PubMed Health⌕ Search

Biomedical subjects

C Fossum

Publications and source records attributed to C Fossum.

At least 55 records · Page 3Linked to original sources

Antigen-specific increases in the number of splenocytes expressing MHC class II molecules following restimulation with antigen in various physical forms.

To understand how a presentation system for antigens initiates an immune response and why it has a strong adjuvant activity, a number of parameters need to be analysed. In this study the frequency of spleen cells expressing MHC class II (Ia antigen) was determined after immunization of mice and restimulation of their spleen cells, in vitro, with influenza virus envelope proteins in different physical forms, namely iscoms, micelles and virus particles. All three forms of the antigen stimulated, in an antigen-specific manner, an increased proportion of spleen cells expressing MHC class II in the restimulation experiments. The induction of increased MHC class II expression was at least partly dependent on antigen-specific induction of IFN-gamma since an antibody to IFN-gamma partly inhibited the increase of MHC class II+ cells induced by iscom or by Concanavalin A. The iscom-borne antigens were superior to micelles to prime the immune response in vitro, indicating a capacity to induce memory cells. This primed immune response was readily recalled in vitro, as measured by IFN-gamma production and an increased number of MHC class II positive cells.

Animals↗

2-Mercaptoethanol influences the in vitro function of bovine peripheral blood mononuclear cells.

The effects of 2-mercaptoethanol (2-ME) on some in vitro functions of bovine peripheral blood mononuclear cells (PBMC) were examined. It was shown that 2-ME enhanced, in a dose-dependent manner, the production of antibodies to bovine coronavirus. In this test the optimal concentration of 2-ME was 50 microM. This molarity of 2-ME was also optimal for the pokeweed mitogen (PWM)-induced proliferation of PBMC obtained from the 7 cattle tested. Similarly, the spontaneous proliferation of PBMC from 4 out of these cattle was enhanced. Thus, 2-ME evoked an increase (up to 2.5 times) or a decrease (at most 10 times) of the quota between the PWM-induced and the spontaneous proliferation (stimulation index). In general, the presence of 50 microM 2-ME enhanced the in vitro production of interferon by bovine PBMC. On the contrary, the highest proliferative response of PBMC to stimulation with bovine virus diarrhoea virus was achieved in cultures without 2-ME or in cultures with 0.5 or 5 microM 2-ME. Since the effects of 2-ME varied, for different tests as well as for cattle tested, attention should be paid to the use of 2-ME in cultures of bovine PBMC.

Animals↗

Effect of estradiol-17 beta treatment of gilts on blood mononuclear cell functions in vitro.

Porcine blood mononuclear cells (BMC) were exposed to prepartum concentration of estrogen in gilts before acquisition (in vivo), and their subsequent reactivity (in vitro) was explored. In a cross-over experimental designed study, 6 ovariectomized gilts were injected once with 3.75 mg of estradiol-17 beta benzoate in arachidic oil or with arachidic oil only during 2 experiments. The ability of their BMC to proliferate in response to stimulation with phytohemagglutinin, concanavalin A, and pokeweed mitrogen was assayed in cultures of blood and in cultures of purified BMC. After 2 days of mitogen stimulation, activity of accessible interleukin 2 was quantified in supernatants obtained from cultures of purified BMC and supernatants of blood cultures stimulated with pokeweed mitogen. Also, production of immunoglobulins by purified BMC in response to polyclonal stimuli was measured. Three days after treatment with estradiol, the proliferative response was suppressed in blood cultures stimulated with concanavalin A (P less than 0.05) and phytohemagglutinin (P less than 0.07). Effects of estradiol treatment were not found in any of the assays performed with purified BMC. We, therefore, assumed that in vivo exposure to estradiol can affect the function of porcine BMC; however, this was only evident when the in vitro assays were performed on blood cultures.

Animals↗

Genetic variation in Con A-induced production of interleukin 2 by porcine peripheral blood mononuclear cells.

Genetic variation in concanavalin A (Con A)-induced proliferation and interleukin 2 (IL-2) production by peripheral blood mononuclear cells (PBMC) was studied in blood collected from 96 piglets, aged 7 weeks. The piglets were the offspring of seven sires and 24 dams. Pronounced differences between litters from various dams were observed in the immune parameters measured. Also, large individual differences in the magnitudes of Con A-induced proliferation and IL-2 production were seen for PBMC collected from individual pigs within each litter. Both the time course and magnitude of IL-2 activity showed genetic variation, as results from the offspring of the seven sires differed significantly. However, only the time course, not the magnitude, of proliferation differed among the offspring groups. It was possible to establish a rank order for the sires based on the IL-2 production of PBMC by their offspring. As IL-2 has a key role in regulating the immune response, mitogen-induced IL-2 activity seems to be a good candidate as a general marker for cell-mediated immunity in pigs.

Animals↗

Detection of porcine MHC class II antigens in different immunoassays.

A monoclonal antibody (Mab), Mab 4-24-11, to human class II major histocompatibility antigens has been tested in commonly used immunoassays for detection of porcine class II major histocompatibility antigens (SLA-D). In a radioimmunoprecipitation assay, Mab 4-24-11 reacted with proteins from mitogen-stimulated porcine mononuclear cells (MNC). The molecular weights of the precipitated proteins were approximately 34 and 28 Kd. In indirect immunofluorescence, approximately 25% of porcine MNC in suspension reacted with Mab 4-24-11. This percentage diminished to 14% when Ig bearing MNC were removed, while it increased to 36% when adherent MNC were enriched. Thus, it was concluded that Mab 4-24-11 cross-reacts with SLA-D. In a immunohistochemical study, Mab 4-24-11 reacted with cells in acetone fixed cryostat sections from the gastric mucosa and endometrium. These properties of Mab 4-24-11 make it useful as a tool for further studies on the porcine immune system.

Animals↗

Primary bovine viral diarrhoea virus infection in calves following direct contact with a persistently viraemic calf.

Six calves, aged 24 to 58 days and not previously exposed to bovine viral diarrhoea virus (BVDV), were infected with this agent by nose-to-nose contact with a persistently BVDV viraemic calf. The study was conducted in two trials, using 3 calves in each. All 6 calves showed a peak interferon level in serum at 4 days post infection (dpi), and they seroconverted to BVDV at 16-21 dpi. The calves in trial 1 had diarrhoea for 2 or 3 days between 2 and 6 dpi and one calf again from 9 to 11 dpi. During the periods of fever, the calves were slightly depressed. Those in trial 2 were more depressed and their oral and nasal mucous membranes were reddened but they never had diarrhoea. In both trials, fever (up to 41.3 degrees C) was a prominent symptom at 8 to 9 dpi and 2 calves showed a diphasic fever course. Respiratory affection was mild and no medical treatment was required. Haematological assessment demonstrated a transient but significant leukopenia and lymphopenia at 4 dpi (P less than 0.01 and P less than 0.05 respectively) and 11 dpi (P less than 0.05 and P less than 0.01 respectively). A significant decrease in thrombocyte count was seen at 4 dpi (P less than 0.05, n = 3). This study has demonstrated that nose-to-nose contact is an effective way of transmitting BVDV from persistently infected to susceptible cattle.

Animals↗

Effect of vitamin E supplementation on weight gain, immune competence, and disease incidence in barley-fed beef cattle.

The aim of the study was to investigate whether vitamin E supplements in larger amounts than recommended could reduce incidence of disease, improve immune competence, and increase rate of weight gain of conventionally barley-fed beef cattle. Mean daily intake of vitamin E by individual calves in the experimental group was 200 mg during the first 2 mo, 400 mg during the next 2 mo, and 600 mg during the rest of the period. Corresponding daily intakes of vitamin E for the control group were 50, 100, and 150 mg. Mean plasma vitamin E of the experimental group increased from .49 mg/L at the start of the trial to 2.03 mg/L at the end, but that of the control group was lower at the end (.36 mg/L) than at the beginning (.53 mg/L). No significant differences were observed between the groups concerning incidence of disease or magnitude of lymphocyte stimulation. The results indicated that there was a surprisingly poor biological availability of the dietary vitamin. Therefore, a comparison in reality was made between calves with inadequate and normal vitamin E status. The differences in daily BW gain and time to reach slaughter weight thus probably were effects of the low vitamin E status rather than positive effects of additional vitamin in the diet.

Animals↗

Effect of iscoms and their adjuvant moiety (matrix) on the initial proliferation and IL-2 responses: comparison of spleen cells from mice inoculated with iscoms and/or matrix.

In the iscom, antigen is attached by hydrophobic interactions to a matrix which is built up by the adjuvant Quil A and lipids. Thus, the iscom presents antigen in multiple copies on a small particle with a built-in adjuvant. By studying the specific antibody response, in vitro proliferation and IL-2 secretion by splenocytes from mice following different in vivo treatments with iscoms and/or matrix, we attempted to distinguish between nonspecific stimulatory effects, caused by the matrix or iscoms, and specific responses to the antigens incorporated into iscoms. The results strongly suggest that matrix and also iscoms exert a nonspecific adjuvant activity by a transient high spontaneous proliferation of cells collected within 2 weeks after administration of iscoms or matrix. This high rate of proliferation was preceded by suppressed proliferation, 3 days after injection with matrix or iscom. The adjuvant component included in iscoms, i.e., the matrix, does not excert a mitogenic stimulation in vitro or influence the levels of specific antibodies in serum. Specific responses to the antigens included in iscoms were recorded both as increasing levels of serum antibodies and as iscom-induced proliferation of immune spleen cells in vitro. The recruitment of IL-2 was only related to the specific stimulation induced by the antigens in iscom.

Adjuvants, Immunologic↗

Monoclonal antibodies to salmonid immunoglobulin: characterization and applicability in immunoassays.

Monoclonal antibodies to rainbow trout (Oncorhynchus mykiss) IgM were prepared and characterized for use in immunoassays. Antibodies produced by the five clones reacted with the heavy chain of the immunoglobulin. No indication of different heavy chain isotype specificity was observed for the MAbs. One clone discerned IgM from rainbow trout while the other four clones cross-reacted with IgM from Atlantic salmon (Salmo salar) and from brown trout (Salmo trutta). The monoclonal antibodies identified a B-cell like lymphocyte population that contributed to approximately 45% of the blood leucocytes in rainbow trout but was absent in the thymus. The proportion of Ig+ cells was higher in blood lymphocyte cultures stimulated with lipopolysaccharide than in nonstimulated cultures or in cultures stimulated with Concanavalin A. Applied in an ELISA for measuring humoral antibodies to Vibrio anguillarum in trout, the monoclonal anti-rainbow trout IgM antibodies discriminated seropositive fish from control fish more efficiently than did polyclonal rabbit antitrout IgM antibodies.

Animals↗

In vitro production of antibodies to bovine virus diarrhoea virus by peripheral blood mononuclear cells from cattle immunized by infection.

A method for in vitro production of antibodies to bovine virus diarrhoea virus (BVDV) by peripheral blood mononuclear cells (PBMC) was developed. The PBMC were cultured in microtitre plates coated with detergent-solubilized BVDV and the supernatants were tested in an enzyme-linked immunosorbent assay which detects IgG antibodies to BVDV. Following incubation of PBMC with an optimal concentration of pokeweed mitogen for 5 days, antibodies to BVDV were detected in culture supernatants of PBMC from immune cattle, but not in supernatants of PBMC from seronegative cattle, from persistently BVDV-infected cattle or from a 5-day-old calf that received BVDV antibodies via colostrum. This antibody-production assay may therefore be used as a tool to discriminate between passively acquired antibodies and those actively induced.

Animals↗

Numerical variations among blood mononuclear cells during the peripartal period in the gilt.

Numerical variations among mononuclear cells (MNC) were studied in blood collected from nine cross-bred primiparous gilts during two weeks before and two weeks after parturition. Simultaneously, the plasma levels of cortisol and oestrone sulphate were measured. The number of polymorphonuclear cells (PMNs) increased significantly (p less than 0.05) at parturition, while the number of MNCs decreased significantly (p less than 0.01) at the same time. Concomitantly with the total number of MNCs, the proportion of Ig-bearing MNC (Ig(+)-cells) also decreased significantly (p less than 0.01). No considerable variations of the proportion of MNCs expressing MHC Class II antigen could be seen throughout the study. The proportion of MNC with receptor for the Fc-part of IgG decreased significantly (p less than 0.01) during the week preceding parturition, but increased (p less than 0.05) again at parturition. The proportion of Ig(+)-cells was positively correlated both to the plasma level of oestrone sulphate and cortisol, while the number of Ig(+)-cells was only positively correlated to the level of cortisol. Thus, the study shows considerable numerical variations within the leukocyte sub-populations in the gilt around parturition.

Animals↗

Inflammatory response and antigen localization following immunization with influenza virus ISCOMs.

The inflammatory response, antigen retention, and antigen localization was studied in mice after immunization with influenza virus glycoproteins presented in two physically defined forms--micelles and ISCOMS (immunostimulating complexes). Two hours after intraperitoneal injection, the proportion of polymorphonuclear leukocytes (PMNs) in peritoneal lavage cells increased from less than 1% to 82% in ISCOM-treated mice and from less than 1% to 41% of the total cell count in micelle-treated mice. For both treatment groups, the proportion of PMNs returned to around zero 24 h postimmunization. Total recovery of radioactive antigen was significantly greater (P less than 0.05) in ISCOM-treated than in micelle-treated mice at one, two, and eight days postinjection. At all times tested, animals given ISCOMs had significantly more radioactive antigen in their spleens than animals given micelles. By electron microscopy ISCOMs were found to attach externally to the plasma membrane or within phagosomes of macrophages in close association with the membranes.

Animals↗

Detection of B and T cells, with lectins or antibodies, in healthy and bovine leukemia virus-infected cattle.

Lectins, polyclonal antibodies and monoclonal antibodies (MAbs) were evaluated as markers for bovine lymphocytes obtained from healthy animals and from cattle infected with bovine leukemia virus (BLV). In the blood from healthy cattle the proportion of cells identified as T lymphocytes with the lectin Helix pomatia (HP) (67.8 +/- 6.2%) using the indirect immunofluorescence technique was similar to the proportion of cells identified by the MAbs P5 (66.1 +/- 3.8%) and BLT-1 (59.8 +/- 7.1%). The proportion of B cells in blood from healthy animals identified with a polyclonal antibody to bovine IgM (18.0%) was similar to that identified with a MAb to bovine IgM (16.2%). However, greater variation between individual values was detected with the MAb (SD = 8.2) than with the polyclonal antibody (SD = 4.0). In the blood from BLV-infected cattle with persistent lymphocytosis, both the polyclonal and the MAb revealed a threefold increase of B cells. A proportion of the B cells had an increased amount of immunoglobulin molecules in their plasma membrane as indicated by flow cytometry. The proportion of T lymphocytes, identified by the MAb P5, was reduced to one-third of that in non-infected cattle. The indirect HP labelling gave inconsistent results and seems not to detect solely T lymphocytes among blood lymphocytes from BLV-infected cattle.

Animals↗

A cellular analysis of immunosuppression in cattle with mucosal disease.

Eleven cattle with mucosal disease (MD) were investigated to determine the constitution and function of the cells participating in the immune response. The total numbers of blood mononuclear cells varied within the normal range but a poor response of the lymphocytes to mitogen stimulation indicated a state of immunosuppression in the cattle with MD. Identification of subpopulations of the mononuclear cells revealed that eight of the cattle with MD had an increased proportion of B cells. However, the proportions of monocytes and T cells were in the normal ranges, resulting in a low proportion of cells not identified as being either B or T cells (null cells). Three of the cattle with MD had an increased proportion of cells with receptor for the Fc part of IgG (Fc gamma + cells). Bovine Fc gamma + cells have previously been shown to exert suppressor activity, as was also demonstrated in the present study by increased mitogen induced lymphocyte stimulation following removal of the Fc gamma + cells. The effect of depletion of Fc gamma + cells was at least five-fold greater in the two cattle with MD tested than the two controls. It appears that a diminished number of null cells and, or, increased suppressor cell activity might contribute to the observed immunosuppression in cattle affected with MD.

Animals↗