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Biomedical subjects

S Ahlstedt

Publications and source records attributed to S Ahlstedt.

At least 19 recordsLinked to original sources

Induction of IgE antibodies and T-cell reactivity to ovalbumin in rats colonized with Escherichia coli genetically manipulated to produce ovalbumin.

The immune response to ovalbumin (OA) and the bacterial antigens, lipopolysaccharide (LPS) and fimbriae were studied in conventional rats colonized from birth with an Escherichia coli strain producing OA. The colonized rats had developed IgE antibodies against OA, but not against the fimbrial or the LPS antigens from the E. coli at 2 months of age. At this time all rats were primed with OA given intracutaneously in Freund's complete adjuvant. Two weeks later the colonized rats showed a 35% greater delayed-type hypersensitivity (DTH) reaction to OA, measured as ear swelling, than the controls. Thus bacteria carrying antigens resembling potential allergens might aggravate, or participate in the induction of allergic symptoms. In addition such bacteria could be efficient vaccine vectors in protection against parasites. The study illustrates the importance of the mode of antigen presentation for the subsequent immune response.

Animals

Antibody-dependent cell-mediated cytotoxicity to beta-lactoglobulin-coated cells with sera from children with intolerance of cow's milk protein.

The capacity of serum antibodies against beta-lactoglobulin to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) was analysed in sera from children with cow's milk protein intolerance (CMPI). The children with CMPI were divided into three groups according to clinical features: delayed-onset CMPI with gastrointestinal symptoms (n = 8); immediate-onset CMPI with gastrointestinal and skin symptoms (n = 8); and immediate-onset CMPI with skin symptoms only (n = 8). The CMPI groups were compared with children with untreated (n = 9) or treated (n = 8) coeliac disease and a control group (n = 22). Sera from the children were examined for cytotoxic effects using lymphocytes from healthy adults as effector cells and radiolabelled beta-lactoglobulin-coated erythrocytes from the same donor as target cells. In addition, IgG and IgA serum antibodies against beta-lactoglobulin were determined with ELISA. Sera from children with CMPI and gastrointestinal symptomatology showed a significantly increased capacity to induce ADCC reactivity as compared with controls. This increased capacity was seen in sera from those with immediate as well as delayed onset of the gastrointestinal symptoms. In contrast, sera from children who had an immediate-onset CMPI with only skin symptoms mediated no such increase in ADCC reactivity. Moreover, children with coeliac disease with a few exceptions, demonstrated low ADCC reactivity, despite the fact that they had high levels of antibodies against beta-lactoglobulin. ADCC may be an immunopathogenic mechanism in certain cases of CMPI with gastrointestinal symptoms.

Antibodies

Enhancement of IgE synthesis in the human myeloma cell line U-266 with an IgE binding factor from a human T-cell line.

An IgE-binding factor(s) (IgE-BF(s] was partially purified from the supernatant of human HTLV-II carrying T-cell line MO. This IgE-BF(s) was shown to increase the IgE synthesis in the human myeloma cell line U-266, but did not affect its viability or growth. The effect of the IgE-BF(s) was dose-dependent and selective for IgE protein synthesis as beta 2-microglobulin synthesis in the U-266 and the immunoglobulin production in the U-1958 IgG-secreting human myeloma cell line were unaffected. The IgE-BF(s) increased the production of the epsilon heavy chain but not the lambda light chain production. The IgE-BF(s) was distinct from IL-1 beta, IL-3, IL-4, IL-5, IL-6, TNF-alpha, IFN-alpha, -beta, -gamma, M-CSF, and fragments of CD23.

Cytokines

Local immune response and bronchial reactivity in rats after capsaicin treatment.

The interaction between the nervous system, immune system and bronchial reactivity was studied in rats by using the neurotoxin capsaicin. Rats were treated with capsaicin at 1-2 days of age or at adult age, before or after sensitization by subcutaneous injections with ovalbumin (OA). The levels of the neuropeptides neurokinin A and calcitonin gene-related peptide were decreased in the lung after capsaicin treatment, as determined with radioimmunoassay, whereas the levels of neuropeptide Y were unaffected. The levels of IgA, IgE and IgG in bronchial lavage were also affected by capsaicin treatment; however, the results were heterogeneous. Capsaicin treatment after sensitization reduced the bronchial reactivity to challenge with OA aerosol and serotonin iv. The results demonstrated that reduction of neuropeptide levels with capsaicin affected both bronchial reactivity and the levels of antibodies in bronchial lavage fluid. However, no correlation between these two parameters was seen, demonstrating the complexity of the system.

Animals

A fluorescence method for the visualization of inflammatory cells in intestinal mucosa.

Treatment of paraffin or cryostat sections from rat, rabbit and man, with fluorescein isothiocyanate (FITC) in buffer (1 microgram/l), resulted in visualization of highly fluorescent leukocytes with a granular staining in light microscopy. FITC staining of paraffin-embedded colonic tissue was compared with three other staining methods, i.e., (1) FITC-conjugated anti-neutrophil antibodies, (2) endogenous peroxidase with 3,3-diaminobenzidine, and (3) FITC-conjugated Lotus tetragonolobus lectin. Good correlations between FITC staining and the three other methods were found (r:0.97, 0.98 and 0.98; P less than 0.001). Inhibition of receptors for the FITC conjugates prior staining, by preincubation with unconjugated anti-neutrophil antibodies or L-fucose, abolished the staining with antibodies and lectin. This study shows that staining with unlabeled FITC represents a quick and convenient method for the estimation of phagocytic cells (polymorphonuclear cells and macrophages) in histological specimens.

Animals

Effects of subinhibitory amounts of ampicillin, amoxycillin and mecillinam on the adhesion of Escherichia coli bacteria to human urinary tract epithelial cells: a preliminary study.

Attachment to mucous surfaces may be a prerequisite for bacteria colonizing these surfaces or invading underlying tissues. Subinhibitory amounts of ampicillin and amoxycillin but not mecillinam decreased the attachment of Escherichia coli bacteria to human uro-epithelial cells in vitro. No significant synergistic effect on the attachment by the antibiotics was obtained. The present report indicates a new parameter for the study of antibacterial actions of drugs.

Adhesiveness

Experimental evidence of a decreased incidence of penicillin allergy through use of pure penicillins.

The results revealed the presence of high molecular weight impurities in commercially available penicillins, measured with a radioimmuno assay. The impurities had penicilloyl specificity and induced antibody formation in mice when the contaminated penicillin was administered in 50 mg/kg body weight daily for ten day periods with a 20 to 30 day interval. Penicillin of high purity similarly administered produced very few antibodies. Furthermore, experimentally contaminated penicillin given according to the same schedule caused IgE antibody formation against the penicilloyl moiety, while pure penicillin did not. These findings were explained by the weak immunogenicity of isologous penicilloylated serum albumin in rabbits both regarding the IgE and the IgG/IgM antibody formation compared to the immunogenicity of heterologous bovine serum albumin similarly penicilloylated.

Animals

Immunotherapy in spring-time hay fever. A clinical and immunological study comparing two different treatment extract compositions.

In a study of the efficacy of two different treatment schedules for perennial immunotherapy, 47 adult patients with spring-time hay fever due to allergy against birch and other deciduous trees were randomly assigned to three treatment groups: one group received birch, alder and hazel allergen in Allpyral, another group received the same Allpyral mixture and in addition all relevant tree pollens in aqueous extract and a control group received no injections. For determination of antibody titres the radioallergosorbent test (RAST) and the ammonium sulphate precipitation (ASP) technique were used. Cellular responsiveness was studied by measuring birch pollen (BP) induced leucocyte histamine release in peripheral blood. The clinical and immunological response was similar in the two treated groups. Treated patients had less symptoms and a lower consumption of antihistamine tablets during the pollen season than the control group. Non-IgE BP antibodies and IgE antibodies recorded with the ASP technique increased after immunotherapy while RAST values did not change significantly. A decrease of RAST values from postseasonal values during the first year to preseasonal values in the following year was seen in all patient groups but was less pronounced in treated than in untreated patients. The decrease was more pronounced in patients with high RAST values of postseasonal sera than in patients with low RAST values. Cellular reactivity increased slightly during the first phase of therapy but returned to the pre-treatment level later. Clinical improvement was positively correlated to the percentage increase of non-IgE antibody titre and to the pre-treatment non-IgE/IgE antibody ratio. Patients with high preseasonal RAST titres or high cellular sensitivity tended to have more severe symptoms during the pollen season. It is concluded that a mixture of birch, alder and hazel is sufficient for immunotherapy in spring-term hay fever. It is obvious that changes of a single immunological variable do not account for the therapeutic results in immunotherapy.

Adult

Immunological aspects of pyelonephritis.

Several virulence factors, such as O and K antigens and capacity to attach to uroepithelial cells, seem to be required for Escheria coli to cause acute pyelonephritis. These factors induce an immune response, however, which can modify the course and clinical expression of the infection. During acute pyelonephritis, autoantibodies to the Tamm-Horsfall protein increase. These antibodies, which probably are evoked by a cross-reaction noted between structures of E. coli LPS and the Tamm-Horsfall protein, may add to the renal tissue engagement in interstitial nephritis caused by bacterial pyelonephritis.

Acute Disease

Antigens in penicillin allergy. III. Antigen and antibody levels in mice treated with pure and contaminated penicillins.

Using a radioimmunoassay, it was shown that commercially available ampicillin preparations often contain penicilloylated high molecular weight impurities. These possess immunological activities and stimulate penicilloyl-specific antibody formation in mice treated according to a therapeutic schedule. Using purified and experimentally contaminated preparations it was also found that exposure of the animals to Escherichia coli and Bordetella pertussis bacteria could increase the antibody formation to small amounts of impurities. In addition, penicilloylated antigen could be recorded in serum from treated animals. The antigen formed by penicilloylation in vivo, however, was very weak and did not induce much antibody formation when injected together with Freund's adjuvant in mice or rabbits.

Ampicillin

Secretory IgA antibodies to enterobacterial virulence antigens: their induction and possible relevance.

1) Milk and salivary s-IgA antibodies are via the homing of IgA producing cells from the Peyer's patches closely connected with antigenic stimuli in the intestine. This explains the presence in human milk of s-IgA antibodies against E. coli O and K antigens, V. cholerae and Shigella O antigens, E. coli and V. cholerae enterotoxins. These secretory antibodies can be induced by intestinal exposure and boosted by parenteral vaccination. 2) Preliminary data suggest that the IgA response in the urinary tract and possibly in the lung may be involved in the homing mechanism as well. 3) The protective role of the milk s-IgA antibodies to enterobacterial virulence antigens is strongly suggested, as is the protection mediated by urinary antibodies against urinary tract infections.

Animals

New knowledge in human milk immunoglobulin.

One of the anti-infection principles of maternal milk is the predominant milk immunoglobulin, secretory IgA. This immunoglobulin contains antibodies against many pathogens and potential pathogens, viruses as well as bacteria, including several members of Enterobacteriacae. The antigenic stimuli for these milk antibodies seem to take place in the Peyer's patches of the intestine. Lymphoid cells leaving the patches after antigenic exposure seem to home to the mammary glands via the lymph and blood circulation. As a result, the milk contains secretory IgA antibodies against, among other things, the intestinal bacteria of the mother. These milk antibodies might reflect the spectrum of bacteria and viruses in the community and may be important for the protection of the breast-fed baby. Via the same homing mechanism the maternal milk obtains antibodies against dietary antigens, including cow's milk proteins. Studies of infants on mixed feeding suggest that the secretory IgA antibodies against the bovine proteins diminish the antigenic exposure, indicating the possibility of an anti-allergic mechanism.

Antibody Formation

Specific antibodies in infants with gastrointestinal intolerance to cow's milk protein.

Antibodies of various immunoglobulin classes against cow's milk proteins were studied in infants and children with cow's milk protein intolerance, gluten-sensitive enteropathy and acute gastroenteritis. Their IgE, IgG, IgM and IgA antibody levels determined with the enzyme-linked immunosorbent assay (ELISA) and the IgE antibodies also determined with RAST, were compared with reference groups of children and adults. IgE, IgT or IgA antibodies against unseparated cow's milk proteins, alpha-lactalbumin, beta-lactoglobulin, alpha-casein and beta-casein were present in many of the studied samples, but did not discriminate between the individuals with and without intolerance symptoms. As a group, the infants with late reactions to cow's milk showed increased levels of IgE and IgG antibodies detected with the ELISA, while patients with gluten-sensitive enteropathy had significantly increased levels of IgG and IgA antibodies of cow's milk proteins compared to the reference group. By combining the findings of antibody increases in various immunoglobulin classes, an individual discrimination could be reached. Thus, 8 of 9 of the patients with late reactions to cow's milk had increased levels of IgE or IgG + IgA antibodies as compared to 3 of 22 in the reference group. Serodiagnosis with the ELISA may, therefore, be of some use in patients with a suspicion of cow's milk protein intolerance.

Animals

Antigens of Escherichia coli, human immune response, and the pathogenesis of urinary tract infections.

Acute pyelonephritis (but not cystitis or "asymptomatic" bacteriuria) due to Escherichia coli induces serum antibodies to O-but rarely to K-antigens, especially not to the most common antigen, K1. Locally produced secretory IgA and IgG antibodies to O-and K-antigens appear in urine during most infections. The E. coli in urine of patients with asymptomatic bacteriuria are different from those in patients with acute pyelonephritis and cystitis and undergo continuous changes, presumably caused by the local antibody response. The E. coli become less virulent and are less able to attach to uroepithelial cells than E. coli causing acute symptomatic infections. Antibodies in urine prevent epithelial adherence. Parenteral and intravesicular injections of killed bacteria can protect against ascending pyelonephritis in rats. A few K-antigens dominate among E. coli that cause urinary tract infections. Vaccination of problem cases is a possibility because of the protective nature of K-antibodies. The mechanism of renal scarring that appears in some patients with urinary tract infections is unknown. Autoantibodies to the Tamm-Horsfall protein that increase after acute pyelonephritis or the cross-reactions noted between certain E. coli and antigens on the kidney may be involved.

Animals

A sensitive method for specific quantitation of secretory IgA.

A method to quantitate specifically secretory IgA (SIgA) has been developed using the enzyme-linked immunosorbent assay. The IgA in the test sample was adsorbed to anti-alpha antibodies attached to plastic tubes via a cost of IgA myeloma protein. The reacted SIgA was determined using anti-secretory component antiserum conjugated with alkaline phosphatase. The technique permitted quantitation of secretory IgA in biological fluids like milk, urine, and saliva with a reproducibility of +/-7%, down to 0.03 mg/l. In contrast to earlier techniques, the presence of up to 157% of serum IgA without secretory component (SC) and free SC did not disturb the measurements of SIgA. Furthermore, variations in pH and osmolarity, within biological ranges in secretions, did not influence the estimations.

Animals

Experimental pulpitis in immunized monkeys.

Bovine serum albumin (BSA) was topically applied to exposed dentin to assess whether inflammatory reactions can be induced in the pulp of monkeys immunized against BSA. Four cynomolgus monkeys received repeated injections of BSA emulsified with Freund's incomplete adjuvant. Pulp challenge was performed by applying BSA in freshly cut dentin cavities prepared on the buccal surface in 34 teeth. In 29 control teeth ovalbumin (OVA) was applied. Control applications of BSA were also performed in 15 teeth prepared in three nonimmunized monkeys. Forty-eight hours after the initiation of the pulp challenge the monkeys were sacrificed and the pulp tissue examined in the light microscope. Topical application of BSA to freshly exosed dentin in immunized monkeys resulted in severe inflammatory lesions in the pulp, characterized by bleeding and extravascular infiltration of large numbers of leukocytes. Extensive tissue damage was an important feature in several pulps. Identical applications of OVA in control teeth and BSA applications in nonimmunized monkeys produced no such reactions. The results indicate that interactions between antigens and antibodies can occur within the dentin-pulp area and following the formation of immune-complexes, severe injury to the pulp can be induced.

Administration, Topical