A morphologic study of bronchus-associated lymphoid tissue in turkeys

Bronchus-associated lymphoid tissue (BALT) in normal turkeys of ages 1 day and 1, 2, 3, 4, 8, and 18 weeks was examined by light microscopy and by scanning and transmission electron microscopy. Turkey BALT resembled other mucosa-associated lymphoid tissues; it was made up of a population of lymphocytes covered by a specialized epithelium different from typical pseudostratified ciliated columnar bronchial epithelium. There were distinct age-related differences in BALT structure. Bronchus-associated lymphoid nodules were larger and more numerous in older turkeys. In 1-day- to 2-week-old turkeys, the primary cell type of BALT epithelium was nonciliated cuboidal; in 2-week old turkeys it was squamous; and in turkeys older than 4-weeks of age, the epithelium was primarily ciliated columnar. In 1- to 4-week old turkeys, large numbers of intraepithelial lymphocytes disrupted the normal organization of the epithelium. In older turkeys, epithelial and lymphoid cells were in discrete compartments separated by connective tissue. Lymphocytes in 1-day-old turkeys were found in loose aggregates around venules and within the epithelium. In 1-week old turkeys, lymphocytes were organized into compartments of morphologically similar cells. By 3-weeks of age, lymphocytes were present in distinct germinal centers. Epithelial cells of BALT did not have large numbers of apical vesicles and thus were not structurally specialized for antigen uptake by endocytosis. However, the epithelial barrier appeared to be disrupted over lymphoid nodules, suggesting that antigen would be readily available to lymphocytes and phagocytes in BALT. Age-related differences in turkey BALT structure may have functional consequences with respect to the respiratory immune response.     

Fine structure of lymphoid cells in the nasal skin of primates.

Electron microscopy of the nasal skin of primates shows that lymphoid cells are normally present in this region. No degeneration or mitosis of these cells or their transformation into other cell types was seen. These intraepidermal lymphocytes contained cytoplasmic fibrils. There was no evidence that they were phagocytic in the epidermis. The present study suggests that these lymphoid cells migrate into the lamina propria but not into the epidermis.     

Histochemical and ultrastructural study of the digestive tract of the tortoise Testudo graeca (Testudines)

The digestive tract of the tortoise Testudo graeca (Testudines) was investigated by means of light and electron microscopy. The esophagus of T. graeca was lined by two types of epithelium: non-keratinized stratified squamous in the upper portion and stratified columnar in the lower. The lamina propria of the esophagus contained tubular or tubuloacinar glands. The mucosa of the stomach showed similar characteristics to those of other reptiles. The small intestine exhibited longitudinal folds lined by absorptive and goblet cells. The large intestine was lined by columnar mucous cells. Within the lamina propria of the large intestine, masses of 10–15 epithelial cells connecting with the surface epithelium by means of slender cytoplasmic processes were observed. A battery of six lectins (Con-A, PNA, WGA, DBA, SBA, and LTA) was used to identify the epithelial mucins. WGA and DBA reacted with all types of mucous cells throughout the alimentary canal. PNA was only unreactive in the intestine, LTA in the esophagus, and SBA in the large intestine. These results indicate a similar lectin-binding pattern throughout the gut of T. graeca.     

A Study of the Structure and Gliding Movement of Gregarina garnhami

SYNOPSIS The structure and gliding movement of Gregarina garnhami Canning, a eugregarine found in the midgut of the desert locust, Schistocerca gregaria , have been studied by light microscopy and transmission and scanning electron microscopy (EM). Ultrastructural studies revealed that the cytoplasm of G. garnhami is separated from the epicyte folds by a basal lamina. The pellicle consists of 3 membrane layers. At the tips of the epicyte folds there are 2 sets of longitudinally oriented filaments. An ectoplasmic network is present in the ectoplasm and the endoplasm contains numerous paraglycogen granules. The effect of cytochalasin B on G. garnhami was studied. Examination of scanning EM preparations of gliding and stationary gregarines yielded inconclusive results. In some instances the epicyte folds were thrown into waves; in others the folds were straight, regardless of treatment before fixation. Gregarina garnhami glides through its environment without any apparent deformation in shape. As it moves, a mucus trail is left behind it. Phase-contrast observations were made of centrifuged gregarines in which the endoplasm was displaced. Centrifuged gregarines continued to glide. Displacement of the endoplasm allows visualization of the epicyte folds in gliding animals. No lateral waves were seen in the epicyte folds of gliding centrifuged animals.     

Histopathological effect ofClostridium perfringens 8-6 enterotoxin on rabbit intestine

Intestinal damage caused by an enterotoxin from a coatless spore mutant ofClostridium perfringens type A (8-6) was identified by both light and scanning electron microscopy. Under the light microscope, damage to the epithelial layer of the villus and to the lamina propria was evident. Whole tissue viewed under the scanning electron microscope confirmed the two distinct forms of damage seen by light microscopy and showed that the action of the enterotoxin on an individual villus appears to occur in a specific sequence. The gross tissue damage observed contrasts with that found in previous studies of the action ofClostridium perfringens enterotoxin on rabbit ileal tissue; this suggests that the 8-6 enterotoxin may have a different mode of action on the cell, which subsequently leads to death and lysis.     

The early postnatal development of the primary immune response in TNP-KLH-stimulated popliteal lymph node in the rat

Summary The popliteal lymph nodes were removed from young rats of various ages five days after a single immunization with TNP-KLH in the hind footpads. Cryostat sections of the lymph nodes were investigated by means of enzyme and immunohistochemical techniques at the light-microscopical level.The presence and localization of anti-TNP antibody-containing cells were examined using a new technique to visualize specific antibodies. Moreover, the development of the lymph nodes following exogenous antigenic stimulation was compared with that of unstimulated lymph nodes.Specific antibody-containing cells could not be found before day 15 after birth, in rats immunized at day 10. From that time these lymphoid cells were located primarily at the border between cortex and medulla. Younger popliteal lymph nodes showed only aspecific immunoglobulin-containing lymphoid cells. With age, the number of specific antibody-containing cells tended to increase. These cells were more mature, according to morphological criteria and were located nearer the medulla.The first primary follicles were seen at day 19, as was the case in unstimulated animals. The first secondary follicles, containing germinal centers, were detected at day 23, whereas in unstimulated popliteal lymph nodes they were never found.Trapping of immune complexes could not be demonstrated before day 33 after birth. The later appearance of this phenomenon might be a consequence of the techniques applied to demonstrate specific antibody-containing cells.Abbreviations PLN popliteal lymph node - FDC follicular dendritic cell - IDC interdigitating cell - HEV high endothelial venule - TNP trinitrophenyl - KLH keyhole limpet hemocyanin - PBS phosphate-buffered saline - GCPC germinal center precursor cell - sIg surface immunoglobulin - cIg cytoplasmic immunoglobulin     

Targeted deletion of the IgA constant region in mice leads to IgA deficiency with alterations in expression of other Ig isotypes

A murine model of IgA deficiency has been established by targeted deletion of the IgA switch and constant regions in embryonic stem cells. B cells from IgA-deficient mice were incapable of producing IgA in vitro in response to TGF-beta. IgA-deficient mice expressed higher levels of IgM and IgG in serum and gastrointestinal secretions and decreased levels of IgE in serum and pulmonary secretions. Expression of IgG subclasses was complex, with the most consistent finding being an increase in IgG2b and a decrease in IgG3 in serum and secretions. No detectable IgA Abs were observed following mucosal immunization against influenza; however, compared with those in wild-type mice, increased levels of IgM Abs were seen in both serum and secretions. Development of lymphoid tissues as well as T and B lymphocyte function appeared normal otherwise. Peyer's patches in IgA-deficient mice were well developed with prominent germinal centers despite the absence of IgA in these germinal centers or intestinal lamina propria. Lymphocytes from IgA-deficient mice responded to T and B cell mitogens comparable to those of wild-type mice, while T cells from IgA-deficient mice produced comparable levels of IFN-gamma and IL-4 mRNA and protein. In conclusion, mice with targeted deletion of the IgA switch and constant regions are completely deficient in IgA and exhibit altered expression of other Ig isotypes, notably IgM, IgG2b, IgG3, and IgE, but otherwise have normal lymphocyte development, proliferative responses, and cytokine production.     

Structure and composition of the oral mucous membrane on the lips and cheeks of the monkey,Macaca fascicularis

Summary In seven monkeys (6 Macaca fascicularis, 1 M. mulatta; 2.4±0.6 kg in weight) the labial and buccal mucosae were studied morphologically and quantitatively. Following fixation by perfusion, the upper and lower lips and entire cheeks were dissected free and processed for light-, scanning and transmission electron microscopy. Established programs (HISTOMEP, MUMANA II) and appropriate morphometric techniques were used to estimate, at the light-microscopic level, the epithelial thickness, the width of the combined lamina propria/submucosa, and the volumetric composition of the gland-containing portions of lip and cheek mucosae. The cheek epithelium was more than twofold thicker than the lip epithelium, on the average 0.46±0.04 and 0.21±0.02 mm, respectively, with no differences related to sex or topographical sites. The combined lamina propria/submuscosa was 1.32 ±0.19 and 1.50±0.26 mm in width in cheeks and lips, respectively. The main mucosal constituents at both sites were glandular and connective tissue, and lymph follicles associated with secretory ducts. In lips, the volume of plasma cells around gland acini correlated positively with the amount of lymphoid tissue present around topographically related ducts. It is suggested that the duct/ lymph follicle assembly may serve as a local antigen-recognition system.

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Zusammenfassung Die Lippen- und Wangenmukosa wurde an 7 Affen (6 M. fascicularis, 1 M. mulatta; 2,4±0,6 kg schwer) morphologisch und quantitativ untersucht. Nach einer Perfusionsfixation wurden Ober- und Unterlippen sowie ganze Wangen herauspräpariert und für Untersuchungen im Licht-, Raster und Transmissionselektronenmikroskop vorbereitet. Bestehende Programme (HISTOMEP, MUMANA II) und morphometrische Standardmethoden wurden verwendet, um lichtmikroskopisch die Epitheldicke, die Breite der kombinierten Lamina propria/Submukosa und die volumetrische Zusammensetzung der drüsenhaltigen Abschnitte der Lippen- und Wangenschleimhaut zu ermitteln. Das Wangenepithel war mehr als zweifach dicker als das Lippenepithel (0,46±0,04 und 0,21±0,02mm); die Epitheldicke war unabhängig von Geschlecht und Topographie. Die kombinierte Lamina propria/ Submukosa war im Wangenbereich 1,32±0,19, im Lippenbereich 1,50± 0,26 mm breit. Die Hauptelemente der Lippen- und Wangenschleimhaut sind Drüsen- und Bindegewebe sowie mit Ausführungsgängen assoziierte Lymphfollikel. In der Lippen-, nicht aber in der Wangenschleimhaut, ist das Volumen der die Drüsenacini umlagernden Plasmazellen positiv korreliert mit der Menge der Lymphfollikel an den benachbarten Ausführungsgängen. Es wird vermutet, daß das Ausführungsgang/Lymphfollikel-System zur lokalen Erkennung von Antigenen dienen könnte.

     

Postnatal development of the tubular lamina propria and the intertubular tissue in the bovine testis

Summary The postnatal development of intertubular cells and vessels and of the tubular lamina propria was studied in three locations of perfusion-fixed bovine testes from 31 animals ranging from 4 to 78 weeks. The postnatal morphological differentiation of the testis is not uniform, regional differences have to be considered. The intertubular cell population is composed of mesenchyme-like cells, fibrocytes, Leydig cells, peritubular cells and mononuclear cells. In 4 and 8-week-old testes mesenchyme-like cells are the dominating element. These pluripotent cells proliferate by frequent mitoses and are the precursors of Leydig cells, contractile peritubular cells and fibrocytes. Morphologically differentiated Leydig cells are encountered throughout the entire period of postnatal development. In 4-week-old testes degenerating fetal and newly formed postnatal Leydig cells are seen in juxtaposition to each other. From the 8th week on, only postnatal Leydig cells are present. Between 16 and 30 weeks large-scale degeneration of prepuberal Leydig cells is observed. The Leydig cells that survive this degenerative phase constitute the long-lasting adult population. 20–30% (numerically) of all intertubular cells at all ages are free mononuclear cells. These are found as lymphocytes, plasma cells, monocytes, macrophages and light intercalated cells (LIC). The latter are monocyte-derived, Leydig cell-associated typical cells of the bovine testis. The differentiation of the two main components of the tubular lamina propria, (i) basal lamina and (ii) peritubular cell sheath, seems to be effected rather independent from each other and also from hormonal signals important for the development of the germinal cells. The laminated basal lamina reaches nearly 3 m at 16 weeks and is later on continuously reduced. At 25 weeks the peritubular cells have transformed into contractile myofibroblasts. At this period the germinal epithelium is still in a prepuberal state.To Dr. E. Schilling, Mariensee, on the occasion of his 65^th birthday     

A fluorescent antibody investigation of sites of gamma-globulin formation in the guinea-pig

Summary Fluorescein-labelled antibody was used to study sites of gamma-globulin formation in the normal guinea-pig. Gamma-globulin staining was confined to plasma cells and cells of germinal centres of lymphoid nodules. The significance of these findings and their possible relation to nuclear protein synthesis and germinal centre function are discussed.     

Regeneration of Vocal Fold Mucosa Using Tissue-Engineered Structures with Oral Mucosal Cells

Objectives

Scarred vocal folds result in irregular vibrations during phonation due to stiffness of the vocal fold mucosa. To date, a completely satisfactory corrective procedure has yet to be achieved. We hypothesize that a potential treatment option for this disease is to replace scarred vocal folds with organotypic mucosa. The purpose of this study is to regenerate vocal fold mucosa using a tissue-engineered structure with autologous oral mucosal cells.

Study Design

Animal experiment using eight beagles (including three controls).

Methods

A 3 mm by 3 mm specimen of canine oral mucosa was surgically excised and divided into epithelial and subepithelial tissues. Epithelial cells and fibroblasts were isolated and cultured separately. The proliferated epithelial cells were co-cultured on oriented collagen gels containing the proliferated fibroblasts for an additional two weeks. The organotypic cultured tissues were transplanted to the mucosa-deficient vocal folds. Two months after transplantation, vocal fold vibrations and morphological characteristics were observed.

Results

A tissue-engineered vocal fold mucosa, consisting of stratified epithelium and lamina propria, was successfully fabricated to closely resemble the normal layered vocal fold mucosa. Laryngeal stroboscopy revealed regular but slightly small mucosal waves at the transplanted site. Immunohistochemically, stratified epithelium expressed cytokeratin, and the distributed cells in the lamina propria expressed vimentin. Elastic Van Gieson staining revealed a decreased number of elastic fibers in the lamina propria of the transplanted site.

Conclusion

The fabricated mucosa with autologous oral mucosal cells successfully restored the vocal fold mucosa. This reconstruction technique could offer substantial clinical advantages for treating intractable diseases such as scarring of the vocal folds.     

On structural patterns of the lamina propria of human seminiferous tubules

Summary The ultrastructure of the lamina propria of human seminiferous tubules was analyzed in normal specimens and compared to biopsies showing great thickenning of this area in light microscopy.The contractile cells are stellate in shape, the intercellular gaps between their branchings being less than 150 Å. The cytoplasmic features of these cells are similar to those described by Ross and Long (1966) and do not differ significantly in the pathological cases examined.The intercellular components, namely collagen fibers, microfibrils and an incomplete basement membrane-like coating of the contractile cells, are strikingly increased in the thickenned lamina propria, although the number of layers making up this structure needs not be increased. Occasionally, the intercellular space is occupied by only one of these materials.The distribution of collagen permits identification of two main patterns in the thickenned lamina propria: a) one where the basement membrane of the seminiferous epithelium is separated from the first layer of contractile cells by a wide collagen zone, and b) another case where the layer displaying greater thickness because of increased collagen deposition is located further away from the germinal epithelium.The functional activity of the contractile cells, the physiological implication of structural alterations of the lamina propria and the necessity to correlate these observations to andrological findings, are discussed.Presented in part at the 68. Meeting of the Anatomische Gesellschaft, Lausanne, April 1973.Fellow of the Alexander von Humboldt Foundation, on leave of absence from Depto. de Biología y Genética, Sede Norte, Universidad de Chile, Santiago.Supported by Grants from the Deutsche Forschungsgemeinschaft.     

Distribution of SIV infection in the gastrointestinal tract of rhesus macaques at early and terminal stages of AIDS

Intestinal tissues from SIV-infected rhesus macaques with subclinical as well as with terminal SIV disease were evaluated for SIV infection. SIV-infected mononuclear cells were detected throughout the gastrointestinal tract in all animals. In early infection, SIV-infected cells were diffusely distributed in lymphoid tissue and lamina propria, whereas in late stages of the disease, the viral infection was more severe and widely disseminated. Lymphoid cells of the lamina propria and gut-associated lymphoid tissue were the primary targets of SIV.     

Age-related decline of lymphoid-tissue components in the oral mucosa of the lip,cheek and soft palate of Macaca fascicularis

Summary In three mature monkeys (Macaca fascicularis; 3.5±0.3 kg in weight), the labial, buccal and soft-palate mucosae were examined morphologically and stereologically. Using fixation by perfusion, standardized methods of tissue preparation and morphometric analysis at the light-microscopic level, the gross dimensions (i.e., epithelial thickness, width of combined lamina propria/submucosa) and the volumetric composition of the oral mucosae were estimated and compared with those of young animals examined previously. The data show (1) an age-related decline in the volume and prevalence of organized lymphoid tissue (i.e., lymphoid follicles associated with secretory ducts), (2) a stable plasma-cell density in the interglandular connective tissue, and (3) an increase of glandular tissue in mature versus young animals. It is suggested that the lymphoid follicles associated with secretory ducts, providing for plasma-cell generation, mirror the tonsillar lymphoid tissue declining after puberty.     

Videostroboscopic and morphological aspects of voice disturbances in patients with larynx atrophy and coexisting hypopharynx cancer

Vocal folds play a crucial role in voice production. The physiological vibrations of vocal folds depend on the unchanged multilayered structure of the vocal folds mucosa. Morphological changes of mucosa are the cause of voice quality disorders - dysphonia. The aim of this study was to determine the morphological base of dysphonia in patients with vocal folds atrophy. A group of 24 patients with larynx atrophy confirmed by endoscopic (VLS) and stroboscopic (VLSS) examination of the larynx was included in the study. The morphological assessment of the larynx mucosa was carried out with the use of the transmission electron microscopy (TEM). Ultramorphological examinations revealed changes in the epithelium, basal membrane and lamina propria of the vocal folds mucosa. An increased number of collagenous fibers, fibroblasts with signs of vacuolar degeneration inflammatory cells and a decreased number of blood vessels and pericytes were observed. Morphological changes found in the epithelium, basal membrane and lamina propria of the vocal folds mucosa were the cause of disorders of vocal folds vibrations registered in the stroboscopic examination of the larynx (VLSS).     

Enzymhistochemical Localization of Alkaline Phosphatases in Germinal Cysts of the Testis of the Frog Rana temporaria (Anura)

Alkaline phosphatases have been demonstrated enzymhistochemically in the testis of the common frog Rana temporaria, caught in the month of January. The follicle cells in the cyst walls and the lamina propria of the seminiferous tubules showed strong enzymatic activity; also a weak activity was observed in the peripheral region of some of the germinal cells lying within the cysts. The possible interaction of alkaline phosphatases in the transport processes across the cyst walls has been discussed.     

Distribution of Leu 7 (HNK-1) antigen in human digestive organs: an immunohistochemical study with monoclonal antibody

Summary Leu 7 immunoreactivity was demonstrated with the indirect peroxidase-labelled antibody method on frozen and paraffin-embedded tissue sections of human digestive organs. Anti-Leu 7 monoclonal antibody, which allegedly detects mononuclear cells with natural killer or killer activity, recognized lymphoid cells among intestinal epithelial cells and in the germinal centres of solitary lymphoid follicles of small and large intestine, and a few in gallbladder, liver and the lamina propria of the intestine. In addition, peripheral nerve fibres, endocrine cells in the gut and pancreas and carcinoid and islet cell tumours were also positively stained. At the ultrastructural level, Leu 7 antigen was localized on the plasma membrane of granulated lymphoid cells in the gut mucosa and on the secretory granules of intestinal endocrine cells. In normal pancreas, Leu 7 immunoreactivity was demonstrated in most cells containing pancreatic polypeptide and in many cells containing somatostatin or glicentin. Insulin-containing cells, however, lacked Leu 7 immunoreactant. These findings were obtained in both frozen sections and paraffin-embedded sections. The possible cross-reactivities of monoclonal antibodies are discussed as they raise an important caveat in immunohistochemical studies using these antibodies.     

Development of alveolar septa and formation of alveolar pores during the early postnatal period in the rat lung

In order to investigate the formation of alveolar pores, lungs of rats, after intratracheal perfusion of glutaraldehyde, are processed at postnatal days 1, 7, 14, 16 and 21 for light and transmission electron microscopy and at days 7 and 16 for scanning electron microscopy. The initial low secondary crests of day 1 rapidly elongate to pleats subdividing the primary saccules. The ledges of some pleats partly grow toward each other as ring like diaphragms, leaving openings whose boundary is composed of alveolar epithelium separated by a basal lamina from a connective tissue sheath with capillaries. At day 7, in scanning electron microscopy the lumina of some rudimentary alveoli communicate by apertures of different sizes, as a result of the outgrowth of curved alveolar pleats which narrow to a ring-like aperture. The interalveolar openings observed in scanning electron microscopy resemble those investigated by light and transmission electron microscopy. The number of interalveolar pores increases from day 7 on; they become more and more frequent at days 14, 16 and 21, respectively. It appears that alveolar multiplication in newborn rats proceeds not only by segmentation of terminal respiratory units but also by compoundment of septa. The difference between genuine pores and transsections of folds in transmission electron microscopy will be given closer attention in this study. Also, the incidence and location of type II pneumocytes during rapid enlargement of the alveolar surface area is discussed.     

Immunohistochemical identification of T- and B-lymphocytes delineated by the unlabelled antibody enzyme method

Summary T-lymphocytes and B-lymphocytes are identified in tissue sections of human tonsils by applying the unlabelled antibody enzyme method. The epithelium of the tonsils contains a majority of immunoglobulin-positive cells and fewer T-lymphocytes. In the subepithelial zones, areas composed of B-cells predominate, however, regions containing T-lymphocytes are also present. The latter are mainly arranged in the lamina propria around high-endothelial venules and often include plasma cells containing immunoglobulin. Follicles containing germinal centres display a complex structure which changes during development. The lymphocytic cap consists of densely packed lymphocytes, labelled heavily by anti-IgM and anti-IGD, and of individual T-lymphocytes. Germinal centres show a framework of immunoglobulin-positive dendritic reticular cells; they contain some heavily labelled lymphoid cells and several cells weakly labelled by anti-IgM and anti-IgA, as well as a small number of T-lymphocytes. Furthermore, the total areas of T- and B-lymphocytes measured by planimetry may differ considerably between different tonsils. Especially total areas of germinal centres vary to a great extent. The quantitative data on amounts of T- and B-cells achieved by planimetry are comparable to those reported in cellular suspensions of tonsils.