Ultrastructural analysis of normal and immunized spleen of the snapping turtle,Chelydra serpentine

In the ultrastructural comparison of normal, unimmunized spleens with immunized spleens at key intervals after antigenic stimulation with keyhole limpet hemocyanin (KLH), we noted cellular and cytological features which reflect the cellular kinetics of the primary immune response, particularly with respect to plasma cell production. Although lymphoblasts and mature plasma cells are present in the white and red pulp, respectively, intermediate stages of the plasma cell line are rarely found in normal spleen. Following antigenic challenge, we found a marked increase in lymphoblasts in the white pulp, most of them containing short segments of rough endoplasmic reticulum suggesting initial differentiation toward plasma cells. Following an apparent migration of cells from the white to the red pulp, we found plasma cells in various stages of maturation in the red pulp cords and sinuses. The ultrastructural features of these cells reflect 'the differentiation of lympho blasts into mature plasma cells. Both immature and mature plasma cells usually possess dilated cisternae of rough endoplasmic reticulum, suggesting that they are capable of producing and storing a secretory product, presumably antibody. We also noted a large number of immature macrophages and monocytes in immunized spleens. These cellular events and their cytological characteristics are compared to those described in other vertebrate classes.     

Marginal zone bridging channels as a pathway for migrating macrophages from the red towards the white pulp in the rat spleen

The spleen of (PvG/c X DA)F1 rats, intravenously injected with carbon, was investigated. Large heavily carbon-laden (LHC) macrophages, which were found only in the red pulp at 30 min, appeared along marginal zone bridging channels (MZBC) from the red pulp towards the white pulp side successively during 1-6 h after carbon injection. After this time, they appeared in the periarterial lymphatic sheaths (PALS) near MZBC and then in the deeper PALS along the arteries by 5-10 days. Frequently, they were found in rows from MZBC into the white pulp. These findings suggest migration of LHC macrophages from the red towards the white pulp trough MZBC. Possible migration of LHC macrophages through MZBC was observed for a long period--at least 3 months examined. LHC macrophages came together preferentially in PALS and in and around the germinal centers consisting of large pyroninophilic lymphoblastoid cells. Occasionally, possible migration of LHC macrophages from regions around sinuses crossing the marginal zone vertically (vertical sinus) was also observed. Sinuses accompanied by LHC macrophages often ran parallel in close association with MZBC, particularly at sites of MZBC near the red pulp.     

Plasmodium chabaudi chabaudi infection in mice induces strong B cell responses and striking but temporary changes in splenic cell distribution

B cells and Abs play a key role in controlling the erythrocytic stage of malaria. However, little is known about the way the humoral response develops during infection. We show that Plasmodium chabaudi chabaudi causes major, but temporary changes in the distribution of leukocytes in the spleen. Despite these changes, an ordered response to infection develops, which includes vigorous extrafollicular growth of plasmablasts and germinal center formation. Early in the response, the lymphocytes in the T zone and follicles become widely spaced, and the edges of these compartments blur. This effect is maximal around the peak of parasitemia. Germinal centers are apparent by day 8, peak at day 20, and persist through day 60. Extrafollicular foci of plasmablasts are visible from day 4 and initiate a very strong plasma cell response. Initially, the plasma cells have a conventional red pulp distribution, but by day 10 they are unconventionally sited in the periarteriolar region of the white pulp. In this region they form clusters occupying part of the area normally filled by T cells. B cells are absent from the marginal zone for at least 30 days after the peak of infection, although flow cytometry shows their continued presence in the spleen throughout infection. Relatively normal splenic architecture is regained by day 60 of infection. These results show that the changes in splenic cell distribution are linked to the presence of parasites and do not seem to interfere with the development of the humoral response.     

The effect of primary and secondary immunization on the lymphoid tissues of the carp, Cyprinus carpio L

Mirror carp immunized with human gamma globulin (HGG) in Freund's complete adjuvant (FCA) show a proliferative response involving cells whose cytoplasm stains deep red with methyl green-pyronin (pyroninophilic cells). This response occurs particularly in the haemopoietic parenchyma of the pronephros and mesonephros. It peaks at week 3, with the formation of clusters of pyroninophilic cells in the pronephros. Immunization with Aeromonas salmonicida elicited a less intense pyroninophilic response but caused a larger increase in pigment-containing cells. After a secondary immunization with HGG in FCA, a distinct response was observed in the spleen: Pyroninophilic cells collected within the ellipsoid sheaths in large numbers and formed nodules. The reticulum of such nodules acquired spherical proportions and resembled the white pulp reticulum of the tetrapod spleen. The roles of such pyroninophilic cells and the possibility that aggregations of them may be functionally analogous to homoiotherm germinal centres are discussed.     

Ultrastructural changes in the spleen of the natterjack,Bufo calamita,after antigenic stimulation

Summary In the present study comparative aspects of the ultrastructure of the spleen were analyzed in non-immunized and T-dependent antigen-challenged natterjacks, Bufo calamita. Special attention is focused on the role of the non-lymphoid components in the splenic immunoreactivity. Ten days after primary immunization with sheep erythrocytes, splenic lymphoid follicles increase considerably in number and size. By that time, lymphoblasts, medium and large lymphocytes abound in the periphery of the white pulp near the marginal zone. Meanwhile, in the red pulp numerous monocytes migrating across the sinusoidal walls apparently transform into giant, dendritic-like cells. Twenty days after immunization the splenic lymphoid follicles decrease in number, although certain reactivity persists and numerous plasma cells occur in the cell cords and sinusoids of the red pulp. These results are discussed comparatively with those reported in other lower vertebrates.     

Structure and histochemical organization of the spleen of Agama stellio (Sauria: Agamidae) and Chalcides ocellatus (Sauria: Scincidae)

The spleen of Agama stellio is composed mainly of red pulp; the white pulp is poorly developed, and its clusters are scattered throughout the organ and contain lymphocytes, reticular cells, and some plasma cells. The red pulp consists of clear reticular cells intermingled with blood cells, sinusoids, and pigment cells. The spleen of Chalcides ocellatus is encapsulated by connective tissue and is composed of white and red pulp. The white pulp consists of lymphoid tissue that surrounds the central arterioles, forming the periarteriolar lymphocyte sheath (PALS). The red pulp is composed of a system of venous sinuses and cords. The results of various histochemical procedures designed to demonstrate mucosubstances, proteins, and nucleic acids indicate that the spleen in these species resembles the mammalian spleen. © 1993 Wiley-Liss, Inc.     

The proliferative response of the spleen in cryosurgery.

Using ³H-thymidine autordiography, we studied the cellular proliferation of the spleen of rats after cryolesions in liver, kidney, spleen, and stomach.In the germinal centers at first, a dissociation develops, followed by a hyperplasia with high labeling indices of the germinal center cells with a maximum between the second and third postoperative day. In the surrounding lymphatic mantle zone of the white pulp, as well as in the marginal zone, an increased labeling index of the cells can be observed between the first and second day. The highest percentages of labeled cells in the red pulp are seen on the fifth postoperative day.These cell kinetic results correspond very well with those after antigenic stimulation, for instance, after intravenous injection of sheep erythrocytes. Therefore, these findings suggest that an immunologic reaction occurs in the spleen after cryolesions on parenchymal organs.     

Morphometric study of the spleen in C57BL strain mice during pregnancy with syngeneic or allogeneic embryos

A morphological study of the spleen in the C57BL line female mice on the 16th day of pregnancy showed a significant increase in the relative area of generative centres and red pulp and a decrease in the white pulp relative area as well as a rise in the amount of plasmatic cells and blast forms in red pulp during allogenic pregnancy (BALB embryos). Such changes are supposed to favour both the preservation of the intact mother-extrafetal organs-fetus system and the improvement of allogenic embryos trophics.     

Ultrastructure of phagocytic cells in the spleen of Psammophis sibilans (serpentes: Colubridae)

The spleen of Psammophis sibilans is composed mainly of red pulp, the white pulp being poorly developed. The white pulp lymphoid clusters are scattered throughout the organ and contain lymphocytes, reticular cells, and some plasma cells. The red pulp consists of reticular cells intermingled with blood cells, sinusoids, and melanomacrophage centers (MMCs). Filtering of particulate matter from the blood occurs in the red pulp by phagocytes of the pulp cord. MMCs are formed by the association of free macrophages that have phagocytosed some blood cells. Early filtering of particulate matter by the phagocytes of the pulp cords may allow for more efficient phagocytosis of erythrocytes by the MMCs. © 1994 Wiley-Liss, Inc.     

Splenic white pulp as a thymus-independent area in the African clawed toad,Xenopus laevis

Summary An indirect immunofluorescence study of the frozen sections of the spleen of an anuran amphibian, Xenopus laevis, showed that lymphocytes bearing a small amount of immunoglobulin (Ig) were localized mostly in the white pulp of non-immunized toads. There were fewer fluorescent cells in the red pulp. In the toads hyperimmunized with human gamma globulin (HGG), cells with strong cytoplasmic fluorescence increased significantly in the outer part of the white pulp. Electron microscopy of spleens from these toads showed that plasma cells at different stages of maturation were abundant in the white pulp, whereas in the red pulp, a smaller number of maturer plasma cells were observed. These results indicate that, in contrast with its mammalian counterpart, the splenic white pulp of this anuran is the site where thymusin-dependent lymphocytes commence blast formation and transformation into plasma cells.     

The spleen and skin wound healing in Xenopus adults

In most vertebrates, the regenerative capacity to restore lost/damage tissues to original structure and functionality decreases at some time during ontogenesis. To evaluate the role of the acquired immunity in the decline of regenerative potential, we examined the cellular responses elicited in the spleen during skin repair in Xenopus adults. Modifications in the architecture were found to be induced and were remarkable 14 days postinjury when the spleen increased significantly in size. In white pulp, the periarteriolar lymphoid sheaths were associated with follicles having central light zones, morphologically similar to germinal centers. With the progress of healing, pigment‐containing cells were seen to accumulate in both white and red pulp regions. Moreover, compared to controls, the cells immunoreactive to anti‐cytokines (TNF‐α, TGF‐β1) and ‐iNOS increased from the first days after wounding. The 14th day, the positive cells formed a dense network of reticular cells in central regions of lymphoid follicles and more frequent reactive leukocytes were detected within the red pulp. A higher number of lymphoid cells immunostained with anti‐CD3ε were also observed in the perifollicular zone. The results suggest that the spleen of adult frogs is involved in skin wound healing with the expansion of lymphoid compartments. J. Morphol. 277:888–895, 2016. © 2016 Wiley Periodicals, Inc.     

Variation of lymphoid activity in the spleen of a migratory bird,the pied flycatcher (Ficedula hypoleuca; Aves,passeriformes)

The size and microscopic structure of the spleen of the migratory pied flycatcher (Ficedula hypoleuca) show marked changes during the reproductive cycle. Upon the spring return to their northern breeding sites, the birds have a small spleen with little lymphoid activity and a poorly developed red pulp. During the breeding period the volume of red and white pulp increases, the number and distinctness of lymphoid follicles (germinal centres) in the white pulp increase, and groups of cells with intensely basophilic cytoplasm, probably B cells (plasma cells), appear. The findings suggest that the immune system of the adult pied flycatcher is activated during periods when it is bound to the nest. Young flycatchers beginning their autumn migration also show a marked increase of lymphoid activity in the spleen.     

Immune proteasomes in the developing rat spleen

Changes in the structure of the rat spleen and the distribution of immune proteasomes in it during early postnatal development have been studied using double immunofluorescent staining of tissue sections with antibodies to the LMP7 immune proteasome subunit and to specific markers of T and B lymphocytes. It has been shown that the white pulp on postnatal day 5 is not yet colonized by lymphocytes and contains a smaller amount of immune proteasomes than the red pulp. At this stage, T and B lymphocytes concentrate mainly in the red pulp. On day 8, B lymphocytes occupy the marginal zone, while T lymphocytes aggregate into dense strands close to the white pulp. By day 18, T lymphocytes form periarteriolar sheaths in the white pulp, and the contents of immune proteasomes in the red and white pulp become equally high. An increase in the total content of immune proteasomes in the spleen on the third postnatal week was revealed in our previous study by Western blotting. In addition to T and B lymphocytes, immune proteasomes have also been revealed in other spleen cell types, probably in macrophages and reticular cells of the white pulp. Thus, the postnatal development of the spleen is associated with an increase in the contents of immune proteasomes in it.     

Cell proliferation in vivo and in vitro in visceral organs of the adult newt, Triturus cristatus carnifex

The mitotic and labelling incidence of intestine, liver, spleen and pancreas cells of Triturus cristatus carnifex adults kept at 15°C, 20°C, 25°C and 30°C were examined. Intestine mitotic and labelling incidences were highest at 25°C and lowest at 30°C. There was no significant difference between 15°C and 20°C. No such relationship could be shown for liver, spleen or pancreas, which had very much lower mitotic and labelling incidences. In culture, intestine mitotic and labelling incidences fell significantly within the first four hours, and maintained these low levels for the next five days. In contrast, liver mitotic and labelling incidences rose for 9–11 days, and then began to fall, while pancreas mitotic and labelling incidences reached peak values at day 5, and were kept in good condition for up to 14 days.     

Ultrastructure of splenic white pulp of the turtle,Mauremys caspica

Summary The ultrastructure of splenic tissue of non-immunized turtles, Mauremys caspica, shows two areas, namely, the white pulp which is lymphoid in nature, and the red pulp which is formed by cell cords and sinusoids. Between both areas there is always a marginal zone with gaps through which cells leak. In the white pulp, there are two blood vessel types; one with muscled walls, and the other showing thinner walls sheathed by reticular cells. Reticular cells constitute a network where there occur dendritic macrophages, lymphoblasts and small and medium lymphocytes. Mature plasma cells are scarce in the white pulp.     

Recovery processes in rabbit spleen after hydrocortisone administration

The effect of hydrocortisone on the rabbit spleen and the recovery processes after the cessation of the hormone injection was studied. A decrease of the absorption function of RES, and spleen atrophy, expressed in a decrease of the organ weight and size followed prolonged hydrocortisons use. Most of the spleen nodes were reduced to an irregular accumulation of lymphocytes. The pyroninophilic cells disappeared in the white pulp. Cessation of the hormone effect was followed by an improvement in the general condition of the animal; the development of the spleen atrophic processes was suspended, the spleen nodes were restored with an increase of pyroninophilic cell count in them. The use of a stimulant under these conditions accelerated the recovery of the animal weight, as well as that of the atrophic spleen, with a normalization of the organ structure.     

Studies on halimeda. V. effect of temperature on chloroplast migration in this Siphonous green alga&*

Abstract

The effect of temperatures between 15 and 30°C on the daily cycle of chloroplast migration in Halimeda distorta and H tuna was determined from changes in segment pigmentation recorded by time‐lapse videography throughout the experiments. An un‐named opuntioid species was also tested between 20 and 35°C. Both the daily pattern and the amplitude of change in surface pigmentation, which were sustained for at least 5 days at 25°C, were unchanged at the higher temperatures. At 20°C the amplitude was considerably reduced but cyclical changes in surface pigmentation continued to occur throughout the 3‐day low temperature period. However, at 15°C even greater reduction in amplitude was observed together with a reduced rate of paling at the onset of darkness and absence of pre‐dawn re‐greening. Furthermore, at 15°C all daily changes in surface pigmentation had ceased by the second day in H tuna and by the third day in H distorta. These effects of lower temperatures were reversed when the plants were returned to 25°C, although after the 15°C treatment of H tuna the amplitude of the change in surface pigmentation in two of the three replicate plants was small on the first night back at 25°C whilst the third plant lost pigmentation progressively and was completely white, and apparently dead, two days later.     

An ultrastructural study of the spleen of the ranid frog Rana perezi

The spleen of Rana perezi is encapsulated by connective tissue and shows by light microscopy two areas with no obvious border: the white pulp and the red pulp. The white pulp-lymphoid clusters are scattered throughout the organ and contain lymphocytes, reticular cells, and some plasma cells. The red pulp displays two different portions. The predominant region consists of reticular cells, lymphocytes, a variety of other leucocytes, and cells undergoing division. This area possibly performs a haemopoietic function. The smaller portion of the red pulp is characterized by reticular-phagocytic cells and may be haemocaretic in its function. Macrophages and pigmented cells occur in both white and red pulp. The organization of the spleen of R. perezi can be considered as a transitional or intermediate state between the primitive condition seen in certain fishes and amphibians and the more complex organ of ammiotes.     

Blood and Spleen Response to Isolation and Cold Stress in Little Brown Bats,Myotis lucifugus

Female bats were housed singly or in groups and maintained at 5° or 23 °C. Red blood cell life spans were determined utilizing the disappearance of labeled sodium chromate (Cr⁵¹). Red cell counts, hematocrits, reticulocyte percentages and diameters, white cell differentials, and spleen histology were determined for bats subjected to three weeks of stress treatment. Isolated animals (warm and cold) had significantly lower hematocrits and red cell counts and significantly higher reticulocyte diameters and percentages than grouped bats. Random destruction of erythrocytes was initially higher in isolated bats. In cold and isolated animals the white pulp of the spleen was most reduced in quantity and in proliferative activity. Red pulp became highly engorged with erythrocytes and showed a hypertrophy of reticuloendothelial elements. Higher rates of random destruction are closely correlated with increased splenic erythrophagocytosis. Gold or isolation imposed separately are mild stress factors in little brown bats, while a severe stress response occurs when the two stressors are imposed simultaneously.     

The generation of large pyroninophilic cells in the lymphoid tissues of rats infused with cell-free lymph.

The thoracic duct of Wistar strain rats was cannulated during 5 days for studying the effect of selective lymphocyte depletion on the lymphoid tissue. A technique for the continuous infusion of cell-free lymph, whole lymph of Eagle's medium to the rat with the thoracic duct fistula is described in detail. The prolonged drainage of lymph from rats was followed by lymphopenia, sever atrophy of lymphoid tissues and the depletion of small lymphocytes in the thymus-dependent areas of spleen and lymph nodes. The infusion of cell-free lymph into the drained rat resulted in the recovery of the weight of lymphoid tissues and in the massive proliferation and accumulation of large cells with prominent nucleoli and intensely pyroninophilic cytoplasm in the lymphocyte depleted areas of the peripheral lymphoid tissues and thymic cortex. There was histological evidence that the large pyroninophilic cells developed well in the spleen and tended to localize preferentially around the periarteriolar region through the marginal zone bridging channels to the red pulp. The infusion of Eagle's medium was found ineffective in restoring the weight of the lymphoid tissues and in bringing about the proliferation of lymphoid cells. The rats infused with whole lymph showed almost similar findings biologically and histologically to those of sham-operated rats.