John W. Osebold

Gamma globulins of bovine lacteal secretions

Abstract Gamma globulin components of bovine colostrum and dry secretion were compared with their counterparts in serum by means of immunoelectrophoresis, anion exchange chromatography, and double-diffusion antigenic analysis. Although comparable components appeared immunochemically and physicochemically similar no matter where they originated, the proportional amount of each component varied greatly according to the character of the secretion. Thus, electrophoretically fast and slow 7S gamma globulin components were found to selectively accumulate in dry secretion relative to other serum proteins. Colostrum formation involved the same discrimination relative to other serum proteins and also a virtually complete discrimination against slow 7S gamma globulin in favor of a fast component. Bovine gamma-1M and gamma-1A globulins were found in lacteal secretions.

Immunoglobulin E-Containing Cells in Mouse Lung Following Allergen Inhalation and Ozone Exposure

Cells containing immunoglobulin E (IgE) were enumerated and their location in mouse lungs was determined by direct immunofluorescence. Lungs were studied from mice that had been immunized with aerosolized ovalbumin as well as from normal mice and from mice that were exposed to ozone (0.5 or 0.8 ppm) prior to receiving aerosolized antigen. In addition, some mice were immunized intraperitoneally with ovalbumin precipitated in alum. IgE-containing cells were primarily airway-related in normal mice and in mice immunized by the intraperitoneal route. Lungs from aerosol-immunized, and aerosol-immunized ozone-exposed mice showed a more disseminated distribution of IgE-containing cells. Fluorescent cells were counted and numbers were expressed as total cells per square millimeter of lung tissue and as airway-associated cells per millimeter of airway. Total IgE cells increased 9.4-fold in mice that received aerosolized ovalbumin as compared to normal mice. When ozone exposure was added to the effects from aerosolized ovalbumin, the increase of IgE cells over normal was 34.2-fold. IgE cell counts correlated well with anaphylactic sensitivity to intravenous challenge with ovalbumin. The observed enhancement of allergic sensitization by ozone exposure has important implications for human health.

Enhancement of allergic lung sensitization in mice by ozone inhalation

Abstract Inhaled ozone was found to exert an enhancing effect for allergic lung sensitization when mice contacted an aerosolized allergen. The animals were exposed to ozone concentrations of 0.24, 0.16, 0.13, and 0.10 ppm. After 4 days of continuous ozone exposure, the mice had allergen contact from an aerosolized solution of ovalbumin. The animals were then maintained in ambient air for several days before the cycle of ozone and aerosolized allergen was repeated over four allergen contact cycles. Mice were rested in ambient air for a week after the last allergen contact, and they were then tested for allergic sensitization by the intravenous injection of 2 mg of ovalbumin to induce anaphylactic shock in allergic individuals. The control groups of mice were maintained in ambient air throughout the experiment, but they experienced identical allergen contact with the ozone-exposed mice. The phenomenon of allergic enhancement from ozone inhalation was detected at 0.24, 0.16, and 0.13 ppm of ozone. The enhancing effect disappeared at 0.10 ppm of ozone. The study indicated a potential for increasing the number of allergically sensitized individuals when various allergens are inhaled during periods of high ozone exposure with the consequent adverse changes on respiratory membranes. The significance to human health of the allergic enhancement phenomenon by ozone needs investigation.

Isolation and characterization of ovine gamma globulins

Abstract Fractions of sheep serum were obtained by anion-exchange column chromatography on DEAE Sephadex, and gel filtration on Sephadex G-200 which contained only one of the following ovine gamma globulins: 7S gamma-2, 7S gamma-1, and gamma-1M. The gamma globulins were identified by immuno-electrophoresis, starch-gel electrophoresis, analytical ultracentrifugation, and by their content of antibody activity. By these physical criteria they were identical to their counterparts in human serum. Circumstantial evidence indicated that ovine serum also contains a protein analogous to the gamma-1A globulin of human serum.

Physical heterogeneity of bovine γ-globulins: Characterization of γM and γG globulins☆

Abstract The γG globulins of bovine serum were characterized by anion exchange chromatography, immunoelectrophoresis, zone electrophoresis, ultracentrifugation, and analysis of the products of papain digestion. Their properties were similar to those of analogous components of human serum. Fast and slow γG globulins were distinguished on the basis of differences in electrophoretic migration rates, chromatographic elution positions, and biological activities (complement fixation). Antigenic differences were detectable only in immunoelectrophoretic patterns (spur formation). Bovine γM was found to have properties quite similar to those of the analogous protein in human serum as determined by the methods of gel filtration, immunoelectrophoresis, anion exchange chromatography, ultracentrifugation, and reduction with mercaptoethanol. Bovine γA was provisionally recognized in immunoelectrophoretic analyses of serum and chromatographic fractions.

An in vitro system for studying the effects of ozone on mammalian cell cultures and viruses

A unique in vitro system was developed for exposing mammalian cell cultures, viruses, or both to ozone under conditions mimicking those of the respiratory tract. The system used borosilicate glass roller culture bottles equipped with specially designed caps to permit the flow of humidified gas (ozone or filtered air) through the rotating vessels. The system was designed to allow two test cultures and one control culture to be simultaneously exposed to different precisely defined concentrations of ozone. The input and exhaust concentrations of ozone were sequentially monitored with an ultraviolet photometric ozone analyzer. The system was used to determine the reactivity of ozone with several tissue culture media at different flow rates. The reaction rate of ozone with media was shown to be a function of the input concentration and increased as the gas flow rate was increased. Input ozone concentrations measured during 48-hr test exposures remained stable, yielding standard deviations of less than 4%. Exposure of Madin-Darby bovine kidney cells to ozone concentrations of 0.16 and 0.64 ppm for 24 hr resulted in a slight but significant decrease in the synthesis of RNA as measured by the incorporation of [3H]uridine into TCA-precipitable material. The synthesis of protein and DNA was not significantly affected by identical treatments. Vesicular stomatitis virus exposed to ozone at concentrations of 0.16 and 0.64 ppm for 12 hr showed marked loss of biological function when compared to identical controls exposed to filtered air. The feasibility of using enveloped viruses as a model for investigation of ozone-induced damage to cellular membranes and membrane proteins is discussed.

Immunological alterations in the lungs of mice following ozone exposure: changes in immunoglobulin levels and antibody-containing cells.

Ozone was added to the air of the environmental chambers containing specific pathogen-free mice. At levels of 0.5 and 0.8 ppm the oxidant was seen to have inflammatory effects, as shown by rising serum albumin levels in lung lavage fluid. Fluorescein conjugated anti-heavy chain sera were used to detect cells containing IgM, IgG, and IgA in measured lung areas termed Pulmonary Units. Antigenic stimuli occurred along the airways, with significant increases of IgA-containing cells in the bronchus-associated lymphoid tissue. The numbers of IgM- and IgG-containing cells did not increase. Immunodiffusion analyses for immunoglobulins in lung lavage fluid indicated increases of IgG1, IgG2, and IgA in lung secretions. The calculation of changing Ig/Alb ratios suggested that the IgA present was largely the result of local synthesis, while IgG molecules were mainly of serum origin. Possible sources of antigenic stimuli to ozone-exposed lungs are discussed.

The biological effects of ozone on representative members of five groups of animal viruses

Abstract In an effort to establish the biological relevance of the reactions of ozone with soluble proteins and lipid bilayer membrane systems, representative viruses from five major virus groups were exposed to moderate concentrations of ozone. The virus suspensions were exposed at 37°C to 0.00, 0.16, and 0.64 ppm ozone in the gas phase. The ozone reacted with the virus suspensions as a thin film of fluid on the surface of a rotating culture bottle as the gas was drawn through the bottle at a flow rate of 2 liters/min. The three enveloped viruses tested exhibited different susceptibilities to ozone inactivation which correlated with their thermolability in the absence of ozone. The order of susceptibility to ozone inactivation of the enveloped viruses was vesicular stomatitis virus (VSV) (Rhabdoviridae) > influenza A virus (WSN strain) (Orthomyxoviridae) > infectious bovine rhinotracheitis virus (IBRV) (Herpesviridae). The inactivation reactions of the enveloped viruses with ozone showed pseudo-first-order kinetics. A simple reaction model was used to derive a reaction rate expression from which rate constants and reaction stoichiometry were estimated. In contrast to the enveloped viruses, the two nonenveloped viruses examined were relatively resistant to ozone inactivation. Polio virus type I (Picornaviridae) was found to be completely resistant to ozone inactivation after 60 hr exposure to either ozone concentration, while infectious canine hepatitis virus (Adenoviridae) showed only slight inactivation after exposure to 0.64 ppm ozone for 66 hr. The significance of these results with regard to the reactions of ozone with cell membranes and other components is discussed.

The Effects of Ozone on the Respiratory Epithelium and Alveolar Macrophages of Mice. I. Interferon Production

Summary The effects of 0.8 ppm ozone on the capacity of the tracheal epithelium and alveolar macrophages of mice to produce interferon in vitro was studied. Exposure of mice to ozone for a period of 11 days or more affected the capacity of the tracheal epithelial cells in vitro to produce interferon. The inability of the tracheal epithelium in vitro to produce interferon was not due to the inhibition in the release of intracellular interferon but to an inhibition in the production of interferon. There was a complete recovery of the ability of tracheal epithelium to respond to interferon inducers after the mice were returned to ambient air 24 days post ozone exposure. However, ozone did not seem to have any affect on the capacity of the alveolar macrophages to produce interferon in vitro.

PHYSICAL HETEROGENEITY OF BOVINE GAMMA GLOBULINS: GAMMA-1M GLOBULIN ELECTROPHORETIC HETEROGENEITY.

Summary In the bovine species, gamma-1M globulin occurs as an electrophoretic continuum with most molecules of near mean mobility. No electrophoretic sub-groups of particularly high concentration, as described for 7S gamma globulins, were noted. Complement-fixing antibody activity was associated with bovine gamma-1M globulin molecules whose mean electrophoretic mobility was slower than the mean mobility of the protein as a whole. The marginal extent of this manifestation argues against anticipation of as heterogeneous a distribution of particular biological activities within the gamma-1M globulin spectrum as has been shown to be the case with 7S gamma globulins.