Human spleen microanatomy: why mice do not suffice

The microanatomical structure of the spleen has been primarily described in mice and rats. This leads to terminological problems with respect to humans and their species‐specific splenic microstructure. In mice, rats and humans the spleen consists of the white pulp embedded in the red pulp. In the white pulp, T and B lymphocytes form accumulations, the periarteriolar lymphatic sheaths and the follicles, located around intermediate‐sized arterial vessels, the central arteries. The red pulp is a reticular connective tissue containing all types of blood cells. The spleen of mice and rats exhibits an additional well‐delineated B‐cell compartment, the marginal zone, between white and red pulp. This area is, however, absent in human spleen. Human splenic secondary follicles comprise three zones: a germinal centre, a mantle zone and a superficial zone. In humans, arterioles and sheathed capillaries in the red pulp are surrounded by lymphocytes, especially by B cells. Human sheathed capillaries are related to the splenic ellipsoids of most other vertebrates. Such vessels are lacking in rats or mice, which form an evolutionary exception. Capillary sheaths are composed of endothelial cells, pericytes, special stromal sheath cells, macrophages and B lymphocytes. Human spleens most probably host a totally open circulation system, as connections from capillaries to sinuses were not found in the red pulp. Three stromal cell types of different phenotype and location occur in the human white pulp. Splenic white and red pulp structure is reviewed in rats, mice and humans to encourage further investigations on lymphocyte recirculation through the spleen.     

Origin of the chicken splenic reticular cells influences the effect of the infectious bursal disease virus on the extracellular matrix

The effects of infectious bursal disease virus (IBDV) (strain F52/70) infection were studied by immunohistochemical methods on the splenic extracellular matrix (ECM). The major fibrillar components of the ECM, the type I and type III collagens and the main ECM organizing glycoproteins (laminin, tenascin and fibronectin) were monitored up to 11 days post-infection (d.p.i.). By 3 d.p.i., the collagens that form the basic scaffold of the antigen-trapping region of the spleen are destroyed, which is followed by deterioration of the glycoproteins. The ECM in the red pulp and the other regions of the white pulp (periarteriolar lymphatic sheath and germinal centre) seem to be normal. The reason for the significantly different pathological alterations in the ECM between the two regions of the spleen may be explained by the origin of the reticular cells. The reticular cells in the antigen-trapping zone and other splenic regions are of haemopoietic and mesenchymal origins, respectively. Possibly, the reticular cells of the haemopoietic origin are more susceptible for the IBDV infection than the mesenchymal ones. Development of the antigen-trapping, B-cell-dependent zone of the splenic white pulp precedes that of the periarteriolar lymphatic sheath and germinal centre, which suggests that this region may contribute to B-cell maturation. Damage of the ECM in the antigen-trapping zones results in impairment of tissue organization, which may contribute to the permanent immunosuppression.     

Butyrylcholinesterase-positive innervation of the spleen in rats

Innervation of the spleen in rats was studied. Butyrylcholinesterase-(BuChE)-positive nerve components of the organ were visualized by the direct thiocholine method. BuChE-positive nerve components enter the spleen in a common bundle with arteries. In the organ they form characteristic periarterial and periarteriolar plexiform arrangements, which are especially conspicuous around the aa. centrales running through the white pulp. Then, nerve fibres extend away from these plexuses into adjacent layers of trabeculae further into marginal layers of periarterial lymphatic sheath (PALS) as well as into the mantle zone of follicles. Several scattered periarteriolar and solitary nerve fibres can be seen in the marginal sinuses and cords of the red pulp. In the fibrous capsula BuChE-positive nerve fibres can also be seen which have an evident connection with trabecular and parenchymal nerves of the organ. Microscopic findings support the notion that BuChE-positive nerve profiles supply not only the vasculature, but also the parenchymal components of the spleen, and they may participate, to a great extent, in the regulation of the immune processes in this organ.     

大鼠脾脏的组织发生

应用光镜、扫描电镜和透射电镜对大鼠脾脏的组织发生进行了形态学观察。脾原基发生在妊娠第16天,继则血管出现,血窦形成,脾脏造血开始。妊娠第19天,网状细胞围绕着发生中的小动脉形成动脉周围鞘。随之淋巴细胞聚集于小动脉周围。出生后,各种血细胞造血相继终止,红、白髓不断发育成熟,生后第5天观察到淋巴树突细胞。边缘带、边缘窦和脾小结相继出现。生后第25天观察到原始生发中心。第40天后,脾脏各部结构基本发育完善。     

Splenic white pulp and associated vascular channels in chicken spleen

We have studied the chicken spleen by light and transmission electron microscopy. Germinal centers are located at the beginning of the central artery which is surrounded by the periarterial lymphatic sheath (PALS). The central artery has no branch crossing the PALS, and there is no histologically identifiable marginal zone in the chicken spleen. The central artery continues as penicilliform capillaries. The mid-portion of the penicilliform capillary is surrounded by the ellipsoid or Schweigger-Seidel sheath. The endothelial lining of this part of the pencilliform capillary contains small channels, formed between neighboring endothelial cells, which enter the ellipsoid. These channels allow circulating substances to accumulate in the ellipsoid. The cells of the ellipsoid are reticular cells, round or ovoid in shape, exhibiting a limited number of cell junctions. At the surface of the ellipsoid are ellipsoid-associated cells (EAC) which have an affinity for toluidine blue. After perfusion fixation, the majority of the cells in the ellipsoid are lost; this suggests weak cellular connections between the ellipsoid cells. The ellipsoid-associated cells remain in loco. On the basis of their shape and cytological features, two stages of EACs can be distinguished. The round or ovoid EACs elaborate active Golgi zones surrounded by numerous small vesicles containing an electron-dense substance. Occasionally, mitotic figures can be seen among them which may suggest that they are immature forms. In the second or more mature stage, the EACs assume an elongated spindle shape. The spindle-shaped EACs reveal inactive Golgi cis-ternae and fewer but larger granules than the round or immature EACs. Unmyelinated nerve fibers and nerve ending were observed in the ellipsoid. Perfusion fixation reveals that the distal portions of the penicilliform capillaries extend into the red pulp and become the sinuses. Therefore, the circulation of the chicken spleen is anatomically closed except for the channels in the mid-portion of the penicilliform capillary. The periellipsoid white pulp (PWP) possesses a few small lymphocytes, macrophages, and plasma cells with the majority of cells being young blastlike cells which may replenish the EACs and ellipsoidal cells. Carbon, injected intravenously, appears on the EAC membrane by 30 minutes, while Brucella abortus is completely phagocytized at this time. Binding the carbon triggers the EAC to detach from the ellipsoid and migrate to the periellipsoidal white pulp (PWP). The EACs phagocytize carbon by 2 hours, and by 5 and 8 days are present in the red pulp and surround the germinal centers at the border of the periarterial lymphatic sheath, respectively. Secretory cells which have been identified in germinal centers might originate from EACs.     

Architecture of the Blood‐Spleen Barrier in the Soft‐Shelled Turtle,Pelodiseus Sinensis

We investigated the structure of the soft‐shelled turtle, Pelodiseus sinensi, spleen and demonstrated that there were several microanatomical peculiarities by light and transmission electron microscopy. In the spleen, the white pulp of the spleen was composed of two compartments, the periarteriolar lymphatic sheath (PALS) and periellipsoidal lymphatic sheath (PELS). No lymph nodules and marginal zones were found. The spleen‐blood barrier stood in the PELS and the ellipsoid. The high endothelial lining of penicilliform capillary contained small channels. These channels allowed circulating substances or lymphocytes to enter the ellipsoid. The distal portion of the penicilliform capillaries directly opened to pulp cords. The ellipsoid‐associated cell (EAC) was located at the surface of the ellipsoid. Reticular fibers were mainly distributed in ellipsoid and the outer PELS. Both reticular cells and macrophages were distributed in the outer layers of PELS. S‐100 protein positive dendritic cells were mainly distributed in out cells layer of the PELS and all over the PALS. Forty minutes after injection, carbon particles of Indian ink were mainly observed in the ellipsoid. Few carbon particles were observed in the outer PELS and fewer carbon particles in the red pulp. These findings suggested that a blood‐spleen barrier indeed existed in the soft‐turtle, P. sinensi, and it was a complex composed of an ellipsoid (including supporting cells, EAC, and reticular fibers) and the outer compartments of PELS (including dendritic cells, reticular fibers and cells, macrophages). Anat Rec, 2009. © 2009 Wiley‐Liss, Inc.     

High-dose interferon-gamma alters the distribution of B lymphocytes and macrophages in rat spleen and lymph nodes.

Continuous intravenous infusion of rat interferon-gamma (IFN-gamma) for 3 days provokes profound alterations of splenic architecture in LEW rats. The marginal zone of the white pulp is almost totally depleted of B lymphocytes and the follicles are reduced to small remnants. IgM kappa + plasmablasts and plasma cells increase substantially in the outer periarteriolar lymphatic sheath (PALS) and in the splenic red pulp. In addition, marginal metallophilic and marginal zone macrophages are augmented, partially by proliferation. It is discussed whether the activation and proliferation of these macrophages prevent replenishment of the marginal zone and follicles with recirculating B cells. Changes in B lymphocyte and medullary macrophage distribution are also present in submandibular and mesenteric lymph nodes.     

The differentiation of white pulp and red pulp in the spleen of human fetuses (72–145 mm crown-rump length)

The development of the white and red pulp in spleen from thirteen human fetuses measuring from 72 mm to 145 mm in crown-rump length (CRL) was studied using the electron microscope. This period follows the development of the primary vascular reticulum (Weiss, '73). The white pulp appears first as a periarterial sheath with variable numbers of large and medium-sized lymphocytes, monocytes, macrophages, and some granulocytes and erythrocytes. It is always rich in macrophages. At 90 to 100 mm CRL, reticular cells closely associated with collagen and having a distinctive dark hyaloplasm appeared first in the endothelium and close about blood vessels and then out in the pulp. In the white pulp they became circumferentially arranged about the central artery while in the red pulp they formed a branching reticulum. Small lymphocytes were present in increasing number in the periarterial lymphatic sheath after the development of the circumferential reticulum. The venous sinuses developed and the marginal zone stood out as an erythrocyte-rich and macrophage-rich shell of reticulum surrounding the periarterial sheath.     

Hodgkin's disease in the spleen

Summary 140 spleens involved by untreated Hodgkin's disease were studied utilizing conventional histological methods. Regardless of the sub-type of Hodgkin's disease, infiltrates of neoplastic cells were present either in the periarteriolar lymphoid sheath, the marginal zone or in both locations. Initially, infiltrates were confined to the splenic white pulp, later larger nodular foci of Hodgkin's disease developed by coalescence of several infiltrates. Neoplastic cells in Hodgkin's disease may reach the spleen by both retrograde lymphatic spread or the splenic artery; the presence of neoplastic cells in both T- and B-cell areas of the splenic white pulp implies a preference for Hodgkin's disease in the spleen with regard to a suitable microenvironment. This may be provided by certain macrophage subpopulations.     

The human spleen; a histological study in splenectomy specimens embedded in methylmethacrylate

In a series of 316 surgically removed spleens, a histological and supportive immunohistological study was performed on methylmethacrylate sections. The structure of the human white and red pulp differs from the rat spleen in many respects, e.g. the human lacks the marginal sinus and the architecture of the periarteriolar lymph sheath seen in the rat. In man, the lymphoid compartment is in both white and red pulps. In the white pulp separate periarteriolar T-cell areas contain a large lymph-vessel plexus, which was reconstructed in serial sections. The circulation in the red pulp is discussed. The area between the red and white pulp, the perifollicular zone, is not the equivalent of the marginal sinus in the rat. Its anatomy in man suggests that it is an area formed from red pulp during the expansion of new follicles. The micro-anatomy was analysed in 119 controls. In cases of traumatic rupture the white pulp showed evidence of stimulation. A pathognomonic histological picture was not found in idiopathic thrombocytopenic purpura. In haemolytic anaemia the pulp cords were engorged by erythrocytes accompanied by a decreased B/T cell ratio in autoimmune haemolytic anaemia and by an increased B/T cell ratio in congenital spherocytosis.     

中华鳖脾脏椭球的微细结构及其循环特征

目的 观察和分析12只中华鳖脾脏椭球的显微和亚显微结构,阐明其循环特点。 方法 应用光镜和透射电镜技术,结合心脏注射墨水悬液方法,显示脾脏椭球的组成、结构与循环。 结果 中华鳖脾脏白髓由动脉周围淋巴鞘（PALS）和椭球周围淋巴鞘（PELS）两种结构组成,缺乏淋巴小结。红髓包括脾索和脾窦,未发现边缘区。中央微动脉穿出PALS后,呈笔毛状分成数条椭球毛细血管,后者周围由PELS包绕。椭球毛细血管的末端直接开口于红髓的脾索,血流注入脾索,而后穿过脾窦内皮间隙进入脾窦。不同于普通血管内皮,椭球毛细血管内皮一般为立方状,基膜不完整,其外即为椭球结构。椭球壁由支持细胞、椭球相关细胞和网状纤维等构成。常见淋巴细胞和红细胞穿过椭球壁。注射墨水悬浮液后40min,可见整段椭球壁上分布着大量墨水碳粒。 结论 中华鳖脾脏椭球毛细血管相当于哺乳动物的高内皮后微静脉,是淋巴细胞和血细胞进出淋巴组织的重要通道。中华鳖脾脏循环属于开放式循环。     

The spleen of the one humped camel (Camelus dromedarius) has a unique histological structure

The histology and structure of 38 spleens of the dromedary (aged 0.5–15 y) were studied in relation to age. The spleen was found to have a thick capsule (292±106 mm) divided into an outer layer (113±39 mm) composed mainly of connective tissue and an inner layer (180±81 mm) consisting mainly of smooth muscle cells. Vascular and avascular trabeculae extend from the capsule, the former containing arteries and nerves but no trabecular veins, the latter being divided structurally into primary and secondary trabeculae. Subcapsular and peritrabecular blood sinuses around primary and vascular trabeculae are unique to the camel spleen. The central artery emerges from the periarterial lymphatic sheath and branches into up to 4 penicilli which extend as sheathed arterioles (42±8 μm). These are found near or surrounded by blood sinusoids of the red pulp. A wide marginal zone surrounds the white pulp and contains sheathed arteries but no marginal sinuses. The red pulp is characteristically divided into cords by secondary trabeculae and contains venous sinusoids of different sizes. The camel spleen is of a sinusal type that can store blood. The thick muscular capsule and trabeculae pump the stored blood according to the body's need. Both closed and open circulations are found. The venous return is unique as the blood flow is from the venous sinusoids of the red pulp to the peritrabecular sinuses to the subcapsular sinuses to the splenic vein. No significant structural differences related to age were found.     

Migration of macrophages from the marginal zone to germinal centers in the spleen of mice

Histological observations of the mouse spleen were carried out at different times after intravenous carbon injection. Large carbon-laden macrophages appeared in great numbers in the marginal zone soon after injection. They came together favorably around the germinal centers. Possible migration of these cells toward the germinal centers diffusely from the periphery of the white pulp or through the periarterial lymphoid sheath was suggested. These macrophages entered the germinal centers on a large scale and clustered for a long period--at least 180 days. Since the same type of macrophages were observed persistently in the marginal zone, it was thought that some of them might arise from the blood stream. Possible migration of these cells from the marginal zone toward the germinal centers was also persistently observed. A second type of much smaller carbon-laden macrophages was seen in the white pulp. However, they never showed any favorable localization in the germinal centers as did large carbon-laden macrophages.     

Cellular composition of the spleen after human allogeneic bone marrow transplantation

The cellular composition of the spleen has been assessed in 18 patients who died 15-326 days after receiving allogeneic marrow for leukaemia. The white pulp showed marked lymphocyte depletion with no germinal centres, very few B cells, and rare plasma cells. The marginal zone was unrecognizable but there were moderate numbers of T cells in the periarteriolar lymphatic sheaths (PALS), showing great variation in CD4/CD8 ratio. The percentage of CD4+ cells decreased with time post transplant. CD8+ cells were reduced in patients with graft-versus-host disease (GvHD) who also showed no increase in cells staining for activation markers. No T cells were detected expressing immature phenotypes and no differences were detected between patients who received marrow purged or unpurged of T cells. Macrophage numbers appeared normal. Extramedullary haemopoiesis (EMH) was predominantly in the red pulp greater than 30 days after transplantation but more commonly in the white pulp before then. Pyknotic cells were common in seven cases and appeared to be associated with EMH rather than GvHD. Chimaeric studies demonstrated small numbers of donor cells in the PALS at 26 days and larger numbers at 56 days.     

Antigens in immunity: XVI. A light and electron microscope study of antigen localization in the rat spleen*

This paper describes the ultrastructural location of labelled antigens and carbon in the spleens of rats from 4 minutes to 5 days after injection. Particular attention was focused on the sites of deposition 4 minutes after intra-arterial injection of microgram quantities of ¹²⁵I-labelled Salmonella flagellar antigens, crayfish haemocyanin and BSA, using colloidal carbon for comparison. The combination of radioautography with both light and electron microscopy showed the importance of antigen binding by lymphocytes in the marginal zone of the spleen. Macrophage sequestration of antigens was not prominent in the spleen, although it occurred in the liver with the flagellar antigens and haemocyanin.

In the spleen marginal zone, avid antigen-binding cells were found in situ 4 minutes after the injection of labelled haemocyanin. These appear to be the counterpart in vivo of antigen-binding lymphocytes prepared in vitro. Such cells also occurred infrequently after the injection of labelled polymerized flagellin, but were not found with either BSA or carbon.

The apparent movement of flagellar antigen from the marginal zone to the white pulp between 1 and 2 hours after injection was seen to involve lymphocyte-associated antigen. The follicular antigen localization occurring from 1 day onwards after injection was on the dendritic reticular cells of germinal centres, as has been described in lymph nodes after subcutaneous injection.

Carbon particles were rapidly sequestered in macrophages of the spleen and liver, although some particles were found between cells in the marginal zone for as long as 2 hours after injection. By 2 and 5 days, however, all the carbon was in phagocytes, even in the white pulp. Differences between the localization of antigens and carbon were clear, even in the ultrastructural sites of their location in tingible body macrophages of germinal centres.

The unexpected emphasis of lymphocyte association with labelled antigens in the spleen marginal zone has allowed a revison of the mechanism previously proposed for the movement of antigens within the microenvironments of the spleen.

     

Periarterial lymphoid sheath in the rat spleen: A light,transmission, and scanning electron microscopic study

The periarterial lymphoid sheath (PALS) in the rat spleen was studied by light, transmission, and scanning electron microscopy. The PALS was divided into three regions: the central region, peripheral region, and marginal zone bridging channel. In the central region, lymphocytes were easily washed away by perfusion. Large spaces were found between flat reticular cells or in large meshworks of stellate reticular cells; these may be deep lymphatic vessels. True lymphatic vessels were found in the central region near the hilus. In the marginal zone bridging channel, flat reticular cells surrounded the central artery in a circumferential pattern and formed channel-like spaces between the flat reticular cells. These spaces were connected with the meshwork of the red pulp reticular cells and may be a route for lymphocytes between the deep lymphatic vessels and the red pulp. In the peripheral region of the PALS, it was usually difficult to wash away free cells by perfusion, and free cells were found among the reticular cells. In places in the peripheral region, however, free cells were washed away. It is suggested that the lymph flow may start from the region surrounding the PALS, and that the peripheral region of the PALS may also be another route for lymphocyte migration.     

Species variation in the structure and function of the marginal zone—an electron microscope study of cat spleen

The marginal zone in the cat spleen consisted of a characteristic mixture of lymphocytes and other blood cells located mainly between the several layers of circumferential reticulum around white pulp. A region of fine-meshed reticulum between white pulp and red pulp, as present in some species, was absent from the cat spleen. Arterial capillaries to the marginal zone were few. Some were continuations of white pulp capillaries, whereas others were red pulp capillaries that likely were continuations of axial capillaries of periarterial macrophage sheaths (PAMS) (ellipsoids). Blood cells deposited in the marginal zone could reach red pulp by passing through the numerous openings in each layer of circumferential reticulum. Lymphocytes appeared to migrate across the marginal zone both toward and away from white pulp. Macrophages lying on the circumferential reticulum of the marginal zone phagocytized cells but did not ingest Thorotrast, although it coated their surfaces. Because of the scarcity of arterial endings and the lack of a macrophage-charged reticular meshwork, the marginal zone in cat spleen is not a major site of blood clearance and phagocytosis. These functions are better served in PAMS and red pulp.     

Periarterial lymphoid sheath in the rat spleen: a light, transmission, and scanning electron microscopic study.

The periarterial lymphoid sheath (PALS) in the rat spleen was studied by light, transmission, and scanning electron microscopy. The PALS was divided into three regions: the central region, peripheral region, and marginal zone bridging channel. In the central region, lymphocytes were easily washed away by perfusion. Large spaces were found between flat reticular cells or in large meshworks of stellate reticular cells; these may be deep lymphatic vessels. True lymphatic vessels were found in the central region near the hilus. In the marginal zone bridging channel, flat reticular cells surrounded the central artery in a circumferential pattern and formed channel-like spaces between the flat reticular cells. These spaces were connected with the meshwork of the red pulp reticular cells and may be a route for lymphocytes between the deep lymphatic vessels and the red pulp. In the peripheral region of the PALS, it was usually difficult to wash away free cells by perfusion, and free cells were found among the reticular cells. In places in the peripheral region, however, free cells were washed away. It is suggested that the lymph flow may start from the region surrounding the PALS, and that the peripheral region of the PALS may also be another route for lymphocyte migration.     

神经营养素在小鼠脾的定位研究

为了解神经营养素与免疫系统的关系，用免疫组织化学方法对神经营养素包括神经生长因子（ＮＧＦ）、脑源性神经营养因子（ＢＤＮＦ）、神经营养素３（ＮＴ－３）进行小鼠脾的定位研究。结果表明：３种神经营养素的免疫反应均存在于脾内，但分布特点各不相同。ＮＧＦ主要分布于白髓动脉周围淋巴鞘（ＰＡＬＳ）外层、边缘区（ＭＺ）和红髓（ＲＰ）的巨噬细胞样和淋巴细胞样细胞；ＢＤＮＦ除具有与ＮＧＦ相似的分布特点外，还见于脾淋巴小结生发中心的淋巴细胞样细胞；ＮＴ－３则存在于白、红髓的网状细胞样细胞。这一结果提示，脾的免疫和非免疫细胞均可能产生神经营养素，并提示不同类型的神经营养素对免疫系统有不同的作用。     

Migration pathways of recirculating murine B cells and CD4+ and CD8+ T lymphocytes

Migration pathways of B cell and CD4⁺ and CD8⁺ T cell subsets of murine thoracic duct lymphocytes (TDL) were mapped. Per weight, the spleen accumulated more TDL than any other organ, regardless of lymphocyte subset. Spleen autoradiographs showed early accumulations of TDL in marginal zone and red pulp. Many TDL exited the red pulp within 1 hr via splenic veins. The remaining TDL entered the white pulp, not directly from the adjacent marginal zone but via distal periarterial lymphatic sheaths (dPALS). From dPALS, T cells migrated proximally along the central artery into proximal sheaths (pPALS) and exited the white pulp via deep lymphatic vessels. B cells left dPALS to enter lymphatic nodules (NOD), then also exited via deep lymphatics. T cells homed to lymph nodes more efficiently than B cells. Lymphocytes entered nodes via high-endothelial venules (HEV). CD4⁺ TDL reached higher absolute concentrations in diffuse cortex than did CD8⁺ T cells. However, CD8⁺ TDL moved more quickly through diffuse cortex than did CD4⁺ TDL. B cells migrated from HEV into NOD. Both T and B TDL exited via cortical and medullary sinuses and efferent lymphatics. A migration pathway across medullary cords is described. All TDL subsets homed equally well to Peyer's patches. T TDL migrated from HEV into paranodular zones while B cells moved from HEV into NOD. All TDL exited via lymphatics. Few TDL entered zones beneath dome epithelium. All subsets were observed within indentations in presumptive M cells of the dome epithelium.