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Biomedical subjects

C J van Oss

Publications and source records attributed to C J van Oss.

At least 19 recordsLinked to original sources

The effect of hydrophilicity-hydrophobicity and solubility on the immunogenicity of some natural and synthetic polymers.

The general effects of solubility and hydrophobicity-hydrophilicity on the immunogenicity of synthetic and natural polymers are presented. The degree of hydrophobicity or hydrophilicity was determined from contact angle measurements. The surface tension components, obtained from the contact angles, were then correlated with the degree of immunogenicity for each substance. The results indicate that highly hydrophobic and highly hydrophilic polymers are not immunogenic. Moderate hydrophobicity as well as moderate hydrophilicity, and solubility in water favor immunogenicity (provided the molecular mass be at least 10,000 Da). For example, the solubilization of zein (a hydrophobic insoluble maize protein) prior to immunization causes zein to become immunogenic.

Animals

The generation of antibody in mice to tuftsin: a naturally occurring phagocytosis stimulating tetrapeptide.

Tuftsin (Thr-Lys-Pro-Arg) is a naturally occurring tetrapeptide that stimulates most known functions of the polymorphonuclear leukocyte and macrophage cell lines. We previously reported our unsuccessful attempts to generate antituftsin antibodies by conjugating tuftsin to several carrier proteins and by polymerizing the peptide with glutaraldehyde. To render tuftsin antigenic the following modifications were made to native tuftsin: three glycine residues were added to the N terminus of tuftsin (Gly3-tuf) and cysteine was added to the N terminus (Cys-tuf) and to the C terminus (tuf-Cys). Native tuftsin was covalently conjugated to sheep red blood cells (SRBC). In a separate experiment Balb/c mice primed with SRBC were immunized with 10(7) SRBC peptide conjugate. Native tuftsin and Gly3-tuf were also conjugated to keyhole limpet hemocyanin (KLH). In another experiment KLH and cationized bovine serum albumin (cBSA) were activated with sulfo-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (s-SMCC), which was used to control orientation of tuf-Cys and Cys-tuf when conjugated to each carrier protein. All conjugates were administered in complete Freund's adjuvant (CFA) except for cBSA conjugates which were administered in alum. Antibody response was determined by solid phase radioimmunoassay. Results showed that specific antituftsin antibodies were elicited only by Cys-tuf, conjugated to KLH. This study reaffirms that tuftsin is weakly antigenic and confirms the previous work by Gottlieb et al. that antibody to tuftsin can only be elicited when tuftsin is conjugated to the carrier protein KLH in a manner that leaves the peptide carboxyl end free.

Amino Acid Sequence

Aspecific and specific intermolecular interactions in aqueous media.

Aspecific as well as specific interactions involve the same noncovalent forces, consisting of Lifshitz-van der Waals, Lewis acid/base, electrostatic, and thermal or Brownian movement interactions. In vivo, aspecific interactions between, e.g., cells and/or biopolymers usually are repulsive, while specific interactions are always attractive. The differences between the two classes of interactions can be shown to lie in the fact that aspecifically interacting bodies are large, while specifically interacting sites are small, or have a small radius of curvature, and in the fact that aspecifically interacting surfaces are homogeneous, whereas specific sites have a heterogeneous composition.

Binding Sites

Surface properties of fibrinogen and fibrin.

By contact angle measurements on layers of fibrinogen and fibrin, it can be shown that the transformation from fibrinogen to fibrin is accompanied by a change in surface properties from very hydrophilic (fibrinogen) to moderately but definitely hydrophobic (fibrin). It is also shown that, contrary to serum albumin and gamma globulin, fibrinogen does not become more hydrophobic upon drying.

Animals

Depletion flocculation and depletion stabilization of erythrocytes.

At dextran (Mw approximately 500,000) concentrations from 2 to approximately 10%, suspensions of normal human erythrocytes flocculate in small convex agglutinates. At dextran concentrations greater than 10%, the erythrocytes resegregate in a stable monodisperse suspension. At all these dextran concentrations, the erythrocytes are coated with considerable amounts of dextran. It can be argued that at dextran concentrations from 2 to 10%, as well as at dextran concentrations greater than 10%, there is a thin layer, which is depleted of dextran, between the dextran layer adsorbed onto the erythrocytes and the bulk dextran solution. It can also be shown that there is a repulsive interaction between the two layers of dextran: one adsorbed and one free. When the adsorbed dextran layer is the most concentrated, stability must ensue, and when the dextran in free solution is the most concentrated, flocculation should occur. Below 7% dextran, the concentration of free dextran is higher than the adsorbed concentration; above 10% dextran that situation is reversed. These data correlate well with the depletion flocculation predicted for the lower concentration and the depletion stabilization predicted for the higher dextran concentration.

Biophysical Phenomena

On the mechanism of the cold ethanol precipitation method of plasma protein fractionation.

Given the negligible difference in the value of the dielectric constant of water at 20 degrees C and that of ethanol solutions at low temperatures, the often advanced explanation for the precipitation of plasma proteins by the cold ethanol process, as being due to a reduction of the dielectric constant and the resulting increase in interprotein charge interactions, is not tenable. It is shown by a surface-thermodynamic approach that, upon dehydration by ethanol, isoelectric serum albumin molecules as well as isoelectric serum gamma globulin molecules will attract each other to a sufficient degree by van der Waals forces to become insoluble in the ethanol-water mixtures used.

Blood Proteins

Energetics of cell-cell and cell-biopolymer interactions.

The energy vs distance balance of cell suspensions (in the presence and in the absence of extracellular biopolymer solutions) is studied, not only in the light of the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory (which considered just the electrostatic (EL) and Lifshitz-van der Waals (LW) interactions), but also by taking electron-acceptor/electron-donor, or Lewis acid-base (AB) and osmotic (OS) interactions into account. Since cell surfaces, as well as many biopolymers tend to have strong monopolar electron-donor properties, they are able to engage in a strong mutual AB repulsion when immersed in a polar liquid such as water. The effects of that repulsion have been observed earlier in the guise of hydration pressure. The AB repulsion is, at close range, typically one or two orders of magnitude stronger than the EL repulsion, but its rate of decay is much steeper. In most cases, AB interactions are quantitatively the dominant factor in cell stability (when repulsive) and in "hydrophobic interactions" (when attractive). OS interactions exerted by extracellularly dissolved biopolymers are weak, but their rate of decay is very gradual, so OS repulsions engendered by biopolymer solutions may be of importance in certain long-range interactions. OS interactions exerted by biopolymers attached to cells or particles (e.g., by glycocalix glycoproteins), are very short-ranged and usually are negligibly small in comparison with the other interaction forces, in aqueous media.

Acid-Base Equilibrium

The lack of antigenicity of tuftsin: a naturally occurring phagocytosis stimulating tetrapeptide.

Tuftsin (Thr-Lys-Pro-Arg) is a naturally occurring tetrapeptide that stimulates all known functions of the polymorphonuclear leukocyte and macrophage cell lines. Tuftsin is located in the FC region of IgG between the 289 and 292 amino acid sequence of the CH2 domain. We describe unsuccessful attempts to generate antituftsin antibodies. In separate experiments tuftsin was chemically conjugated to methylated bovine serum albumin (CH3BSA), BSA, keyhole limpet hemocyanin (KLH) and purified protein derivative (PPD). Tuftsin was also polymerized with glutaraldehyde. Animals used for immunization were rabbits, roosters, and dogs. All experiments failed to produce antituftsin antibody. Probable reasons for the lack of antigenicity include: I) Lack of "foreignness" of tuftsin in mammal species. II) The small size of the tetrapeptide. III) Tuftsin may be exerting an adjuvant effect when coupled to foreign antigens and is therefore not recognized by the host immune system.

Animals

Orientation of the water molecules of hydration of human serum albumin.

Through contact-angle measurements with a number of liquids, on layers of hydrated human serum albumin (HSA), built on anisotropic ultrafilter membranes, the apolar, Lifshitz-van der Waals surface tension component, as well as the polar, electron-acceptor and electron-donor parameters of the hydrated layers could be determined. From these data, it was found that the degree of orientation of the water molecules of hydration of HSA is approximately 98% in the first layer of hydration and approximately 30% of the second layer. The water molecules of hydration are oriented with the H atoms closest to, and the O atoms farthest from, the protein surface.

Humans

Mechanisms of successive modes of erythrocyte stability and instability in the presence of various polymers.

Upon examination in real time of the adhesion of human erythrocytes by observing cells suspended by ultrasonic radiation force in solutions of dextran, polylysine, and polyethylene glycol, it was reported earlier that concave-ended cell pairs and rouleaux are seen in low (0.5-2.0% w/v) concentrations of Dextran T500. At concentrations of 5-7%, dextran spherical cell doublets and convex-ended cell agglutinates are formed. When adhesion occurs in polylysine (MW 14,000) or in polyethylene glycol (MW 8,000) only spherical cell doublets or convex-ended cell clumps occur. The final cell movement completing the formation of these adhesion products takes place over time scales of the order of 1s. In this work, quantitative consideration is given to the extent to which repulsion between adhesion-inducing macromolecules associated with the glycocalyx and those free in solution can influence adhesion through a phase separation effect. It is shown for cells in dextran and in polylysine that the forces associated with this repulsion are of the same order of magnitude as the electrostatic interactions between cells.

Cell Adhesion

Monopolar surfaces.

Following the development of a methodology for determining the apolar components as well as the electron donor and the electron acceptor parameters of the surface tension of polar surfaces, surfaces of a number of quite common materials were found to manifest virtually only electron donor properties and no, or hardly, any electron acceptor properties. Such materials may be called monopolar; they can strongly interact with bipolar materials (e.g., with polar liquids such as water); but one single polar parameter of a monopolar material cannot contribute to its energy of cohesion. Monopolar materials manifesting only electron acceptor properties also may exist, but they do not appear to occur in as great an abundance. Among the electron donor monopolar materials are: polymethylmethacrylate, polyvinylalcohol, polyethyleneglycol, proteins, many polysaccharides, phospholipids, nonionic surfactants, cellulose esters, etc. Strongly monopolar materials of the same sign repel each other when immersed or dissolved in water or other polar liquids. The interfacial tension between strongly monopolar surfaces and water has a negative value. This leads to a tendency for water to penetrate between facing surfaces of a monopolar substance and hence, to repulsion between the molecules or particles of such a monopolar material, when immersed in water, and thus to pronounced solubility or dispersibility. Monopolar repulsion energies can far outweigh Lifshitz-van der Waals attractions as well as electrostatic and "steric" repulsions. In aqueous systems the commonly observed stabilization effects, which usually are ascribed to "steric" stabilization, may in many instances be attributed to monopolar repulsion between nonionic stabilizing molecules. The repulsion between monopolar molecules of the same sign can also lead to phase separation in aqueous solutions (or suspensions), where not only two, but multiple phases are possible. Negative interfacial tensions between monopolar surfactants and the brine phase can be the driving force for the formation of microemulsions; such negative interfacial tensions ultimately decay and stabilize at a value very close to zero. Strongly monopolar macromolecules or particles surrounded by oriented water molecules of hydration can still repel each other, albeit to an attenuated degree. This repulsion was earlier perceived as caused by "hydration pressure". A few of the relevant colloid and surface phenomena are reviewed and re-examined in the light of the influence of surface monopolarity on these phenomena.

Colloids

Characterization of non-T effector cell subpopulations involved in production of human alpha interferon.

The phenotype of the human effector leukocyte subset involved in production of alpha interferon was examined using positive and negative selection techniques including murine monoclonal antibodies. The data suggest that the effector cell responsible for the elaboration of human alpha interferon is surface immunoglobulin positive, lacks either the E-rosette receptor or any monocyte determinants and is also negative for expression of the surface antigens identified by Leu-10 and Leu-11b monoclonal antibodies. Neither monocytes nor polymorphonuclear leukocytes were shown to play a role in regulation or production of alpha interferon. The data further imply, however, that the Leu-7+, large granular lymphocyte subpopulation may play a critical role in regulating human alpha interferon production by surface immunoglobulin positive B cell effectors.

Antibodies, Monoclonal

A disquisition on the energetics of immunoglobulin binding to receptors in vivo and in vitro.

The binding constant of Fc-moieties of IgG and their receptors (R), derived via the law of mass action, yields values that are of the order of 10(6) to 10(8) L/M. In circulating blood, phagocytic R must be bound rather strongly to IgG, which is normally present in high concentrations, so that it is unlikely that Fc-R mediated interactions between rather sparse sensitized particles and phagocytes take place to any significant degree in the blood stream. However, in the spleen, where Fc-R mediated interactions do play a more important role, the situation is different, due to: a) an increased cell concentration; b) a decreased relative IgG concentration; c) a locally very high macrophage concentration, with large numbers of R per cell. It can be shown that these changed conditions in the spleen cause a shift in the equilibrium of the Fc-R interaction in favor of sensitized particle Fc-R binding, with diminished involvement of freely circulating IgG. The law of mass action can also be used to predict the degree of washing of phagocytic cells needed to remove bound immunoglobulin. Conversely, measurement of the concentrations of free and bound immunoglobulin at different dilutions allows the determination of Ka as well as of the number of R per cell.

Adult

Surface thermodynamics of leukocyte and platelet adhesion to polymer surfaces.

Adhesion of leukocytes and platelets to solid substrates of different surface tensions and hence different wettability is studied from a thermodynamic point of view. A simple thermodynamic model predicts that a cellular adhesion should increase with increasing surface tension of the solid substrate if the surface tension of the medium in which the cells are suspended is lower than the surface tension of the cells. If the surface tension of the suspending medium is higher than that of the cells, the opposite behavior is predicted. These predictions are borne out completely by neutrophil adhesion tests, where the surface tension of the aqueous suspending medium is varied by addition of dimethyl sulfoxide (DMSO). Platelet adhesion experiments also confirm these predictions, the only difference being that surface tensions of the suspending medium above that of the platelets cannot be realized, owing to exudation of surface active solutes from the platelets. Utilization of the thermodynamic prediction that cellular adhesion should become independent of the surface tension of the substrate when the surface tensions of the cells and that of the suspending medium are equal leads to a value of the surface tension of neutrophils of 69.0 erg/cm(2), in excellent agreement with the value obtained from contact angles measured on layers of cells.

Cell Adhesion

Preparative cell electrophoresis with D2O as a stabilizing agent.

The method of ascending preparative cell electrophoresis in a nonstabilized vertical column can be greatly improved by layering the cells on a starting cushion of buffer prepared in D2O and by stabilizing the liquid column above it with a D2O gradient. D2O/H2O mixtures have no apparent biochemical effects on the cells, and their physiochemical effects are short-lived and reversible. It was possible, by this method, to separate a mixture of glutaraldehyde-fixed erythrocytes of three species (rabbit, horse and chicken) into three distinct zones after 15 minutes of electrophoresis. It was also possible to obtain an enriched human lymphocyte fraction containing 96% T-cells, after 1 hour of electrophoresis.

Animals

Thermodynamic studies of cellular adhesion.

Cellular adhesion of granulocytes and of platelets to solid substrates of different surface tensions has been studied from a thermodynamic aspect. A simple thermodynamic model predicts that cellular adhesion should increase as the surface tension of the solid substrate increases provided that the surface tension of the liquid medium in which the cells are suspended is lower than the surface tension of the cells themselves. If, however, the surface tension of the liquid medium is higher than the surface tension of the cells, then a decrease in cell adhesion with increasing substrate surface tension can be predicted. These predictions are completely substantiated by granulocyte adhesion tests in which the surface tension of the suspending liquid medium is varied through the addition of different volumes of dimethyl sulfoxide (DMSO). Platelet adhesion experiments also confirmed these predictions, the only difference being that it is not possible to obtain a suspending liquid medium with a surface tension higher than that of platelets themselves, as a consequence of the exudation of surface active substances by the platelets.

Blood Platelets

Simplified cell microelectrophoretic method applied to the macrophage electrophoretic mobility test for cancer diagnosis.

Simplified agarose-coated capillary microelectrophoresis was adapted to the Macrophage Electrophoretic Mobility (MEM) test for cancer detection. Guinea pig alveolar macrophages gave superior reproducibility to peritoneal macrophages. As good or better reproducibility was obtained with cryopreserved, as with fresh macrophages. The electrophoretic mobilities of patients' lymphocytes themselves, after incubation with Encephalitogenic Factor (EF) showed a significant increase in electrophoretic mobility, while in control lymphocytes a decrease occurred. Thus, for the detection of the results of the interaction of EF on human lymphocytes, a direct lymphocyte electrophoretic mobility test may suffice, and guinea pig macrophages may no longer be required at all.

Animals