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.

Isolation and characterization of low molecular weight Ia-like antigens from normal human serum

1981 ◽  
Vol 18 (6) ◽  
pp. 513-519 ◽  
Author(s):  
Mauro S. Sandrin ◽  
Ian F.C. McKenzie ◽  
Terry J. Higgins ◽  
Christopher R. Parish
Keyword(s):  
Molecular Weight ◽  
Human Serum ◽  
Normal Human Serum ◽  
Low Molecular Weight ◽  
Isolation And Characterization ◽  
Normal Human

BIOCOLLOIDS IN NORMAL HUMAN URINE: II. PHYSICOCHEMICAL AND IMMUNOCHEMICAL CHARACTERISTICS

1958 ◽  
Vol 36 (11) ◽  
pp. 1167-1175 ◽  
Author(s):  
T. Webb ◽  
B. Rose ◽  
A. H. Sehon
Keyword(s):  
Molecular Weight ◽  
Normal Serum ◽  
Low Molecular Weight ◽  
Free Diffusion ◽  
Albumin Fraction ◽  
Serum Component ◽  
Normal Human ◽  
Normal Urine ◽  
Immunochemical Characteristics ◽  
Serum Components

The biocolloids of normal urine were separated by electrophoresis on starch and compared with similarly prepared fractions of serum by ultracentrifugal, free diffusion, and immunochemical techniques. The albumin fraction of urine was indistinguishable from the serum component. The urinary γ2-globulins were shown to consist of low molecular weight (10,000) fragments of the normal serum components. The other globulins of the urine were antigenically related to some of the serum components but appeared to contain lower molecular weight materials. Some of the components of normal serum could not be detected in the urine and the urine contained at least two components which were not present in the serum.

BIOCOLLOIDS IN NORMAL HUMAN URINE: II. PHYSICOCHEMICAL AND IMMUNOCHEMICAL CHARACTERISTICS

1958 ◽  
Vol 36 (1) ◽  
pp. 1167-1175 ◽  
Author(s):  
T. Webb ◽  
B. Rose ◽  
A. H. Sehon
Keyword(s):  
Molecular Weight ◽  
Normal Serum ◽  
Low Molecular Weight ◽  
Free Diffusion ◽  
Albumin Fraction ◽  
Serum Component ◽  
Normal Human ◽  
Normal Urine ◽  
Immunochemical Characteristics ◽  
Serum Components

The biocolloids of normal urine were separated by electrophoresis on starch and compared with similarly prepared fractions of serum by ultracentrifugal, free diffusion, and immunochemical techniques. The albumin fraction of urine was indistinguishable from the serum component. The urinary γ2-globulins were shown to consist of low molecular weight (10,000) fragments of the normal serum components. The other globulins of the urine were antigenically related to some of the serum components but appeared to contain lower molecular weight materials. Some of the components of normal serum could not be detected in the urine and the urine contained at least two components which were not present in the serum.

scholarly journals Complement Interaction with Trypanosomatid Promastigotes in Normal Human Serum

2002 ◽  
Vol 195 (4) ◽  
pp. 451-459 ◽  
Author(s):  
Mercedes Domínguez ◽  
Inmaculada Moreno ◽  
Margarita López-Trascasa ◽  
Alfredo Toraño
Keyword(s):  
Human Serum ◽  
Binding Capacity ◽  
Normal Human Serum ◽  
Leishmania Infection ◽  
Classical Pathway ◽  
Classical Complement Pathway ◽  
Ethylenediaminetetracetic Acid ◽  
Human Sera ◽  
Normal Human

In normal human serum (NHS), axenic promastigotes of Crithidia, Phytomonas, and Leishmania trigger complement activation, and from 1.2 to 1.8 × 105 C3 molecules are deposited per promastigote within 2.5 min. In Leishmania, promastigote C3 binding capacity remains constant during in vitro metacyclogenesis. C3 deposition on promastigotes activated through the classical complement pathway reaches a 50% maximum after ∼50 s, and represents >85% of total C3 bound. In C1q- and C2-deficient human sera, promastigotes cannot activate the classical pathway (CP) unless purified C1q or C2 factors, respectively, are supplemented, demonstrating a requirement for CP factor in promastigote C3 opsonization. NHS depleted of natural anti-Leishmania antibodies cannot trigger promastigote CP activation, but IgM addition restores C3 binding. Furthermore, Leishmania binds natural antibodies in ethylenediaminetetracetic acid (EDTA)-treated NHS; after EDTA removal, promastigote-bound IgM triggers C3 deposition in natural antibody-depleted NHS. Serum collectins and pentraxins thus do not participate significantly in NHS promastigote C3 opsonization. Real-time kinetic analysis of promastigote CP-mediated lysis indicates that between 85–95% of parasites are killed within 2.5 min of serum contact. These data indicate that successful Leishmania infection in man must immediately follow promastigote transmission, and that Leishmania evasion strategies are shaped by the selective pressure exerted by complement.

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.

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 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 ”).

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