Monoclonal antibodies directed against Brucella abortus cell surface antigens

A panel of monoclonal antibodies (MAb) has been raised against Brucella abortus cell surface antigens from mice immunized with either heat/phenol treated or UV killed bacterial suspensions of B. abortus. The hybridomas were screened by either a microagglutination procedure or by an indirect enzyme immunoassay (EIA) on sonicated bacterial preparations. From a large number of MAb generated by various procedures, two distinct types of MAb emerged. The most numerous type was capable of agglutinating B. abortus and reacting with a soluble preparation of lipopolysaccharide (LPS). A second type was not capable of agglutinating the bacterial suspensions or of binding to the soluble LPS preparation but reacted with an antigen present in bacterial sonicates. Two MAb of this type react differentially with sonicates prepared from virulent and avirulent strains of B. abortus. There appeared to be sufficient evidence from our analysis of the relative degree of cross reaction with antigens present on a range of B. abortus strains and Brucella and xenogenic bacterial species to conclude that each of the seven MAb was recognising a separate antigenic site on the B. abortus cell surface.     

Developmental changes in cell proliferation and apoptosis in the normal duck bursa of Fabricius

The aim of this work was to investigate developmental changes in cell proliferation and apoptosis in normal duck bursa of Fabricius using flow cytometry and immunohistochemistry. Studies were carried out on Tianfu ducks on days 24 and 27 of embryogenesis (E24 and E27) along with days 20, 70, and 200 of postnatal development (P20, P70, and P200). Results showed that the percentage of G₀/G₁ bursa cells significantly increased between E24 and P200 while the percentage of cells in the S phase or G₂ + M phase as well as the proliferating index obviously decreased during the same period. Proliferation cell nuclear antigen was detected in lymphocyte and interfollicular epithelium. The proliferative lymphocyte density tended to decrease from E24 to P200. Apoptotic bodies in macrophages, free apoptotic bodies, or nuclei with condensed chromatin in lymphocytes in follicles were identified by transferase-mediated dUTP nick-end labeling. Both flow cytometry and microscopic analysis reveal that the proportion of apoptotic cells and apoptotic lymphocyte density increased from E24 to P20, fell on P70, then rose again on P200. Our foundings demonstrate that cell proliferation decreases and apoptosis increases with age. These changes may account for duck bursa development and involution.     

Differential expression of immune-related genes in the bursa of Fabricius of two inbred chicken lines following infection with very virulent infectious bursal disease virus

Among different inbred chickens’ lines, we previously showed that lines P and N of Institute for Animal Health, Compton, UK are the most susceptible and the least affected lines, respectively, following infection with very virulent infectious bursal disease virus (vvIBDV). In this study, the differential expressions of 29 different immune-related genes were characterized. Although, birds from both lines succumbed to infection, line P showed greater bursal lesion scores and higher viral copy numbers compared to line N. Interestingly, line N showed greater down-regulation of B cell related genes (BLNK, TNFSF13B and CD72) compared to line P. While up-regulation of T-cell related genes (CD86 and CTLA4) and Th1 associated cytokines (IFNG, IL2, IL12A and IL15) were documented in both lines, the expression levels of these genes were different in the two lines. Meanwhile, the expression of IFN-related genes IFNB, STAT1, and IRF10, but not IRF5, were up-regulated in both lines. The expression of pro-inflammatory cytokines (IL1B, IL6, IL18, and IL17) and chemokines (CXCLi2, CCL4, CCL5 and CCR5) were up-regulated in both lines with greater increase documented in line P compared to line N. Strikingly, the expression of IL12B was detected only in line P whilst the expression of IL15RA was detected only in line N. In conclusion, the bursal immunopathology of IBDV correlates more with expression of proinflammatory response related genes and does not related to expression of B-cell related genes.     

Monoclonal antibodies as probes for defining cellular subsets in the bone marrow, thymus, bursa of fabricius, and spleen of the chicken

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.     

Comparative studies on the secondary lymphoid tissue areas in the chicken bursa of Fabricius and calf ileal Peyer's patch

The chicken bursa of Fabricius and calf ileal Peyer's patch are thought to be the primary lymphoid organs of B cell development. In the bursa, the existence of secondary lymphoid tissue, called the diffusely infiltrated area, has been recognized. Recently, we have found the presence of a region of secondary lymphoid tissue in the ileal Peyer's patch at the period of the most rapid growth of this organ. In this study, we compared the development of these secondary lymphoid tissue regions in the bursa and ileal Peyer's patch histologically. Before hatching, lymphatic follicle formation occurred in the bursa, but not in the diffusely infiltrated area, where only a small number of lymphoid cells were found. However, during fetal calf development, lymphatic follicle formation occurred not only in the primary lymphoid organ but also in the secondary lymphoid tissue regions. Therefore, the prenatal development of the secondary lymphoid tissue regions of the bursa and ileal Peyer's patch were distinct. After hatching, formation of the germinal center, which contained many CD4⁺ cells, was observed in the diffusely infiltrated area of the bursa. After birth, many CD4⁺ cells and IgG mRNA expression were observed in the lymphatic follicle of the secondary lymphoid tissue regions in the ileal Peyer's patch, but rarely in the ileal Peyer's patch lymphatic follicles. The change of character of these secondary lymphoid tissue regions at the postnatal stage might be dependent on external antigens.     

Isolation and immunomodulatory activity of bursal peptide,a novel bursal peptide from the chicken bursa of Fabricius

The bursa of Fabricius (BF), which is unique to birds, serves as the central humoral immune organ and plays a significant role in B lymphocyte differentiation. In this study, a new bursal peptide (BP-IV) was isolated from BF, which promoted colony-forming unit pre-B formation and regulated B cell differentiation. BP-IV also exerted immunomodulatory effects on antigen-specific immune responses via both humoral and cellular immunity in chicken and mice that had been immunized with inactivated avian influenza virus (AIV; H9N2 subtype), including enhancing AIV-specific antibody and cytokine production. The results of this study provided novel insights into the use of a potential candidate reagent for B cell development and future immuno-pharmacological use.     

Cryptosporidiosis of the bursa of Fabricius of chickens.

Light-microscope and electron-microscope studies of a coccidial organism found in the bursa of Fabricius from 3 chickens clearly established the parasite as belonging to the family Cryptosporiidae. Hyperplasia and heterophil infiltration were associated with the presence of organisms attached to the microvillus border of epithelial cells lining the plicae of the bursa of Fabricius. Although there were no clinical signs or gross lesions common to the 3 cases described, all had similar histologic lesions in the epithelium lining the bursa of Fabricius.     

Effects of aflatoxin B1 on the development of the bursa of Fabricius and blood lymphocyte acid phosphatase of the chicken

1. In this study, embryotoxicity and effects of in ovo administration of aflatoxin B1 (AFB1) on the embryonic development of the bursa of Fabricius were determined in fertilised chicken eggs by histological methods and histochemical demonstration of acid phosphatase (ACP-ase) enzyme. Peripheral blood lymphocyte percentages were also estimated. 2. Embryonic stages were determined according to the Hamburger-Hamilton (H-H) scale. Mortality rates increased in AFB1-injected groups in a dose-dependent manner. Embryonic deaths were concentrated at H-H 31 in the 5 ng AFB1/egg group while the deaths were highest at H-H 25 in the 10 and 20 ng AFB1/egg groups. In the 40ng AFB1/egg group, embryonic deaths mostly occurred during the first 70 to 72 h of incubation (H-H 20). 3. Bursal development was quite similar at d 7 of incubation in control and all experimental groups although development had been substantially impaired and retarded at d 10 of incubation in 10, 20 and 40 ng AFB1/egg groups. A gradual decrease of ACP-ase positive lymphocyte numbers was observed with increasing AFB1 doses. The chicks in AFB1-treated groups hatched with poorly developed bursae compared with those of the controls. However, proportions of ACP-ase positive lymphocytes substantially increased at hatch and at 21 d of incubation all groups had similar numbers. 4. The results revealed that breeder diets should be investigated for aflatoxins and specifically AFB1, in cases of low hatchability and flock immunity, because low concentration of AFB1 transferred into the fertilised eggs might be the cause of serious problems.     

B cell differentiation in the bursa of Fabricius and spleen of embryos and chicks immediately after hatching

Flow cytometric analysis and immunohistochemical observation were used to qualitatively and quantitatively clarify the nature of B cell differentiation in the bursa of Fabricius of chick embryos and to determine the timing of antibody class switching in chicken spleens based on positivity of IgM and IgG on and in the cells. In the bursa, the sIgM‐positive cell population formed from the 12^th to 15^th day of embryogenesis. The proportion of sIgM‐high expressing (sIgM^high) cells was lower among bursacytes than splenocytes of hatched chicks, suggesting that the sIgM^high bursacytes are to be released to peripheral sites. The proportion of sIgM^high cells was higher at 0 days old than at any other examined stage of development. Colonization of the spleen by B cells occurred between the 18^th day of embryogenesis and 0 days old. Antibody class switching was thought to start in the spleen between 1 and 2 weeks of age, because IgG‐positive cells were present in the spleen of 2‐week‐old chicks, but not 0‐day‐old or 1‐week‐old chicks.     

雏鸭腔上囊的生长及组织发育

通过对1～49日龄雏鸭腔上囊绝对质量和生长指数测定以及组织学观察,探讨雏鸭腔上囊生长及组织发育规律。结果:随日龄增长,腔上囊绝对质量逐渐增高;腔上囊生长指数21日龄达最高;黏膜大皱褶高、宽度、淋巴滤泡面积、皮质宽度均不断增加;小结相关上皮向腔面形成突起,并不断增高增宽。结果表明,雏鸭腔上囊1～14日龄生长较为缓慢,14～35日龄生长较快,35～49日龄发育基本趋于稳定,其中21～28日龄是雏鸭腔上囊生长发育的高峰时期,28日龄时,雏鸭腔上囊组织结构基本发育成熟。     

Studies on the pathogenesis of Escherichia coli in the bursa of Fabricius (cloacal bursa) of the turkey.

Turkey poults were inoculated with avirulent or virulent strains of Escherichia coli by direct application to anal lips and were killed at postinoculation hours (PIH) 0.1, 2, 5, 10, 24, 48, 72, and 96. Bursae of Fabricius (cloacal bursae) were collected, cultured, and examined by light, fluorescent, and electron microscopy. The virulent strain of E coli was not recovered from the bursae after PIH 24, although the avirulent strain was recovered up to PIH 96. The E coli strains neither localized at nor associated with the bursal fold epithelium, passed through the follicular pad epithelium, nor caused cytopathologic changes in the lymphoid follicle. A mild catarrhal bursitis was observed at PIH 48 with the avirulent strain of E coli.     

鸡传染性法氏囊病的防制

鸡传染性法氏囊(又称腔上囊)病,是由法氏囊病毒引起的一种急性接触性传染病,仅鸡发病,发病日龄以14日龄多见,近年来有所提前,最早的有7～5日龄发病的,鸡场里,初次暴发本病通常危害最严重,后代再发生本病时就轻得多.     

Histologic characterization of the involuting bursa of Fabricius in single-comb white Leghorn chickens

Bursas from specific-pathogen-free white leghorn chickens of both sexes were examined at several intervals from hatching to 28 weeks of age. No histologic alterations other than scattered atrophic or cystic follicles were observed through 20 weeks. Obvious involution, first noted at 24 weeks, was in early stages in females and quite advanced in males. Involution was essentially complete by 26 weeks, and only cicatrized vestiges of bursas were present at 28 weeks of age. Gross manifestations included bursal atrophy, variable yellowish discoloration of the mucosa, and matting or total loss of identity of the mucosal plicae. Histologic characteristics of involution are summarized by the following approximate sequence: atrophy and exfoliation of plica epithelium; subepithelial stromal fibrosis; fusion and ultimate collapse of plicae; liquefactive necrosis of first medullary then cortical elements of follicles, which seemed to progress from basal to apical portions of the plicae; progressive proliferation of stromal connective tissue and infiltration of macrophages into areas occupied by necrotic follicles; and, finally, complete fibrous organization of luminal debris, leaving a firm nodule formed by a contracted muscularis surrounding the cicatrized remains of the mucosa.     

Functional restoration of the bursa of Fabricius following in ovo infectious bursal disease vaccination

The primary role of the avian bursa of Fabricius is to provide an essential microenvironment for B-lymphocytes to diversify their immunoglobulin genes by gene hyperconversion. Infectious bursal disease (IBD) vaccination using intermediate plus vaccine strains can temporarily deplete the bursal follicles and interrupt the normal B-cell development, which is generally followed by B-cell repopulation and histological regeneration. To find evidence that functional restoration of the bursa of Fabricius occurs in addition to the histological regeneration, we have analysed the chB1 gene expression, which indicates active bursal B-lymphocytes, and also the surface expression of a carbohydrate structure Lewis(x), a marker which identifies those bursal B-lymphocytes that are undergoing gene hyperconversion. In ovo vaccination with an immune complex vaccine (IBDV-BDA) caused transient bursal destruction in both the SPF and the maternally protected broiler groups with differences evident in the starting time, the severity and the duration of the effect. After the depletion phase, signs of histological regeneration appeared together with chB1- and Lewis(x) expression indicating that B-lymphocytes were functionally active and the bursa of Fabricius was serving again as an efficient primary lymphoid organ providing an appropriate microenvironment for B-cell development.     

Developmental process of Cryptosporidium in the intestine and bursa of Fabricius of chickens.

The developmental process of a Cryptosporidium isolated in Japan in the chicken intestine was investigated by scanning (SEM) and transmission electron microscopies (TEM). The parasites were detected in the ileum, cecum, colon, cloaca and bursa of Fabricius (BF). The intensity of infection tended to peak later in the BF than ileum. Trophozoites and schizonts were detected in all the portions of intestine, and were dominant in the developmental stages. Although macrogamonts were the secondary dominant stage, they were absent in the ileum and cecum at 60 hr postinoculation (PI). A few microgamonts were detected in the ileum at 36 hr PI and in the BF on day 19 PI. Oocysts were observed in the ileum at 48 hr PI and in the BF on day 19 PI.