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Biomedical subjects

E M Janse

Publications and source records attributed to E M Janse.

At least 19 recordsLinked to original sources
PubMed1999

Prenatal development of hematopoietic and hormone-producing cells in the chicken adenohypophysis.

The developmental sequence of markers for hematopoietic, hormone-producing, and folliculo-stellate cells in the chicken adenohypophysis is described using immunohistochemical staining techniques. Hematopoietic cells are detected in cryosections of the adenohypophysis starting from 10- or 12-day embryos, using chicken-specific monoclonal antibodies against the leukocyte common antigen (CD45) and the macrophage antigen CVI-ChNL-68.1, respectively. During the second half of embryonic development, CVI-ChNL-68.1-positive macrophages, which are also found in several lymphoid and nonlymphoid tissues of the embryo, progressively populate the adenohypophysis, simultaneously with the maturation of the different hormone-producing cell types in their characteristic topographical location. In cryosections of embryonic chicken adenohypophyses, from day 10, distinct cell populations gradually become immunoreactive to chicken-specific monoclonal antibodies against proopiomelanocortin, the beta-subunit of luteinizing hormone, growth hormone, and prolactin. At hatching, these pituitary hormones are immunohistochemically detectable in a topographical pattern corresponding to the known distribution of hormone-producing cells in the adult chicken adenohypophysis. However, in the hatchling, there is no immunoreactivity to the S100 protein, a marker characteristic for the non-hormone-producing folliculo-stellate (FS) cells in the adult adenohypophysis, although FS cells in the 4-week-old chicken show a strong immunoreactivity to a polyclonal antiserum against bovine S100. Immunoreactivity to the major histocompatibility complex class II (MHC-class II of the chicken) is also absent in the embryonic adenohypophysis, thereby corroborating the absence of these characteristic markers of dendritic cells (MHC class II) and FS cells (S100) in the perinatal adenohypophysis, as in the rat. It is concluded that, whereas early macrophages populate the adenohypophysis simultaneously with the maturation of hormone-producing cells (i.e., during the second half of embryonic development), the FS cell-specific expression of S100 protein does not take place before hatching, and neither does the expression of MHC-class II antigens in the embryonic chicken adenohypophysis.

Animals↗
PubMed1999

Further characterization of M cells in gut-associated lymphoid tissues of the chicken.

M cells are considered to be the most effective cells for the transport of antigens from the intestinal lumen into the gut-associated lymphoid tissue. M cells are characterized by their ultrastructural appearance, the selective uptake of antigens, the binding of lectins, and the presence of underlying lymphocytes. Little attention has been paid to the interaction of intra-epithelial leucocytes and M cells in chickens; therefore, we have investigated both cell types separately and using double immunocytochemical staining in cecal tonsils and Meckel's diverticulum. In the follicle-associated epithelium (FAE), cells were present that differ from their neighbors by short, irregular microvilli. Ferritin was absorbed by these putative M cells, but also by other epithelial cells. The lectins of Triticum vulgaris (WGA) and Glycine max (SBA) showed a patchy staining of the FAE. The numbers of intra-epithelial leucocytes (IEL) increased rapidly after hatch, reaching innumerable at 6 wk of age. Most IEL were T lymphocytes expressing CD8 and only about 30% of them were B lymphocytes. Nevertheless, double staining of M cells (WGA/SBA) and IEL showed that M cells were much fewer than IEL. These results indicate that M cells are not solely induced by the intra-epithelial localization of leucocytes. Because the phenotype of IEL reflected the content of the adjacent underlying lamina propria, IEL immigrate the FAE locally and do not migrate along with the epithelial cells from the crypts. In conclusion, M cells exist in the chicken, but their phenotype and function are less well demarcated from neighbor epithelial cells than is seen in mammals.

Animals↗
PubMed1998

Induction of a local and systemic immune response using cholera toxin as vehicle to deliver antigen in the lamina propria of the chicken intestine.

In this study, the humoral mucosal immune response to a recombinant Eimeria antigen (Ea1A) was enhanced using cholera toxin (CT). Chickens were primed intra-intestinally with Ea1A either conjugated or not to CT. The local and systemic antibody responses to both Ea1A and CT were determined to find out whether the chickens could respond to CT and whether both antigens had reached the lamina propria. In addition the effects of CT on lamina propria leukocytes were examined. The results showed that chickens had receptors on the caecal epithelium that could bind CT. At day 7 after administration, the number of CD4+ and CD8+ T lymphocytes in the lamina propria of the caecum had increased, indicating that CT had a specific immunological effect. At this timepoint, anti-CT antibody containing cells were detected locally in the lamina propria of the caecum. In serum all antigen preparations containing CT induced IgM and IgG antibody titres specific for CT within 10 days after priming. In addition, the recombinant Ea1A antigen also induced serum responses when administered together with CT or conjugated to CT, thus both CT and the antigen had reached the lamina propria. Nevertheless, the Ea1A specific response was much higher in the primary response and after booster immunization when the antigen was conjugated to CT than when only mixed with CT. Therefore, we conclude that CT is a suitable adjuvant for intra-intestinal application in chickens, especially when the antigen is conjugated to it.

Adjuvants, Immunologic↗
PubMed1998

Inadequate anti-polysaccharide antibody responses in the chicken.

Chickens are notorious for the fact that they carry bacteria such as Salmonellae and Campylobacter, which can cause zoonoses by contamination of the end product, without hampering growth and development of the chicken itself. This carrier status can only been explained by the inability of the chickens immune system to eliminate the pathogen, this in turn being due to insufficient humoral responses towards the polysaccharides of the bacterial capsule. In a previous study, we demonstrated that in chickens a model thymus-independent type 2 (TI-2) polysaccharide antigen, trinitrophenylated Ficoll (TNP-Ficoll), hardly evokes a humoral immune response. Furthermore this TI-2 antigen was shown to exhibit a very specific initial localization pattern after intravenous injection, i.e. in the periellipsoidal lymphocyte sheaths (PELS) and the surrounding ring of macrophages. The functional equivalent of these macrophages in mammals, the marginal zone macrophages, were shown to suppress the humoral responses against TI-2 antigens. Therefore we investigated whether other standard TI-2 antigen models also induce low antibody responses, whether this low response is dose-dependent, and whether macrophages are responsible for this low response. It was found that other TI-2 antigens, such as hydroxyethyl starch and detoxified lipopolysaccharides, also induced very low IgM and IgG responses, indicating a general phenomenon that could not be overcome by using a higher dose of antigen. In addition, selective depletion of splenic macrophages with liposomes containing dichloromethylene diphosphonate prior to immunization increased the specific humoral response to TD and TI-1 antigens, but failed to do so for TI-2 antigen. This result indicates that the low humoral responses are not (only) due to a macrophage suppressive activity but also to other yet unknown mechanisms, for example the lack of responsive B cells in the splenic PELS.

Animals↗
PubMed1998

The working mechanism of an immune complex vaccine that protects chickens against infectious bursal disease.

The role of immune complexes (Icx) in B-cell memory formation and affinity maturation allow for their potential use as vaccines. Recently, a new immune complex vaccine has been developed that is currently under field trials conducted in commercial poultry. This immune complex vaccine is developed by mixing live intermediate plus infectious bursal disease virus (IBDV) with hyperimmune IBDV chicken serum (IBDV-Icx vaccine). Here we have investigated the infectivity of this vaccine as well as the native IBDV (uncomplexed) vaccine in terms of differences in target organs, in target cells and speed of virus replication. At various days after inoculation on day 18 of incubation (in ovo) with either one dose of virus alone or the IBDV-Icx vaccine, the replication of IBDV and the frequency of B cells and other leucocyte populations were examined in the bursa of Fabricius, spleen, and thymus using immunocytochemistry. With both vaccines, IBDV was detected associated with B cells, macrophages and follicular dendritic cells (FDC) in bursa and spleen, although complexing IBDV with specific antibodies caused a delay in virus detection of about 5 days. Most remarkable was the low level of depletion of bursal and splenic B cells in IBDV-Icx vaccinated chickens. Furthermore, in ovo inoculation with the IBDV-Icx vaccine induced more germinal centres in the spleen and larger amounts of IBDV were localized on both splenic and bursal FDC. From these results we hypothesize that the working mechanism of the IBDV-Icx vaccine is related to its specific cellular interaction with FDC in spleen and bursa.

Animals↗
PubMed1998

The use of chicken-specific antibodies in veterinary research involving three other avian species.

In this study we investigated a panel of 20 polyclonal and monoclonal antibodies specific for chicken cells and tissues for cross-reactivity with other avian species, i.e. turkey, duck and quail, using immunoperoxidase staining on cryostat sections. The number of cross-reacting antibodies was highest for turkey (12/20) and quail (10/20) and lowest for duck (5/20), reflecting the phylogenetic distance between these birds. In ducks only antibodies specific for mononuclear phagocytes and for stromal molecules in mesenchym, muscle cells, endothelial, and epithelial cells cross-reacted. In turkeys and quails cross-reaction was seen with antibodies against T-helper lymphocytes, IgM- and IgG-positive B cells and plasma cells, and mononuclear phagocytes. In addition, most antibodies specific for stromal molecules reacted against turkey or quail molecules. The panel of reacting antibodies can be used for research into the natural and specific defence mechanisms of turkeys and quails.

Animals↗
PubMed1996

Eimeria tenella infections in chickens: aspects of host-parasite: interaction.

Intestinal coccidiosis, caused by various species of Eimeria, has become an economically important disease of poultry and livestock throughout the world. Infection of chickens starts after ingestion of oocysts when sporozoites penetrate the epithelium of the villi. After passage through the lamina propria, they enter crypt epithelial cells where they undergo several rounds of asexual and sexual proliferation, thus forming merozoites and later, gametocytes. When macrogametes are fertilized by microgametes, oocysts are formed that are shed in the faeces. Nowadays, coccidiosis is prevented by anticoccidial drugs that are added to food, but the prolonged use of these drugs leads inevitably to the emergence of resistant Eimeria strains. During infection, there are three stages when the chicken immune system can inhibit parasitic development. The first is when the sporozoite searches for a site of penetration and binds to the epithelium. The second is when the sporozoite is in the villus epithelium amongst intra-epithelial leucocytes. The third is during its passage through the lamina propria to the crypt epithelium. To investigate this, the decisive factors in the induction and effector phase of immunity against coccidiosis have been investigated in situ. Our studies have revealed that three phenomena are responsible for immunity against Eimeria infections. First, the actual passage and presence of parasites in the lamina propria to induce immunity. Second, the sporozoite seems to be the most important parasite stage for immunity, and third, cytotoxic T cells are necessary to inhibit parasites.

Animals↗
PubMed1992

Histological and functional differentiation of non-lymphoid cells in the chicken spleen.

The phenotypes and functions of various populations of non-lymphoid cells in the chicken spleen were investigated with monoclonal antibodies and after in vivo administration of antigens. Monoclonal antibody CVI-ChNL-68.1 was used to detect red pulp macrophages, interdigitating cells, and monocytes, whereas CVI-ChNL-68.2 was used to detect ellipsoid-associated non-lymphoid cells (EANC). Two new monoclonal antibodies were developed: CVI-ChNL-74.2, which specifically recognized red pulp macrophages and a ring of macrophages surrounding the peri-ellipsoid lymphocyte sheath; and CVI-ChNL-74.3, which specifically recognized follicular dendritic cells (FDC) in germinal centres and small clusters of stromal cells in T-cell areas. After intravenous injection of lipopolysaccharide (LPS), the number of 68.1+ and 74.2+ macrophages decreased dramatically, whereas 68.2+ EANC and 74.3+ FDC were unaffected. After intravenous injection of heat-inactivated Brucella abortus, the numbers of both macrophages and EANC decreased. In contrast, a significant increase of 74.3+ cells was observed in the T-cell areas outside the germinal centres. As expected, intravenous injection of non-mitogenic antigens, such as keyhole limpet haemocyanin and Ficoll, and carrageenan did not affect the non-lymphoid cell populations. At least six subpopulations of non-lymphoid cells in the chicken spleen can now be discriminated with monoclonal antibodies. Our results show that mononuclear phagocytes are sensitive for mitogenic stimulators such as LPS and Brucella abortus. In contrast, stromal non-lymphoid cells are only sensitive to the particulate mitogen Brucella abortus. We conclude that the complex formed by ellipsoid cells, the peri-ellipsoid B-cell sheath, and the surrounding macrophages, is the functional equivalent of the mammalian marginal zone.

Animals↗
PubMed1991

Ontogeny and function of two non-lymphoid cell populations in the chicken embryo.

The purpose of the study was to determine what type of non-lymphoid cells develop in chicken embryos during ontogeny, and whether these cells are functional. To detect these cells, we used monoclonal antibodies specific to two groups of non-lymphoid cells: CVI-ChNL-68.1, specific for mononuclear phagocytes, and CVI-ChNL-68.2, specific for a subpopulation of reticulum cells in spleen, liver and bursa. Monoclonal antibodies HIS-C7, HIS-C1, and HIS-C12, which are specific to leukocytes, B lymphocytes, and IgM respectively, were used to correlate the ontogeny of non-lymphoid cells and lymphoid cells. Mononuclear phagocytes and reticulum cells were detected in the liver, spleen, yolk sac, bursa, gut, and thymus at about the same time as leukocytes, but earlier than B lymphocytes. To determine whether mononuclear phagocytes and reticulum cells in spleen and liver absorb antigen, we injected embryos intravenously with colloidal carbon and the antigen FITC-Ficoll. In addition, acid phosphatase was used as a marker for phagocytic activity. Reticulum cells in the liver and spleen were functional from the first point of detection, whereas mononuclear phagocytes in the liver and spleen started to absorb antigen a few days after their development.

Animals↗
PubMed1989

Postnatal development of mucosa-associated lymphoid tissues in chickens.

The postnatal development of chicken mucosa-associated lymphoid tissues of the eyes, lungs, and intestines were investigated with monoclonal antibodies specific for either all leucocytes, B lymphocytes, mononuclear phagocytes, IgM, IgG, or IgA. Attention has been paid to the relation of lymphoid infiltrates with their surrounding mucosae, the segregation into B-cell and T-cell areas, development of germinal centers, and secretory immunoglobulins. Abundant secretory IgM and IgA was detected in the epithelium of the Harderian glands in the orbits, even though they lacked large leucocyte infiltrates with germinal centers. Lymphoid tissues in the mucosae of lungs and intestines developed separate B-cell and T-cell areas. The proventriculus, Meckel's diverticulum, and Peyer's patches generally contained germinal centers from 12 weeks of age on. Because chickens as young as 2 weeks old had germinal centers in bronchus-associated lymphoid tissue and cecal tonsils, these areas were probably highly stimulated by antigens. Isotype-specific monoclonal antibodies were used to detect IgM-, IgG-, and IgA-bearing follicular cells in the same germinal center.

Animals↗
PubMed1989

Distribution and function of non-lymphoid cells positive for monoclonal antibody CVI-ChNL-68.2 in healthy chickens and those infected with Marek's disease virus.

Immuno-enzyme histochemistry was used to study the staining pattern and tissue distribution of monoclonal antibody CVI-ChNL-68.2 that specifically reacts with a subset of non-lymphoid cells in healthy chickens and those infected with Marek's disease virus (MDV). Functional characteristics of CVI-ChNL-68.2-positive cells, e.g. antigen uptake, are determined. In the liver CVI-ChNL-68.2 recognizes reticulum cells, whereas in the bursa of Fabricius it detects single cells in the interfollicular connective tissue. In the spleen CVI-ChNL-68.2 reacts selectively with the reticulum cells of the ellipsoid. In some MDV-infected chickens the splenic reticulum cells show a different staining and distribution pattern. Furthermore, the proliferative lesions associated with Marek's disease contain many CVI-ChNL-68.2-positive cells. The possible role of CVI-ChNL-68.2-positive cells in disseminating Marek's disease virus is discussed.

Animals↗
PubMed1988

The monoclonal antibody CVI-ChNL-68.1 recognizes cells of the monocyte-macrophage lineage in chickens.

The characteristics of monoclonal antibody CVI-ChNL-68.1, which specifically reacts with a group of chicken non-lymphoid cells, are described. Both tissue distribution shown on cryostat sections using immuno-enzyme histochemistry, and quantitative data obtained on cell suspensions are presented. Functional characteristics of CVI-ChNL-68.1-positive cells, such as antigen uptake and glass adherence, are determined. Results show that CVI-ChNL-68.1 reacts with monocytes, macrophages, and interdigitating cells. Possible relationships between the various non-lymphoid cells are discussed.

Acid Phosphatase↗
PubMed1988

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.

Animals↗
PubMed1988

The ontogenetic development of macrophage subpopulations and Ia-positive non-lymphoid cells in gut-associated lymphoid tissue of the rat.

The ontogenetic development of macrophage subpopulations and Ia-positive non-lymphoid cells was studied in gut-associated tissue in fetal and neonatal Wistar rats. A two-step immunoperoxidase method was carried out on cryostat sections, a panel of monoclonal antibodies being applied and aimed specifically at rat macrophages (ED1, ED2 and ED3) and at Ia antigen (Ox4). The first ED1-positive macrophages appeared in the liver on Day 15 (gestational age), and they did not express Ia. In developing mesenteric lymph nodes and in the gut wall, macrophages were found for the first time on Day 17 and 18, respectively, of gestation. These early macrophages were also ED1-positive. Until birth, ED2 recognized few cells in the gut-associated tissue; on the day of birth this subpopulation showed a sudden and considerable increase. The distribution pattern of ED3-positive macrophages appeared to be the same in fetal and in adult rats; it was confined to lymph nodes and Peyer's patches. Ia-positive non-lymphoid cells appeared in the abdomen early in ontogeny. The first Ia-positive cells displayed dendritic features and were found on Day 15 of fetal life in the mesenchymal tissue between intestinal loops. A few days later, many Ia-positive cells with a dendritic appearance were demonstrable in the gut wall and in developing mesenteric lymph nodes. Their number increased rapidly during the following days. Based on these results, the existence of two differentiation lines for dendritic cells and classical macrophages is discussed. The function of early Ia-positive cells in the abdomen is suggested as not being an antigen-presenting one.

Animals↗