Lymphoid organ development in Xenopus thymectomized at eight days of age

This paper examines the effect of early thymectomy on the subsequent development of lymphoid tissues in the toad, Xenopus laevis. At the time of thymic removal (8 days post-fertilization) all the lymphoid organ anlagen are at a rudimentary state of differentiation and contain few, if any, small lymphocytes. Despite the absence of any thymic tissue all thymectomized animals grew normally. Thymectomized larvae developed relatively normal lymphoid organs. However, lymphoid depletion was apparent in the splenic red pulp and in the pharyngeal ventral cavity bodies. Examination of the lymphoid organs of post-metamorphic Xenopus revealed reduction in spleen size following thymectomy. Lymphoid depletion was evident in the splenic red pulp of many thymectomized toadlets and reduction in proportion of white to red pulp was also noted in a few of these animals. Absence of the thymus had no apparent effect on the histology of the other lymphoid organs examined.     

Electron microscopic study on the early histogenesis of thymus in the toad,Xenopus laevis

Summary Sequential electron microscopic observations of thymic histogenesis in the toad, Xenopus laevis, reveal that the thymus arises as epithelial buddings of the visceral pouches at Nieuwkoop-Faber stage 40, and acquires its basic histological features at stages 48–49. In the rudiments and the surrounding mesenchyme at stages 43–45, there are non-epithelial cells with pseudopodia, abundant ribosomes, and marginated heterochromatin. These cells, possible precursor cells of thymic lymphocytes, are frequently observed to attach and pass through the basal lamina which coats the thymic rudiment. The proliferation and differentiation of large lymphocytes are evident at stage 47. During stages 48–49 the small lymphocytes, lymphoid cortex and epithelial medulla including the thymic cysts, differentiate, and vascularization occurs.The results provide an ultrastructural basis for recent experimental evidence that the thymus exerts its essential function at stages 47–48. The possibility of non-epithelial derivation of thymic lymphocytes is discussed.The author wishes to express his thanks to Asst. Prof. Ch. Katagiri for his helpful advice during the course of this study     

Lympho-myeloid organs of Amphibia. I. Appearance during larval and adult stages of Rana catesbeiana

The bullfrog, Rana catesbeiana, possesses lympho-myeloid and epithelial structures that are morphologically similar in some respects to lymph nodes of mammals. These organs are present during the entire life cycle of the frog, however, the structures that are present during larval stages do not appear to be morphological precursors of adult organs. According to certain terms used previously by other investigators, two major organs are present throughout the larval stages: the lymph gland and the ventral cavity body. In the adult, the jugular body, the epithelial body, the precoracoid and propericardial bodies are found in the ventral neck region in contrast to the lateral and ventral arrangement of the lymph gland and ventral cavity body in larvae. The function of these organs is not known but it is believed that they play a role in the production of certain blood cells, particularly lymphocytes, and they may be involved in some aspects of the differentiation and maintenance of the immune response capacity.     

Development of T lymphocytes in Xenopus laevis: appearance of the antigen recognized by an anti-thymocyte mouse monoclonal antibody

Development of T lymphocytes in Xenopus laevis was studied using a mouse monoclonal antibody (mAb), XT-1, that was produced against surface determinants on thymocytes of J strain frog. Ontogenic studies, employing immunofluorescence, showed that cells positive for the determinant recognized by XT-1 mAb (XT-1⁺ cells) were first detected in the thymus of J strain Xenopus by Nieuwkoop and Faber stage 48 (7 days postfertilization) and then in the spleen, liver and kidney by stage 52 (20 days postfertilization). Percentages of XT-1⁺ cells in the thymus increased rapidly by stage 49 (10 days postfertilization) and reached adult levels by stage 52, and those in the spleen, liver, and kidney reached adult levels by stage 56 (40 days postfertilization). Electron microscopic immunohistochemistry revealed that most XT-1⁺ cells in thymuses from stage 56 larvae were typical small lymphocytes (4–7 μm in diameter). In contrast, many XT-1⁺ cells in larval thymuses at stage 49 are large (8–10 μm in diameter) lymphoblastoid cells. Thymectomy at stage 46 (5 days postfertilization) depleted XT-1⁺ cells in larval and adult lymphoid organs to background levels. These results suggest that the XT-1⁺ cells are differentiated from the lymphoid precursor cells in the thymus before the appearance of small lymphocytes and migrate into peripheral lymphoid organs. The cell surface determinant recognized by the XT-1 mAb may provide an important marker for the differentiation of T lymphocytes in Xenopus.     

The thymus microenvironment in regulating thymocyte differentiation

The thymus plays a crucial role in the development of T lymphocytes providing an inductive microenvironment in which committed progenitors undergo proliferation, T-cell receptor gene rearrangements and thymocyte differentiation into mature T-cells. The thymus microenvironment forms a complex network of interaction that comprises non lymphoid cells (e.g., thymic epithelial cells, TEC), cytokines, chemokines, extracellular matrix elements (ECM), matrix metalloproteinases and other soluble proteins. The thymic epithelial meshwork is the major component of thymic microenvironment, both morphologically and phenotypically limiting heterogeneous regions in thymic lobules and fulfilling an important role during specific stages of T-cell maturation. The process starts when bone marrow–derived lymphocyte precursors arrive at the outer cortical region of the thymic gland and begin to mature into functional T lymphocytes that will finally exit the thymus and populate the peripheral lymphoid organs. During their journey inside the thymus, thymocytes must interact with stromal cells (and their soluble products) and extracellular matrix proteins to receive appropriate signals for survival, proliferation and differentiation. The crucial components of the thymus microenvironment and their complex interactions during the T-cell maturation process with the objective of contributing to a better understanding of the function of the thymus as well as assist in the search for new therapeutic approaches to improve the immune response in various pathological conditions are summarized here.     

Kinetics of transfer of immunity by immune leucocytes and PFC response to HRBC in isogeneic ginbuna crucian carp

Transfer of immunity to horse erythrocytes (HRBC) by immune lymphoid cells was performed to analyze the kinetics of adoptive immunity in the clonal ginbuna crucian carp, Carassius auratus langsdorfti . Immune lymphoid cells were intravascularly transferred to the unprimed recipients and then recipients were evaluated by measuring the antibody titre of the plasma. Antibody productivity was most successfully conferred by splenic cells, followed by pronephric and mesonephric cells, taken from immune donors 7 days post-immunization, while transferability by thymic cells was lacking or very low, even if possible. Peak response of plaque-forming cells (PFCs) was observed at 5–7 days after the first injection, and the maximum number of PFCs at peak response was almost the same in all organs examined, such as the pronephros, mesonephros, spleen and the thymus. Direct correlation between transferability and number of PFCs was not observed on individuals, although the peak of transferability corresponded to that of the PFC response. Preliminary experiments of cell transfer by separated pronephric cells showed that the lymphocyte-rich fraction was more effective than the bottom fraction containing fewer lymphocytes in transferring immune reactivity. These results suggest that cells involved in transferring immune reactivity are B lymphocytes, composed of different developmental stages and distributed differently in the different lymphoid organs.     

Ultrastructure of the developing thymus of the leopard frog (Rana pipiens)

Summary A study of the ultrastructure of the developing thymus of the leopard frog (Rana pipiens) revealed that the thymus had undergone all of the major changes which would persist through larval life and metamorphosis by the time that the animals had reached larval stage IV of Taylor and Kollros (1946). These changes included development of an outer, lymphoid cortical region and an inner, essentially nonlymphoid medulla; mitotic activity among lymphoid cell precursors and the formation of the first small lymphocytes; development of complex cysts containing PAS-positive material and the appearance of other signs of secretory activity among epithelial cells of the medulla; and differentiation of large myoid cells containing bundles of striated muscle fibrils. The changes are particularly noteworthy because they first appear during a period in which the animals are known to be developing the capacity to respond immunologically to allografts.Supported by a grant from the National Institutes of Health number GM-11782 to E.P.V.     

The thymus microenvironment in regulating thymocyte differentiation

The thymus plays a crucial role in the development of T lymphocytes by providing an inductive microenvironment in which committed progenitors undergo proliferation, T-cell receptor gene rearrangements and thymocyte differentiate into mature T cells. The thymus microenvironment forms a complex network of interaction that comprises non lymphoid cells (e.g., thymic epithelial cells, TEC), cytokines, chemokines, extracellular matrix elements (ECM), matrix metalloproteinases and other soluble proteins. The thymic epithelial meshwork is the major component of the thymic microenvironment, both morphologically and phenotypically limiting heterogeneous regions in thymic lobules and fulfilling an important role during specific stages of T-cell maturation. The process starts when bone marrow-derived lymphocyte precursors arrive at the outer cortical region of the thymic gland and begin to mature into functional T lymphocytes that will finally exit the thymus and populate the peripheral lymphoid organs. During their journey inside the thymus, thymocytes must interact with stromal cells (and their soluble products) and extracellular matrix proteins to receive appropriate signals for survival, proliferation and differentiation. The crucial components of the thymus microenvironment, and their complex interactions during the T-cell maturation process are summarized here with the objective of contributing to a better understanding of the function of the thymus, as well as assisting in the search for new therapeutic approaches to improve the immune response in various pathological conditions.Key words: thymus, T-cell maturation, thymic microenvironment, thymocyte differantiation, chemokines, extracellular matrix, thymic nurse cells, metalloproteinases     

Ontogeny of IgM-producing cells in the lymphoid organs of rainbow trout, Salmo gairdneri Richardson: an immuno- and enzyme-histochemical study

The ontogenetic development of IgM-containing cells is described as demonstrated by immunoperoxidase staining with a mouse anti-trout IgM monoclonal antibody and the differentiation of enzyme-histochemical markers in the non-lymphoid cells forming the stroma of the thymus, spleen and kidney of the rainbow trout. The first lymphoid cells staining with the monoclonal antibody occurred at day 4-5 after hatching in the renal lympho-haemopoietic tissue. By 1 month after hatching IgM-positive cells also appeared in the spleen and thymus. Enzyme-histochemical demonstration of the alkaline and acid phosphatase and non-specific σ-naphthyl acetate esterase enzymatic activities in the non-lymphoid cells indicated that a certain degree of maturation of the cellular stroma of the developing lymphoid organs of trout was reached before or at the time when IgM-expressing cells could be observed. The relationships of the stromal components of the various lymphoid organs to the development of IgM-positive cells, and the possible role of the renal lympho-haemopoietic tissue as a primary lymphoid organ for B-cell differentiation in the trout are discussed.     

Ontogeny of the Immune System in Amphibians

SYNOPSIS. Experiments with amphibians have revealed that tissuesallografted in embryonic and early larval life may later succumbto a host immunological attack. In Xenopus, the ontogeny ofthis alloimmune response is correlated with the lymphoid maturationof the thymus. This article describes experiments primarilydesigned to ascertain if such a correlation exists in Rana pipiens. Initially, a histological study of the differentiation of thelymphomyeloid complex of the leopard frog was undertaken. At18–21°C lymphoid histogenesis is well under way inthe thymus and is beginning in many other organs during thethird week of larval life. Extensive growth and differentiationof these organs follow. Observations are also presented on thestructure, function and development of the lymphoid and Iymphomyeloidorgans from late larval life throughmetamorphosis to adulthood. Experiments were then performed to determine the onset of thealloimmune response to embryonically transplanted neural foldmaterial. At 18–21°C incompatibility phenomena, albeitslight, are first detected in these grafts as early as 17 daysafter fertilization, i.e. 15 days after transplantation. Thus,in the leopard frog, the alloimmune response develops soon afterlymphoid maturation of the thymus. At later stages of development,a more vigorous response is witnessed, concomitant with a rapidphase of lymphoid organ differentiation.     

Lymphoid Organs and Blood Cells of the Caecilian Ichthyophis kohtaoensis

The present work analyzes by light microscopy circulating blood cells and lymphoid organs of the caecilian, ichthyophis kohtaoensis. Peripheral blood contains erythrocytes, thrombocytes, lymphocytes, monocytes, heterophils, easinophils and basophils. Thymus, liver and spleen are the major lymphomyeloid organs in this caecilian and there is not a haemopoietic functioning bone marrow. The results are discussed on the basis of the possible phylogenetic positions of the Apoda emphasizing their relationships to the Urodela.     

Lymphomyeloid organs of amphibia. II. Vasculature in larval and adult Rana catesbeiana

There were no lymphatic vessels and lymph nodes demonstrable in adult and larval Rana catesbeiana by a method that adequately demonstrated the same in mammals. Although the parenchymal arrangement in the lymphomyeloid organs is not exactly the same as in mammalian hemal nodes, nonetheless the vascular patterns of the lymph glands and jugular bodies are prima facie evidence that they function as blood-filtering organs among other probable functions. The vascular pattern of the lymph gland is that of a rete mirabile, particularly a venous portal system, inasmuch as the afferent and efferent vessels are venous in character and interposed between them is a labyrinth of sinusoids. This is not the case, though, in the adult organs. The vascular pattern of the jugular bodies is very much like the spleen, viz., artery-capillary-sinusoid-vein sequence. It is doubtful, however, if the propericardial and procoracoid bodies ever filter blood, because the smallest blood vessels in them are capillary in type Because of the absence of a well-defined capsule in some parts of the propericardial body, similarly to lymphoid follicles, especially in the mammalian gastrointestinal tract, it is probable that it filters tissue fluid. The last two organs are apparently mainly blood cell-forming organs. It is inferred from the vascular connections of the larval and the adult lymphomyeloid organs that they are not genetically related. This aspect was analyzed from earlier developmental data, but actual follow-up of the larval organs to the adult stage is still in progress.     

Suppression of the allograft response by implants of mature lymphoid tissues in larval Xenopus laevis

One hundred and twenty-two larvae of Xenopus laevis, the South African clawed toad, at developmental stages 48, 50, 52 and 54, were implanted in the tail with two allografts from adult tissues. In each case, one allograft was from kidney, while the other was either from kidney, thymus, spleen, or liver. In any particular host the two implants were always from the same donor and the implants were all visually matched in size. The experimental period was a maximum of nine days, so as to minimize the large numbers of changes normally accompanying larval progress from stage to stage. We are concerned with the timing of allograft response initiation under the implant conditions of each experimental group at a particular point in development. An allograft response was defined as an infiltration and accumulation of small lymphocytes in the “test” kidney allograft. Larvae of all stages developed allograft responses within one week post-implantation when the variable implant was from kidney, but implants from spleen and thymus suppressed both the timing of initiation and the subsequent intensity of the response. Spleen was more effective in this regard than thymus and both were more effective in the earlier larval stages. Liver proved to be toxic to the larvae. The relationship between the maturation of the lymphomyeloid tissues and external morphological staging is also discussed.     

On the Origin of Thymic Lymphocytes

In the leopard frog (Rana pipiens), thymic lymphocytes do notoriginate from blood-borne stem cells that migrate into thethymus anlage; rather they arise in situ from elements in thethymic rudiment itself. After thymic differentiation, the lymphocytes(or their descendants) leave the thymus and extensively seedthe peripheral lymphoid organs. Indeed, virtually all the lymphocytesin the spleen, kidney, and bone marrow are ontogenically derivedfrom thymic cells. In postmetamorphic life, the thymus representsan organ in which lymphopoiesis is genuinely self-sustaining.Throughout the juvenile life of the frog, there is no indicationof an inward afferent stream of cells entering the intact thymus.     

Ontogeny of the immune system inSebastiscus marmoratus: Histogenesis of the lymphoid organs and effects of thymectomy

Synopsis The ontogenetic development of the immune system in a marine teleostSebastiscus marmoratus was studied by histological examination and removal of the thymus. The pronephros and the spleen had been differentiated at the time of birth and contained small numbers of haemopoietic cells. In contrast to most vertebrates, the rudiments of the thymus were first visible 1 week post-birth in the dorsoposterior part of the pharynx, the same location as in the adults. However, small lymphocytes first appeared in the thymus of fish at 3 weeks of age, followed by the pronephros at 4 weeks and the spleen at 6 weeks. Complete or partial suppression of the antibody response to sheep red blood cells (SRBC) occurred in fish that were thymectomized at 1.5 months of age and immunized 2 weeks later, and a marked decrease in lymphocytes was observed in the pronephros and spleen. The thymectomy of adult fish also caused reduced serum antibody titres in fish immunized 1 month after the operation. These results suggest that the thymus plays an essential role in the development of the immune system and its functions continue into adult life.     

Changes in the rat lymphoid organs in acute hypoxia

Changes occurring in the rat thymus, spleen, mesenteric lymph nodes have been studied by means of some histological and cytofluorimetrical methods under the effect of an acute hypoxia that imitates the rise to 7,000 m above the sea level for 1 h and to 6,500 m for 6 h. Under the effect of hypoxia, migration of differentiated lymphocytes out of the lymphoid organs is increasing, certain essential shifts in temporal parameters take place in the mitotic cycle of the lymphocytes, contents of nucleic acids in the lymphoid cells change. The phenomena mentioned demonstrate that under the acute hypoxic stress, intensified differentiation processes of the lymphocytes in the thymus, spleen and the lymph node take place and the lymphoid tissue functional activity increases.     

The development of the lymphoid organs of flounder, Paralichthys olivaceus, from hatching to 13 months

The growth of the lymphoid organs, such as head kidney, spleen and thymus were studied in flounder, Paralichthys olivaceus Temminck & Schlegel, from hatching to 13 months of age. Except for the thymus, all organs grew as the fish grew. By 2 months of age the lymphoid organs attained their maximum relative weight. The organ weight showed a closer correlation to body weight than they did to age. The total number of leucocytes in the lymphoid organs increased with age, but the number per milligram of lymphoid organ remained constant. A micro and ultrastructural study of the lymphoid organs showed that the full development of the lymphoid organs was not achieved until the juvenile stage. The spleen and head kidney had mixed populations of "red" and "white" cells. The head kidney was more lymphoid than the spleen. The thymus involuted quickly during the first 6 months. The blood components had no obvious relationship with age or season during the period studied.     

Functional lymphocytes in the chicken pineal gland

The primary antibody response of lymphoid tissue occupying the pineal gland of 6-wk-old chickens was studied subsequent to injection of the carotid artery with sheep red blood cells (SRBC) or bovine serum albumin (BSA). Injection of SRBC did not produce plaque-forming cells (PFC) among pineal lymphocytes whereas BSA stimulated synthesis of anti-BSA immunoglobulin in pineal lymphoid tissue. A cytotoxic assay using appropriate anti-lymphocyte sera indicated that single-cell suspensions of pineal lymphocytes were composed of 42% B lymphocytes and 51% T lymphocytes. Bursal and thymic lymphocytes labeled with tritiated thymidine migrated into pineal lymphoid tissue when injected into 4- and 5-wk-old naive chicks. These observations indicate that the bursa and thymus make equivalent contributions to the lymphoid mass in the chicken pineal gland. Challenge of pineal-established lymphocytes by antigen introduced via the blood vascular system suggests that soluble antigens--rather than particulate ones--stimulate antibody production in the pineal gland. Collectively, these studies indicate that the pineal gland should be considered as a functional component of the chicken's lymphomyeloid system.     

鳜鱼头肾的组织发生及成鱼头肾B淋巴细胞的分布

通过整体连续切片,研究了鳜鱼不同发育时期的头肾结构,并利用原位PCR方法检测了B淋巴细胞在鳜鱼头肾中的分布。在孵化后第1d观察到了肾组织,主要由肾小管组成。尔后头肾的发育经历了三个结构和功能的转变。第一个阶段为孵化后第1d到第7d,头肾作为滤过性器官存在,由肾小管及少量淋巴细胞组成。第二个阶段从第8d到第36d,是一个功能混合型阶段,头肾中既有肾小管,又有造血组织;随时间推移,肾小管数量减少,淋巴细胞数量剧增。紧接着进入第三个阶段：肾小管完全消失,头肾中开始出现大量的嗜铬细胞,头肾作为淋巴-肾上腺组织而存在。肾上腺首先出现在头肾前端,随发育成熟,集中分布于头肾门静脉周围。IgM在鳜头肾中大量表达,IgM分泌细胞分布于整个头肾组织,在血管周围有集中趋势[动物学报51(3)：440—446,20051。