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J W Scholten

Publications and source records attributed to J W Scholten.

14 recordsLinked to original sources

Long-term effects of social stress on antiviral immunity in pigs.

Mixing of unfamiliar pigs is common practice in intensive pig husbandry. Since pigs maintain a dominance hierarchy, mixing often leads to vigorous fighting. Apart from the negative impact that fighting has on welfare, there is evidence that the social stress associated with fighting suppresses immune function. In the present experiment, we investigated the impact of mixing on specific long-term immune responses and protection against challenge infection after vaccination with pseudorabies virus (PRV). Specific pathogen-free (SPF) pigs were mixed pairwise with an unfamiliar same-gender conspecific or left undisturbed with a same-gender littermate at 3 days after vaccination with PRV. Half of the pigs were females (gilts) and half were castrated males (barrows). Mixing increased agonistic behavior to the same degree in gilts and barrows. Cortisol concentrations in saliva and catecholamine excretion in urine were increased in mixed pigs, and these effects were independent of dominance status and gender. Subsequently, the effects of mixing, gender, dominance status and interactions between these factors on immune response parameters were studied. The main result was that mixed barrows showed suppressed immune responses after vaccination and increased clinical symptoms after challenge infection compared to control barrows. Mixed gilts however did not differ from control gilts. It also appeared that mixed dominants were more seriously affected than mixed subordinates were. We conclude that, in some pigs, social stress after mixing suppresses the immune response to a viral vaccine and consequently impairs protection against challenge infection.

Agonistic Behavior↗

Development of the natural response of immunoglobulin secreting cells in the pig as a function of organ, age and housing.

We analysed the development of the natural immunoglobulin-secreting cell (Ig-SC) response in systemic- and mucosal-lymphoid tissues of specified pathogen free pigs between 1 and 40 weeks of age. As antigen exposure may influence the development of the Ig-SC repertoire we also compared the frequencies of Ig-SC in various lymphoid tissues of 40 weeks old specified pathogen free pigs and conventional pigs. A procedure to isolate lamina propria cells from porcine intestine was adapted for this study. The frequencies of IgM-, IgG-, and IgA-secreting (spot forming) cells were determined with a reversed enzyme linked immunospot assay, which was also adapted for detection of Ig-SC in pigs. The Ig-SC frequencies were calculated as percentage of the mononuclear leukocytes isolated from the various organs. The observations till 40 weeks of age were as follows: Splenic IgM-SC predominated at all ages and reached a plateau of 0.1-0.2% of the mononuclear leukocytes already at 4 weeks of age. The IgM-SC of mesenteric lymph node (MLN) predominated up till 12 weeks of age and reached an optimum of 0.15% reached at 4 weeks of age. The frequencies of IgG-SC of spleen and MLN had dips around 4 weeks of age and increased thereafter till 40 weeks of age (spleen 0.025%, MLN 0.05% at 40 weeks of age). The frequencies of IgA-SC were low in the spleen (< or =0.003%) and moderate in the MLN (0.01-0.02%) at all ages tested. In peripheral lymph node (PLN) and bone marrow (BM), the frequencies of IgM-SC (0.03-0.05%) were much lower than in the spleen. The IgG-SC frequencies of BM and MLN also had dips around 4 weeks of age and increased thereafter. The IgG-SC frequency of BM reached a plateau at 12 weeks of age (0.15%) and for PLN the highest frequency was observed at 40 weeks of age (0.05%). The frequencies of IgA-SC were low in BM and PLN (<0.003%). High frequencies of IgA-SC were observed in mucosa associated tissue like Peyer's patches (PP) and intestinal lamina propria (till 20% of the mononuclear leukocytes in intestinal lamina propria of 12-40 weeks of age). IgM and IgA are both important isotypes in mucosal lymphoid organs in the pig. The shift from IgM to IgAas predominant, mucosal isotype was first observed in duodenum and jejunum (12 weeks) and later in ileum (40 weeks). The influence of ageing on the frequency of Ig-SC in PP was only observed in jejunal PP. whereas in ileal PP the frequencies of Ig-SC did not vary over time. We combined our data about the frequencies of IgM-, IgG-, and IgA-SC in various organs with data obtained by others about the distribution of lymphocytes over porcine lymphoid organs at about 12 weeks of age. Based on these calculations we concluded that the small intestine, with more than 80% of all Ig-SC, is fair most the major site of Ig production in the pig. We also concluded that the small intestine is the major site of IgA and IgM production cells in the pig. Although IgA becomes predominant along the intestine, the results demonstrated that in the pig IgM is more a mucosal isotype compared with other species. With 40% of all IgG-SC the porcine BM appeared to be the major site of IgG production. Unexpected results were obtained for IgG-SC in the systemic lymphoid organs. In these organs the frequencies of IgG-SC dropped firstly from 1 to 4 weeks of age and steadily increased thereafter till 40 weeks of age. This observation is discussed in relation to the possibility that systemic IgG-SC at one week of age were passively acquired from maternal colostrum. The influence of housing/antigenic load at 40 weeks of age was mainly expressed by an increase (2-8x) of the frequency of IgG-SC in spleen, PLN, BM, and intestinal lamina propria, whereas the typical mucosal IgA-SC frequencies in the lamina propria were hardly affected.

Aging↗

Silent memory induction in maternal immune young animals.

Maternal immunity was shown to be an effector mechanism which does not include transfer of memory. 'Boosting' of maternal immunity by vaccination was not effective. Transferred maternal immunity negatively interfered with the induction of optimal protection by vaccination. Antibody formation was not observed after vaccination of maternally immune piglets. In contrast, induction of memory had occurred in animals under maternal immune suppression. Vaccination in young animals negatively interfered with or abrogated, effective maternal immune protection. There was no correlation between specific serum antibody titres in piglets and protection to PRV. Thus apart from protection provided by antibodies contributions of other soluble factors and the cellular immune compartment as represented in colostrum and/or milk were important for protection.

Animals↗

A comparison of natural and recombinant cholera toxin B subunit as stimulatory factors in intranasal immunization.

Cholera toxin B (CTB) is often envisaged and used as an immune stimulating agent in protocols for mucosal immunization. However, the nature of the CTB used (natural vs recombinant) is frequently not taken in consideration. This is important since the usage of natural CTB in mucosal immunization regimen and the mucosal response resulting from such an immunization can be effected by the presence of the CTA subunit in commercial CTB preparations. To clarify this, we have compared natural vs recombinant CTB in an intranasal (i.n.) mucosal immunization procedure using ovalbumin (OVA) as antigen. The results show that recombinant CTB induces similar immune responses like natural CTB. Furthermore, our experiments show that covalent coupling of OVA to CTB is not required for the induction of OVA specific mucosal and systemic immune responses upon i.n. immunization.

Adjuvants, Immunologic↗

Parenteral vaccination of mice and piglets with F4+ Escherichia coli suppresses the enteric anti-F4 response upon oral infection.

We studied with a mouse model and in piglets the requirements to prime for a secondary, mucosal B-cell response against Escherichia coli F4 fimbriae, an important virulence factor of enterotoxigenic E. coli, the agent associated with postweaning diarrhoea in piglets. The major observation obtained with the mouse model was verified for piglets. Mice and piglets were primed orally or parenterally with purified F4ac antigen or whole bacterial cells carrying the F4ac antigen and were later orally infected with live F4ac+ E. coli bacteria. Cell suspensions of murine spleen or porcine serum were used to study the systemic B-cell response. Cell suspensions were also made of murine and porcine enteric lamina propria and were used to study the mucosal B-cell response. Enzyme-linked immunospot assays and enzyme-linked immunosorbent assays specific to E. coli F4ac antigen were used to quantify either the antibody-secreting cells or antibody titres in serum. Results showed that in mice only primary oral immunization with live bacteria induced an enteric immune response against the E. coli F4ac+ fimbriae. Oral immunization with killed bacteria induced hardly any mucosal immune response. Parenteral immunization induced a state of suppression that was reflected by the lack of an enteric immune response upon a subsequent oral infection with live bacteria. A comparable induction of suppression was observed in piglets using the same protocol. We conclude that parenteral vaccination of piglets with the E. coli F4ac antigen is ineffective to induce protective immunity at the mucosal level against postweaning diarrhoea and is possibly detrimental.

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↗

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↗

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↗

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↗

Basic defect in the expression of adenosine deaminase in ADA-SCID disease. II. Deficiency of ADA-CRM detected in heterozygote human-Chinese hamster cell hybrids.

A specific competitive radioimmunoassay (RIA) was employed to quantify human adenosine deaminase molecules produced in human-Chinese hamster somatic cell hybrids. Studies on a set of hybrids in which the normal and aberrant expressions of adenosine deaminase (assigned earlier to human chromosome 20) were segregating, have demonstrated that in the patient with ADA-SCID disease reported by Herbschleb-Voogt et al. (1981 a), the deficiency of ADA activity was associated with a comparable deficiency of adenosine deaminase specific immuno-crossreacting material (ADA-CRM).

Adenosine Deaminase↗