Adult Human Serum Research Articles

Importance of complement source in bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide.

The bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide was investigated as a function of the complement source. The immunoglobulin M murine monoclonal antibody 2-2-B was shown by several different methods to be highly specific for meningococcal group B and Escherichia coli K1 capsular polysaccharides. It had strong bactericidal activity in conjunction with either rabbit or human complement, but gave a higher titer with rabbit complement. A strong prozone was observed in each case. Human postvaccination antibody to meningococcal group B polysaccharide was strongly bactericidal with rabbit complement, but had little or no bactericidal activity in conjunction with human complement. Antibodies in adult normal human sera that were bactericidal with rabbit complement were also found to be predominantly directed against the meningococcal group B capsular polysaccharide. Human antibodies that were bactericidal with human complement appeared to be primarily directed against noncapsular antigens.

Effect of lactate, pyruvate, acetone, acetoacetate, and β-hydroxybutyrate on albumin binding of bilirubin

Lactate, pyruvate, acetone, acetoacetate, and beta-hydroxybutyrate were tested for their bilirubin-displacing effect on human serum albumin. Only lactate had a significant effect at levels found in asphyxiated infants (up to 20 mM). The reserve albumin equivalent for binding bilirubin was determined, using the deputy ligand monoacetyldiaminodiphenyl sulfone (MADDS), in adult human serum albumin solution, neonatal serum, and neonatal albumin solution. Twenty mM lactate caused a 23% decrease of reserve albumin when adult albumin was used, but did not cause any change of binding when neonatal serum or neonatal albumin solution was used. It is unlikely that endogenous substances, acting as competitive ligands, cause the low binding affinity of albumin for bilirubin in sick, premature infants.

The effect of human peripheral lymph on cell growth in vitro

Peripheral lymph is similar in composition to the interstitial fluid that surrounds most cells in vivo. Gel filtration is used to show that the protein composition of such lymph is considerably different from that of plasma. Primary cell cultures fail to survive in adult human plasma or serum but grow well in adult peripheral lymph collected from the dorsum of the foot. The different effects on cell cultures may be because toxic components such as low-density lipoproteins are partially filtered out by the capillary endothelium.

Ontogeny of cortisol-cortisone interconversion in human tissues: A role for cortisone in human fetal development

While in adult human serum, cortisol (F) is present in higher concentrations than cortisone (E), the reverse has been recognized in fetal serum for two decades. To clarify the relationship between biologically active F and its inert metabolite E, the interconversion of these steroids was investigated in various tissues of the human fetus, neonate, child and adult. Tissues obtained within 48 h of death or surgery were incubated with tritiated F and E at 37°C for 2 h with and without added substrate in the absence of cofactors. Conversion of F to E was highest in placenta > fetal kidney and lung > adult testis and ovary > other fetal tissues, while that from E to F was greatest in adult liver > membranes > adult ovary and testis > term uterus. Activity in either direction was greatly decreased at the time of delivery of both mature and premature infants. It is postulated that inactivation of F to E in the hemochorial placenta and in the fetus itself protects the fetus against the growth-inhibiting effects of F, while production of F from E in the membranes and uterine wall contributes to the maintenance of the fetal allograft.

C-reactive protein as a mediator in the lysis of human erythrocytes sensitized by brown recluse spider venom.

AbstractHemolytic reactions, frequently associated with envenomization by the brown recluse spider, Loxosceles reclusa, are poorly understood. Using an in vitro model some fundamental observations on venom-induced hemolysis have been characterized which indicate that C-reactive protein could be a mediator in the natural toxic process. Washed human type O erythrocytes, when added to small amounts of brown recluse spider venom, show no direct lysis; however, when added to normal fresh, blood group-compatible adult human serum, these venom-sensitized cells are destroyed. The ability to lyse cells is complement dependent, but individual sera show quantitative differences which are not related to the levels of functional complement. Fresh human cord sera contain adequate levels of complement but are unable to lyse venom-sensitized erythrocytes. Experiments were done in which reactive fresh adult human serum was immunoabsorbed with specific rabbit antibodies to human C-reactive protein. This absorbed human seru...

Effect of serum concentration on the chromosome number of a rat liver epithelial cell line

The dispersion of the chromosome number was studied in a rat liver cell line which was grown up to the 65th passage in our standard culture medium (Ham F10 containing 10% fetal calf and 10% adult human sera) and thereafter cultured in the same chemical medium supplemented with decreasing serum concentrations. Until the serum concentration changes, the parental cell line exhibited for several passages a very wide range of chromosome numbers, from 35 up to 83. This range narrowed down to 42 when the cells were passed in a 5% fetal calf serum medium. Then, after several passages in this 5% serum medium, when subculturing again in 20% serum, a second cell population, hypotetraploid, progressively appeared until a compound population reached a 45–80 bimodal state. Another subline grown in 1% fetal calf serum exhibited similar chromosome evolution but returned to a slight aneuploid state (43 chromosomes) in media supplemented with 20% serum of different types.

Activation of the alternative complement pathway by Naegleria fowleri.

Naegleria fowleri amoebae were lysed by adult fresh human serum, and their multiplication was inhibited in culture medium supplemented with 10% fresh human serum. Heat inactivation (56 degrees C, 30 min) of serum abrogated these lytic and inhibitory effects. Absorption of human serum with amoebae failed to reduce immunoglobulin levels, and no specific antibody was detected in untreated or treated sera by counterimmunoelectrophoresis. Conversion of C3 and C3i occurred after incubation of n. fowleri with serum which had been treated with ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid, indicating activation of complement via the alternative pathway.

The role of specific antibody in alternative complement pathway-mediated opsonophagocytosis of type III, group B Streptococcus.

The native capsular polysaccharide antigen of type III, group B Streptococcus contains a terminal sialic acid residue on each repeating unit that masks all end-group galactopyranose residues and prevents alternative pathway complement activation by adult human sera in the absence of type-specific antibody. The critical role of the sialic acid residues in allowing the organism to evade activating the alternative complement pathway was shown when neuraminidase treatment of the organism converted the bacteria to activators of the alternative pathway as assessed in agammaglobulinemic serum. The requirement for specific antibody in permitting alternative pathway activation by the fully sialated bacteria was shown when sera that contained low levels of specific antibody failed to activate this pathway, and when prior absorption of serum that contained higher type-specific antibody levels with the capsular antigen failed to activate this pathway. The use of C2-deficient sera showed that the calssical pathway was not required for antibody-dependent alternative pathway activation. The use of isotonic, pH 7.5, veronal-NaCl buffer that contained 1% gelatin and that was supplemented to 4 mM Mg++ and 16 mM EGTA and adjusted to pH 7.5 (MgEGTA) ruled out the participation of the C1-bypass pathway. The presence of sialic acid on the bacterial surface is one means of evading an important mechanism of natural immunity, namely activation of complement by the alternative pathway. Only specific antibody, i.e., acquired immunity, can overcome this virulence factor.

Growth Stimulation of BHK Cells in Culture by Free Biotin in Serum<xref ref-type="fn" rid="FN2">2</xref>

A dialysate of fetal bovine serum stimulated the growth of BHK cells in a medium containing 1% dialyzed serum. The growth-stimulating activity of the dialysate was separated in a single peak with Biogel P-2 chromatography, and a single active component was further fractionated with paper and thin-layer chromatography. The component was identified as biotin. Pure biotin behaved the same as the active component in the dialysate and had the same growth-stimulating activity. The dialysate contained only the free biotin that was present in serum. Free biotin was found in relatively large amounts in fetal (bovine and human) sera, and only a very small percentage of dialysates of adult (bovine, horse, swine, and human) sera had traces of growth-stimulating activity. The possible role of free biotin in the body is discussed.

A simple microassay for detection of antibodies to fetal calf serum and related antigens and its application to the serological definition of human tumor antigens

The use of cultured cells for the production and testing of anti-tumor antisera is complicated by the fact that cultured cells can adsorb macromolecules derived from the serum supplement used in the culture medium onto their surface membrane. The need to monitor antisera for the presence of this unwanted reactivity has led us to adapt the mixed hemadsorption assay for the detection of antibodies to antigens present in fetal calf serum (FCS), the most widely used serum supplement. Our improved microassay involves the fixation of FCS to the bottom of plastic microplates with glutaraldehyde for the production of an antigen substrate which is stable for at least 16 weeks. The microassay is simple, reproducible, and capable of detecting nanogram quantities of FCS antigens. Anti-FCS reactivity is detectable with this microassay in approximately 90% of sera from both normal individuals and melanoma patients. Similarly, anti-tumor antisera raised in monkeys, rabbits and patients with melanoma against cultured human melanoma material contain high titers of anti-FCS reactivity. But this reactivity can be completely removed by absorption with insolubilized FCS. Xenoantisera raised against melanoma cells grown in human adult serum still show strong reactivity against FSC. This is due to immunologic cross-reactivity between FCS and normal human serum antigens.

Binding of diuretics to human serum albumin and to human serum from healthy adults and patients with renal failure.

It was found by in vitro equilibrium dialysis method that there is a small difference in the binding percentage of three diuretics, hydrochlorothiazide, furosemide, and ethacrynic acid, to bovine serum albumin and that to human serum albumin (between two animal species). The binding of three diuretics to human serum was examined in healthy adults and in patients with renal failure. In the healthy adults, binding of three diuretics was dependent on the degree of dilution of human serum. The same type of binding percentage curves was obtained when crystalline human serum albumin and healthy adult serum were diluted with M/15 phosphate buffer. This fact may suggest the possibility that the albumin level in human serum was closely related to the diuretics binding process. In the renal failure, the binding of hydrochlorothiazide and furosemide was correlated with the albumin concentration in human serum. However, the renal failure had reduced the binding of ethacrynic acid due to decreased serum binding capacity. Calculation of binding percentages would be required to assess the clinical importance of decreased binding of diuretics to serum of patients with renal failure.

The occurrence of human wart-virus antibodies in dogs, pigs and cattle

Using the immunodiffusion method, antibodies against human wart-virus were detected in dog, pig and cattle sera but not in horse and reindeer sera. Antibodies were found in 25 percent (28/114) of the dog sera, the prevalence of antibodies being fairly similar to that in children of the same age. By electron microscopy the antibodies in dog sera were shown to attach to and precipitate the human wart-virus particles, and in immunodiffusion the precipitation lines of human and dog sera were shown to be identical. Of the cattle sera studied, 10 percent (10/97) and of the pig sera 13 percent (5/40) gave a faint precipitation line, having identity with that given by the human sera, against human wart-virus. Conversely 18 percent (9/50) of adult human sera contained precipitating antibodies against a bovine papilloma antigen, probably bovine papilloma virus.

An Endogenous Competitor of Estrogen‐rat AFP Interaction Present in Embryo and Adult Rat and Human Sera: Demonstration and Preliminary Characterization

A highly active competitor of the binding of estrogens to rat alpha‐fetoprotein (AFP) is demonstrated in rat and human sera of various age, sex and hormonal conditions. This new high affinity physiological ligand of the estrophilic AFP is characterized as a low molecular, low polar, non‐phenolsteroid compound with negligible cross‐reactivity towards anti‐estrogen antibodies.

Ferritin: structure, biosynthesis, and role in iron metabolism.

Ferritin: structure, biosynthesis, and role in iron metabolism.

Inhibition of human complement components by Loxosceles reclusa venom.

Venom from the brown recluse spider, Loxosceles reclusa, was capable of inactivation human C1-C7 in vitro. This inactivation occurred if venom was added to fresh adult human serum, human cord serum, or functionally pure specific human components. Optimal incubation conditions for the inactivation of each component were determined and were found generally to be in the range of 25 or 37 degrees C for 30-60 min. The alternative complement pathway did not appear to be involved, since C1, C4, and C2 were readily inactivated, and inactivation took place in sera depleted of factor B of the properdin system. Venom-induced inactivation appeared to require few, if any, serum cofactors, because, with the possible exception of C2, functionally pure components, as well as those in sera, were readily inactivated.

Rat and human embryo and post-natal sera contain a potent endogenous competitor of estrogen-rat alpha-fetoprotein interactions

A highly active inhibitor of the binding of estrone and estradiol-17β to rat alpha-fetoprotein is demonstrated for the first time in embryo, immature and adult rat sera as well as in fetal and adult human sera. The competitive character and the narrow specificity of this inhibition effect is shown. The major compound responsible for this activity is isolated by successive column Sephadex LH 20 and thin layer chromatography : it is characterized as a nonpolar, nonphenolic, dialysable and thermostable substance, unreactive towards anti-estrone and anti-estradiol-17β anti-bodies. The possible biological role of an endogenous non-estrogen ligand of rodent fetoproteins is discussed.

The Mitogenic Effect of the Lymphocytosis Promoting Factor from Bordetella Pertussis on Human Lymphocytes

The purified lymphocytosis promoting factor (LPF) from Bordetella pertussis was found to be a potent mitogen for peripheral blood lymphocytes (PBL) from normal adults as well as for cord blood lymphocytes. Proliferation occurred in autologous plasma or fetal calf serum, regardless of previous exposure to pertussis infection or immunization. Only one adult human serum, from a physician constantly working with B. pertussis, inhibited the mitogenic response to LPF and this serum was shown to contain precipitating antibody against LPF. The proliferative effect of LPF was characteristic of a "nonspecific" mitogen and not of antigen stimulation of sensitized cells.LPF, phytohemagglutinin, and concanavalin A were approximately equal in potency although variation occurred depending upon the cell donor. Experiments with lymphocyte subpopulations obtained by rosetting techniques employing sheep erythrocytes, mouse erythrocytes, and sheep erythrocytes coated with antibody and complement suggested the requirement of a multicellular system for LPF mitogencity.PBL from most patients with chronic lymphatic leukemia and lymphosarcoma cell leukemia were even less responsive to LPF than to phytohemagglutinin, whereas PBL from patients with lymphosarcoma usually responded to both mitogens. It can be inferred from the results of experiments with both normal and leukemic cells that LPF, which is a murine thymus-derived (T)-cell mitogen, is also a T-cell mitogen for human PBL. The exact cell requirement and mode of action, however, are as yet unknown.

Fluorescence assay for dopamine β-hydroxylase activity in human serum by high-performance liquid chromatography

As a model for the assay of enzyme activity by high-performance liquid chromatography, a simple, sensitive and specific fluorescence assay for dopamine β-hydroxylase (DBH) activity was devised using o-phthalaldehyde as reagent. Tyramine was used as a substrate and was incubated under optimal conditions. Endogenous inhibitors in the enzyme preparation were completely inactivated by addition of N-ethylmaleimide and a small amount of Cu 2+. After isolating the amines on a small Dowex 50 column, the eluate was analyzed fluorometrically by high-performance liquid chromatography. The product, octopamine, was eluted prior to and completely separated from tyramine, and the height of the fluorescence peak gave the amount of enzymatically formed octopamine. DBH activity in serum obtained from the umbilical vein was much lower than the activity in adult human serum. Normal values of serum DBH activity from 153 subjects (1–74 years) showed a mean and standard deviation of 42.5 ± 30.9 and 34.3 ± 18.2 μmol/min/liter of serum at 37°C, before and after computation by the method of Hoffmann. Male serum had a slightly higher activity than female serum. Serum DBH activity rapidly increased during the first 10 years after birth, gradually increased up to an optimum at 40–50 years, and then decreased.

Electroradioimmunoassay: A sensitive method for the quantitation of alpha-fetoprotein

A sensitive technique, electroradioimmunoassay has been adapted for the estimation of α 1 -fetoprotein in normal adult human plasma and serum. Commercially available antisera were used and a 10-fold increase in sensitivity was obtained in comparison with two conventional radioimmunoassays. Plasma α 1 -fetoprotein levels of 2.6 ± 1.2 μg/l (mean and standard deviation) were found in a normal population as compared with levels of 4.1 ± 2.6 μg/l in a group of 49 patients with alcoholic liver disease.

Cellular and humoral components of monocyte and neutrophil chemotaxis in cord blood.

Monocyte and polymorphonuclear neutrophil (PMN) chemotaxis was studied in cord blood from healthy term infants. Monocyte chemotaxis was normal to increased (115-126%) whereas PMN chemotaxis was decreased (79%) in comparison with that of healthy adult control subjects. Generation of chemotactic factors from cord sera was impaired, being 55% of that generated by pooled normal human serum (PNHS). Cord serum was less inhibitory than pooled adult human serum for adult monocytes when the cells were suspended in 10% serum and tested for chemotaxis. No inhibition of chemotactic factors by either cord or adult sera was observed. The dissociation of chemotactic response of the two different phagocytic cells may represent a protective mechanism whereby one cell can compensate for a defect in the response of the other.