scholarly journals ELECTROPHORETIC STUDIES ON ELEMENTARY BODIES OF VACCINIA

1940 ◽  
Vol 72 (5) ◽  
pp. 511-521 ◽  
Author(s):  
Theodore Shedlovsky ◽  
Joseph E. Smadel
Keyword(s):  
Moving Boundary ◽  
Serum Proteins ◽  
Vaccine Virus ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Buffer Solution ◽  
Buffer Solutions ◽  
Heat Stable ◽  
Heat Stable Antigen ◽  
Sufficient Magnitude

Electrophoretic studies were made on vaccine virus, collodion particles, and glass particles suspended in 0.01 molar buffer solutions at pH 7.9, in which the moving boundary method was used. In some experiments, uncoated particles were used; in others, particles were coated with proteins and then resuspended in the buffer solution after a washing; in still others, an excess of protein which had been used to coat the particles was included in the buffer medium. Streaming boundaries were obtained with all dilute suspensions of particles in solutions containing no soluble protein instead of the flat ones usually observed with the Tiselius moving boundary technique. This boundary artifact was suppressed by maintaining a density gradient of sufficient magnitude in association with the moving boundary to counteract the tendency of endosmotic flow. This was done partially by increasing the concentration of the particles in the suspensions, and almost completely by retaining an excess of soluble-coating substance in the solutions containing the particles. The mobility of elementary bodies of vaccinia corresponds to that found for the heat-stable (S) antigen. This value was not altered by drying, heating, ether extraction, or simple washing, but was materially increased by treatment with the surface active detergent (duponol) which presumably altered the nature of the surface of the virus particles. Collodion particles coated with the heat-stable antigen of vaccinia had the same mobility as elementary bodies under comparable conditions. Glass particles coated with normal rabbit serum moved at the rate of albumin, the fastest serum component in the buffer solutions used. However, both collodion particles and vaccine virus moved at a somewhat slower rate when they were similarly coated and measured in the presence of an excess of serum in the solutions. This was probably due to adsorption of a small amount of one of the slower components (globulin) of rabbit serum on the surface of the particles. Simple washing after treatment seemed to remove the coating of serum proteins, at least in part, from both collodion particles and elementary bodies of vaccinia.

scholarly journals ULTRACENTRIFUGATION STUDIES ON THE ELEMENTARY BODIES OF VACCINE VIRUS

1938 ◽  
Vol 68 (4) ◽  
pp. 607-627 ◽  
Author(s):  
Joseph E. Smadel ◽  
Edward G. Pickels ◽  
Theodore Shedlovsky
Keyword(s):  
Sedimentation Rate ◽  
Vaccine Virus ◽  
Normal Rate ◽  
Buffer Solution ◽  
Buffer Solutions ◽  
Original Volume ◽  
Sucrose Solutions ◽  
Rate Of Sedimentation

Ultracentrifugal studies of the CL dermal strain of vaccine virus warrant the following conclusions: 1. When suspended in increasing concentrations of sucrose, glycerol, or urea solutions, elementary bodies of vaccinia show variations in sedimentation rate which indicate changes in the density or size of the particles. For a given change in the density of the medium these changes are smallest with sucrose and most marked with urea. The normal rate of sedimentation of Paschen bodies may be restored by resuspending them in dilute buffer solution. 2. The density of elementary bodies of vaccinia suspended in dilute buffer solutions is estimated to be 1.16 gm. per cc. Higher values for the density are found if the particles are suspended in solutions containing sucrose, glycerol, or urea. In 53 per cent sucrose, for example, the density is 1.25 gm. per cc. 3. Paschen bodies appear to be quite permeable to water and urea, less so to glycerol, and only slightly, if at all, to sucrose. 4. The increased density of the elementary bodies of vaccinia in sucrose solutions may be accounted for by an osmotic extraction of water from the particles. On this basis the water which can be thus extracted corresponds to at least a third of the original volume of the particles.

Partial purification of antigens collected during in vitro cultivation of the larval stages of Taenia pisiformis

Parasitology ◽  
1976 ◽  
Vol 72 (3) ◽  
pp. 269-279 ◽  
Author(s):  
M. D. Rickard ◽  
J. C. Katiyar
Keyword(s):  
Protective Immunity ◽  
Culture Medium ◽  
Serum Proteins ◽  
Skin Tests ◽  
Rabbit Serum ◽  
Partial Purification ◽  
In Vitro Cultivation ◽  
Normal Rabbit Serum ◽  
Skin Reactions

SummaryLarvae of Taenia pisiformis were cultured in vitro in medium containing 2·5, 5 or 20% (v/v) of normal rabbit serum (NRS). Greatest development occurred in 20% NRS, and the potency of antigens collected in medium from each culture tested by intradermal (i/d) skin tests in infected rabbits paralleled the in vitro growth rate of larvae. ‘Culture’ antigens from 5% NRS stimulated good immunity in rabbits to a challenge infection with T. pisiformis eggs, although they were poorly reactive in skin tests.T. pisiformis larvae were also cultured in 10% (v/v) of nitrates of serum reduced to one-half of its volume by passage through 300 000 MW cut oif (XM300F) or 100000 MW cut off (XM100F) ultrafiltration membranes. Larvae cultured using XM300F had growth rates comparable with those cultured in 20% NRS, and the antigens released into the culture medium had equal potency in i/d tests and in stimulating protective immunity in rabbits. Larvae did not develop in XM100F orproduce skin-reactive or protective antigens.Crude ‘culture’ antigen from cultures in 20% NRS was separated into 4 fractions by nitration on Sephadex G200. All of these fractions gave i/d skin reactions in infected rabbits. Fraction 3 (F3) was the most active, but was shown by acrylamide gel electrophoresis and immunoelectrophoresis to be highly contaminated with rabbit serum proteins. F3 was separated into fractions on DEAE-Sephadex A50, and the third fraction from this was as active as the original culture medium in i/d skin tests, but had only 5% of the original protein concentration. Electrophoresis demonstrated few serum contaminants, and 2 indistinct protein bands that were not present in a similar fraction of NRS.Neither Sephadex G200 F3 nor DEAE-Sephadex F3 stimulated protective immunity in rabbits, suggesting that antigens stimulating immunity against the establishment of T. pisiformis in rabbits and those provoking cell-mediated immune reactions may be different.

scholarly journals OBSERVATIONS ON MIXTURES OF ELEMENTARY BODIES OF VACCINIA AND COATED COLLODION PARTICLES BY MEANS OF ULTRACENTRIFUGATION AND ELECTROPHORESIS

1940 ◽  
Vol 72 (5) ◽  
pp. 523-529 ◽  
Author(s):  
J. E. Smadel ◽  
E. G. Pickels ◽  
T. Shedlovsky ◽  
T. M. Rivers
Keyword(s):  
The Other ◽  
Rabbit Serum ◽  
Soluble Antigen ◽  
Normal Rabbit Serum ◽  
Electrical Field ◽  
Heat Stable ◽  
Other Hand ◽  
Single Boundary

It has been shown experimentally that mixtures of two types of particles, namely, elementary bodies of vaccinia and collodion particles coated with protein, sediment with a single boundary in the analytical centrifuge. Such mixtures have been shown to develop one or two boundaries on electrophoresis in the Tiselius apparatus, depending on the type of coating on the surface of the collodion particles. When covered with the heat-stable soluble antigen of vaccinia, collodion particles migrate in the electrical field at the same rate as elementary bodies. On the other hand, if they are coated with a component of normal rabbit serum, they migrate at a different rate. The estimation of purity of preparations of virus by means of data obtained by ultracentrifugation and electrophoresis is discussed.

scholarly journals THE CHEMOTACTIC EFFECT OF MIXTURES OF ANTIBODY AND ANTIGEN ON POLYMORPHONUCLEAR LEUCOCYTES

1962 ◽  
Vol 115 (3) ◽  
pp. 453-466 ◽  
Author(s):  
Stephen Boyden
Keyword(s):  
Chemotactic Activity ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Polymorphonuclear Leucocytes ◽  
Heat Stable ◽  
Motile Cells ◽  
Fresh Serum ◽  
In Vitro Technique ◽  
Chemotactic Effect

An in vitro technique is described for assessing the chemotactic activity of soluble substances on motile cells. Antibody-antigen mixtures when incubated (37°C) in medium containing fresh (i.e. non-inactivated) normal rabbit serum exert a strong chemotactic effect on rabbit polymorphonuclear leucocytes. Results are described which indicate that, when antibody-antigen complexes are incubated (37°C) in fresh serum, a heat-stable (56°C) substance (or substances) is produced which acts directly as a chemotactic stimulus on the polymorphs. This heat-stable chemotactic substance is not produced when antibody-antigen complexes are incubated in serum which has been heated at 56°C for 30 minutes.

scholarly journals THE EFFECT ON SUBSEQUENT AGGLUTINATION OF THE EXPOSURE OF BACTERIA TO HEATED ANTISERUM

1928 ◽  
Vol 47 (2) ◽  
pp. 245-254 ◽  
Author(s):  
F. S. Jones
Keyword(s):  
Serum Proteins ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Bacterial Cells ◽  
Hog Cholera ◽  
Serum Agglutination

It is shown that when dilute rabbit serum rich in agglutinin for the hog cholera bacillus is heated at 75°C. for 20 minutes and the bacilli incubated with the heated serum, agglutination fails to result on the addition of unheated immune serum. When the immune serum is first heated to 80°C., it no longer greatly inhibits secondary agglutination when the organisms are exposed to fresh agglutinin. The abortion bacillus agglutinin acts in a similar manner except that the immune serum must be heated above 80°C. for 20 minutes to prevent the second agglutination. The reactions are specific since control experiments with normal rabbit serum heated at various temperatures failed to influence further agglutination. It has also been shown by precipitation tests that there is definite fixation of serum proteins and bacterial cells with the heated sera which would prevent subsequent agglutination. Furthermore, heated antiserum which would prevent the secondary agglutination still possessed the property of deviating complement in a hemolytic series.

scholarly journals EFFECTS OF SERUM ON MEMBRANE TRANSPORT

1973 ◽  
Vol 137 (2) ◽  
pp. 359-368 ◽  
Author(s):  
Phyllis R. Strauss ◽  
Richard D. Berlin
Keyword(s):  
Membrane Transport ◽  
Total Radioactivity ◽  
Transport Systems ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Rabbit Lung ◽  
Heat Stable ◽  
Transport Studies ◽  
Adenosine Transport ◽  
High Concentrations

The effects of normal rabbit serum (NRS) on two transport systems in rabbit lung macrophages have been examined. A 20 min preincubation with serum was required for the effects, which were retained for at least 40 min after serum was removed. No serum was present during the transport studies. (a) Preincubation with 0.5 or 1.0% NRS resulted in depression of lysine transport to 59 ± 2.6% (SE, 31 observations) of control levels. The activity was heat stable to 100°C for 30 min and lost after dialysis. Pretreatment with serum did not alter the intracellular concentration of lysine attained when cells were then incubated with 10 mM lysine for 30 min. The relative depression of lysine transport by serum was unaltered by preloading with such high concentrations of lysine. (b) Preincubation with 5% NRS resulted in enhancement of adenosine transport by 35 ± 2.3% (SE, 60 observations). Activity was stable to heating at 65°C for 40 min but lost at 100°C for 20 min. It was nondialyzable. Total radioactivity accumulated after 30 min incubation with 1 mM adenosine was unaffected by serum pretreatment. The two activities were separable by passage over Sephadex G25.

scholarly journals THE INTERACTION IN VITRO BETWEEN GROUP B MENINGOCOCCI AND RABBIT POLYMORPHONUCLEAR LEUKOCYTES

1967 ◽  
Vol 126 (5) ◽  
pp. 795-818 ◽  
Author(s):  
Richard B. Roberts
Keyword(s):  
Bactericidal Activity ◽  
Polymorphonuclear Leukocytes ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Opsonic Activity ◽  
Heat Stable ◽  
Group B

The interaction in vitro between group B meningococci and rabbit polymorphonuclear leukocytes has been described. Phagocytosis did not occur in the presence of normal rabbit serum. Antiserum collected 12–21 days following one subcutaneous inoculation of living log phase meningococci exhibited opsonic activity with type specificity; this opsonic action depended on both heat-labile and heat-stable factors. Following ingestion by granulocytes, meningococci were rapidly killed. These studies suggest that group B meningococcal strains contain specific antiphagocytic surface factors of an as yet unknown chemical nature. Antisera obtained 4 or more wk after immunization showed bactericidal activity with the same type specificity as opsonic activity. This bactericidal activity was also lost after heating and restored by the addition of normal serum. Further studies on opsonins and bactericidins for meningococci may shed light on virulence factors in these microorganisms, and may prove useful for a more precise classification of meningococci according to type rather than group specificity.

scholarly journals INFLUENCE OF EXTRANEOUS PROTEIN AND VIRUS CONCENTRATION ON THE INACTIVATION OF THE RABBIT PAPILLOMA VIRUS BY X-RAYS

1941 ◽  
Vol 74 (5) ◽  
pp. 463-487 ◽  
Author(s):  
William F. Friedewald ◽  
Rubert S. Anderson
Keyword(s):  
Rabbit Serum ◽  
Virus Concentration ◽  
Normal Rabbit Serum ◽  
X Rays ◽  
Papilloma Virus ◽  
Egg Albumin ◽  
Buffer Solutions ◽  
Dilute Suspensions ◽  
Antiviral Antibody ◽  
Extraneous Material

The pronounced resistance to the x-rays manifested by the papilloma virus in ordinary suspensions is due to the protecting influence of extraneous matter and also in considerable degree to the amount of virus present in the preparation. Two to 4 million r were required to inactivate the virus contained in the crude papilloma extracts prepared for the present work, whereas 100,000 r or less was enough to inactivate comparable concentrations of virus after extraneous matter had been excluded by repeated differential centrifugation. The addition of normal rabbit serum or crystalline egg albumin to purified suspensions of virus was found to increase greatly the amount of irradiation required to inactivate the virus. Furthermore the percentage destruction of virus by a given amount of irradiation increases as the concentration is decreased by dilution with saline or buffer solutions. As little as 3,000 r will inactivate much of the virus in very dilute suspensions. The complement-binding antigen of papilloma virus suspensions is also inactivated by x-rays, but requires a somewhat larger amount of irradiation than necessary to destroy the infectivity of the suspensions. The effects of irradiation on the antiviral antibody present in the blood of animals which have become immune to the virus—an antibody that specifically fixes complement in mixture with the papilloma virus—are also conditioned by extraneous material. 250,000 to 500,000 r had only a slight effect on the antibody in whole serum, while this amount of irradiation completely inactivated comparable amounts of antibody in preparations partially purified by precipitation with ammonium sulfate. As a whole the findings indicate that under certain conditions of purity and concentration most of the radiation does not act by direct hits on virus or antibody particles, but indirectly by ionizing or exciting some other molecules present in the exposed suspension, which then react with the virus or antibody molecules.

NEUTRALIZATION OF ACTIVITY OF CIRCULATING GONADOTROPHIC HORMONES BY ANTISERUM TO RAT PITUITARY

1963 ◽  
Vol 25 (4) ◽  
pp. 451-456 ◽  
Author(s):  
A. N. CONTOPOULOS ◽  
T. HAYASHIDA
Keyword(s):  
Serum Proteins ◽  
Follicular Development ◽  
Inhibitory Effect ◽  
Female Rats ◽  
Male Rats ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Hormone Activity ◽  
Immature Female ◽  
Rat Pituitary

SUMMARY Antiserum prepared by the immunization of rabbits, with homogenates of rat anterior pituitary gland according to a procedure previously outlined, was absorbed with homologous serum proteins and tissues to remove various non-specific antibodies. Varying levels of plasma from gonadectomized male rats, containing high levels of gonadotrophic hormone activity, were injected into hypophysectomized, immature female rats. The simultaneous injection of antiserum resulted in complete neutralization of gonadotrophic hormone as judged by the inhibition of ovarian and uterine weight responses and the extent of follicular development in the ovaries of the rats used for the bioassay. The degree of inhibition was dependent upon the relative amount of antiserum employed. Normal rabbit serum did not have any inhibitory effect. No detectable non-specific or toxic effects were noted in the animals injected with antiserum.

scholarly journals A HEMOLYTIC SYSTEM ASSOCIATED WITH ENTERITIS IN RABBITS

1963 ◽  
Vol 117 (4) ◽  
pp. 647-661 ◽  
Author(s):  
Robert S. Evans ◽  
Margaret Bingham ◽  
Russell S. Weiser
Keyword(s):  
Red Cells ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Red Cell ◽  
Heat Stable ◽  
Heat Labile

A disease characterized by frequent association of enteritis and polyagglutinable cells often develops in weanling rabbits. The red cell lesion renders the cells susceptible to agglutination and hemolysis in normal rabbit sera. The degree of red cell abnormality varies among different animals and disappears when the animals recover. The abnormality of the red cells responsible for their polyagglutinability and susceptibility to hemolysis was resistant to the action of trypsin or papain and persisted in heated stroma preparations derived from polyagglutinable cells. The factors necessary for agglutination and hemolysis of the polyagglutinable cells are present in normal rabbit sera but are lacking in the sera of affected rabbits. These factors returned to normal levels as the polyagglutinable cell lesion disappeared. The sera of rabbits with polyagglutinable cells contained normal levels of complement and properdin. Whereas the agglutinating factor in normal sera is heat-stable at 56°C for 30 minutes, the hemolytic factor is heat labile. The hemolytic factor is apparently distinct from complement and properdin since it was adsorbed from normal rabbit serum by zymosan or by polyagglutinable cells at 0°C. However, complement was fixed when normal rabbit serum was reacted with stroma from polyagglutinable cells. Hemolysis of polyagglutinable cells by normal rabbit serum at 25°C was inhibited by preliminary incubation of the mixture at 0°C prior to incubation at 25°C. Evidence was obtained which indicated that this inhibition was due to progression of a reaction involving Ca++ independent of a reaction involving Mg++.

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