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The entry of environmental antigens into the lumen of the bursa of Fabricius was prevented by ligating the bursal duct prior to hatching (BDL: bursal duct ligation). The development of `natural' serum agglutinins for bacteria and heteroerythrocytes was markedly suppressed by BDL. The antigen-specific recovery was observed by simultaneous administration of sterile antigens into the bursal lumen at the time of BDL. These results strongly suggest that the bursa of Fabricius is a major channel through which environmental antigens stimulate the immune system and induce the formation of `natural' serum agglutinins. [ABSTRACT FROM AUTHOR]

Neonatal chickens were primed by multiple or a single injection with sheep red blood cells (SRBC) into the cloacal lumen and then were challenged by intravenous injection with the same antigen. Direct plaque-forming cells (PFC) in the spleen were markedly increased shortly after intracloacal priming. This enhancing effect was antigen specific and was abolished by surgical bursectomy 24 h after intracloacal priming. A similar effect was also found in mature adult chickens, although the degree of enhancement was far less than that found in neonatal chickens. These results suggest that a certain stage of B-cell differentiation in the bursa of Fabricius (BF) is regulated by antigenic stimulation. [ABSTRACT FROM AUTHOR]

Describes the suppression of immune response by isolation of the bursa of Fabricius from environmental stimuli. Gradual increase in lymphopoiesis in the bursa and the immune reactivity of the chickens with age; Importance of environmental stimuli across the bursal epithelium in the amplification mechanisms for B-cell differentiation in the bursa.

We had an opportunity to perform a general autopsy of a case with chronic organic mercury toxicosis in 2017. He had been engaged in synthesizing a variety of organic mercury compounds throughout the four years from 1966 and developed chronic organic mercury poisoning in 1969. Almost forty years on, he still remained to complain of persistent paresthesia at finger tips and tongue, and of narrowed visual field. Neurological examinations clarified a rise of two-point discrimination thresholds, a systemic increase of touch thresholds, constriction of the visual field caused by general visual depression, and sensorineural hearing loss while primary modalities of his somatic, visual, and auditory sensations were preserved. These symptoms and signs are characteristic of human organic mercury poisoning. Furthermore, he had difficulty in processing a lot of visual and auditory information at a time. His two-point discrimination thresholds and systemic elevation of touch thresholds were comparable to those of mild organic mercury poisoning cases. He had slight sensory ataxia, but not cerebellar ataxia. Brain [ 18 F]-2-fluorodeoxyglucose positron emission tomography analysis exhibited marked hypometabolism at bilateral postcentral gyrus, striate cortex, and superior temporal gyrus, but not the cerebellum. Histopathological studies revealed considerable decrease of granular neurons and neuronal networks in bilateral primary somatosensory, visual, and auditory cortices. Those characteristic brain lesions fairly explain increase of thresholds of somatic, visual, and auditory sensations, and degradation of integrating sensory information. It is noted that damages to the peripheral nervous system and the cerebellum were not detected and that his intellectual faculties were preserved., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

IgM(+)IgG(+) B cells were detected, by immunofluorescence staining of single cells, in the bursa of Fabricius after hatching. To study the role of maternal IgG (MIgG) in this emergence of IgM(+)IgG(+) B cells, MIgG-free chicks were established from surgically bursectomized hens. Deprivation of MIgG in chicks completely prevented the appearance of IgM(+)IgG(+) B cells in the bursa 1 week after hatching. However, introduction of fluorescein-isothiocyanate-labeled MIgG to MIgG-free chick embryos on day 18 of incubation retrieved IgM(+)IgG(+) B cells in the bursa 1 week after hatching. Thus, IgM(+)IgG(+) B cells are induced by the binding of MIgG to IgM(+) B cells in the bursa after hatching. Nevertheless, no binding of MIgG to IgM(+) B cells was observed in the bursa of chick embryos in which B-cell proliferation and differentiation were independent of external antigens (Ags). Additionally, the binding of MIgG to IgM(+) B cells after hatching was prevented by the isolation of the bursa from environmental stimuli by bursal duct ligation. Therefore, Ag stimulation from the external environment to the bursa is indispensable for the binding of MIgG to IgM(+) B cells in the bursa. Taken together, the data demonstrate that IgM(+)IgG(+) B cells are generated by Ag-dependent binding of MIgG to IgM(+) B cells in the bursa after hatching.

The bursa of Fabricius is a primary lymphoid organ for B-cell development and gut-associated lymphoid tissue. After hatching, IgG-containing cells with reticular branches are found in the medulla of bursal follicles on frozen sections stained with anti-Cγ antibody, and IgM(+)IgG(+) B cells are detected in single-cell suspension of the bursa. IgG-containing cells in the medulla do not biosynthesize IgG and are composed of aggregated maternal IgG and environmental antigens. Then, those cells in the medulla are acknowledged as follicular dendritic cells retaining immune complexes. Also, it is presumed that IgM(+)IgG(+) B cells are generated by the attachment of immune complexes to IgM(+) bursal B cells because IgM(+)IgG(+) B cells are induced by antigen-dependent attachment of maternal IgG. Therefore, it is reasonable to suppose that immune complexes exert further B-cell differentiation in the medulla., (© 2014 Elsevier Inc. All rights reserved.)

The bursa of Fabricius of the chicken is known to be both a primary lymphoid organ and a secondary lymphoid tissue. Bursal follicles are equipped with antigen-trapping follicle-associated epithelium. However, bioactive antigens such as protein and bacteria have not been detected in the bursal parenchyma. By immunoperoxidase staining with a polyspecific antibody (Ab) against Escherichia coli, we detected aggregated E. coli antigens in the medulla of bursal follicles after hatching. The distribution of aggregated E. coli antigens is restricted to the medulla of bursal follicles. The antigens are not found in the spleen or the parenchyma of the caecal tonsil. The bursa is thus a trapping site for E. coli antigens from the external environment. Furthermore, two-color immunostaining clarified that these antigens form immune complexes with maternal IgG (MIgG) and are retained by reticular cells. Additionally, immune complexes in the bursa were shown to induce the rapid development of serum IgM Ab for indigenous E. coli. Our results suggest that immune complexes of MIgG and environmental antigens in the medulla of bursal follicles exert positive effects on B-cell differentiation in the bursa in situ.

The bursa of Fabricius of the chicken is known as a primary lymphoid organ for B-cell development. Morphologically, the origin of IgG-containing cells in the bursa has not been clear until now, because abundant maternal IgG (MIgG) is transported to the chick embryo and distributed to the bursal tissue around hatching. Thus, it has been difficult to find out whether these cells themselves biosynthesize IgG or if they acquire MIgG via attachment to their surface. Our present study employing in situ hybridization clarified that IgG-containing cells in the medulla of bursal follicles did not biosynthesize IgG. To study the role of MIgG in the development of those IgG-containing cells, MIgG-free chicks were established from surgically bursectomized hen (SBx-hen). We found that, on the one hand, deprivation of MIgG from chicks completely inhibited the development of IgG-containing cells in the medulla after hatching. On the other hand, administration of MIgG to MIgG-free chicks recovered the emergence of those cells. In addition, we observed that those cells did not bear a B-cell marker and possessed dendrites with aggregated IgG. These results demonstrate that IgG-containing cells in the medulla are reticular cells that capture aggregated MIgG. Moreover, we show that the isolation of the bursa from environmental stimuli by bursal duct ligation (BDL) suppressed the development of IgG-containing cells after hatching. Thus, it is implied that environmental stimulations play a key role in MIgG aggregations and dendritic distributions of aggregated MIgG in the medulla after hatching.

The chicken DT40 B lymphocyte line diversifies its immunoglobulin (Ig) V genes through translesion DNA synthesis-dependent point mutations (Ig hypermutation) and homologous recombination (HR)-dependent Ig gene conversion. The error-prone biochemical characteristic of the A family DNA polymerases Polnu and Pol led us to explore the role of these polymerases in Ig gene diversification in DT40 cells. Disruption of both polymerases causes a significant decrease in Ig gene conversion events, although POLN(-/-)/POLQ(-/-) cells exhibit no prominent defect in HR-mediated DNA repair, as indicated by no increase in sensitivity to camptothecin. Poleta has also been previously implicated in Ig gene conversion. We show that a POLH(-/-)/POLN(-/-)/POLQ(-/-) triple mutant displays no Ig gene conversion and reduced Ig hypermutation. Together, these data define a role for Polnu and Pol in recombination and suggest that the DNA synthesis associated with Ig gene conversion is accounted for by three specialized DNA polymerases.

We established the method of isolating individually encapsulated germinal centers (GCs) from immunized spleen and analyzed single cell suspension of GCs by flowcytometry. In GCs, the high frequency of sIgG+ cells (29%) and sIgA+ cells (5%) was detected. Two-color flowcytometry analysis showed that GCs contained 27% of sIgM-IgG+ cells, in which isotype switch from IgM to IgG had occurred, and 5% of Bu1-IgG+ cells, which were differentiating into plasma cells. On the other hand, sIgM-IgG+ and Bu1-IgG+ cells were not detected in the bursa, which contained 95% of B cells and only 1% of T cells. CD4+ but not CD8+ T cells were detected in the light zone of GCs and these CD4+ T cells are supposed to play a key role in isotype switching and differentiation into plasma cells in GCs. These results clearly demonstrate that GCs provide a site for isotype switching and differentiation into plasma cells.

The first well-documented outbreak of acute methyl mercury (MeHg) poisoning by consumption of contaminated fish occurred in Minamata, Japan, in 1953. The clinical picture was officially recognized and called Minamata disease (MD) in 1956. However, 50 years later there are still arguments about the definition of MD in terms of clinical symptoms and extent of lesions. We provide a historical review of this epidemic and an update of the problem of MeHg toxicity. Since MeHg dispersed from Minamata to the Shiranui Sea, residents living around the sea were exposed to low-dose MeHg through fish consumption for about 20 years (at least from 1950 to 1968). These patients with chronic MeHg poisoning continue to complain of distal paresthesias of the extremities and the lips even 30 years after cessation of exposure to MeHg. Based on findings in these patients the symptoms and lesions in MeHg poisoning are reappraised. The persisting somatosensory disorders after discontinuation of exposure to MeHg were induced by diffuse damage to the somatosensory cortex, but not by damage to the peripheral nervous system, as previously believed.

The first acute case of methylmercury (MeHg) poisoning by the consumption of fish arose in Minamata, Japan, in 1953. It was officially recognized and called Minamata disease (MD) in 1956. There are still arguments about the definition of MD in terms of its associated clinical symptoms and lesions even 50 years after the initial recognition of MD. Studies on this MD epidemic are reviewed along with its historical background. Since MeHg dispersed from Minamata to the Shiranui Sea, residents living around the sea had been exposed to low-dose MeHg through fish consumption for about 20 years (at least from 1950 to 1968). These chronic MeHg poisoning patients complained of paresthesia at the distal parts of their extremities and around the lips even 30 years after the cessation of exposure to MeHg of anthropogenic origin. The persisting somatosensory disorders after the discontinuation of exposure to MeHg were induced by diffuse damage to the somatosensory cortex, but not by damage to the peripheral nervous system, as previously believed. Based on these findings, symptoms and lesions in MeHg poisoning are reappraised.

The first well-documented methylmercury (MeHg) poisoning by consumption of fish arose in Minamata, Japan in 1953. MeHg had dispersed from Minamata to the Shiranui Sea. The temporal changes in MeHg in the umbilical cords indicate that residents living around that Sea had been exposed to low-dose MeHg through fish consumption for about 20 years (at least from 1950 to 1968). They have complained of paresthesia at the distal parts of the extremities and around the lip even 30 years after the cessation of exposure to anthropogenic MeHg. The thresholds of touch and two-point discrimination of those residents and Minamata disease (MD) patients were examined using the quantifiable instruments. They could perceive the stimulation of touch although their touch thresholds significantly increased in comparison to those of the control people. Their touch thresholds increased at the proximal extremities and the trunks as well as at the distal extremities. The evenly distributed increases at both distal and proximal parts revealed that the persistent somatosensory disturbances were not caused by the injuries to their peripheral nerves. The thresholds of two-point discrimination, which are associated with the function of the somatosensory cortex, increased at both forefingers and the lip in both groups. Taking into consideration that, the apraxia limb kinetics, astereognosis and disorder of active sensation, which are all associated with damage to the somatosensory cortex, were detected, it is proposed that the persisting somatosensory disorders after discontinuation of exposure to MeHg were induced by diffuse damage to the somatosensory cortex.

The germinal center (GC) develops after antigenic stimulation and is thought to occur at the site of various immune responses. We separated a single GC from chicken spleen after antigenic stimulation. Flow cytometric analysis of the cells derived from a single GC and RT-PCR analysis of Ig mRNA expression in GC was performed. Direct evidence indicates that: (1) there was a considerable difference in the cell population of each GC, (2) the ratio of CD3(+) cells in a GC remains constant at 10-20%, (3) the highest proportion of sIgY(+) cells in a GC occurs 1 week after the time of highest proportion of sIgM(+) cells, and (4) RT-PCR analysis was used to detect IgY mRNA expression in a GC. The continuous existence of CD3(+) cells, the alterations in sIgM(+) and sIgY(+) cell ratios, and the expression of IgY mRNA strongly suggest that Ig class switching occurs in the GC during an immune response.

In chickens, a single set of unique functional segments of both Ig H and L chain genes is rearranged during early embryogenesis to generate a pool of B cell progenitors that will be diversified in the bursa by gene conversion, forming the preimmune repertoire. After hatching, bursal cells are exposed to environmental Ags in the bursal lumen. We prepared B cells from each single bursal follicle and used PCR-directed Ig L chain gene analysis to study the differentiation of B cells and the effect of antigenic stimulation from the bursal lumen on the neonatal chicken B cell repertoire formation. Selective amplification of B cell clones with a productive V-J joint was observed during the late embryonic stage, possibly by the interaction with ligands expressed on the bursal stroma and further accelerated in the neonatal chicken. Administration of the artificial Ags into the bursal lumen before the isolation of bursa by bursal duct ligation in the embryo caused a significant increase in lymphocytes with a productive V-J joint in the neonatal chicken bursa compared with the isolated bursa. Intra- and interclonal diversity of a complementarity-determining region measured by an evolutionary distance increased during bursal development. Clonal diversification did not require stimulation by artificial Ags from the bursal lumen. Thus, the preimmune repertoire in the bursa is generated by gene conversion during Ag-independent B cell proliferation, and antigenic stimulation from the bursal epithelium to bursal B cells plays roles in the selection of clones with a productive V-J joint.

The calf contains two types of Peyer's patches (PPs): jejunal and ileal. The ileal PP has been thought to be equivalent to the bursa of Fabricius (BF) as a central lymphoid organ. The morphologies of ileal and jejunal PPs in the calf were compared with those of the BF and the caecal tonsil (CT) in the chicken. Immunoglobulin G-positive (IgG(+)) cells appear in the follicles of them all and exhibited a dendritic appearance after birth. We investigated whether the IgG in these follicles was produced in situ. IgG-producing cells were detected in the follicular medullas of the jejunal PP and the CT, but not in those of the ileal PP and the BF. CD4(+) cells were distributed in the follicular medullas of the jejunal PP and the CT, but not in those of the ileal PP and the BF. The data suggest that Ig class switching occurs in both jejunal PP follicles and CT follicles, but does not occur in either the ileal PP follicles or the bursal follicles. Because CD4(+) T cells would be prerequisite for Ig class switching in these follicles, IgG(+) cells of the follicular medullas in the ileal PP and the BF would trap immune complexes from the gut lumen. The primary B-cell repertoire might be selected by gut-derived antigens in the ileal PP and the BF before seeding the periphery., (Copyright 2002 Wiley-Liss, Inc.)

Chicken single germinal centers enable us to analyze the postbursal diversifications of B cells due to their easy isolation. Germinal center formation has peaked by day 7 of primary responses and begins to wane 14 days after immunization. To detail the kinetics of Ig mutation and selection, we analyzed Ig light chain sequences recovered from single germinal centers at 7 and 11 days postimmunization with an artificial Ag. Our observations show that multiple, Ag-activated B cells migrating into single germinal centers are diversified by gene conversion in the very early phase of the germinal center reaction and are subsequently subjected to point mutations and selection for oligoclonality.

IgG+ cells were detected in the bursa of Fabricius after hatching by immunofluorescence staining of single cells and immunohistologic studies using mAb anti-Ig gamma-heavy chain. The frequency of IgG+ cells in the bursa increased rapidly immediately after hatching. In histologic studies, most of the IgG+ cells were found in the medullary areas of the bursal follicle. Isolation of the bursa from the gut by bursal duct ligation before hatching suppressed the development of IgG+ cells in the bursa after hatching. In addition, administration of Ags into the bursal lumen just before bursal duct ligation caused a significant increase of IgG+ cells in the bursa compared with the isolated bursa. However, parenteral administration of Ags had no effect on the frequency of IgG+ cells in the isolated and normal bursa. These results suggest that Ags in the bursa are a prerequisite for the development of IgG+ cells in the bursa. Although the majority of IgG+ bursal cells were IgM+ IgG+ double-positive cells, sorted Bu1+ bursal cells, which contained 99.9% of IgG+ cells but not plasma cells, synthesized de novo IgM but little or no IgG. Therefore, it is likely that surface IgG on the bursal cells is not produced locally, but is trapped by IgM+ bursal cells. We speculate that Ags derived from the bursal lumen form immune complexes within the bursa, which are subsequently trapped on the surface of IgM+ bursal cells mediated either by Fc-receptors, C3 receptors, or surface Ig.

Effect of environment antigens on S phase entry rate of bursa cells was studied by utilizing in vivo incorporation of bromodeoxyuridine (BrdUrd). S phase entry rate of bursa cells in chick embryo was determined to be 6.4%/h, this rate increased to 10.0%/h at neonatal stage and decreased to 6.4%/h 6 weeks after hatching. Evidence is presented that isolation of the bursa from the gut before hatching suppressed S phase entry rate of bursa cells after hatching, whereas S phase entry rates of thymocytes were not affected. On tissue sections, a decrease of BrdUrd+ cells was observed in medullary areas of the isolated bursa. It is proposed that in situ stimulation of bursa cells by environmental antigens to the bursa accelerates S phase entry rates, and that medullary areas are the major sites for antigen-driven B cell proliferation after hatching.

Using immunohistochemistry, the distribution and characteristics of cells detected by the newly developed monoclonals HIS-CI (B lymphocytes), HIS-C7 (leucocytes), HIS-C12 (IgM), CVI-ChIgM-59.7 (IgM), CVI-ChIgG-47.3 (IgG), and CVI-ChIgA-46.5 (IgA) are described in bone marrow, thymus, bursa of Fabricius, and spleen of chickens of different ages. Furthermore, quantification of cells positive with the described monoclonal antibodies was performed on cytocentrifuge preparations. The specificities of the monoclonal antibodies are discussed.

The fine distribution of lymph vessels in the bursa of Fabricius and the efferent lymph vessels coming from the bursa were demonstrated. The lymph vessels never entered lymphoid follicles, whereas the blood vessels pierced the cortex of lymphoid follicles and formed a network at the boundary between the cortex and medulla of lymphoid follicles. The lymph vessels commenced in the immediate exterior of lymphoid follicles. A large number of lymphoid cells were found in the lumen of the lymph vessels. Migration of lymphoid cells from the cortex of lymphoid follicles to the lymph vessels was also observed. Therefore, it was assumed that the lymph vessels of the bursa are important in the transport of lymphoid cells from the bursa to the blood stream.

Rats of inbred DA strain were given 20 intraperitoneal injections of typhoid-paratyphoid bacilli at 10 day intervals. Thymic lymph follicles with germinal centres were formed in females but not in males. However, orchidectomy permitted the formation of large numbers of thymic lymph follicles with germinal centres in males. The formation of lymph follicles with germinal centres in the thymus was not affected by ovariectomy. These results would appear to suggest an inhibitory role of androgens on the formation of thymic lymph follicles with germinal centres. In addition, large numbers of plasma cells, mast cells, and reticular cells, or macrophages with haemosiderin in their cytoplasm, appeared in the parenchma of the thymus in close association with the formation of thymic germinal centres.

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.