Complement Fixation Inhibition Research Articles

Rabbit, guinea pig, rat and human antibodies to human growth hormone: immunological reactions with human and nonhuman primate growth hormones.

The relatedness of owl monkey, squirrel monkey, marmoset and human pituitary growth hormone was measured immunologically by quantitative complement fixation with rabbit antiserum to human growth hormone. Complement-fixation curves obtained with antihuman growth hormone and owl monkey, squirrel monkey and marmoset pituitary extracts were similar. However, the complement-fixation curves obtained with the 3 nonhuman primate growth hormones were qualitatively different from complement-fixation curves obtained with human growth hormone. Serologic specificities of antibodies to human growth hormone produced in the rabbit, guinea pig, rat and human were also examined. Guinea pig, rat and human antibodies, which did not fix complement directly with primate growth hormones, were detected by inhibition of complement fixation. The specificities of these antisera were measured by the extent of their cross-reactions with 8 growth hormones obtained from 3 different superfamilies of primates: Hominoidea, Cercopitheco idea and Ceboidea. All the antisera (rabbit, guinea pig, rat and human) to human growth hormone cross-reacted less effectively with growth hormones from the Ceboidea than with growth hormones from the other 2 groups of primates. The extent of the cross-reactions of guinea pig and rat antibodies, measured by inhibition of complement fixation, was similar to the extent of the cross-reactions of rabbit antibodies, measured by direct complement fixation. Human antibodies, however, showed a considerably greater specificity for growth hormones of the Hominoidea. The degree of relatedness of nonhuman primate growth hormones to human growth hormone correlated with primate phylogeny when measured by direct complement fixation with rabbit antihuman growth hormone or by inhibition of complement fixation with guinea pig, rat or human antihuman growth hormone. (Endocrinology79: 615, 1966)

Agglutinating Antigen from an Ornithosis Agent. I. Preparation Method and Tests with Turkey Serums

The serologic diagnosis of ornithosis in fowl depends upon use of the complement fixation inhibition (CFI) test (5) or the implemented direct complement fixation (IDCF) test (3). There is need for a reliable, more rapid, and simplified test usable for testing large numbers of birds. This is a report of the production of an agglutinating antigen from an ornithosis agent grown in cell cultures. The tests were done with turkey serums since explosive outbreaks of ornithosis have occurred in these birds.

Immunochemical studies of angiotensin

Water-soluble carbodiimides and a benzisoxazolium salt were used to synthesize conjugates containing angiotensin coupled to albumin or poly- l-lysine. antibodies to angiotensin were induced with a conjugate containing the hormone bound to rabbit serum albumin. The antigenic determinants of angiotensin were investigated by inhibition of complement fixation by analogues of the polypeptide. Among the structural features of angiotensin tested, those which had greatest importance for serologic activity were conformation as determined by an optical isomer in the middle of the hormone molecule: the presence of the two amino acids at the N-terminus; and the single phenolic hydroxyl. The structural features shown to be important for serologic activity were those previously shown to be important for biological activity.

Inhibition of complement fixation by rheumatoid sera.

Inhibition of complement fixation by rheumatoid sera.

PARTIAL HYDROLYSIS OF BOVINE PARATHYROID HORMONE BY DILUTE ACID AND PROTEOLYTIC ENZYMES: EFFECTS ON IMMUNOLOGICAL AND BIOLOGICAL ACTIVITIES.

Phenol-extracted bovine parathyroid hormone was partially hydrolyzed with hot dilute HC1 and with trypsin, chymotrypsin and carboxypeptidase A. The effects of each treatment on the immunological and biological activities and on the electrophoretic mobility of the hormone were measured by quantitative complement fixation and complement fixation inhibition, by the calciummobilization assay method of Munson, and by vertical starch gel electrophoresis. Hydrolysis of the hormone with hot 0.2N HC1 did not alter the biological activity despite changes in the structure of the hormone shown immunochemically and by electrophoresis. These results indicate that the entire polypeptide chain is not essential for full calcium-mobilizing activity in the rat. Digestion of the hormone with trypsin was rapid and produced peptide fragments that were devoid of antigenic or biological activities. Chymotryptic digestion was less marked, substantially decreasing, but not eliminating, the immunological and biological activities. Neither the biological activity nor the electrophoretic mobility of the polypeptide chain was altered following removal of a COOH-terminal fragment by carboxypeptidase A. However, the COOH-terminal portion of the molecule contributed to the immunological activity of the hormone, as shown by a decrease in the amount of complement fixed at optimal antigen concentration. (Endocrinology76: 979, 1965)

SPECIFIC FRACTIONATION OF HUMAN ANTIDEXTRAN ANTIBODIES. II. ASSAY OF HUMAN ANTIDEXTRAN SERA AND SPECIFICALLY FRACTIONATED PURIFIED ANTIBODIES BY MICROCOMPLEMENT FIXATION AND COMPLEMENT FIXATION INHIBITION TECHNIQUES.

Human antidextran of one individual, absorbed specifically on sephadex, was fractionated into two populations of antibody molecules by successive elution with oligosaccharides of the isomaltose series of increasing size. The purified antibody fractions and some whole antidextran sera were found to fix complement with dextrans of molecular weight of 195,000 and above. It could be demonstrated by quantitative microcomplement fixation inhibition assays that the antibody eluted with isomaltotriose had a higher affinity for smaller oligosaccharides relative to isomaltohexaose, indicating a high content of antibody molecules with smaller combining sites, while with the second fraction, eluted with isomaltohexaose, the small haptens were very poor inhibitors and the larger oligosaccharides inhibited readily, presumably due to a higher proportion of molecules with larger combining site size. Assays of similarly prepared fractions, obtained from earlier bleedings of the same individual (1), with inhibition of complement fixation were in good agreement with those obtained by inhibition of precipitation. The two purified antidextran fractions were shown to differ with respect to their complement-fixing capacity. The fraction with molecules with smaller size-combining sites fixed only about half as much complement per unit antibody N as did the fraction containing largely molecules with larger combining sites suggesting that the strength of complement fixation is affected by the strength of the antigen-antibody interaction.

Studies on a Common Bedsonia-Group Antigen (CBA) Found in the Yolk of Hens′ Eggs

Summary In the course of investigation of antigens of the psittacosis - lymphogranuloma - trachoma group (Bedsonia) it was noted that uninoculated hen's egg yolk elicited the production, in rabbits, of high titers of antibodies, reacting against Bedsonia antigens in complement fixation (CF) tests. Similar cross-reactions were obtained in guinea pigs, but were demonstrable only in CF inhibition (CFI) tests. Untreated yolk contained low titers of antigen reacting with anti-Bedsonia sera, but this titer was enhanced by treating the yolk with ethanol. This antigen, called common Bedsonia antigen or CBA, is ethanol soluble and soluble in ether only after treatment with ethanol. The possibility has been excluded that the above-described cross-reactions were due to contamination with yolk of Bedsonia antigens used in CF tests or for the immunization of animals. CBA was detected in unincubated eggs or in the yolk of embryos 1 to 15 days old. Its similarity to Bedsonia antigens was further illustrated in CF tests in which titers obtained by primary incubation at 37°C for 90 min and at 4°C for 18 hr were compared. Temperature of incubation affected differently the reactions between anti-yolk serum against CBA and the major antigens of yolk, but did not discriminate between the reactions against CBA and Bedsonia antigens. When this test was applied to anti-psittacosis serum, in this case also, CBA and Bedsonia antigens could not be differentiated.

Inhibition of Complement Fixation by Human Serum

Summary Kinetic and static analyses of complement fixation (CF) reactions were carried out using EA, EA (human), solutions of human thyroglobulin-rabbit anti-thyroglobulin and latex particles coated with human γ-globulin (LPF II). Heated rheumatoid arthritis (RA) sera and N sera, rheumatoid factor (RF) and inhibitor (Ag-Ab)—a γ1M-globulin isolated from RA sera—were tested for their effect on CF reactions. Addition of RA serum and N serum to EA and gpC′ delayed immune hemolysis. Preabsorption of the sera with solid heat denatured γ-globulin did not affect their inhibitory activity. Inhibitor (Ag-Ab) was shown to delay lysis of EA by gpC′. The substance also inhibited CF by soluble antigen-antibody complexes, but CF reactions involving LPF II remained unaffected. RF did not interfere with the lysis of EA by gpC′. This substance was also inactive in CF reactions involving soluble antigen-antibody complexes. RF, however, inhibited CF by LPF II, especially when preincubated with LPF II for 90 min at 56°C. There was also a decrease in the latex fixation titer of RA sera when LPF II was preincubated with gp serum, suggesting some competition between RF and gpC′ for sites of attachment on the γ-globulin molecule.

Serology of Eimeria tenella Oocysts with Rabbit Serum

The antigenicity of Eimeria tenella preparations was studied by means of capillary-tube flocculation and indirect fluorescent-antibody tests. Oocyst extracts reacted weakly and somewhat specifically in flocculation tests. Reactions of oocyst walls, sporocysts, and sporozoites were as great with normal serum as with antiserum. Marked fluorescence was seen in oocyst walls tested by the indirect fluorescent-antibody test while sporocyst walls fluoresced weakly. Nonspecific reaction was much reduced by absorption with liver powder and by dilution, but was not completely eliminated. Apparently the only reports in the literature on the use of rabbit serum in serology of fowl coccidiosis are concerned with the agglutination of Eimeria tenella merozoites (McDermott and Stauber, 1954; Itagaki and Tsubokura, 1955), complement-fixation inhibition (Moore, 1959), and precipitation of E. tenella oocyst extracts (Burns, 1959). The present study reports the application of capillary-tube flocculation and indirect fluorescent-antibody tests to the serology of E. tenella. MATERIALS AND METHODS Sporulated oocysts of E. tenella and E. acervulina were obtained through the kindness of Dr. W. M. Reid, Poultry Department, University of Georgia. These were freed of tissue debris by the pepticdigestion technique of Rikimaru et al. (1961), further cleaned by the density-gradient technique (Sharma et al., 1962), and sterilized by antibiotics (Doran and Farr, 1961). This material was inoculated on Difco blood agar and in Difco thioglycollate broth, found to be culturally sterile, and handled aseptically thereafter. The oocyst suspensions were next divided into two portions, one part being treated with 0.5 mg pronase/ml of suspension, the remaining part being left untreated. The pronase-treated susension was buffered with 0.5 M sodium barbital at pH 8.6. Treated and untreated oocysts were next ruptured in glass homogenizers, the pestles of which were attached to a low-speed motor. Supernate was separated from sediment by centrifugation in an angle-head centrifuge for 40 min at 12,000 g and 0 C and filtered through a 0.45-A millipore filter. Oocyst walls and sporocysts from the sediment Received for publication 12 March 1963. * Present address: Jalaun, Dist. Jalaun, U. P., India. were separated by density-gradient centrifugation in 50% glycerol (Sharma et al., 1962) for 10 min at 2,200 g and 0 C. The sporocysts formed a band about two-thirds of the way down the tube and were collected by a syringe fitted with a 4-inch cannula, placed in about 10 vol of water at 0 C. and concentrated by centrifugation in an angle centrifuge for 10 min at 5,000 g and 0 C. This process was repeated three or four times until microscopic examination showed the suspension to consist almost entirely of sporocysts. Oocyst walls were recovered from a layer just above the bottom of the centrifuge tube and processed further by the same method used on the sporocysts. Live sporozoites were obtained from day-old chicks 1.5 hr after feeding large numbers of cleaned oocysts to fasting chicks. Fifteen-cm pieces of small intestine were removed aseptically and flushed with 0.85% saline at 37 C. The washings were filtered through 16 to 32 layers of gauze and the sporozoites collected from the filtrate by repeated cycles of washing and centrifugation at 800 g at room temperature for 3 or 4 min. Merozoites were obtained as follows. Five 11-day-old chicks were given 500 thousand sporulated oocysts orally. Feed was withdrawn 2 days later and 5 days later three chicks were dead; the other two were necropsied. The contents of the cecum were flushed out with warm saline and the lining scraped. Scrapings and cecal contents, suspended in saline, were mixed thoroughly and filtered through 16 layers of gauze. The filtrate was centrifuged for 6 min at 800 g at room temperature and the supernate discarded. The sediment was reconstituted in saline and centrifugation repeated. The supernate was discarded again. The remaining sediment contained many merozoites and some tissue debris. The preparations described above were administered to rabbits as shown in Table I. The New Zealand white rabbits used had been raised in a coccidia-free environment. Blood samples were collected from each animal before inoculation and

Inhibition of Complement Fixation by Human Serum

Summary Assays for inhibition of fixation of gpC′ were described. The inhibitory activity of human sera was found to vary, but inhibitory activity was significantly elevated in sera of individuals having rheumatoid arthritis. Fractionation of a number of sera of healthy donors by DEAE cellulose chromatography revealed two inhibitors of CF. In addition to these substances, a third compound occurred in the sera of individuals with rheumatoid arthritis. The compound was purified 400-fold and was shown to be a γ-macroglobulin and was distinct from RF.

Inhibition of Complement Fixation by Rabbit Serum

Summary The fixation of guinea pig complement by aggregates of rabbit antibody against ovalbumin and ovalbumin is impeded by normal rabbit serum. The inhibitory effect of rabbit serum is a function of the γ-globulin fraction. Prolonged storage in the cold, or heating at 56–65°C for 1 hr or less have no significant effect on this activity of rabbit serum or its γ-globulin; exposure to 85°C results in the loss of activity within 30 min. The inhibitor can be removed from serum by absorption with bentonite, celite or Zymosan, or by “decomplementation” with large doses of specific precipitate. Preliminary kinetic studies show that the inhibitor retards the rate of complement fixation and that its action can be overcome, in part or entirely, by suitable extension of the incubation period. The mode of action of the inhibitor has been discussed in terms of two hypotheses, namely: a) the possibility that the inhibitor combines with the antigen-antibody aggregates in a manner which blocks fixation of complement, or b) that it combines reversibly with complement.

トキソプラズマ症の補体結合阻止反応に関する研究

トキソプラズマ症の補体結合阻止反応に関する研究

The complement fixation inhibition test with sera from chickens experimentally infected with toxoplasms.

The complement fixation inhibition test with sera from chickens experimentally infected with toxoplasms.

Antigenic Studies on the Psittacosis-Lymphogranuloma Venereum Group of Viruses

Summary The time of onset of cutaneous hypersensitivity in chickens infected with ornithosis virus was related to the LD50 used for infection. Hypersensitivity developed within 4 weeks of infection and was detected for 1 year after infection. Complement-fixation inhibition antibody appeared earlier in the infection than detectable allergy, although after 7 weeks of infection the serological and skin-test methods agreed in approximately 80% of the infected chickens. The skin-test antigen was resistant to periodate, trypsin, desoxyribonuclease, and exposure to 90°C. The complement-fixing and allergenic activities were separated by ultracentrifugal sedimentation of the complement-fixing antigen. The skin-test antigen was standardized on a nitrogen basis in guinea pigs and chickens.

Complement fixation studies in ragweed allergy: II. Determination of antibody in human sera to ragweed antigen by means of a complement fixation inhibition test; The relationship of antibody so determined to the passive transfer for blocking antibody

Complement fixation studies in ragweed allergy: II. Determination of antibody in human sera to ragweed antigen by means of a complement fixation inhibition test; The relationship of antibody so determined to the passive transfer for blocking antibody

Inhibition of complement fixation by certain serums when tested against psittacosis and lymphogranuloma venereum antigens.

Journal Article Inhibition of Complement Fixation by Certain Serums When Tested Against Psittacosis and Lympho-Granuloma Venereum Antigens Get access Nathalie Schmidt, Nathalie Schmidt From the Departments of Bacteriology and Pathology, Northwestern University Medical School, Chicago, Illinois Search for other works by this author on: Oxford Academic PubMed Google Scholar Harry B. Harding, Harry B. Harding From the Departments of Bacteriology and Pathology, Northwestern University Medical School, Chicago, Illinois Search for other works by this author on: Oxford Academic PubMed Google Scholar Opal E. Hepler, Opal E. Hepler From the Departments of Bacteriology and Pathology, Northwestern University Medical School, Chicago, Illinois Search for other works by this author on: Oxford Academic PubMed Google Scholar Edna Murmann Edna Murmann From the Departments of Bacteriology and Pathology, Northwestern University Medical School, Chicago, Illinois Search for other works by this author on: Oxford Academic PubMed Google Scholar The Journal of Infectious Diseases, Volume 91, Issue 2, July 1952, Pages 173–179, https://doi.org/10.1093/infdis/91.2.173 Published: 01 July 1952 Article history Received: 13 March 1952 Published: 01 July 1952

The complement fixation inhibition test and its application to the diagnosis of ornithosis in chickens and in ducks; principles and technique of the test.

For the last 15 years, laboratory diagnosis of psittacosis and similar diseases has been sucessfully aided by the complement fixation test. For clinical use it is a practical device because among other reasons, isolation of the virus is often difficult and expensive. It has gained in importance in the last few years when diagnosis has provided a direction for effective therapy. In epidemiological and experimental work the test has effectively detected antibodies in the serum of infected psittacine birds (Meyer and Eddie, 1939; Meyer et al, 1939), pigeons (Meyer, 1941; Meyer et al, 1942; Eddie and Francis, 1942), seashore birds (Pollard, 1947), guinea pigs (Bedson, 1932; Meyer and Eddie, 1939), goats (Meyer and Eddie, 1939), monkeys, rabbits and man. Since psittacine birds were considered to be the reservoir

The complement fixation inhibition test and its application to the diagnosis of ornithosis in chickens and in ducks; confirmation of the specificity and epidemiological application of the test.

Journal Article The Complement Fixation Inhibition Test and its Application to the Diagnosis of Ornithosis in Chickens and in Ducks: II. Confirmation of the Specificity and Epidemiological Application of the Test Get access H. Karrer, H. Karrer From the George Williams Hooper Foundation, University of California, San Francisco, California Search for other works by this author on: Oxford Academic PubMed Google Scholar K. F. Meyer, K. F. Meyer From the George Williams Hooper Foundation, University of California, San Francisco, California Search for other works by this author on: Oxford Academic PubMed Google Scholar B. Eddie B. Eddie From the George Williams Hooper Foundation, University of California, San Francisco, California Search for other works by this author on: Oxford Academic PubMed Google Scholar The Journal of Infectious Diseases, Volume 87, Issue 1, July 1950, Pages 24–36, https://doi.org/10.1093/infdis/87.1.24 Published: 01 July 1950 Article history Received: 24 January 1950 Published: 01 July 1950

Zone Phenomenon in Complement Fixation. Relation to Greater Sensitivity of Ice Box Type of Wassermann Reaction

Two hypotheses were recently presented for the explanation of the mechanism of zoning occurring in complement fixation—one by Levine, the other by Eagle. The two hypotheses rest on principles which possess no element in common. Therefore, I undertook a critical study of the subject matter. In this paper are presented in brief the results of the study of the theory of Eagle. The latter can be briefly summarized as follows: Serum in concentrations greater than 1:25 effects an inhibition of complement fixation through the adsorption of fixation-preventing substances presumably present in negative serum. These inhibiting substances are adsorbed by the antigen-antibody complexes after they had become fully organized. The adsorption is more intense the more negative a serum is, and corresponds roughly to the concentration of the serum protein. Hence, in weakly positive serums a more pronounced fixation of complement will occur in the higher serum dilution, and hence, the zone phenomenon. This same inhibition, Eagle states, is less marked at lower temperatures, and hence, in an indirect manner it is also responsible for the greater sensitivity of the icebox test. Experimental studies were carried out as follows: (1) Complement and amboceptor titrations in the presence of high concentrations of negative serum; (2) complement fixation tests with Kahn antigen precipitated from positive and negative reactions in the presence of high negative serum concentrations; (3) the effect of the removal of the native antisheep amboceptor from negative serum and the bearing of this procedure on the assumption that the results of Eagle's original experiments were influenced by natural amboceptor.

STUDIES IN THE SEROLOGY OF SYPHILIS

Serum, in concentrations greater than 1:25, causes a marked inhibition of complement fixation in general and of the Wassermann reaction in particular. The serum protein is probably adsorbed by the colloidally dispersed lipoid-reagin complexes, forming a protective film which prevents the fixation (adsorption) of complement. This inhibition explains the zone phenomenon in complement fixation: a weakly positive serum may give a completely positive reaction in e.g., 1:5 dilution, and yet, because of this serum inhibition, may appear completely negative when tested as whole serum. The greater sensitivity of the ice box test is due (1) to the fact that the serum inhibition just described is less marked at lower temperature; (2) to the prolonged incubation time, making for greater specific fixation; (3) to a more marked non-specific destruction of complement by antigen; and (4) a spontaneous deterioration in the longer ice box test. Because of the inhibition by serum protein in high concentration, a quantitative Wassermann technique involving the use of graded quantities of serum is worthless when carried out at 37 degrees C. Even the ice box test, which is less susceptible to this inhibiting effect, will yield a positive reaction with whole serum only when the circulating reagin exceeds a surprisingly high threshold (six to ten times the quantity which could be detected in dilute serum). It is well known that a negative Wassermann, even by a very sensitive test, does not exclude syphilis: it now appears that a negative Wassermann does not exclude circulating reagin.