scholarly journals ON THE MECHANISM OF OPSONIN AND BACTERIOTROPIN ACTION

1929 ◽  
Vol 49 (5) ◽  
pp. 797-813 ◽  
Author(s):  
Balduin Lucké ◽  
Morton McCutcheon ◽  
Max Strumia ◽  
Stuart Mudd
Keyword(s):  
Surface Properties ◽  
Surface Potential ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Polymorphonuclear Leucocytes ◽  
Bacterial Surface ◽  
Wetting Properties ◽  
Physical Chemical ◽  
Human Sera ◽  
Immune Rabbit

The work reported in this and in previous papers (1, 7) demonstrates the following relations for acid-fast bacteria and rabbit polymorphonudear leucocytes: 1. The combination of a substance or substances present in fresh immune rabbit serum, heated or unheated, or in fresh unheated normal rabbit serum, with a substance or substances in the bacterial surface causes an increase in cohesiveness, decrease in surface potential difference and characteristic alteration in wetting properties of the bacteria, and prepares the bacteria for phagocytosis. 2. (a) The effective substance or substances in the serum may become so altered as the result of heating or aging that combination with the bacterial surface, while causing changes in bacterial surface properties indistinguishable by the present physical-chemical tests from these just mentioned, may not lead to phagocytosis, or may lead to phagocytosis with a prezone not paralleled by a prezone in the changes in surface properties. (b) Sensitization of bacteria with human sera causes changes in surface properties similar to those caused by rabbit sera, but does not lead to phagocytosis by rabbit leucocytes. The spreading requirements of rabbit polymorphonuclear leucocytes are evidently highly selective.

scholarly journals Detection of Leukocyte Antibodies by Means of I131-Labelled, Purified Anti—Human-Globulin Antibody. Problems of Nonspecific Adsorption of Globulin by Leukocytes

Blood ◽  
1960 ◽  
Vol 16 (5) ◽  
pp. 1523-1534 ◽  
Author(s):  
ROBERT E. ANDERSON ◽  
ROY L. WALFORD
Keyword(s):  
Blood Cells ◽  
Radioactive Isotope ◽  
Quantitative Method ◽  
White Blood Cells ◽  
Final Concentration ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Nonspecific Adsorption ◽  
Human Sera ◽  
Vexing Problem

Abstract Initial studies of a quantitative method for detecting sensitization of white blood cells by antileukocyte antibodies are presented. The method entails isolating I131-labelled, anti—human-globulin antibody (prepared in rabbits) by adsorption and elution from insoluble antigen, globulinazobenzylcellulose. The degree of adsorption of the labelled antibody by presumably sensitized as opposed to nonsensitized leukocytes is measured by radioactive isotope counting technics. The vexing problem of nonspecific adsorption of human or rabbit globulin by leukocytes is considered, and procedures for partially circumventing this problem are set forth. Seligmann's solution with normal rabbit serum added to a final concentration of 10 per cent appeared to be the most effective washing fluid in most instances for removing nonspecifically adsorbed globulins from leukocytes. The use of purified anti—human-globulin antibody, as opposed to whole Coombs serum, also very favorably affected the ratio between immunologically specific adsorption and nonspecific adsorption. Results of the over-all I131 method as applied to selected positive and negative human sera are detailed.

scholarly journals INTERACTIONS BETWEEN RABBIT POLYMORPHONUCLEAR LEUCOCYTES AND STAPHYLOCOCCI

1959 ◽  
Vol 110 (3) ◽  
pp. 419-443 ◽  
Author(s):  
Zanvil A. Cohn ◽  
Stephen I. Morse
Keyword(s):  
Bacterial Species ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Polymorphonuclear Leucocytes ◽  
Bacterial Inactivation ◽  
Intracellular Killing ◽  
Positive Strain ◽  
Simultaneous Evaluation ◽  
Coagulase Positive Staphylococci

A method has been described for the study in vitro of leucocyte-bacteria interactions which permits the simultaneous evaluation of both phagocytosis and intracellular bacterial inactivation. Employing this technique, the fate and localization of staphylococci in homogeneous suspensions of rabbit polymorphonuclear leucocytes have been studied. Coagulase-positive strains of S. aureus were not efficiently ingested in the presence of normal rabbit serum. In contrast, coagulase-negative strains of S. albus were rapidly engulfed and inactivated. Immune sera prepared against a coagulase-positive strain enhanced the the ingestion of the homologous organism as well as of three heterologous strains of S. aureus. Following phagocytosis, prompt intracellular killing of S. aureus occurred. The thermostable opsonins in immune sera reacted only with strains of S. aureus. A comparison between polymorphonuclear leucocytes obtained from normal and immune animals revealed no differences in their ability either to ingest or kill coagulase-positive staphylococci. Studies with other bacterial species are presented to illustrate: (a) phagocytosis followed by intracellular inactivation; (b) phagocytosis followed by intracellular survival; and (c) the absence of phagocytosis.

scholarly journals ON THE MECHANISM OF OPSONIN AND BACTERIOTROPIN ACTION

1929 ◽  
Vol 49 (5) ◽  
pp. 779-795 ◽  
Author(s):  
Stuart Mudd ◽  
Balduin Lucké ◽  
Morton McCutcheon ◽  
Max Strumia
Keyword(s):  
Electric Potential ◽  
Experimental Error ◽  
Chemical Properties ◽  
Active Immunization ◽  
Bacterial Suspension ◽  
Polymorphonuclear Leucocytes ◽  
Intrinsic Properties ◽  
Bacterial Surface ◽  
Physical Chemical ◽  
Serum Component

Methods are described for investigating the relation between phagocytosis of bacteria by polymorphonuclear leucocytes, and certain physical-chemical properties of the bacterial surface. Serum sensitization causes the following changes in the properties of acid-fast bacteria: (a) increased cohesiveness, (b) decrease in surface electric potential difference, (c) decrease in wettability of the bacteria by oil, and (d) increased phagocytosis. Tests have been conducted periodically with the sera of 4 rabbits under active immunization with as many strains of acid-fast bacteria; the parallelism between the alteration in bacterial surface properties and the promotion of phagocytosis by these sera has been, within the experimental error, complete. The percentage of phagocytosis of a given bacterial suspension has been found to depend both upon the sensitizing serum component or components deposited upon the bacterium and upon the intrinsic properties of the unsensitized bacterial surface.

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 IMMUNITY STUDIES OF ROCKY MOUNTAIN SPOTTED FEVER

1923 ◽  
Vol 38 (5) ◽  
pp. 605-626 ◽  
Author(s):  
Hideyo Noguchi
Keyword(s):  
Experimental Infection ◽  
Guinea Pigs ◽  
Spotted Fever ◽  
Rocky Mountain ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Rocky Mountain Spotted Fever ◽  
Refrigerator Temperature ◽  
Considerable Loss ◽  
Immune Rabbit

Freshly prepared mixtures of spotted fever virus and immune rabbit serum in neutral or superneutral proportions confer complete immunity on guinea pigs. The mixtures undergo a considerable loss in immunizing power when heated to 60°C. for 20 minutes, but are still capable, if used in sufficient quantity, of conferring a degree of immunity on the vaccinated animal such that a subsequent experimental infection is rendered less severe and non-fatal. Unheated mixtures which had been preserved in the refrigerator at 4°C. for a period of 32 days still retained a certain degree of immunizing property. The virus alone, or mixed with normal rabbit serum, when allowed to die out by prolonged preservation at refrigerator temperature, or when killed either by heating at 60°C. for 20 minutes or by chemicals (chloroform, ether, xylene) does not induce immunity in guinea pigs.

scholarly journals STUDIES OF PHAGOCYTOSIS OF GROUP A STREPTOCOCCI BY POLYMORPHONUCLEAR LEUCOCYTES IN VITRO

1960 ◽  
Vol 111 (3) ◽  
pp. 309-322 ◽  
Author(s):  
James G. Hirsch ◽  
Alice B. Church
Keyword(s):  
Hyaluronic Acid ◽  
M Protein ◽  
Rabbit Serum ◽  
Polymorphonuclear Leucocytes ◽  
Group A Streptococci ◽  
Test Conditions ◽  
Human Sera ◽  
Group A ◽  
Hyaluronic Acid Capsule

Studies have been made on phagocytosis and killing of Group A streptococci during mixing with suspensions of leucocytes in vitro. Under appropriate test conditions an anti-phagocytic effect can be demonstrated for the streptococcal hyaluronic acid capsule as well as for its M protein. The results obtained suggest an explanation for the suitability of human, but not rabbit, blood for opsonophagocytic tests designed to measure type-specific streptococcal antibodies. Human sera contain a factor which counteracts the anti-phagocytic effects of streptococcal hyaluronic acid capsules, and hence human blood serves well for detection of antibodies which combine with the only other phagocytosis-resisting component of this microorganism, namely M protein. In contrast, rabbit sera contain none of this factor, and addition of antibody to M protein to phagocytic test systems employing rabbit serum does not necessarily render the streptococci susceptible to engulfment by white cells, since the hyaluronic acid capsule may continue to interfere with phagocytosis. The nature of the human serum factor which opsonizes encapsulated streptococci is unknown. It does not appear to be an antibody or an enzyme capable of depolymerizing hyaluronic acid.

scholarly journals ANTIBODY FORMATION AGAINST TREPONEMA PALLIDUM—AGGLUTINATION

1915 ◽  
Vol 21 (6) ◽  
pp. 576-582 ◽  
Author(s):  
Hans Zinsser ◽  
Joseph Gardner Hopkins
Keyword(s):  
Quantitative Difference ◽  
Treponema Pallidum ◽  
Diagnostic Value ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Beings ◽  
Human Sera ◽  
Normal Human

It has been shown by our experiments that the serum of rabbits treated with emulsions of Treponema pallidum contains agglutinating substances. Normal rabbit serum also possesses agglutinating power for this organism, but, as in the case of normal bacterial agglutinins, to an extent very much inferior to that possessed by the sera of immunized animals. Normal human sera will agglutinate similar pallidum emulsions, as will the sera of certain syphilitic patients with positive Wassermann reactions. Whether or not there is a quantitative difference of diagnostic value between the sera of normal human beings and those of syphilitics remains to be seen. The sera of rabbits immunized with strain A agglutinate Noguchi's strain 9 in dilutions as high as 1 to 500. We regard as the most important result of these experiments the demonstration of definite antibodies in the circulation of animals treated with dead emulsions of Treponema pallidum. Since it is our belief that the agglutinating effect is due to an antibody essentially the same as that which produces bactericidal, precipitating, and opsonic effects, i. e., that there is probably one type of antibody only, we believe that the demonstration of agglutinins establishes the fact that in syphilis as in bacterial diseases the host responds by the formation of antibodies or sensitizers specific for the treponema. Spirocheticidal experiments with these sera, both in vitro and in vivo, are in progress.

Artifact-free electron microscopic immunocytochemistry on rat brain tissue

1991 ◽  
Vol 49 ◽  
pp. 272-273
Author(s):  
Veronika Burmeister ◽  
N. Ludvig ◽  
P.C. Jobe
Keyword(s):  
Microscopic Examination ◽  
Free Electron ◽  
Electron Microscopic Examination ◽  
Ultrastructural Localization ◽  
Rabbit Serum ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Phosphate Buffered Saline ◽  
Rat Brain Tissue ◽  
Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

Identification of Maize Rayado Fino Virus by Immunoelectron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 636-637
Author(s):  
O. E. Bradfute
Keyword(s):  
Electron Microscopy ◽  
Zea Mays ◽  
Costa Rica ◽  
Severe Disease ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Virus Particles ◽  
Far North ◽  
Maize Rayado Fino Virus ◽  
Antibody Coating

Maize rayado fino virus (MRFV) causes a severe disease of corn (Zea mays) in many locations throughout the neotropics and as far north as southern U.S. MRFV particles detected by direct electron microscopy of negatively stained sap from infected leaves are not necessarily distinguishable from many other small isometric viruses infecting plants (Fig. 1).Immunosorbent trapping of virus particles on antibody-coated grids and the antibody coating or decoration of trapped virus particles, was used to confirm the identification of MRFV. Antiserum to MRFV was supplied by R. Gamez (Centro de Investigacion en Biologia Celular y Molecular, Universidad de Costa Rica, Ciudad Universitaria, Costa Rica).Virus particles, appearing as a continuous lawn, were trapped on grids coated with MRFV antiserum (Fig. 2-4). In contrast, virus particles were infrequently found on grids not exposed to antiserum or grids coated with normal rabbit serum (similar to Fig. 1). In Fig. 3, the appearance of the virus particles (isometric morphology, 30 nm diameter, stain penetration of some particles, and morphological subunits in other particles) is characteristic of negatively stained MRFV particles. Decoration or coating of these particles with MRFV antiserum confirms their identification as MRFV (Fig. 4).

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