scholarly journals A Study of Non-specific Complement-fixation with particular reference to the Interaction of Normal Serum and certain Non-antigenic substances

1928 ◽  
Vol 28 (2) ◽  
pp. 172-197 ◽  
Author(s):  
T. J. Mackie ◽  
M. H. Finkelstein
Keyword(s):  
Human Serum ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Normal Serum ◽  
Fixation Test ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Considerable Proportion ◽  
Normal Human ◽  
Fixation Reaction

1. When a solution of commercial peptone is substituted for antigen in a complement-fixation test with the unheated normal serum of certain species (man, ox, sheep, horse, rabbit, white rat), a definite fixation reaction occurs both at 37° C. and at 0° C. In the ox, sheep, horse and rabbit this property of serum is partially stable at 55° C., but normal human serum and the serum of the white rat are inactive after heating at this temperature. The property is resident mainly in the carbonic-acid-insoluble globulins of the serum.2. The same results are obtained when ethyl alcohol diluted with several volumes of normal saline solution is substituted for antigen in a complement-fixation test with normal serum.3. Analysis of these reactions shows a close correspondence with complement-fixation by the sera of normal animals plus the Wassermann “antigen”—the Wassermann reaction of normal animals.4. Marked complement-fixation effects are also obtained with heated normal serum of the rabbit, ox, sheep, horse plus cholesterol suspension, and particularly cholesterolised-peptone, these effects occurring in parallel with those produced by serum plus alcohol-saline, peptone solutions and the Wassermann “antigen.” The heated normal serum of the pig, white rat and guinea-pig do not exhibit these reactions, and the same applies to heated normal human serum. Unheated pig serum fails to react. Such results also elicit a close relationship between these non-specific reactions and the Wassermann reactions of normal animals.5. The reacting property is absent from the serum (heated and unheated) of young rabbits during the first 2 to 3 weeks of life, but appears soon after this (e.g. by the 37th day) and is progressive in development. Its development in early life runs parallel to that of the natural haemolytic property of the serum for sheep's blood (due to a natural antibody-like substance). The two properties are, however, independent as illustrated by absorption tests.6. Besides the agents referred to above as capable of fixing complement along with normal sera, other substances possess a similar property, e.g. certain alcohols, sodium oleate, tissue proteins, certain amino-acids and sodium nucleate. Commercial peptone purified by precipitation with alcohol is equally active with the original material. Cholesterolisation of these agents may yield a product whose activity is greater than that due to summation of effects.7. Wassermann-positive and -negative human sera have been tested in the complement-fixation reaction with certain of these “pseudo-antigens,” viz. alcohol-saline, peptone, cholesterol, and cholesterolised-peptone, but a uniform parallelism has not been demonstrated between the reactions with these agents and the Wassermann effect. Some Wassermann-positive sera react also with alcohol-saline, peptone, cholesterol and cholesterolised-peptone, while sera from selected normal persons are quite inactive. A considerable proportion of Wassermann-positive sera yields definite complement-fixation with cholesterol and cholesterolised-peptone; a small proportion of Wassermann-negative sera reacts with these agents.8. The thermolability of the serum principles acting with various “pseudoantigens” has been studied by testing unheated serum and serum heated at temperatures ranging from 46° to 60° C. Two types of thermolability curve have been demonstrated with different specimens of rabbit serum: (1) a more or less progressive weakening of the various reactions with inactivation at 60° C.; (2) inactivation of the effects with Wassermann “antigen,” alcoholsaline and cholesterol at 50–52° C., activation of the effects with the Wassermann “antigen” and cholesterol at 54–56°C. and inactivation again above 60° C.; in this case the curves for peptone and cholesterolised-peptone do not show such double inactivation. Unheated normal human serum yields reactions with the various agents (including the Wassermann “antigen”) but inactivation occurs at 50° to 54° C. whereas certain syphilitic sera yield thermolability curves somewhat similar to type (1) of rabbit serum, with inactivation at 60° C. or over.

State of iron(III) in normal human serum: low molecular weight and protein ligands besides transferrin

1970 ◽  
Vol 48 (12) ◽  
pp. 1339-1350 ◽  
Author(s):  
Bibudhendra Sarkar
Keyword(s):  
Molecular Weight ◽  
Human Serum ◽  
Binding Capacity ◽  
Normal Serum ◽  
Normal Human Serum ◽  
Void Volume ◽  
Antibody Concentration ◽  
Low Molecular Weight ◽  
Protein Ligands ◽  
Normal Human

A fraction of Fe(III) in normal human serum is bound to both low molecular weight as well as protein ligands besides transferrin. Citrate was shown to be the major Fe(III)-binding substance in the low molecular weight fraction. Amino acids, sugars, and organic acids, such as ascorbate, pyruvate, and lactate, showed very little or no binding to Fe(III) in normal serum. Iron(III)-binding proteins other than transferrin were shown to be present in normal serum when the native serum with [59Fe(III)] was fractionated by (NH4)2SO4 and Sephadex G-150. The presence of these proteins was observed when trace amounts of Fe(III) were added to the normal serum and when the iron-binding capacity was saturated with Fe(III) to 50% and 100%. These proteins were eluted in the void volume of Sephadex G-150 and none of them corresponded electrophoretically to transferrin. The results of the gel filtration of a mixture of [131I]-transferrin and the proteins eluted in the void volume of Sephadex G-150 were strongly in favor of the Fe(III)-proteins as being neither transferrin aggregates nor transferrin adducts with other proteins. Immunoelectrophoresis of the Sephadex G-150 void volume proteins on agar gel against the antibody to transferrin revealed the absence of transferrin. The presence of at least six proteins in this fraction was shown by immunoelectrophoresis. Positive precipitin reactions were obtained with the antibodies to α2-macroglobulin, γG-globulin, γA-globulin, and γM-globulin. At least two more proteins in this fraction remained unidentified. When the same fraction containing [59Fe(III)] was treated with the whole antisera and the precipitates were counted for radioactivity, a typical antigen-antibody reaction curve was obtained as the antibody concentration was increased. Similar experiments with this fraction and antibodies to α2-macroglobulin, γG-globulin, γA-globulin, and γM-globulin failed to show any significant radioactivity in the precipitate. Since this fraction did not contain any transferrin, it was concluded that there are proteins besides transferrin which can act as ligands for Fe(III) in normal blood plasma.

ESR Spectra o f Normal Human Serum after Treatment with Complement Activating Agents*

1980 ◽  
Vol 35 (9-10) ◽  
pp. 844-847
Author(s):  
Melchiorre Brai ◽  
Antonio Bartolotta ◽  
Maria Brai ◽  
Marcello Messina ◽  
Pietro L. Indovina
Keyword(s):  
Free Radical ◽  
Human Serum ◽  
Complement Activation ◽  
Esr Spectra ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Esr Spectrum ◽  
Normal Human ◽  
Radical Signal

Abstract We describe the appearance of a free-radical signal in the ESR spectrum of normal human serum incubated with several complement activating agents. The intensity of this signal is dependent of dose of activating agents, time and temperature. Signals elicited by different complement activators differ in morphology and kinetics. Inhibition by treatment with EDTA and the presence of the signal in activated C 6-deficient rabbit serum suggest that the con-vertase forming steps of complement activation (C2 to 5) could be the source of free-radical containing molecules.

scholarly journals AN IMMUNOLOGICAL STUDY OF THE A SUBSTANCE OR ACETYL POLYSACCHARIDE OF PNEUMOCOCCUS TYPE I

1934 ◽  
Vol 60 (2) ◽  
pp. 127-147 ◽  
Author(s):  
John F. Enders ◽  
Chao-Jen Wu
Keyword(s):  
Human Serum ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Type I ◽  
Immunological Study ◽  
Protective Capacity ◽  
Similar Treatment ◽  
Active Immunity ◽  
Elementary Analysis ◽  
Normal Human

1. The A carbohydrate isolated from Type I pneumococcus by Pappenheimer and Enders, on the basis of elementary analysis, the presence of the acetyl group and its immunological properties, appears to be identical with the acetyl polysaccharide described by Avery and Goebel. 2. The A carbohydrate possesses a greater anti-opsonic action than either the deacetylated substance obtained by boiling in alkali or the soluble specific substance of Type I pneumococcus prepared according to the procedure of Heidelberger, Goebel and Avery. The opsonic titre of normal human serum is practically eliminated upon the addition of the A carbohydrate—an effect not observed with equivalent amounts of either the deacetylated material or the specific soluble substance. In immune serum, the A carbohydrate brings about a quantitatively greater reduction in opsonic activity than its derivatives, but it has not been possible to demonstrate complete inhibition of phagocytic action by the method of absorption of antibody. 3. In a system of normal human serum and leucocytes capable of destroying Type I pneumococcus, the bactericidal effect maybe entirely removed upon the addition of the A carbohydrate. It proved impossible to inactivate the bactericidal action with the deacetylated substance in equivalent proportions. In this system, the A carbohydrate was about 64 times more effective as an antibactericidal agent than the deacetylated compound. Essentially similar results were obtained in a study of the antibactericidal properties of the A carbohydrate and the deacetylated derivative in the presence of anti-Type I pneumococcus rabbit serum added to a mixture of exudative leucocytes and the defibrinated blood of the rabbit. 4. The mouse-protective titre of anti-Type I pneumococcus rabbit serum is lowered to a greater degree after absorption with the A carbohydrate than it is by similar treatment with the deacetylated compound. Absorption with the A carbohydrate does not, however, completely remove the protective antibody. 5. As Avery and Goebel have shown in the case of the acetyl polysaccharide, so the A carbohydrate, when administered in very small quantities, protects mice against an otherwise fatal dose of Type I pneumococcus. Active immunity in mice has been obtained with as little as 0.00005 mg. of the A carbohydrate administered in a single dose. Doses larger than 0.005 mg. confer no protection on these animals. Deacetylization of the A carbohydrate after boiling in N/10 sodium hydroxide destroys its protective capacity while similar treatment in N/50 alkali does not completely remove its immunizing property. Active immunity may arise within 3 days following a single injection of the A substance. It appears to be at its height from 6 to 25 days thereafter, and is retrogressive by the 49th day following vaccination. Injection of the A carbohydrate into immunized mice immediately before infection deletes the state of resistance. 6. The immunity actively induced as a result of injection of the A carbohydrate may be passively transferred to normal mice with the serum of vaccinated animals. 7. Since the evidence obtained in the course of this study indicates that the A carbohydrate of Type I pneumococcus and the acetyl polysaccharide of Avery and Goebel represent the same chemical substance, it is suggested that the designation "A carbohydrate" or "A substance" be relinquished in favor of the more exactly descriptive term "acetyl polysaccharide."

scholarly journals BLOOD FACTOR IN ACUTE DISSEMINATED LUPUS ERYTHEMATOSUS

Blood ◽  
1950 ◽  
Vol 5 (8) ◽  
pp. 718-722 ◽  
Author(s):  
JOHN R. HASERICK ◽  
LENA A. LEWIS
Keyword(s):  
Bone Marrow ◽  
Human Serum ◽  
Lupus Erythematosus ◽  
Gamma Globulin ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Supernatant Fluid ◽  
Distinct Component ◽  
Normal Human ◽  
Human Gamma Globulin

Abstract The gamma globulin fractionated by the Tiselius electrophoretic technic from the plasma of patients with acute disseminated lupus erythematosus was used as an antigen to induce antibodies in rabbits. Similarly, antibodies were induced in control rabbits with normal human serum and with normal human gamma globulin. The antisera developed in these three groups of rabbits were added to the L. E. gamma globulin solution and, after precipitation, the supernatant fluid was added to bone marrow preparations. The L. E. phenomenon was not demonstrable after the L. E. gamma globulin was precipitated by anti-L. E. gamma globulin rabbit serum. However, after precipitation of the L. E. gamma globulin by anti-normal human serum rabbit serum, or by anti-normal human gamma globulin rabbit serum, the L. E. phenomenon was still apparent. These studies suggest that the L. E. factor is an immunologically distinct component of L. E. gamma globulin.

scholarly journals The inhibitory action upon subsequent Phagocytosis, exerted on active normal serum by inactive normal serum through which Bacilli have been passed

1907 ◽  
Vol 79 (534) ◽  
pp. 482-484 ◽  
Keyword(s):  
Human Serum ◽  
Salt Solution ◽  
Inhibitory Action ◽  
Normal Serum ◽  
Normal Human Serum ◽  
The Other ◽  
Supernatant Fluid ◽  
Other Hand ◽  
Difference Of Opinion ◽  
Normal Human

Considerable difference of opinion still prevails regarding the nature of the opsonic substances present in normal serum. Wright, Bulloch and Atkin, etc., uphold the view that the opsonin of normal serum is a simple thermolabile body. Muir, on the other hand, regards the opsonin as a body which behaves like complement, while Dean holds that it is essentially thermostable and in all probability co-operates in its action with a thermolabile complement. The demonstration of anti-bodies by complement-deviation experiments (Bordet, Gengou, Pfeiffer and Friedberger, etc.) has recently proved fruitful in connection with the bacteriolysins, hæmolysins, precipitins of immune sera, and the following experiments were designed to test whether, by a similar method applied to phagocytosis, the presence in normal serum of opsonic amboceptors could be demonstrated :─ Experiment I. Normal human serum was heated for 30 mins. at 60° C. (denoted “ A ”). A very thick emulsion of tubercle bacilli in 1 : 1000 salt solution was added in equal volumes to “ A ” and kept in contact therewith for 1 hr. 30 mins. at 37° C. The mixture was then centrifugalised (7000 revolutions per minute) for 1 hr., and the supernatant fluid pipetted off (denoted “ B ”).

scholarly journals STUDIES IN THE SEROLOGY OF SYPHILIS

1932 ◽  
Vol 55 (4) ◽  
pp. 667-681 ◽  
Author(s):  
Harry Eagle
Keyword(s):  
Human Serum ◽  
Intravenous Injection ◽  
Human Heart ◽  
Colloidal Suspensions ◽  
Antigenic Property ◽  
Normal Serum ◽  
Foreign Protein ◽  
Rabbit Serum ◽  
Human Sera ◽  
Normal Human

More than one-half of normal rabbits contain complement-fixing or precipitating antibodies against Wassermann antigens (the alcohol-soluble lipoids of beef, rabbit, and human hearts) by a sufficiently sensitive technique. Normal human sera tested by the same technique are uniformly negative. The intravenous injection of colloidal suspensions of beef and human heart lipoids into rabbits occasionally causes a significant increase in this normal Wassermann (antilipoid) titre. This may indicate a certain degree of antibody response to the lipoids as such; it may be due to the presence in such extracts of traces of foreign protein, which would activate the lipoid haptene into a complete antigen; or it may be a non-specific increase in a normal antibody, not due to a specific antigenic stimulus. Confirming the results of Sachs, Klopstock, and Weil, the addition of normal foreign (human) serum to rabbit, beef, and human heart lipoids makes them antigenic for rabbits. The intravenous injection of such lipoid-serum mixtures usually causes a significant increase in the titre of the complement-fixing and precipitating antibody against tissue lipoids. The precipitate which forms upon the addition of tissue lipoids to human syphilitic serum is by far the most efficient antigen for the production, in rabbits, of antibodies to tissue lipoids which we have as yet encountered. Rabbits injected intravenously with such a precipitate regularly develop a Wassermann titre which is many times higher than either the titre observed in human syphilis, or that induced by the injection of a normal serum-lipoid mixture. The very marked antigenic property of the precipitate as compared with that of a normal serum-lipoid mixture is considered to be due to the fact that it contains a foreign protein firmly bound to the lipoid particles, namely, the human reagin-globulin with which they have combined. This interpretation is supported by the observations (1) that heating at 100°C., which does not affect the lipoid constituent of the precipitate, destroys its antigenic power for rabbits, and (2) that a similar precipitate derived from Wassermann positive rabbit serum instead of syphilitic human serum, and therefore containing tissue lipoid in combination with homologous (rabbit) protein, is completely non-antigenic for rabbits.

scholarly journals Dourine and the Complement Fixation Test

Parasitology ◽  
1915 ◽  
Vol 8 (2) ◽  
pp. 156-183 ◽  
Author(s):  
E. A. Watson
Keyword(s):  
Diagnostic Test ◽  
Research Laboratory ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Fixation Test ◽  
Veterinary Research ◽  
Fixation Reaction

This paper is written with the purpose of drawing further attention to the value of the complement fixation reaction as a diagnostic test in dourine and of recommending a method of procedure and technique arrived at with an experience of 15,000 tests for dourine made at the Veterinary Research Laboratory, Lethbridge.

scholarly journals High doses of intravenous immunoglobulin do not affect the recognition phase of the classical complement pathway

Blood ◽  
1991 ◽  
Vol 78 (3) ◽  
pp. 700-702
Author(s):  
M Basta ◽  
LF Fries ◽  
MM Frank
Keyword(s):  
Human Serum ◽  
Intravenous Immunoglobulin ◽  
Normal Serum ◽  
Normal Human Serum ◽  
Complement Pathway ◽  
Classical Complement Pathway ◽  
Recognition Phase ◽  
Normal Human ◽  
Classical Complement

We have recently found that intravenous immunoglobulin (IVIg) prevents deposition of C3 and C4 fragments onto antibody sensitized erythrocytes. To find out if such an effect results from the blockade of the recognition phase of the classical complement cascade, we investigated the ability of human serum containing high concentrations of IVIg to deposit the recognition subunit of the first complement component (C1q) onto targets. Normal human serum supplemented in vitro with IVIg did not demonstrate reduced C1q binding to targets as determined by radiolabeled antihuman C1q antibody uptake. Similarly, methylamine-treated normal human serum to which IVIg was added was equally effective in terms of C1q binding as the same serum without IVIg. At increasing doses of sensitizing antibody, C1q uptake decreased proportionally; however, at all antibody dilution points C1q uptake was not significantly different in the serum with IVIg in comparison with normal serum. Serum from a patient treated with IVIg did not differ in its capacity to deposit C1q from the same patient's serum before therapy. Our data suggest that IVIg does not interfere with the recognition step of classical complement pathway. This is a US government work. There are no restrictions on its use.

scholarly journals High doses of intravenous immunoglobulin do not affect the recognition phase of the classical complement pathway

Blood ◽  
1991 ◽  
Vol 78 (3) ◽  
pp. 700-702 ◽  
Author(s):  
M Basta ◽  
LF Fries ◽  
MM Frank
Keyword(s):  
Human Serum ◽  
Intravenous Immunoglobulin ◽  
Normal Serum ◽  
Normal Human Serum ◽  
Complement Pathway ◽  
Classical Complement Pathway ◽  
Recognition Phase ◽  
Normal Human ◽  
Classical Complement

Abstract We have recently found that intravenous immunoglobulin (IVIg) prevents deposition of C3 and C4 fragments onto antibody sensitized erythrocytes. To find out if such an effect results from the blockade of the recognition phase of the classical complement cascade, we investigated the ability of human serum containing high concentrations of IVIg to deposit the recognition subunit of the first complement component (C1q) onto targets. Normal human serum supplemented in vitro with IVIg did not demonstrate reduced C1q binding to targets as determined by radiolabeled antihuman C1q antibody uptake. Similarly, methylamine-treated normal human serum to which IVIg was added was equally effective in terms of C1q binding as the same serum without IVIg. At increasing doses of sensitizing antibody, C1q uptake decreased proportionally; however, at all antibody dilution points C1q uptake was not significantly different in the serum with IVIg in comparison with normal serum. Serum from a patient treated with IVIg did not differ in its capacity to deposit C1q from the same patient's serum before therapy. Our data suggest that IVIg does not interfere with the recognition step of classical complement pathway. This is a US government work. There are no restrictions on its use.

The C3b Component of Complement Inhibits NK Cell Activation Induced by Rituximab-Coated Target Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2505-2505 ◽  
Author(s):  
Siao-Yi Wang ◽  
Emilian Racila ◽  
Ronald P. Taylor ◽  
George J. Weiner
Keyword(s):  
Human Serum ◽  
Nk Cells ◽  
Cell Activation ◽  
Complement Fixation ◽  
Nk Cell ◽  
Normal Human Serum ◽  
Heat Inactivation ◽  
Target Cells ◽  
Normal Human ◽  
The Relationship

Abstract Rituximab (RTX) is an accepted therapy for B cell malignancies, but there is still much to learn about the mechanisms responsible for the observed responses and the potential interactions between various mechanisms of action. Some studies suggest that complement fixation followed by lysis through the membrane attack complex contributes to the anti-tumor effects of RTX. Other investigations indicate that antibody dependent cellular cytotoxicity (ADCC) mediated by NK cells is central to the response of therapy. In prior studies, we found that RTX-coated target cells activate NK cells as indicated by NK cell modulation of CD16, upregulation of CD54 and CD69, and production of IFNγ. NK activation induced by RTX-coated target cells was dependent on the affinity of multivalent interactions between Fc γ receptors III (CD16) of the NK cell and Fc regions of cell-bound RTX molecules. We used these in vitro assays to assess the relationship between complement fixation, and the ability of RTX-coated target cells to activate NK cells. Normal human serum inhibited the modulation of NK cell CD16, and also blocked upregulation of CD54, induced by RTX-coated target cells. The ability of serum to inhibit NK activation was dose dependent and was abrogated upon heat inactivation. Serum depleted of C1q or C3 also failed to inhibit NK cell activation. The inhibitory activity of serum depleted of these complement components was restored when purified C1q or C3 were added back respectively. In addition, the level of NK cell inhibition was dependent on the amount of C3b deposited on the target cells. An antibody that stabilizes C3b on the target cell surface (3E7, DiLillo et al., Molec Immunol 2006) further enhanced the inhibition of NK cell activation induced by RTX-coated target cells. One possible explanation for these findings is that complement-mediated lysis destroyed the RTX-coated target cells before they had the opportunity to induce activation of the NK cells. To assess this possibility, lymphoma cells were killed, fixed with formaldehyde, and washed prior to their use as target cells. These RTX-coated and fixed target cells were able to induce modulation of CD16 on the NK cells, which was again inhibited by normal human serum. These findings indicate that the observed inhibition of NK activation by complement is unlikely to be a consequence of complement mediated lysis of the target cells. Instead, these data suggest that C3b deposition on RTX-coated target cells inhibits the interaction between the Fc portions of RTX and CD16 on the NK cells, and so limits the ability of RTX-coated target cells to induce NK activation. These results could have implications in our understanding of the relationship between complement fixation and ADCC, and their relative roles in potentiating destruction of malignant cells in the blood and tissues.

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