Complement Fixation Reaction Research Articles

Myocardial Infarction Due to Coronary Thromboarteritis, Following Acute Febrile Mucocutaneous Lymph Node Syndrome (MLNS) in an Infant

A previously healthy 6-month-old male infant was admitted to our clinic for persistent high fever and an erythematous skin rash. Hyperemia of conjunctivae, redness of oral mucosa, and swelling of several cervical lymph nodes were also observed. Soon after admission he developed jaundice which continued for one week. Results of laboratory examinations revealed leukocytosis, elevated ESR, mild proteinuria, leukocytes in urinary sediment, hyperbilirubinemia, increased serum alpha-2-globulin and positive CRP. Serum ASO titer was less than 100 Todd units and complement fixation reactions for various viruses were not significant. Cultures of blood, urine, stool and throat were negative. He was treated with antibiotics and steroids and improved gradually, but then developed signs and symptoms of congestive heart failure. An ECG showed a pattern of extensive myocardial infarction. He died of congestive heart failure at the age of 21 months. Autopsy findings revealed focal aneurysmal dilatation and obstruction of the bilateral coronary arteries. The myocardial infarction involved the septum of the anterolateral wall of the left ventricle. The coronary arteries, distal aorta, iliac arteries and pulmonary arteries were also involved, but very slightly. The symptoms, laboratory findings and the course of this patient were compatible with those of acute febrile mucocutaneous lymph node syndrome (MLNS), MLNS has been accepted as a new syndrome by most Japanese pediatricians in recent years. Its etiology is still unknown. Some of the patients with this syndrome had died suddenly of coronary thromboarteritis after recovering from the acute illness. Although autopsy findings of the fatal cases have been similar to those describing infantile polyarteritis nodosa, the relationship between MLNS and infantile polyarteritis is controversial.

Modulation of cell-surface antigens of a murine neuroblastoma.

Antisera were produced in rabbits to morphologically differentiated cells from the C1300 murine neuroblastoma (i.e., cells in which process formation was induced by maintenance on serum-free medium for 5 days). These antisera reacted more strongly in the complement fixation reaction with such "differentiated" cells than with "undifferentiated" (nonprocess-bearing) neuroblastoma cells. Adsorption of the antisera with undifferentiated cells removed the reactivity to cells without processes, while the reactivity with serum-free cells which possess processes was retained. Indirect immunofluorescence studies confirmed the results obtained by complement fixation and demonstrated that antibodies to the surface antigens of process-bearing cells could be adsorbed by particulate preparations from brain but not liver, spleen, or kidney. This is the first description of neural-associated cell-surface changes that correlate with the morphological differentiation in culture of neuroblastoma cells.

Immunofluorescence methods for detection of influenza and mumps antibodies using infected shedded allantoic cells and impression smears of organ cultures as the sources of antigens.

Indirect immunofluorescence technique was used for detection of specific antibodies against influenza and mumps viruses in the IgM and IgG fractions of human gamma globulins. Infected cells shed into the allantoic fluid from the allantoic membrane of inoculated hen's eggs served as a source of cell-bound influenza antigens. Impression smears of mumps infected chick embryo tracheal organ cultures yielded a sufficient amount of mumps antigens containing cells. Using these techniques, specific antibodies against the two viruses were readily detectable in one or both fractions of human gamma globulins. No cross-reactions between influenza and mumps virus occured, techniques were found to be simple and reliable, the slides prepared in quantities and fixed in acetone were available anytime, when required for the tests. Sera used in this investigation were previously screened by complement fixation reaction and the presence of specific antibodies were thus indicated. Comparisons between findings achieved by complement fixation and immunofluorescence were made.

Immunological Studies on the Envelope Component of Venezuelan Equine Encephalomyelitis Virus

Treatment of purified Venezuelan equine encephalomyelitis virus with the nonionic detergent Triton X-100 permitted spearation of the envelope from the core component. The isolated envelope was a noninfectious immunogen which reacted in hemagglutination, hemagglutination inhibition, complement fixation, and neutralization serological reactions.

A study of reproduction, disease and physiological blood and serum values in Idaho elk.

A study was conducted on elk (Cervus canadensis nelsoni) in the Selway-Bitterroot Wilderness and adjoining primitive area of the Clearwater drainage of Idaho over a 3 year period, 1969-1972. Midwinter examination of cows in this herd indicated the pregnancy rate was high, but game counts showed only 15 to 20 calves per 100 cows indicating that the losses occurred around or after calving. A serologic survey did not indicate the presence of antibodies to the agents of brucellosis, leptospirosis, vibriosis, bluetongue, infectious bovine rhinotracheitis or bovine virus diarrhea. Positive complement fixation reactions for anaplasmosis were present in nearly 50% of the animals tested; however, many of the serums were anticomplementary. Blood chemistry determinations were made on all elk and compared with the values obtained from captive elk. Similar values were noted in the captive and wild elk in calcium phosphorous ratio, glucose and total protein. Dissimilarity occurred with cholesterol, blood urea nitrogen,...

Physical properties and antigenic components of Oriboca virus.

Two complement fixation (CF) components were detected in all of the various preparations (mouse liver, brain, serum, and BHK-21 cell culture) of Oriboca examined by equilibrium density centrifugation. The CF components possessed a buoyant density of 1.21 and 1.18 g/ml, and they were precipitated by treatment with protamine. Analysis of Oriboca virus by rate zonal sedimentation separated three components. The most rapidly sedimenting component was the infectivity which cosedimented with a peak of hemagglutinating (HA) activity designated VHA. A second, more slowly sedimenting, noninfectious HA component (HA-2) was followed by a peak of CF activity which barely moved from the top of the gradient. The single rate zonal CF peak and the two density gradient CF components were broadly cross-reactive serologically with Murutucu antiserum. They could not be identified as Oriboca on the basis of their CF reactions with Murutucu and Oriboca antisera. Oriboca-infective virions possessed an equilibrium density of 1.20 g/ml if derived from BHK-21 cell culture fluid concentrates or mouse serum. Infective virions prepared from mouse brain or liver banded at a buoyant density of 1.17 to 1.18 g/ml. These density differences, along with observed differences in reactivity with protamine and in rate of neutralization, are consistent with the interpretation that these are related to the problem of whether the source preparations of Oriboca represented primarily intracellular or extracellular particles.

The transmission of Sarcocystis tenella in sheep.

1. Experiments, carried out to demonstrate the mode of infection with S. tenella, are described. The complement fixation reaction became positive 42 and 84 days after ingestion of infected material and sarcocysts were detected in the muscle 65 and 93 days later in the same animals.

The Orientation of Serologically Active Phospholipids in Mitochondrial Membranes, II. Studiesin vitrowith a Complement Fixation Reaction

Article The Orientation of Serologically Active Phospholipids in Mitochondrial Membranes, II. Studies in vitro with a Complement Fixation Reaction was published on January 1, 1973 in the journal Biological Chemistry (volume 354, issue 2).

Measurement of Mycoplasma Pneumoniae Mycoplasmacidal Antibody in Human Serum

Abstract Investigation of the conditions which affect the mycoplasmacidal action of antibody and complement upon Mycoplasma pneumoniae indicated the importance of a filtered suspension of predominantly single organisms, and an appropriate concentration of cations. The mycoplasmacidal effect of antibody and complement resembled the hemolytic reaction in that variation in concentration of either antibody or complement yielded an S-shaped curve. The antigens involved in the mycoplasmacidal reaction appeared to be the glycolipids of the organism which had previously been shown to be of major importance in the complement fixation and metabolism inhibition reactions. The standardized mycoplasmacidal antibody assay proved to be considerably more sensitive than the conventional assay systems for measurement of M. pneumoniae antibody in human serum.

Serologic Specificities of Brain Subcellular Organelles

Abstract Antisera were prepared to synaptosomal fractions isolated from rat cerebral cortex. The antisera, at optimal dilutions, reacted in the complement fixation reaction only with the synaptosomal fraction and synaptosomal plasma membrane fractions, but not with myelin, nuclei, soluble protein, mitochondria or synaptic vesicles from rat cortex. Little or no reaction was observed with membrane preparations from other rat organs. The specific antigenic determinants of the synaptosomal membrane were relatively stable to heat, sonication and mild tryptic digestion. These results demonstrate that the plasma membrane of cortical synaptosomes can be serologically distinguished from the plasma membrane of other organs and from the rest of the subcellular and sub-synaptic fractions of brain.

Electrophoretic pattern of sera of normal and cancer patients

Acrylamide disc electrophoresis patterns of serum from cancer patients are compared with those of normal serum. A significant difference in pattern is observed, especially when sera are incubated, prior to electrophoresis, with total yeast RNA. The results obtained with this technique are then compared with a complement fixation reaction using as antigen the same yeast RNA. It was found that the electrophoresis technique has a higher percentage concordance with histological diagnosis. However, certain anomalous results were obtained and these are discussed.

Biochemical characterization and skin test sensitivity of mycelial growth filtrate of Histoplasma duboisii.

Abstract A controlled study of the biochemical characteristics and potency of skin-test antigens was conducted on crude mycelial filtrates (1–24 weeks old) of two isolates of Histoplasma duboisii. Results of the study indicate that mycelial filtrates of H. duboisii may be specific for the detection of allergies created by this organism. A mycelial filtrate of Histoplasma capsulatum, H-42, gave a complement-fixation reaction with serum of guinea pigs infected with H. duboisii. Also, immunoelectrophoretic studies indicate that H. duboisii and H. capsulatum have at least one antigen in common, but that the “M” antigen of H. capsulatum differs from that of H. duboisii. The crude mycelial filtrate may be fractionated by ion-exchange chromatography, producing materials which are useful in skin tests. The characteristic development of severe orchitis in male guinea pigs was also observed following intraperitoneal injections of large doses of mycelial fragments and yeast cells of H. duboisii.

The Complement-Fixation Reaction in Equine Infectious Anemia: Demonstration of Inhibition by IgG(T)

Abstract The role of non-complement-fixing anti-equine infectious anemia (EIA) antibody in the conversion of complement fixation (CF) tests from positive to negative in EIA-infected horses was investigated. Complement-fixation inhibition (CFI) tests demonstrated antibodies in sera that were CF negative. These antibodies would bind to antigen, but would not fix complement. The inhibiting antibodies were isolated and shown to be IgG(T) by immunoelectrophoresis and immunodiffusion against monospecific anti-IgG(T) antisera. Separation of immunoglobulins from affected horse sera by DEAE cellulose chromatography revealed 11 of 13 had complement-fixing IgG and non-complement-fixing IgG(T) antibodies present simultaneously regardless of the reactivity of whole sera. The reactivity of whole sera in CF and CFI tests evidently depended upon the relative concentration and perhaps affinity of the Ig classes. The predominant class fluctuated in serial sera from four of six persistently infected horses.

Characterization of a Soluble Nuclear Ribonucleoprotein Antigen Reactive with Sle Sera

Abstract A soluble nuclear ribonucleoprotein antigen reactive with SLE sera in precipitin and complement fixation reactions is described. The antigen was shown to be quite labile and was not recovered from human autopsy material (liver and spleen) but was easily demonstrated in extracts of surgically removed human thymoma and thymus, as well as in extracts of human chronic myeloid leukemia and normal bone marrow cells. It could also be extracted from calf thymus tissue and the thymus and spleen of the rabbit. It was characterized as ribonucleoprotein because the antigenic activity was destroyed by both ribonuclease and trypsin and the washed immune precipitate contained material which yielded an orcinol reaction product with an absorption spectrum indistinguishable from ribose. The antigen was found to be unstable at relatively moderate temperatures (40°C), at pH below 5.0 and above 9.0, and was destroyed by treatment with sodium metaperiodate. Antibodies to this antigen, designated Mo, were found in a high proportion of SLE sera and were absent from a variety of sera from patients with other connective tissue diseases as well as normal sera. The nuclear fluorescent pattern associated with this system was shown to be “speckled.”

Micro complement fixation assay for type-specific group A streptococcal antibody.

A micro complement fixation assay was devised to measure the type-specificity of anti-M antibody. Unabsorbed sera were from rabbits hyperimmunized with heat-killed streptococci and from humans with naturally occurring antibody or immunized with purified M protein vaccines. These sera fixed complement only in the presence of homologous M proteins (serotypes 1, 3, 6, 12, and 14). The complement fixation reaction paralleled the results obtained with the Lancefield bactericidal (opsonic) assay and did not exhibit the cross-reactions frequently seen with type-specific anti-M sera assayed by passive hemagglutination. All serological activity of the antisera resided in the fraction corresponding to 7S globulin eluted from a Sephadex G-200 gel filtration column. Because of the close correlation between micro complement fixation and the bactericidal assay for type-specific antibody, it is proposed that the micro complement fixation procedure, as we have described it, merits further evaluation as a substitute for the bactericidal test of immunity to group A streptococcal infection.

Immunochemical and physicochemical properties of haptoglobin, abtihemoglobin antibody, and their complexes with hemoglobin

The immunochemical properties of haptoglobin which binds specifically with hemoglobin were compared with those of antihemoglobin A 1 antibody. Hemoglobin-haptoglobin shows neither precipitation reaction nor complement fixation reaction under conditions in which hemoglobin-antihemoglobin antibody shows these immunological reactions. From these observations, the absence of lattive formation between hemoglobin and haptoglobin was suggested. Antihuman hemoglobin antibody binds deoxygenated human hemoglobin and hemoglobin-haptoglobin complex, both of which human haptoglobin does not. On the other hand, although human haptoglobin binds hemoglobins of horse, rabbit, and frog, antihuman hemoglobin antibody does not. And from the data on circular dichroism spectra of hemoglobin-haptoglobin complex and hemoglobin-antihemoglobin antibody complex, the differences in environmental structure around the heme between hemoglobin-haptoglobin complex and hemoglobin-antihemoglobin antibody complex are clarified which may have some connection with the differences in immunochemical reactions between these two complexes.

Biochemical and Immunological Studies onAspergillus

Two kinds of polysaccharides, glucan and glycopeptide (galactomannan-peptide), were obtained from Aspergillus fumigatus (IFO 5840) by extraction with 50% pyridine and were purified by fractional precipitation with acetone, by a column chromatography on Dowex-50 and DEAE-cellulose and by the gel filtration on Sephadex G-50 and G-200. The glycopeptide, designated APSK-66 fraction, showed both an Arthus and delayed type (tuberculin type) skin reactions in sensitized rabbits and guinea pigs. By treatment with proteolytic enzymes, the delayed type skin reactivity of APSK-66 fraction was reduced but the Arthus type skin reactivity was not affected. However, the Arthus type skin reactivity of APSK-66 fraction was completely lost by periodate oxidation, and the delayed type skin reactivity of APSK-66 fraction was retained. The APSK-66 fraction showed precipitation, complement fixation and passive hemagglutination reactions with rabbit antisera against A. fumigatus. Glucan, designated as APSK-33 fraction, did not show any immunological activity when tested in the present experiment. The chemical structures of the glucan and galactomannan were discussed.

鳥類血清の補体結合反応に関する研究

Various test conditions were examined to establish the complement fixation (CF) test procedures of chicken sera for practical application. From the experiment in which fresh duck, goose, and pigeon sera were tested for their cofactor (avian C′1) activity that is essential for CF reaction of avian antibody, duck serum was proved to be most suitable as cofactor. Thus, standard CF procedure was established by employing a given amount of fresh duck serum in the reaction system, and was shown to have higher reproducibility and to be more practical than the other methods previously reported. The procedure was successfully applied for detection of antibody to Newcastle disease virus in convalescent and hyperimmune chicken sera, and antibody to Mycoplasama galisepticum in convalescent chicken sera.

A Poxvirus of Primates. II. Immunology<xref ref-type="fn" rid="FN2">2</xref>

OrTeCa poxvirus shares antigens with Yaba virus, as evidenced by two-way crossreactions in complement-fixation and neutralization tests. Antibodies produced by vaccinia and monkeypox viruses showed weak complement-fixation reactions with OrTeCa viral antigen, but neutralization reactions were negative and cross protection in vivo was not demonstrated. OrTeCa and Yaba virus antisera both contained complement-fixing and neutralizing antibodies against swinepox virus, but swinepox antiserum had no antibodies to any other poxvirus. Long-lasting immunity to the OrTeCa pox disease was produced in monkeys and man by spontaneous infection or vaccination with OrTeCa poxvirus. Complement-fixing antibody titers could be measured in convalescence, but tended to drop to low levels or disappear thereafter. In contrast, neutralizing antibody titers remained high for 3 years, and monkeys that were challenged with OrTeCa virus were reFractory to a second infection.

Soluble components of adenovirus type 8.

Soluble components of type 8 adenovirus have been studied. Four different components were isolated by anion-exchange chromatography and purified by further chromatographic procedures, by zonal centrifugation, and by erythrocyte absorption and elution. The four components exhibited the following characteristics. (i) Fiber antigen was trypsin-resistant and functioned as incomplete hemagglutinin (agglutinated rat and human erythrocytes only in the presence of certain types of adenovirus antisera). (ii) The penton was trypsin-sensitive, exerted a cytotoxic effect, and also showed incomplete hemagglutination, being active in the presence of a majority of heterotypic adenovirus antisera studied. (iii) The group-specific hexon antigen reacted in complement fixation reaction and gel precipitation with sera prepared against other types of adenoviruses, besides showing characteristics indicating the presence of a type-specific antigen component. (iv) The soluble complete hemagglutinin was trypsin-sensitive, displayed cytotoxic effect, adsorbed easily to human and rat erythrocytes, and could be eluted from them by means of receptor-destroying enzyme. The three hemagglutinins of adenovirus type 8 proved to be highly unstable, and their demonstration was only successful by using a large quantity of freshly prepared concentrated virus material. Considering these conditions, a method was developed for their concentration and purification.