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A T Bianchi

Publications and source records attributed to A T Bianchi.

At least 19 recordsLinked to original sources

Development of the B- and T-cell compartments in porcine lymphoid organs from birth to adult life: an immunohistological approach.

Using immunohistological techniques, we studied the development over time of B- and T-cell compartments in the lymphoid organs of specific-pathogen-free pigs. Tissue samples were collected at various time-points, starting 2 days before the pigs were born until the pigs were 10 months old. The samples were collected from the spleen, thymus, peripheral lymph node, mesenteric lymph node, duodenum, jejunum, ileum, jejunal Peyer's patch and ileal Peyer's patch. Monoclonal antibodies specific to B- and T-cells were used to identify where the following cells were localized: IgM-B cells (cells positive to surface immunoglobulin), IgM-, IgG- and IgA-containing cells (cells positive to cytoplasmic immunoglobulin), and CD2-, CD4- and CD8-positive cells. The development of the B- and T-cell subpopulations in each organ was analysed. Two days before birth, most organs contained quantities of IgM-B cells. The spleen, lymph nodes, Peyer's patches and, notably, the thymus, contained some immunoglobulin-containing cells (Ig-CC); this finding indicates that pigs have cells that secrete immunoglobulins before birth. Just after birth, the incidence of Ig-CC increased in most organs; first IgM-CC increased, then either IgG- or IgA-CC increased, depending on the organ. T-cell development was observed clearly in spleen and in the lamina propria of the small intestine, in contrast to other organs, in which the T-cell compartments containing various T-cell subpopulations were well developed before birth. Comparison of the incidence of CD4+ and CD8+ cells showed that the CD4:CD8 ratio of these cells in the spleen, lymph nodes, Peyer's patches and small intestine is low, especially in adult pigs, compared with the CD4:CD8 ratio in other species. Weaning had little influence on the incidence of B- and T-cells in lymphoid organs. This study is the first immunohistological survey to describe the development of the major B- and T-cell subpopulations in various lymphoid organs of pigs, and it should be useful for future immunopathological and comparative immunological studies in pigs.

Aging

Manipulation of intestinal immune responses against ovalbumin by cholera toxin and its B subunit in mice.

We studied the effect of mucosal presentation of ovalbumin (OVA) conjugated to cholera toxin (CT) or cholera toxin B subunit (CTB) on the intestinal immune responses against OVA. Mice were primed intraperitoneally (i.p.) with OVA in a water-in-oil emulsion and boosted intraduodenally (i.d.) with OVA conjugated to CT or CTB in various molar ratios. Responses were evaluated by testing intestinal secretions for OVA-specific antibodies and by quantifying the OVA-specific antibody secreting cells (ASC) in the lamina propria of the small intestine. OVA-CT conjugates were tested in a molar ratio ranging from 1.8:1 to 4500:1. OVA-CTB conjugates were tested in a molar ratio ranging from 0.25:1 to 500:1. The optimum intestinal immune response was reached at a molar ratio of 1.8:1 for OVA-CT and 5:1 for OVA-CTB. The binding capacity of OVA-CTB, but not of OVA-CT, to GM1 ganglioside corresponded with the capacity to enhance the intestinal immune response. The effect of conjugating CTB or CT to OVA on the immune response against OVA was more striking when mice were not only boosted i.d., but also primed i.d. Both OVA-CT and OVA-CTB induced detectable immune responses, whereas free OVA did not. Therefore, the carrier effect of CT or CTB is essential to trigger a mucosal immune response against OVA when presented mucosally only. We conclude that enhancing antigen uptake greatly facilitates mucosal immune responses.

Adjuvants, Immunologic

[Mucosal immunity].

The mucosal immune system protects against infections that enter the body via the mucosae. This article describes how the mucosal immune system functions and how mucosal immune responses can be induced by vaccination. It also discusses various vaccination regimens that maximize the induction of mucosal immune responses against bacterial and viral infections. One paragraph of the article discusses the possibilities of developing vaccines that induce a mucosal immune response against parasite infections.

Animals

The use of monoclonal antibodies in an enzyme immunospot assay to detect isotype-specific antibody-secreting cells in pigs and chickens.

Monoclonal antibodies directed against porcine immunoglobulin isotypes G, G1, G2, M, and A and against chicken immunoglobulin isotopes G, M, and A were tested in an antigen-specific spot-forming cell (SFC) assay based on the principle of the enzyme immunoassay. The SFC assay was used to quantitate ovalbumin (OA)-specific antibody-secreting cells (ASC) in pigs that had been primed and boosted with OA. The SFC assay was also used to quantitate trinitrophenyl (TNP)-specific ASC in chickens that had been primed with TNP-conjugated keyhole lympet haemocyanin (TNP-KLH). Although, the classical plaque-forming cell (PFC) assay cannot reliably detect isotope-specific ASC in pigs and chickens, it can detect these cells in mice. Therefore, we compared the OA- and TNP-specific SFC assays with PFC assays that were specific for these antigens in mice. The study demonstrated that the SFC assay is superior to the PFC assay in detecting both OA-specific ASC and TNP-specific ASC. The frequencies of OA-specific and TNP-specific SFC detected in mice were of the same order of magnitude as those detected in pigs and chickens. We concluded that the SFC assay is the better method for quantitating ASC in pigs, chickens, and probably all domestic animals for which isotype-specific monoclonal antibodies are available.

Animals

Background (spontaneous) immunoglobulin production in the murine small intestine before and after weaning.

The ontogeny of the murine intestinal B-cell compartment before and after weaning was studied by quantitative analysis of immunoglobulin-secreting cells (Ig-SC) in the small intestine (SI). Before weaning, few Ig-SC were detected in the SI, whereas spleen and bone marrow already contained many Ig-SC. The number of Ig-SC in the SI started to increase immediately after weaning. Comparing early-weaned mice with non-weaned mice of the same age clearly demonstrated that weaning brought on the development of Ig-SC in the SI. The influence of a gut flora on the number of Ig-SC in the SI was examined by comparing the number of Ig-SC in the SI of conventionally housed, specific pathogen free (SPF) and germ-free mice. A bacterial flora was apparently needed for the normal development of Ig-SC in the SI. Comparing mice containing an aerobic Gram-negative bacterial flora with mice containing only an anaerobic Gram-positive bacterial flora demonstrated that the type of bacterial flora is relatively unimportant. No evidence was found that circulating maternal antibodies suppressed the development of the "spontaneous" intestinal and systemic B cell response. The results show that bacterial colonization of the intestine plays a pivotal role in the development of the Ig-SC compartment in the SI.

Animals

Quantification of antigen-specific antibody-secreting cells in the small intestine and other lymphoid organs of mice after oral booster immunization.

The intestinal immune response of mice against ovalbumin (OVA) was quantified by isolating lymphoid cells from the small intestine (SI) and testing them for antigen-specific immunoglobulin (Ig) secretion. The isolation procedure for functionally active lymphoid cells from the SI, originally developed to quantify the number of 'background' Ig-secreting cells in the SI, proved to be a useful method for evaluating antigen-specific intestinal immune responses quantitatively. The method was able to detect antigen-specific antibody-secreting cells (ASC) in the SI even when these cells occurred at a minimum frequency of only 0.006%. When mice were primed intraperitoneally (i.p.) with polymerized OVA and given an oral OVA booster immunization, OVA-specific ASC appeared in the SI from Day 3 after booster. After i.p. priming and an i.p. booster these cells could not be detected in the SI. The OVA-specific IgA-ASC responses in various organs after oral booster immunization were compared. From Day 5 after booster, when the response peaked, most OVA-specific IgA-ASC occurred in the SI. This suggested that these cells are mainly responsible for the OVA-specific antibodies demonstrated by ELISA in intestinal secretions from Day 6 after oral booster immunization. It is concluded that the quantitative method used in this study detects antigen-specific ASC in the SI with great sensitivity and could be used to evaluate immunization regimes aimed at inducing intestinal mucosal immune responses.

Animals

Background (spontaneous) immunoglobulin production in the murine small intestine as a function of age.

The development of the intestinal B-cell compartment in C3H/He mice was studied as a function of age by quantification of the number of intestinal immunoglobulin-secreting cells (Ig-SC). Before and at weaning, the number of Ig-SC in the small intestine (SI) was below 10(3) Ig-SC per SI. During the first few weeks after weaning, this number rose steeply and continued to rise until the mice were about 48 weeks old, when a maximum of more than 25 x 10(6) Ig-SC per SI was found. After 1 year of age the number of Ig-SC decreased. At all ages, the great majority of Ig-SC in the SI produced IgA. The increase of the number of IgA-SC in the SI after weaning is reflected in the amount of IgA in intestinal secretions measured by ELISA. The number of Ig-SC in the SI was compared with the number of Ig-SC found in spleen, bone marrow, Peyer's patches and mesenteric lymph nodes. Striking differences were observed between the SI and the other organs tested in total number, isotype distribution and kinetics of the increase of Ig-SC during ontogeny. These differences are discussed in relation to the regulation of the immune response in the SI and the migration patterns of lymphocytes in mucosal tissues.

Aging

In vivo priming of helper and suppressor T cells by alloantigens. Frequency analysis with the use of an in vitro limiting dilution assay.

Earlier studies have demonstrated that T cells activated in mixed lymphocyte reactions can exert positive as well as negative allogeneic effects on B cells expressing the appropriate alloantigens on their surface. We investigated the effect of in vivo priming of T cells with alloantigens on their capacity to help or suppress allogeneic B cell cultures against sheep erythrocytes. We used immunization protocols that have been shown to be optimal for induction of alloantigen-specific delayed-type hypersensitivity (DTH) and alloantigen-specific suppressor T (Ts) cells for DTH. The results show that in vivo stimulation with alloantigens, depending on the immunization route and the lymphoid organ studied, can be as effective as in vitro stimulation in increasing the frequency of alloantigen-specific helper T (Th) cells and Ts cells. Subcutaneous immunization induced a 10-fold frequency raise of Th cells as well as of Ts cells in the lymph nodes. In the spleen the Th cell population was hardly affected by s.c. immunization, whereas the Ts cell population increased by at least a factor 20. Intravenous immunization, on the other hand, selectively expanded the Th cell population in the spleen, whereas the splenic Ts cell population and the Th and Ts cells in the lymph nodes were not affected. Comparison of these results with our previous data concerning characteristics and the requirements of in vivo activation of alloantigen-specific DTH reactive T cells and of alloantigen-specific Ts cells suggest that different Ts cell populations are involved in suppression of alloantigen-specific DTH in vivo and of allogeneic suppression of in vitro induced sheep erythrocytes specific antibody formation.

Animals

Contribution of immunoglobulin-secreting cells in the murine small intestine to the total 'background' immunoglobulin production.

In this study we investigated the contribution of the small intestine of C3H/He mice to the spontaneous ('background') immunoglobulin (Ig) production in terms of the number of Ig-secreting cells (Ig-SC), and compared the results with the numbers of Ig-SC found in various lymphoid organs (spleen, bone marrow, mesenteric lymph nodes and Peyer's patches). The results show that in C3H/He mice of 20 weeks of age, on average 16 x 10(6) Ig-SC can be isolated from the small intestine. Almost all of these Ig-SC produce IgA. Compared to the other lymphoid organs, the small intestine contains more than 80% of all Ig-SC present in adult C3H/He mice. These results are in agreement with the need to maintain relatively high levels of Ig at the mucosal surfaces, especially of secretory IgA, for the prevention of penetration of these surfaces by micro-organisms. In man and mouse most of these Ig are supposed to be produced locally in the underlying mucosal tissues and subsequently transported across the epithelium. Although the IgM and IgG levels in serum are predominantly maintained by non-mucosae associated lymphoid organs, the results of this study clearly indicate that the mucosal tissues are the major site of 'background' Ig-production.

Animals

Suppression of antigraft immunity by preimmunization. III. Characterization of suppressor T cells involved in suppression of the efferent phase of DTH against alloantigens.

Suppressor T (Ts) cells that can suppress delayed type hypersensitivity (DTH) against histocompatibility (H) antigens can be isolated from spleen and lymph nodes a few days after i.v. immunization of mice with irradiated allogeneic spleen cells. In this paper we investigated the suppression of the efferent phase of DTH to characterize the Ts cells involved, and to compare them with the afferent phase Ts cells that have been characterized in a previous paper of this series. The DTH against third party alloantigens that were not used for the i.v. suppressive immunization could be suppressed by presenting the third party alloantigens together with the original alloantigens in the challenge inoculum for eliciting the DTH reaction. Thus the ultimate suppressive effect by the Ts cells that are active during the efferent phase of DTH is nonspecific. This non-specific suppression of DTH to alloantigens has previously been found for the afferent phase Ts cells as well. For suppression of the efferent phase of DTH to alloantigens, a population of Lyt-1+2+ Ts cells appeared to be essential, just like in the suppression of the afferent phase of DTH to alloantigens. We did not find evidence for the involvement of cyclophosphamide-sensitive auxiliary Ts cells in suppression of the efferent phase of DTH. Also no evidence was found for H-2 or Igh-restricted activation and function of the Ts cells that were active during afferent and efferent phases of the DTH response to H antigens. In view of these similarities between afferent phase and efferent phase Ts cells we conclude that there are no arguments as yet to suppose that there is more than one type of T cells involved in the suppression of the afferent and efferent limb of DTH against H antigens.

Animals

The influence of a water-in-oil emulsion on humoral immunity.

A strategy for research of immunostimulants is best served choosing a way by which the immune response can be studied without being complicated by a combination of effects originating both in adjuvant and antigen. This means that adjuvant and antigen preferably are applied with an interval and by different routes. In our model an adjuvant (a W/O emulsion) was applied intraperitoneally, whereas the antigen was injected intravenously. The stimulatory effect on the splenic plaque forming B-cell response depended on the dose of antigen, on the interval between adjuvant and antigen application, on the mouse strain used, and on the quality of the antigen with respect to the intrinsic adjuvanticity of the antigens.

Adjuvants, Immunologic

Research of the application of a water-in-oil emulsion for the prevention of post-weaning diarrhoea and oedema disease in piglets.

A stable water-in-oil emulsion was injected intraperitoneally (i.p.) in piglets about 5 days before weaning to prevent post-weaning diarrhoea (PWD) and oedema disease (OD). So far more than 200,000 piglets have been treated with this adjuvant on a number of farms. On these farms the mortality rate due to PWD and OD decreased, whereas the need for antibiotic treatment declined. Experiments involving alternate application of adjuvant and physiological saline, or adjuvant treatment and no treatment at all, showed a statistically significant positive effect of adjuvant application. The effect of i.p. adjuvant application on specific and non-specific defence mechanisms were examined in well defined rat- and mouse-models, to throw light upon the mechanisms behind the observed adjuvant effect in piglets.

Adjuvants, Immunologic

Decreased in vivo functional T cell capacity in the murine autoimmune strains MRL/Mp-lpr/lpr and male BXSB/Mp.

We have studied the functional and proliferative capacity of the T cells of systemic lupus erythematosus (SLE)-prone MRL and BXSB mice during aging. The study was performed in vivo, in the delayed-type hypersensitivity (DTH) assay, and in vitro, in the mixed lymphocyte reaction (MLR). Both assays showed depressed T cell responses in MRL/l and male BXSB mice at 4 to 5 and 9 to 10 months of age, respectively, when a significant proportion of the animals showed clear-cut disease. At younger ages, the proliferation-dependent DTH was not affected in MRL/l and male BXSB mice. This is in agreement with the reported primary B cell defect of male BXSB mice, but not with the reported early T cell abnormalities in MRL/l mice. A subnormal reactivity of the highly proliferating T helper cells or the existence of a T cell subset with normal DTH reactivity might account for the relatively long-lasting normal DTH reactivity in MRL/l mice.

Animals

Separate application of adjuvant and antigen: the effect of a water-in-oil emulsion on the splenic plaque-forming cell response to sheep red blood cells in mice.

The effect of a non-immunogenic adjuvant on the murine splenic plaque-forming cell (PFC) response against sheep red blood cells (SRBC) was studied. The adjuvant, a stable water-in-oil (W/O) emulsion, was injected intraperitoneally at the same time as or prior to the intravenous (i.v.) injection of SRBC. Enhancement of the SRBC-specific IgM-, but not IgG- and IgA-responses was observed. The stimulatory effect depended on the dose of both adjuvant and antigen and on the interval between their application. The minimal dose of adjuvant needed to induce maximal stimulation increased with the interval between the injections. Administration of an optimal adjuvant dose one week before antigen application still resulted in a clear stimulation of the response to the antigen. In adjuvant-treated animals, the primary PFC response did not exceed the maximum level reached after i.v. injection of a high dose of SRBC. Adjuvant therapy also resulted in polyclonal B cell-activation, since the number of spontaneous Ig-secreting cells in the spleen was increased. The kinetics and isotype distribution of the SRBC-specific and polyclonal responses, however, were different. Therefore, the observed stimulatory effect on the SRBC-specific PFC-response cannot be explained by the polyclonal activation of the immune system. From this study it appears that injection of a W/O emulsion provokes an active stimulation of the immune system, which demonstrates that the adjuvant effect of W/O emulsions is not only passively obtained by prolonged antigen presentation by depot formation.

Adjuvants, Immunologic