Decreased numbers of circulating plasmablasts and differences in IgA1-plasmablast homing to skin in coeliac disease and dermatitis herpetiformis.

The two clinical phenotypes of gluten enteropathy, coeliac disease (CD) and dermatitis herpetiformis (DH), were characterized for numbers and homing profiles of circulating final effector B cells, plasmablasts, identified as immunoglobulin (Ig)-secreting cells (ISC). In CD, the numbers of ISC were approximately 50% lower than in DH or controls. ISC expressed peripheral lymph node homing receptor (HR), L-selectin, less frequently in CD (54%) and DH (52%) patients than in controls (70%). The expression of gut mucosal HR, alpha(4)beta(7), was less frequent in CD (42%) than in DH (65%) or controls (60%). In DH, but not in CD or controls, a higher proportion of IgA1-ISC (40%) than IgA2-ISC (25%) expressed the skin HR, cutaneous lymphocyte-associated antigen. In gluten enteropathy circulating plasmablasts are more mature, but decreased in number, and have distorted homing profiles. Differential IgA1-plasmablast homing could be associated with the development of skin rash with IgA1-deposits in DH but not in CD.

Probiotics: effects on immunity.

The gastrointestinal tract functions as a barrier against antigens from microorganisms and food. The generation of immunophysiologic regulation in the gut depends on the establishment of indigenous microflora. This has led to the introduction of novel therapeutic interventions based on the consumption of cultures of beneficial live microorganisms that act as probiotics. Among the possible mechanisms of probiotic therapy is promotion of a nonimmunologic gut defense barrier, which includes the normalization of increased intestinal permeability and altered gut microecology. Another possible mechanism of probiotic therapy is improvement of the intestine's immunologic barrier, particularly through intestinal immunoglobulin A responses and alleviation of intestinal inflammatory responses, which produce a gut-stabilizing effect. Many probiotic effects are mediated through immune regulation, particularly through balance control of proinflammatory and anti-inflammatory cytokines. These data show that probiotics can be used as innovative tools to alleviate intestinal inflammation, normalize gut mucosal dysfunction, and down-regulate hypersensitivity reactions. More recent data show that differences exist in the immunomodulatory effects of candidate probiotic bacteria. Moreover, distinct regulatory effects have been detected in healthy subjects and in patients with inflammatory diseases. These results suggest that specific immunomodulatory properties of probiotic bacteria should be characterized when developing clinical applications for extended target populations.

Selenium metabolism and platelet glutathione peroxidase activity in healthy Finnish men: effects of selenium yeast, selenite, and selenate.

The mean dietary selenium intake in Finland increased from 40 to 100 micrograms/d in 1987 because of the addition in 1985 of selenium to fertilizers. A selenium-supplementation study was performed in 1987 on the same men as were followed in a 1981 study that had a similar design (200 micrograms Se/d). Selenite and selenate, but not selenium yeast increased platelet glutathione peroxidase (GSHPx) activity by 30% compared with placebo, much less than the 70% found in the previous study. Selenium yeast and selenite increased plasma selenium after 11 wk from 1.39 mumol/L to peak values of 2.15 and 1.58 mumol/L, respectively. Only yeast selenium was incorporated into red cells. From a regression plot based on present and literature data, it was estimated that the plasma selenium concentration needed to achieve maximal platelet GSHPx activity was 1.25-1.45 mumol/L. At the present selenium intake in Finland, 100 micrograms/d, GSHPx activity is saturated in plasma and red cells and almost saturated in platelets.

Bioavailability of selenium to Finnish men as assessed by platelet glutathione peroxidase activity and other blood parameters.

Three groups of 10 men of low selenium status were given 200 micrograms Se/day as Serich wheat, Se-rich yeast, or sodium selenate for 11 wk. Twenty unsupplemented subjects served as controls. Plasma Se levels increased steadily in the wheat and yeast groups for 11 wk without plateauing, whereas in the selenate group, plasma Se plateaued around 110 ng/ml after 4 wk. Platelet glutathione peroxidase (GSH-Px) activities increased rapidly in the wheat and selenate groups for 4 wk and then plateaued. Platelet GSH-Px increased more slowly in the yeast group. Ten weeks after the supplements were discontinued, platelet GSH-Px was higher in the wheat and yeast groups than in the selenate group. Assessment of Se bioavailability requires a short-term platelet GSH-Px measurement to determine immediate availability, a medium-term plasma Se measurement to estimate retention, and a long-term platelet GSH-Px measurement after supplements are discontinued to determine the covertibility of tissue Se stores to biologically active Se.

A method for simultaneous determination of rosette formation and phagocytosis by cells.

A fluorescent agent insoluble in water was readily phagocytosed by 5-20% of human blood mononuclear cells, mainly monocytes. These cells were easily detected even amidst abundant rosetting erythrocytes. When rosette assays for the detection of complement or SRBC receptors on the surface of the cells were performed, only phagocytic cells exhibited the complement receptor with the method used in the study, whereas mainly nonphagocytic cells formed SRBC rosettes. The effect on the calculation of the percentage of T lymphocytes (SRBC receptor lymphocytes) of varying amounts of monocytes present in mononuclear cell population is demonstrated. The method presented here might prove useful particularly in studies on Fc or complement receptors on white blood cells.

The effect of phenytoin in vitro on normal human mononuclear cells and on human lymphoblastoid B cell lines of different Ig isotype specificities.

The anticonvulsant drug phenytoin, in less than cytotoxic concentrations, caused significant reductions in Ig secretion by unstimulated or EBV-stimulated normal MNC, as measured by PFC or secretion of Ig into the culture medium. Isotype-specific LBL varied in their sensitivity, the secretion of IgA (1 line) and IgG (3 lines) being reduced by phenytoin near therapeutic concentrations, whereas that of IgM (1 line) was resistant. Six-day exposure of MNC to phenytoin caused no selective depletion of or enrichment for B cells, monocytes or T cell subsets. The results suggest that the reduction in serum Ig levels reported in phenytoin-treated epileptic patients is, at least in part, due to a direct effect of the drug on the B lymphocyte. However, among EBV-activated normal MNC, those secreting IgA were no more sensitive to the drug than those secreting IgG or IgM, and other factors may, therefore, operate to cause the preferential reduction in serum IgA in phenytoin-treated patients.

ELISPOT for detecting antibody-secreting cells in response to infections and vaccination.

Based on immunoassay principles, methods have been developed for the analysis of secreted products at the cellular level. This approach offers substantial methodological advantages compared to traditional immunoassays. In a number of applications cell-based methods have proved able to overcome many of the problems inherent to immunoassays of biological fluids. This review focuses on applications of ELISPOT in natural infections and vaccinations of human individuals. The studies reviewed here have contributed to our understanding of the B-cell responses in infections and the independence of mucosal and systemic immune responses. Whilst diagnostic applications are rare, enzyme immunospot assays have been extensively used in testing the immunogenicity of vaccines. In particular, B-cell responses to mucosal vaccines are better covered with this cellular assay.

Cell-mediated immune responses to antigens of Bordetella pertussis and protection against pertussis in school children.

BACKGROUND: Increasing evidence suggests that cell-mediated immunity (CMI) is involved in immune response against Bordetella pertussis. However, there are practically no studies evaluating the significance of pertussis-specific CMI in relation to protection against clinical pertussis. METHODS: An outbreak of pertussis was studied prospectively in 13-year-old pupils in a rural school. B. pertussis infection was diagnosed by culture, microagglutination and enzyme immunoassay serology with the use of pertussis toxin, filamentous hemagglutinin and pertactin as antigens. Pertussis-specific CMI responses were assessed by in vitro proliferation assay of peripheral blood mononuclear cells. RESULTS: At the initial sampling 7 of 22 children had symptoms suggestive of pertussis and 15 were asymptomatic. Of the latter 3 remained healthy, 8 were later confirmed to have had asymptomatic infection, 3 developed laboratory-confirmed pertussis and 1 developed cough without laboratory evidence of pertussis. Initial in vitro proliferations of peripheral blood mononuclear cells induced by pertussis toxin, filamentous hemagglutinin and/or pertactin were positive in all 3 healthy children, in 6 of 8 children who had asymptomatic infection, but in none of the 3 children who later developed pertussis. Although some children who remained healthy had high values of antibodies, no clear association was found between initial serum antibody values and clinical outcome. CONCLUSIONS: These preliminary data suggest that CMI may have an important role in protection against clinical pertussis but do not exclude a role for antibodies. Furthermore the results stress a multifactorial nature of the immune protection against B. pertussis.

Modulation of human humoral immune response through orally administered bovine colostrum.

Eighteen healthy volunteers were randomized into two treatment groups and consumed liquid prepackaged bovine colostrum whey and placebo for 7 days. On days 1, 3 and 5, an attenuated Salmonella typhi Ty21a oral vaccine was given to all subjects to mimic an enteropathogenic infection. The circulating antibody secreting cells and the expression of phagocytosis receptors of the subjects before and after oral immunization were measured with the ELISPOT assay and flow cytometry. All subjects responded well to the vaccine. No significant differences were observed in ELISPOT values for IgA, IgG, IgM, Fcgamma and CR receptor expression on neutrophils and monocytes between the two groups. There was a trend towards greater increase in specific IgA among the subjects receiving their vaccine with bovine colostrum. These results suggest that bovine colostrum may possess some potential to enhance human special immune responses.

Importance of intestinal colonisation in the maturation of humoral immunity in early infancy: a prospective follow up study of healthy infants aged 0-6 months.

AIM: To evaluate the role of intestinal microflora and early formula feeding in the maturation of humoral immunity in healthy newborn infants. STUDY DESIGN: Sixty four healthy infants were studied. Faecal colonisation with Bacteroides fragilis group, Bifidobacterium-like, and Lactobacillus-like bacteria was examined at 1, 2, and 6 months of age, and also the number of IgA-secreting, IgM-secreting, and IgG-secreting cells (detected by ELISPOT) at 0, 2, and 6 months of age. RESULTS: Intestinal colonisation with bacteria from the B fragilis group was more closely associated with maturation of IgA-secreting and IgM-secreting cells than colonisation with the other bacterial genera studied or diet. Infants colonised with B fragilis at 1 month of age had more IgA-secreting and IgM-secreting cells/10(6) mononuclear cells at 2 months of age (geometric mean (95% confidence interval) 1393 (962 to 2018) and 754 (427 to 1332) respectively) than infants not colonised (1015 (826 to 1247) and 394 (304 to 511) respectively); p = 0.04 and p = 0.009 respectively. CONCLUSIONS: The type of bacteria colonising the intestine of newborns and the timing may determine the immunomodulation of the naive immune system.

High-performance liquid chromatographic determination of tocopherols and tocotrienols and its application to diets and plasma of Finnish men. II. Applications.

The composed one-day diets and plasma of 40 Finnish men screened for a selenium supplementation study were analyzed for tocopherols and tocotrienols. The men were divided into a low-Se group (in the screening phase plasma Se levels less than 70 micrograms/l and plasma alpha-tocopherol levels less than 1.2 mg/100 ml) and a high-Se group (plasma Se greater than 70 micrograms/l, plasma alpha-tocopherol not determined before the study). In the low-Se group plasma levels of alpha-tocopherol averaged 0.97 +/- 0.18 mg/100 ml. The daily dietary intake of alpha-tocopherol was 6.1 +/- 2.7 mg and that of total vitamin E 7.3 +/- 3.1 mg of alpha-tocopherol equivalents. In the high-Se group the corresponding average values were 1.16 +/- 0.21 mg of alpha-tocopherol/100 ml of plasma, 8.8 +/- 4.3 mg of alpha-tocopherol/day and 10.3 +/- 5.1 mg of alpha-tocopherol equivalents/day. The overall average for the contribution of alpha-tocopherol to the total dietary tocopherols was 44.6 +/- 11.0%. In the plasma samples alpha-tocopherol accounted for 92.0 +/- 2.1%, beta-tocopherol for 2.7 +/- 0.7% and gamma-tocopherol for 5.3 +/- 2.1% of the total amount of tocopherols.

Grouping of beta-haemolytic streptococci by using coagglutination, precipitation or bacitracin sensitivity.

This study was made to compare coagglutination to precipitin test in grouping beta-haemolytic streptococci from clinical specimens and to investigate the accuracy of the bacitracin test in identification of group A streptococci. Results of grouping 126 strains with coagglutination and precipitation were identical in all except two cases. These two strains were nongroupable with precipitation but appeared as group B and C by coagglutination. When the distribution of group A, B, C and G streptococci in various clinical sources was investigated it was found that group B strains were the most frequently (41 per cent) isolated streptococci and even in isolates from pharyngeal swabs their proportion was 33 per cent. The accuracy of the bacitracin test in identification of group A streptococci was unsatisfactory as 26/62 (42 per cent) strains reported as group A by using this test were in fact group B, C or G streptococci. One of the reasons for this high number of false positives appeared to be the medium used for the preparation of the blood agar plates. In view of the frequent occurrence of non-A-streptococci in clinical specimens and high incidence of false positive in the bacitracin test it is suggested that this test should be replaced by a more efficient method of serological grouping.

Cell walls, peptidoglycans, and teichoic acids of Gram-positive bacteria as polyclonal inducers and immunomodulators of proliferative and lymphokine responses of human B and T lymphocytes.

In this study the mitogenic and immunomodulating effects of bacterial cell wall preparations were investigated. Cell walls, peptidoglycans, and teichoic acids from Bacillus subtilis and Staphylococcus aureus Wood 45 activated both human T cells (supplemented with 10% monocytes) and B cells to proliferate and to produce leukocyte-inhibitory factor. Similar results were obtained with adult and umbilical cord blood cells, suggesting that these bacterial preparations were acting as mitogens. Cell walls and peptidoglycans had a modulating effect on purified protein derivative- or protein A-induced proliferation. In the presence of suboptimal concentrations of these stimulants, bacterial components enhanced the proliferative response. However, at optimal concentrations of purified protein derivative or protein A, bacterial components suppressed lymphocyte proliferation. Peptidoglycans solubilized by lysozyme activated B lymphocytes but not T cells. Solubilization had no effect on the immunomodulating capacity.

Cellular interactions in spontaneous or autologous cell-induced proliferative and lymphokine responses of human lymphocytes.

Purified human B cells were activated in cultures in the absence of any intentional stimulants as judged by lymphokine synthesis and proliferation. These responses were not augmented by monocytes. Lymphokine production (LIF) was increased in the presence of T cells. Autologous mixed lymphocyte reaction of T cells against B cells (AMLR) did not include LIF production in spite of the proliferative response. We would suggest that activated B cells present in the population of purified B cells are the stimulators in AMLR. In our interpretation these results support the hypothesis that AMLR reflects a mechanism by which T cells regulate lymphocyte function.