, cecum and colon and isolated single follicles in the large intestine suggest similar functional properties.     

Effects of mimosine and 2,3-dihydroxypyridine on fiber shedding in Angora goats.

The effects of intravenous infusion of mimosine or 2,3-dihydroxypyridine (2,3-DHP) and the effects of oral dose level of mimosine on fiber shedding in Angora goats were determined. In one experiment, 20 mature Angora wethers (36+/-1.9 kg BW) were infused for 2 d with 79, 102, or 135 mg/(kg BW.d) of mimosine, 90 mg/(kg BW.d) of 2,3-DHP, or saline. At 7 d after infusion began, fiber shedding was observed in all goats receiving mimosine but not in any goats infused with 2,3-DHP or saline. Fiber shedding varied among goats; in some goats, fiber shedding was complete and occurred without hand-plucking, whereas in others fiber was retained by nonshed fibers but could be removed by hand-plucking. Nonshed fibers were larger in diameter and more likely to be medullated (P < .05) compared with hand-plucked fibers. Mean plasma mimosine concentration at 24 and 48 h after infusion began was 79 and 98 micromol/L (P < .05), respectively, and greater (P < .05) for mimosine infused at 135 than at 102 mg/(kg BW.d) (89, 68, and 108 micromol/L for mimosine infused at 79, 102, and 135 mg/[kg BW.d], respectively; SE 9.5). In another experiment, oral dosing of eight Angora bucks (23+/-.5 kg BW) with 400 or 600 mg/kg BW of mimosine rapidly increased plasma mimosine concentration, which reached approximately 100 and 160 micromol/L at 5 h after dosing; however, periods of time during which plasma mimosine concentrations were comparable to those in the first experiment were considerably shorter. Oral mimosine dosing did not induce fiber shedding in 7 d. After 31 d, fiber was retained by nonshed fibers but could be removed by hand-plucking or could only be partially removed with difficulty by hand-plucking. There were no toxic effects of mimosine or 2,3-DHP administration; only minor, short-term inhibitions of feed intake by mimosine were noted in some goats. In conclusion, mimosine holds promise as a safe means to remove fiber of Angora goats; further research is necessary to characterize the seasonality of follicle activity and to develop convenient means of mimosine delivery.     

Electron microscopic cytochemical studies of anionic sites in the rat spleen

The distribution patterns of the intensity of negative charge on the free surfaces (glycocalyx of the plasma membrane) of endothelial cells (ECs) in blood vessels and reticular cells (RCs) in the splenic cord of the rat spleen were studied by an electron microscopic cytochemical method using polyethyleneimine (PEI) as a cationic probe. Spleens from adult male rats were perfusion-fixed with 0.5% glutaraldehyde -4% paraformaldehyde containing 0.05% cetylpyridinium chloride and then perfused with 0.5% PEI at pH 7.4. On the free surfaces (glycocalyx of the plasma membrane) of the ECs examined, distinct PEI-positive reactions were observed in blood vessels, such as trabecular arteries, central arteries, arterial capillaries, pulp veins and trabecular veins. These PEI-positive electron-dense substances in the trabecular arteries, central arteries, and trabecular veins took the shape of a band of 170-250 nm in thickness. On the other hand, the corresponding ultrastructure of the ECs lining the splenic sinuses and the RCs in the splenic cord showed exceedingly weak PEI reactions. The PEI-reactive deposits were significantly thinner than those in the above blood vessels. As the thickness of the electron-dense substances can be related to the density of the negative charge, these results suggest that there is a high intensity of negative charge on the free surfaces (glycocalyx of the plasma membrane) of ECs in blood vessels where blood cells and plasma pass into the red pulp or are discharged from the red pulp. In contrast, the splenic sinuses and RCs, which are the main components of the red pulp, contain weakly negative-charged sites. This may contribute to the microcirculation of the splenic blood vessels and elucidate the possible physiological functions of the spleen, such as blood storage.