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.     

Enhancement activity of anti-brucella sera in experimental Brucella abortus infection in mice.

The passive transfer of rabbit anti-brucella sera promotes the multiplication of Brucella abortus in the spleen of mice infected intravenously with a low dose of this bacteria. The enhancement activity of the anti-brucella sera is related to their modes of preparation in rabbits. The agglutinin titers of all these antisera were relatively high, while there was some discrepancies in their content in haemagglutinating, immune adherence and complement fixing antibodies.     

Anaphylactic shock following the administration of clindamycin

A case of anaphylactic shock following the administration of clindamycin is reported. The antibody response was studied by the passive haemagglutination reaction and the presence of haemagglutinating antibodies to both clindamycin and lincomycin was detected. In the discussion, the authors try to account for the causes of development of the mentioned anaphylactic reaction after a single therapeutic dose. In this connection they mention a possible development of haemagglutinating antibodies after the administration of two doses to the patient in an experiment at a long-term interval.     

Serological identification of infection by verocytotoxin-producing Escherichia coli

One hundred sera from patients with haemolytic ureamic syndrome were screened for antibodies to the lipopolysaccharide (LPS) of Escherichia coli serogroup 05, 026, 0115, 0128, 0145 and 0153, and Shigella dysenteriae 1. Three sera contained antibodies to the LPS of E. coli belonging to serogroup 05.     

Effect of urea and uric acid on the hemolytic activity of Sendai virus

It was found that haemolytic activity of Fushimi strain of Sendai virus multiplied in allantoic cavity of chicken embryos is independent on its haemagglutinating titer and also on allantoic fluid urea and uric acid content. It was shown in experiments with embryonated eggs that these two compounds have no also influence on haemolytic activity induction in Sendai virus. Moreover, the results of an experiment in which allantoic fluid was replaced by Eagle's liquid suggest that most probably the other components present in allantoic fluid do not also influence the appearance of haemolytic activity of this virus.     

Serological Diagnosis of Classical Swine Fever

Antibody levels in post-infection sera from a pig inoculated with a low virulent strain of classical swine fever virus (Hannover 62) and in sera from two pigs inoculated with another low virulent strain (Spielbach 66) and from an in-contact pig were assayed by complement fixation and immunofluorescence using classical swine fever virus (ALD strain) and bovine virus diarrhoea virus (UG 59 strain) as antigens. The complement fixation test used was modified by addition of a preparation of porcine Glq to the complement and by mercaptoethanol treatment of the immune serum before use. The mercaptoethanol treatment of the immune serum resulted in complete elimination of a haemolytic prozone often seen with porcine immune sera. In the sera from the inoculated animals complement-fixing antibodies appeared earlier than neutralizing antibodies. A few weeks after inoculation there was a correlation between the presence of complement-fixing and neutralizing antibodies. During the entire observation period of 13 weeks it was not possible to demonstrate complement-fixing or neutralizing antibodies in serum from a pig exposed to infection by contact with the two pigs inoculated with the Spièlbach 66 strain of classical swine fever virus.     

Carbohydrate specificity of T-cell cytophilic chicken anti-SRBC IgM antibodies.

Fixation of passively sensitized lymphocytes by formaldehyde resulted in amplified erythrocyte rosette formation. The cytophilic anti-SRBC antibodies were of the IgM class and they attached selectively to thymocytes and T lymphocytes. Anti-SRBC sera from approximately 30% of immunized chickens gave rosette counts between 500–1500 per 10⁴ cells whereas the remainder gave values considerably lower. The onset and peak of the cytophilic IgM response were reached later than that of IgM haemagglutinating antibodies and there was little correlation between the count of cytophilic rosettes and haemagglutinjn titres in individual chickens. On the basis of competitive inhibition of rosette formation by a hog blood group substance it is suggested that the cytophilic antibodies have binding site specificity for a saccharide on the surface of SRBC.     

Spontaneous haemolytic activity of Atlantic halibut (Hippoglossus hippoglossus L.) and sea bass (Dicentrarchus labrax) serum

The spontaneous haemolytic (SH) activity of sera was compared in groups of cultured halibut and sea bass. The optimum assay temperature was determined for each species and different red blood cell donors were tested. The effects of heat inactivation, storage temperature and of different agents like EDTA, EGTA, yeast cell components and bacterial LPS were compared. Halibut sera gave optimum lysis with sheep red blood cells (RBC) at 16 degrees C whereas sea bass sera showed optimum lysis with rabbit RBC at 37 degrees C. The haemolytic activity of halibut sera was inactivated at 45 degrees C while sea bass sera were inactivated at 56 degrees C. The haemolytic activity of halibut sera was significantly reduced during short-term storage at -80 degrees C, whereas the sea bass sera maintained fairly good activity after 1-year storage at -80 degrees C. EGTA and EDTA inhibited the spontaneous haemolytic activity of sera from both the species. Zymosan and MacroGard from yeast cells also inhibited the haemolytic activity of the sera of both species, whereas LPS had a very slight effect. Considerable variation in haemolytic activity was observed within both the halibut and sea bass groups studied.     

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.     

The serological response to Verocytotoxigenic Escherichia coli in patients with haemolytic uraemic syndrome

AIMS: To screen sera from 80 patients with clinical haemolytic uraemic syndrome (HUS) and serum antibodies to the lipopolysaccharide (LPS) of Escherichia coli O157, for antibodies to Verocytotoxin-producing Escherichia coli (VTEC) belonging to serogroups O5, O26, O104, O111, O128, O145, O153 and O165. METHODS AND RESULTS: Sera were screened by an LPS-based ELISA and SDS-PAGE/immunoblotting. None of the 80 sera contained antibodies binding to long-chain LPS of any of the LPS types employed; however, nine sera contained antibodies binding to R3 LPS-core epitopes. CONCLUSIONS: The presence of patients' serum antibodies to the LPS of E. coli O157, in the absence of antibodies to the LPS of a range of other VTEC, demonstrated that cases of HUS may be caused by strains of O157 VTEC alone and that concurrent infection with multiple strains of VTEC is not a prerequisite for cases of HUS. SIGNIFICANCE AND IMPACT OF THE STUDY: Antibodies to long-chain LPS of VTEC other than O157 were not detected, and so there was no evidence of infection with VTEC belonging to more than one serogroup. The results of immunoassays such as ELISAs and micro-agglutinations must take into consideration antibodies binding to R3 epitopes located on LPS-core.     

Heterophilic infectious mononucleosis antigen used in the latex diagnostic test. II. Preliminary evaluation of the usefulness of latex reagents for detection of serum heterophile antibodies in patients with infectious mononucleosis

The aim of this study was to evaluate the usefulness of laboratory made reagent of polystyrene latex coated with three preparations of mononucleosis antigen (reagent MZ-I, MZ-II, and MZ-III) for detection of heterophile antibodies in patients sera with clinical symptoms of infectious mononucleosis. These studies were carried out on 153 serum samples taken from persons suspected of being infected with EB virus and on 100 healthy controls--blood donors. The results of latex tests were compared to the results of Paul-Bunnell-Davidsohn (PBD) and hemolytic tests. Out of three latex reagents evaluated only one (reagent MZ-I) showed sensitivity equal to PBD and haemolytic tests. The sensitivity reached 91.1%. Agglutination reaction when appeared in latex test at serum dilution 1:5, is considered positive and diagnostically significant result.     

Genetic variation in haemolytic activity in Atlantic salmon (Salmo salar L.)

Spontaneous and antibody-dependent haemolytic activity against sheep red blood cells (SRBC) has been studied in serum from Atlantic salmon, Salmo salar L. Considerable increase in haemolytic activity was detected when SRBC were sensitized with haemolysin produced in Atlantic salmon. Haemolytic activity was sensitive both to divalent cations and to heat treatment. Significant reduction in haemolytic activity was detected after absorption with SRBC, indicating the presence of natural antibody against SRBC in Atlantic salmon serum. Persistent haemolytic activity of unsensitized SRBC in serum absorbed to remove natural antibodies was found, which suggests the activation of the alternative complement pathway, while the observed increased haemolytic activity in the presence of specific antibody against SRBC suggests activation of the classical complement pathway. Spontaneous and antibody-dependent haemolytic activity were both analysed in absorbed and non-absorbed sera in a family material of Atlantic salmon. The material consisted of 574 fish belonging to 57 fullsib groups within 20 paternal halfsib groups. Fish with signs of sexual maturity generally showed reduced haemolytic activity. Statistically significant effect of sire on the spontaneous haemolytic activity of both absorbed and nonabsorbed serum, and on antibody-dependent activity, provides evidence of significant additive genetic variation in both the alternative and the classical complement activation in Atlantic salmon. Neither on a phenotypical nor on a family basis were the two traits statistically correlated. The estimated heritabilities were 0.2–0.3 with a standard error of approximately 0.1.     

Enhancement of immune response by the proteinaceous crystal of Bacillus thuringiensis var thuringiensis

The proteinaceous crystal of $\text{Bacillus thuringiensis}$ var $\text{thuringiensis}$ was found to enhance humoral immune response in rats and guinea pigs immunised with sheep red blood cells. The enhancement was due to the increased levels of both 19S and 7S antibodies in the sera of the treated animals. A novel synthesis of 7S haemolytic antibodies was observed in case of crystal treated animals.     

Human antibody responses to R3-core epitopes on the lipopolysaccharide of Escherichia coli O157

AIMS: To establish the incidence of serum antibodies binding to the R3-core lipopolysaccharide (LPS) of verocytotoxin-producing Escherichia coli (VTEC) O157, in patients with serum antibodies to E. coli O157 LPS, and to characterize the class(es) of antibodies binding to epitopes on the R3-core. METHODS AND RESULTS: SDS-PAGE profiles of LPS prepared from VTEC O157 were used in combination with immunoblotting to detect and characterize serum antibodies binding to the R3-core LPS of VTEC O157. Of 417 sera, referred to the Laboratory of Enteric Pathogens (LEP) for routine O157 serology and found to have serum antibodies to long-chain VTEC O157 LPS, 31 had antibodies binding to the R3-core of VTEC O157 LPS. The majority of the 31 sera contained IgA-class antibodies to both long-chain and R3-core LPS epitopes. Patients who did not develop haemolytic uraemic syndrome (HUS) produced antibodies of the IgM class to R3-core and IgG-class antibodies to long-chain LPS more frequently than patients with HUS. CONCLUSIONS: Only 7.4% of sera received by the LEP, and shown to have antibodies to VTEC O157 LPS, contained antibodies binding to the R3-core of VTEC LPS. Most sera contained IgA-class antibodies to both long-chain and R3-core LPS epitopes. SIGNIFICANCE AND IMPACT OF THE STUDY: Patients infected with VTEC O157 produced antibodies binding to the R3-core epitopes of VTEC O157 LPS only rarely, and these antibodies are unlikely to interfere with the serodiagnosis of infections caused by these organisms.     

Frequent production of toxins by Escherichia coli strains isolated from human urinary tract infections: relation with haemagglutination

Abstract We have investigated the production of heat-labile enterotoxin (LT), Verotoxin (VT), cytotoxic necrotizing factor (CNF), haemolysis (Hly) and lethal activity for mice in 48 Escherichia coli strains isolated from patients with urinary tract infections. Mannose-resistant and mannose-sensitive haemagglutination in these strains were also studied. Among the total strains investigated, 50% were haemolytic, 50% synthesized CNF and 58% were lethal for mice. A total of 33 (69%) strains were toxigenic, showing positive results at least in one of the tests employed for toxin detection. No strain was positive for LT and VT production. We conclude that, in addition to haemolytic and haemagglutinating activities, the production of CNF was closely associated to virulence in E. coli strains isolated from urinary tract infections.     

Transfer of passive immunity from mother to young in a teleost fish: haemagglutinating activity in the serum and eggs of plaice, Pleuronectes platessa L

Plaice were immunized against rabbit red blood cells (rbc) and the haemagglutinating activity in their sera and eggs was isolated by gel filtration in Sephacryl S300 SF. A "heavy" (greater than 6.69 X 10(5) Daltons) and a "light" (less than 2.32 X 10(5) Daltons) component of haemagglutinating activity were present in plaice immune serum and eggs. The "heavy" haemagglutinating component had the characteristics of immunoglobulin M (IgM) antibody. By contrast the "light" component was suppressed greatly by monosaccharides but not by 2-mercaptoethanol (2-ME) and it was not precipitable against heterologous rabbit anti-plaice Ig immune serum. Since this "light" component was also identified in the serum and eggs of non-immunized plaice the activity was attributed to non-specific lectin-like activity. The presence of an inducible, high molecular weight, haemagglutinating Ig-like protein in plaice serum and eggs suggests the possibility of transfer of passive adaptive immunity from mother to young in this fish.     

Relevance of blood groups in transfusion of sickle cell disease patients

Blood groups are clinically significant in sickle cell disease (SCD) as transfusion remains a key treatment in this pathology. The occurrence of a delayed haemolytic transfusion reaction (DHTR) is not rare and is a life-threatening event. The main cause of DHTR is the production of alloantibodies against red blood cell antigens. The high rate of alloimmunization in SCD patients is mainly due to the differences of red blood groups between patients of African descent, and the frequently Caucasian donors. From an immuno-haematological point of view, DHTR in SCD patients has specific features: classical antibodies known to be haemolytic can be encountered, but otherwise non significant antibodies, autoantibodies and antibodies related to partial and rare blood groups are also frequently found in individuals of African descent. In some cases, there are no detectable antibodies. As alloimmunization remains the main cause of DHTR, it is extremely important to promote blood donation by individuals of African ancestry to make appropriate blood available.     

Immunochemical characterization of human plasma fibronectin.

Human plasma fibronectin has been purified by a non-denaturing affinity chromatography procedure [Vuento & Vaheri, (1979) Biochem.J. 183, 331--337], and antisera have been raised by immunizing rabbits with the native protein. The antisera reacted strongly with native fibronectin, but only weakly with reduced and alkylated fibronectin or with heat-denaturated fibronectin. Denaturation also affected the haemagglutinating and gelatin-binding activities of fibronectin and increased its susceptibility to proteolytic degradation. The antisera reacted with fragments of fibronectin obtained by proteolysis with plasmin. Large fragments (mol.wt. 180000--200000), lacking the region harbouring the interchain disulphide bridges but containing the sites responsible for gelatin-binding and haemagglutinating activity, showed as intense a reaction with the antisera as intact fibronectin. Smaller peptides showed a weaker reaction. All fragments tested showed sensitivity to denaturation in their reaction with the antisera. The results were interpreted as showing that: (1) native fibronectin has an ordered conformation that is easily perturbed by denaturation; (2) most of the antigenic determinants of the protein are dependent on conformation; (3) the region of the fibronectin molecule containing the interchain disulphide bridges has only few antigenic determinants; and (4) covalent interaction of the two subunits does not contribute to the antigenic structure recognized by rabbit antisera. The observed correlation between the antigenic activity and a structural and functional intactness of fibronectin suggests that the antibodies to native fibronectin could be used as a conformational probe in studies on this protein.     

The effects of short-term acute cadmium exposure on blue tilapia,Oreochromis aureus

Synopsis Disease-free blue tilapia,Oreochromis aureus, exposed to cadmium (0.5 ppm and 10 ppm) showed very significant and dramatic decrease in food consumptions. Food consumptions returned to near normal 21 days after fish were returned to cadmium-free water. Along with anorexia there were decreases in body weights during the period when fish were exposed to cadmium. Fish in cadmium-free water that were pair-fed to cadmium-exposed fish (0.5 ppm) did not gain weight but there were no decreases in body weights. Cadmium was still in tissues 34 days after the fish were placed in cadmium-free water and the accumulation was highest in the kidney; this was followed by liver, brain, gill filaments and muscles. The accumulations of the heavy metal (in kidneys and gills) were significantly higher in fish exposed to high than low cadmium levels. There were no differences in complement levels (haemolytic acitivity) in cadmium-exposed and cadmium-free fish. However, cadmium-exposed fish did not produce detectable haemagglutinating antibodies against sheep red blood cells while cadmium-free fish responded well to the antigen. The anorexia in cadmium-exposed fish contributed to the depression in antibody production.     

Biochemical properties of latices from the Euphorbiaceae

Latex sera from 18 Euphorbia species were surveyed for protein, carbohydrate and total solid contents. Protease, esterase, alkaline and acid phosphatase, N-acetyl-β-glucosaminidase, α-mannosidase, α- and β-galactosidase, α- and β-glucosidase, α-amylase, lysozyme, leucine amino peptidase and cellulase activities were also measured. The effects of nine protease inhibitors were assayed as were haemagglutinating (lectin) contents. Two-dimensional (isoelectric focusing and SDS) PAGE maps of the sera were made. The results obtained show that the latices have widely varying biochemical properties.