Increase in the haemolytic activity of anti-erythrocyte antibodies by anti-gamma-globulin serum

It appears that anti-gamma-globulin sera not only affect the haemagglutinating reaction but can also be employed for detecting antibodies at the erythrocyte surface through the haemolytic reaction. The observed increase in the haemagglutinating as well as haemolytic reaction brought about by anti-gamma-globulin sera is due mainly to the effect on the reaction of 7 S anti-erythrocyte antibodies, the reaction of macroglobulin antibodies being little affected.     

Influence of the amount of antigen and the interval on the secondary reaction

The course of the revaccination reaction in mice immunized with different doses of sheep red blood cells was determined at different intervals after the primary stimulus. The maximum level of haemagglutinating antibodies in the secondary reaction was found after a high primary and secondary antigenic stimulus. On the contrary, if the level of haemolytic antibodies was determined, the higher was the primary antigenic stimulus, the lower was the secondary antibody response. Differences between haemagglutinins and haemolytic antibodies were also manifested in the earlier onset of the maximum haemolytic secondary reaction (five months after the first dose of antigen); the maximum haemagglutination response was not attained until eight months after the primary dose of antigen. The results comfirm that the basis of preparation for the secondary reaction is proliferation of immunologically activated Y cells; differences in the haemolytic and haemagglutination response are related to differences in the character of the antigenic determinants of sheep red cells.     

The passive anaphylactic reaction in guinea pigs elicited by whole and cathepsin D-degraded antigen

The systemic anaphylactic reaction and thein vitro anaphylactic contraction of the terminal segment of the ileum performed according to the Schultz-Dale technique, were elicited in guinea pigs passively sensitized with rabbit antibodies to human serum albumin, using whole and cathepsin D degraded antigen. The intensity of the systemic anaphylactic response that was evoked by degraded antigen was lower; a highly significant suppression of the response was obtained provided an antigen degraded more than by 70% was injected. With an increasing degradation of antigen, more animals responded with lower or even with no reaction; the number of animals developing severe or lethal shock, decreased at the same time. The number of animals that developed a medium anaphylactic response remained at the same level. The degraded antigen did not evoke the anaphylactic contraction of the terminal segment of the ileumin vitro, and moreover, it blocked the contraction after addition of the whole antigen.     

Indirect haemagglutination reaction (LH)-the method of choice for the detection of anamnestic antibodies to varicella - zoster (VZ) virus.

The method of indirect haemagglutination is a sensitive quantitative serological reaction for the detection of anamnestic titres of VZ antibodies. The curve obtained by the examination of a representative population sample has shown that the occurrence of VZ antibodies increases with the age so that 60% of children of under-school age and 90% of 15-year-old children have antibodies to varicella-zoster virus. In adult age groups, seropositivity ranges between 90--100%. The incidence and level of antibodies are identical in both sexes and do not decrease in old age groups. The mean titre of indirect haemagglutinating antibodies was 1 : 128.     

Acceleration of intradermal catabolism of guinea pig IgG1 and IgG2 antibodies by challenge with antigen.

The intradermal catabolism of antibodies injected in guinea pigs to provoke skin reactions was studied using 125I-labeled guinea pig IgG1 and IgG2 anti-ovalbumin antibodies. Disappearance of both the IgG1 and IgG2 antibodies from injected sites was accelerated by intravenous injection of the antigen. The antigen-antibody complexes produced in vitro were also catabolized more rapidly than free antibodies, when estimated using 125I-labeled antibodies. On the other hand, the catabolism of normal IgG2 was not influenced by local anaphylactic reaction elicited by IgG1 antiovalbumin antibody coexisting at the sites. Therefore, the enhanced catabolism of antibodies on challenge was not caused by increased vascular permeability due to anaphylactic reactions, but by more rapid elimination of immune complexes formed at the sites. The Fc parts of IgG1 and IgG2 antibodies played an essential role in the enhancement of catabolism since the catabolism of the F(ab')2 fragments was not accelerated by complex formation with ovalbumin, but rather reduced.     

Presence of IgE antibodies to bovine serum albumin in a patient developing anaphylaxis after vaccination with human peptide-pulsed dendritic cells

Dendritic cells are professional antigen-presenting cells that can be generated in vitro either from monocytes or from CD34⁺ peripheral blood progenitor cells by using recombinant cytokines. These cells have potential implications for immunotherapeutic approaches in the treatment of cancer and other diseases. We have conducted a phase I study in melanoma patients using peptide-pulsed dendritic cells cultured in medium supplemented with 10% fetal calf serum (FCS) and a cocktail of cytokines. Peptide-pulsed dendritic cells were injected intravenously at 2-week intervals. Here we report on a case of type I hypersensitivity anaphylactic reaction after repetitive vaccination with autologous peptide-pulsed cells. Pre-vaccination and post-vaccination serum samples were evaluated for the presence of antibodies to FCS and bovine serum albumin (BSA). A retrospective study in 7 patients vaccinated with FCS-cultured dendritic cells demonstrated the presence of IgG and IgM antibodies to FCS and BSA after vaccination in 6 out of 7 patients. However, IgE antibodies were absent in all patients with the exception of the patient developing anaphylaxis. The patient's serum was demonstrated to contain a strong IgE response directed against BSA. In contrast, 2 patients vaccinated with dendritic cells cultured under serum-free conditions developed no antibodies to FCS and BSA after repetitive vaccination. We suggest that patients can be sensitized with an IgE response against BSA leading to anaphylactic reactions. On the basis of these data, dendritic cells cultured in autologous serum or under serum-free conditions are recommended for therapeutic applications in vivo. Received: 25 November 1999 / Accepted: 20 January 2000     

Anaphylactic reactions to cinoxacin.

During 1981 to mid-1988 three cases of anaphylactic shock after treatment with the quinolone derivative cinoxacin were reviewed by the Netherlands Centre for Monitoring of Adverse Reactions to Drugs and 17 cases of an anaphylactic type of reaction notified to the World Health Organisation Collaborating Centre for International Drug Monitoring. In five out of six patients for whom data were available the reaction began shortly after taking a single capsule of a second or next course of treatment. Cinoxacin is related to nalidixic acid, and one patient previously treated with that agent subsequently had an anaphylactoid reaction to cinoxacin and later developed a skin reaction to nalidixic acid. There were no deaths, and patients treated as an emergency with plasma expanders or with adrenaline and corticosteroids generally recovered promptly and uneventfully. In view of the potentially fatal consequences of anaphylactic reactions to cinoxacin and other quinolones doctors should take care when prescribing these drugs.     

Motor activity as an unbiased variable to assess anaphylaxis in allergic rats

The release of mediators by mast cells triggers allergic symptoms involving various physiological systems and, in the most severe cases, the development of anaphylactic shock compromising mainly the nervous and cardiovascular systems. We aimed to establish variables to objectively study the anaphylactic response (AR) after an oral challenge in an allergy model. Brown Norway rats were immunized by intraperitoneal injection of ovalbumin with alum and toxin from Bordetella pertussis. Specific immunoglobulin (Ig) E antibodies were developed in immunized animals. Forty days after immunization, the rats were orally challenged with the allergen, and motor activity, body temperature and serum mast cell protease concentration were determined. The anaphylaxis induced a reduction in body temperature and a decrease in the number of animal movements, which was inversely correlated with serum mast cell protease release. In summary, motor activity is a reliable tool for assessing AR and also an unbiased method for screening new anti-allergic drugs.     

Changes in the level and character of colostral immunoglobulins during the lactation period in sows

A profound decrease in the concentration of colostral proteins (among immunoglobulins primarily IgG) during the first three days of lactation is accompanied by changes in the molecular heterogeneity of IgA. In the course of lactation, the amount of 7S IgA increases in relation to 10S IgA. The total amount of IgA, after the initial decrease during the first three days, maintains a gradual increase. In addition, a transient increase of natural haemagglutinating antibodies toEscherichia coli (serotype O55) was found in fractions corresponding to IgA and IgG during lactation.     

Trichinella spiralis: anaphylactic antibody formation and susceptibility in strains of inbred mice.

The anaphylactic antibody response of various strains of inbred mice of different H-2 specificities was investigated using the passive cutaneous anaphylactic technique (PCA) for the detection of the antibody response. Neither IgC₁ nor reaginic antibody were detected in serum samples obtained at the end of the first week of infection with Trichinella spiralis. Subsequently, all animals had detectable IgG₁ antibodies, although in some strains the titers were very low. Reaginic antibody was detected in relatively high titers in C57L, A, and DBA/1 mice. Two other strains were very poor responders (SJL and AKR). In most strains, reagin and IgG₁ remained detectable for 14 wk or longer. The pattern of response of all strains was very reproducible, indicating genetic control of the anaphylactic antibody production to the infection. In F₁ hybrids obtained from crosses between good and poor anaphylactic antibody responders, intermediate levels of both antibody classes were detected.Adult worm recovery rates were established at various points during the intestinal phase of infection, and no correlation between worm numbers and reaginic antibody titers in the various strains of mice could be demonstrated. There were noticeable differences in larval yields obtained after muscle digestion of mice belonging to the different inbred strains. In fact, we generally observed an inverse relationship between the number of larvae recovered from a given strain and their reaginic antibody titer.The intravenous injection of newborn larvae (NBL), obtained upon in vitro incubation of adult worms, produced detectable antibodies only in mice of the DBA/1 strain. These antibodies were consistently of low titer and became detectable only after the administration of two additional injections of NBL. This contrasted with the results observed after “per os” infection of DBA/1 mice, where high titers of these antibodies were always obtained, in spite of comparable ratios of muscle larval yield.     

Testing the properties of an experimental batch of zoster immune gammaglobulin (ZIG).

The methods of indirct haemagglutination (IH) and precipitation in gel (ID) were employed to test the level of varicella-zoster (VZ) antibodies in an experimental batch of zoster gammaglobulin (ZIG). The titre of indirect haemagglutinating antibodies in ZIG was about 64 times higher than in the ordinary batches of normal immunoglobulin and about 8 times higher in comparison with the level of the initial plasma pool. In the reaction of precipitation in gel, ZIG produced 5 to 6 zones. In comparison with the initial pool of convalescent plasma, ZIG also showed an 8-fold concentration of precipitating antibodies. ZIG was administered preventively to 6 children with risk diagnoses. None of the children fell ill with varicella. According to the results of subsequent serological examination in the reactions of indirect haemagglutination and radioimmunologic analysis, only 3 children were definitely susceptible to VZ infection. In two other children (very low antibody titres) the risk could not be excluded. No substantial increase in the levels of IH and RIA antibodies was observed in the 4 children under serological observation in a period of 6 months following the administration of ZIG. ZIG was administered therapeutically to four children with varicella. The effect of ZIG therapy was very suggestive, especially in two newborn infants lacking maternal antibodies, where the dose of ZIG per 1 kg body weigt was unusually high.     

The unique enzymatic function of field bean (<Emphasis Type="Italic">Dolichos lablab</Emphasis>) <Emphasis Type="SmallCaps">d</Emphasis>-galactose specific lectin: a polyphenol oxidase

The polyphenol oxidase (PPO) of field bean (Dolichos lablab) is a tetramer made up of two subunits of mass 29,000 and 31,000 Da. The amino acid sequence of the tryptic peptides showed approximately 90% sequence identity to the d-galactose specific legume lectins. The haemagglutinating activity of a pure and homogenous preparation of PPO measured using human erythrocytes was 1261 HAU mg⁻¹ protein and was inhibited by d-galactose. Purification by galactose-sepharose chromatography also indicated that the PPO and haemagglutinating activities were associated with a single protein. Crude extracts of other legumes did not exhibit PPO activity, yet cross reacted with anti-PPO antibodies. This dual function protein with PPO and haemagglutinating activity is unique to field bean. The two activities are independent of each other occurring at different loci on the protein. These observations further evidence and strengthen the assumption that galactose specific legume lectins have enzymatic function. Both PPO and lectins are proteins that play a vital role in the defense mechanism of plants. The complementarity of these two simultaneous and independent powerful defense mechanisms exhibited by a single protein renders it a candidate gene for the development of inbuilt plant protection.     

Platelets, lysosomal enzymes and anaphylactic shock in the rat

1. Rat serum levels in beta-glucuronidase and beta-galactosidase are higher than plasma levels. Rat platelets release these lysosomial enzymes during blood coagulation in vitro. 2. After anaphylactic shock, in the sensitized rat, there is no increase in beta-galactosidase and beta-glucuronidase plasma levels. The tissues of the sensitized rat do not release these enzymes during the antigen-antibody reaction. The blood platelet level is diminished after anaphylactic shock and the serum levels of the lysosomial enzymes are decreased. 3. In thrombopenic rat, anaphylactic shock is identical as in control animals. Rat platelets do not play a significant role in the anaphylactic shock.     

Observation of antibody activity against antibiotics in patients with atopic disease

Antibodies to antibiotics were studied in patients hypersensitive to these preparations and in patients with other types of allergic disease. In addition to examining these antibodies, a whole series of other immunological tests were carried out (IgG, IgA, IgM, IgE, C'3 and OPI). Antibodies to antibiotics were found in a high percentage of subjects, not only in those hypersensitive to antibiotics but also in patients with asthma bronchiale and rhinitis pollinosa. The considerable differences in IgG and IgE observed in patients with high titres of haemagglutinating antibodies to antibiotics point out the blocking character of these antibodies.     

Quantification of Moraxella bovis haemagglutinating adhesins with monoclonal antibodies

Six monoclonal antibodies (MAbs) against Moraxella bovis GF 9 were used to quantify haemagglutinating adhesins of 16 strains of this organism. The amount of each MAb necessary to inhibit one haemagglutinating unit of each strain varied between 4 and 0.007 times that required by strain GF 9. Five strains reacted with six MAbs, one with five, two with four, one with three, two with two and three with none. The procedures used enabled to detect dominant strains candidates for vaccines.     

Antibiotic susceptibility, hemolysin production and haemagglutinating activity of uropathogenic Escherichia coli

The hemolysin production, haemagglutinating activity (HA) with human 0 group erythrocytes and antibiotic susceptibility of 130 uropathogenic Escherichia coli strains were studied. 43% of the strains produced hemolysins and 39% showed haemagglutinating activity. In 12% of the haemagglutinating strains HA was inhibited by D-mannose. 45% of the hemolytic strains showed haemagglutinating activity. There was a significant relationship between hemolysin production and haemagglutination activity (p less than 0.05). 85% of the 130 Escherichia coli strains were found to be multiple resistant to antibiotics.     

Histochemical investigations on phosphorylase, branching enzyme and glycogen in guinea pig livers in experimental anaphylactic and histaminic shock

The investigations have been carried out on 116 guinea pigs divided in three groups: the control (first group), the experimental group with animals after acute anaphylactic shock (second group), the animals after histaminic shock (third group). The animals of the experimental (second) group were sensitized with 25% egg white suspension in 0.9% NaCl applied subcutaneously. The same animals were exposed to the action of the antigen in aerosol (second group). The healthy animals were exposed to the action of 1% solution of dihydrochloride histamine (third group). In acute anaphylactic shock a decrease of histoenzymatic activity of phosphorylase A and branching enzyme in liver parenchyma was observed. It has been concluded that in anaphylactic shock there occurred disturbances in the function of the phosphorylase A--branching enzyme system. In histaminic shock the phosphorylase reaction becomes intensified in numerous liver cells. This is possible because the exogenic histamine may lead to the activation of the enzymatic system under studies.     

Effect of myeloma IgE injections on passive and active cutaneous anaphylaxis in rats

The ability of injected rat IgE myeloma protein IR162 to inhibit passive and active cutaneous anaphylaxis in Lewis rats was investigated. IgE injected i.p. 24 hr before the sensitization with IgE anti-ovalbumin (OVA) completely inhibited both IgE- and IgG2a-induced passive cutaneous anaphylactic (PCA) reactions at a dose (2.5 mg/100 g body weight) that resulted in peak serum concentrations of 150 micrograms IgE IR162/ml. Peak IgE IR162 serum concentrations of 20 to 60 micrograms/ml inhibited the PCA reaction in approximately 50% of the rats. Intracutaneous injection of a mixture of myeloma IgE and anti-OVA IgE in a ratio of 100:1 or more also inhibited the PCA reaction. In contrast, the PCA reaction was not inhibited by seven daily doses of IgE beginning 24 hr after passive sensitization. Likewise, the cutaneous anaphylactic reaction elicited in rats 14 days after immunization with OVA and Bordetella pertussis was not prevented by daily injections of myeloma IgE despite a 1000- to 3000-fold excess of the myeloma IgE to anti-OVA IgE serum concentration. The data demonstrate that parenteral administration of myeloma IgE inhibits the PCA reaction only when given before passive sensitization and does not prevent cutaneous anaphylaxis in actively immunized rats. Because myeloma IgE failed to inhibit anaphylactic reactions in actively immunized rats, it is questionable whether administering human IgE-derived synthetic peptides or recombinant DNA-produced IgE fragments will be able to prevent allergic diseases by blocking the IgE Fc receptors on mast cells.     

Purification and characterization of a surface lectin from the nematode-trapping fungus Arthrobotrys oligospora.

Several studies have indicated that the capture of nematodes by the nematophagous fungus Arthrobotrys oligospora is mediated by a lectin on the fungal surface. One of the major surface proteins of this fungus showed haemagglutinating activity and was isolated by affinity chromatography using a mucin Sepharose column. Biochemical analysis showed that the protein was a dimeric glycoprotein with a molecular mass of 36 kDa and an isoelectric point of pH 6.5, and contained no sulphur amino acids. The protein was N-terminally blocked; four internal peptides were sequenced, and showed no significant similarity to sequences in the Swiss-Prot or PIR databases. The haemagglutinating activity of the isolated protein was not inhibited by any of the mono- or disaccharides tested, but it was inhibited by the glycoproteins fetuin and mucin. The haemagglutinating activity changed after incubating the protein in buffers of different pH, with maximal activity at pH 11.0 and no activity at pH 2.8. The lectin was tested for different enzymic activities but none were detected. Analysis of the haemagglutinating activity in various cell fractions indicated that the protein was associated with extracellular polymer layers and with the cell wall of the fungus. About the same amount of the haemagglutinating protein was recovered from samples of vegetative mycelium and of mycelium containing nematode-trapping cells.