scholarly journals ELECTROPHORETIC STUDY OF ANTIVIRAL SERA

1947 ◽  
Vol 85 (5) ◽  
pp. 515-530 ◽  
Author(s):  
Hilary Koprowski ◽  
Gilbert Richmond ◽  
Dan H. Moore
Keyword(s):  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Electrophoretic Study ◽  
Neutralizing Activity ◽  
Electrophoretic Patterns ◽  
Serum Fractions ◽  
Neutralizing Capacity ◽  
Before And After ◽  
Consistent Change ◽  
Γ Globulins

1. Sera of animals immunized against Japanese B encephalitis, Venezuelan equine encephalomyelitis, and Western equine encephalomyelitis viruses were fractionated by electrophoresis. 2. Electrophoretic patterns of rabbit sera before and after immunization against Japanese B virus showed no consistent change traceable to antibody formation. 3. To determine the antibody content, the electrophoretic fractions of the respective sera were mixed in varying dilutions with infected mouse brain suspensions, and the neutralizing titers of the fractions were compared. 4. In all instances serum fractions containing γ-globulin were protective, whereas in no case did serum albumin show any virus-neutralizing activity. The Japanese B encephalitis antibody appeared to be associated entirely with the γ-globulin. The Venezuelan and Western equine encephalomyelitis antibodies were associated with the ß- and γ-globulins and probably possessed an average electrophoretic mobility between that of ß- and γ-globulins. 5. Normal rabbit serum similarly separated electrophoretically showed no neutralizing properties. 6. Chickens, whose electrophoretic serum pattern is markedly different from that of rabbits, were also immunized against the Japanese B encephalitis virus. Their antisera were electrophoretically fractionated and similarly subjected to neutralization tests. The specific neutralizing capacity of chicken serum was considerably lower than that of rabbit serum and no neutralizing activity was found in the fractions containing the faster moving components. The antibody appeared to be associated with component 4 which had a mobility of approximately 2.3 x 10–5 cm.2/volt/sec.

Factors from paraventricular nucleus mediating adrenocorticotropin release in cats

1987 ◽  
Vol 252 (1) ◽  
pp. R109-R121
Author(s):  
D. E. Carlson ◽  
D. S. Gann
Keyword(s):  
Paraventricular Nucleus ◽  
Corticotropin Releasing Factor ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Immunocytochemical Localization ◽  
Before And After ◽  
Alpha Chloralose ◽  
Crf Neurons ◽  
Caudal Area ◽  
Stimulation Of

Experiments were conducted in alpha-chloralose-urethan-anesthetized cats. We stimulated the hypothalamic paraventricular nucleus (PVH) electrically before and after intracerebroventricular (icv) injection of an antiserum to arginine vasopressin (AVP), one to corticotropin-releasing factor (CRF), or one to normal rabbit serum (NRS). Stimulation of the ventral portion of the dorsal PVH led to increases in arterial pressure and heart rate that did not change after any of the icv treatments. However, the effect of each agent on the increases in plasma adrenocorticotropin (ACTH) after stimulation was related to the area of the PVH that was stimulated. The response to stimulation of a rostral area that extended dorsally from the anterior PVH was blocked completely by the anti-AVP but not by anti-CRF or NRS. The response to stimulation of a caudal area located in the dorsal PVH was attenuated after anti-CRF, unchanged after anti-AVP, and augmented after NRS. The antibodies that were given icv were found immunocytochemically to enter the median eminence at sites that include some adjacent to the portal vessels. Immunocytochemical localization of AVP- and of CRF-containing neurons in the PVH showed that the anterior PVH had the highest proportion of AVP neurons in the PVH but had only a few CRF neurons. In contrast, the dorsal PVH contained the highest density of CRF neurons in the PVH as well as some AVP neurons. We suggest that, in the cat, the primary releasing factor for the anterior PVH is AVP and that for the dorsal PVH is CRF.

scholarly journals Fibronectin in artery subendothelium is important for platelet adhesion

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 598-604 ◽  
Author(s):  
WP Houdijk ◽  
JJ Sixma
Keyword(s):  
Shear Rate ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Fluorescein Isothiocyanate ◽  
Wall Shear Rate ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Rabbit Igg ◽  
Before And After ◽  
Wall Shear

Abstract The role of subendothelial fibronectin in platelet interaction with subendothelium was studied. Human umbilical artery subendothelium was exposed to flowing blood containing 111In-labeled platelets in an annular perfusion chamber. Platelet adhesion was determined from the 111In radioactivity on the vessel wall. When perfusions were performed for five minutes at a wall shear rate of 1,800 s-1, platelet adhesion was the same whether normal plasma or fibronectin-free plasma was used. Preincubation of subendothelium with rabbit anti-human fibronectin serum, however, resulted in a marked inhibition of platelet adhesion. Preincubation with normal rabbit serum had no effect. Platelet adhesion was also diminished when the vessel wall was preincubated with anti- fibronectin IgG fraction or F(ab')2 fragment. After the latter preincubations, frozen sections of 4 micron were incubated with fluorescein isothiocyanate-conjugated goat anti-rabbit IgG, F(ab')2 fragment specific. Fluorescence was seen throughout the subendothelium both before and after perfusion. No fluorescence was seen when subendothelium was preincubated with normal rabbit IgG or F(ab')2 or with anti-fibronectin IgG that had been absorbed with purified fibronectin. After absorption of anti-fibronectin IgG with purified fibronectin, the inhibiting effect on platelet adhesion was also no longer present. Preincubation of the vessel wall with anti-fibronectin IgG reduced platelet adhesion significantly at a wall shear rate of 800 s-1. This effect was even greater at 1,800 s-1. At low shear rate (400 s-1), there was no inhibition.

scholarly journals Role of macrophage migration inhibitory factor (MIF) in allergic and endotoxin-induced airway inflammation in mice

2000 ◽  
Vol 9 (1) ◽  
pp. 15-23 ◽  
Author(s):  
M. Korsgren ◽  
L. Källström ◽  
L. Uller ◽  
T. Bjerke ◽  
F. Sundler ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Migration Inhibitory Factor ◽  
Macrophage Migration Inhibitory Factor ◽  
Inhibitory Factor ◽  
Rabbit Serum ◽  
Chronic Obstructive ◽  
Normal Rabbit Serum ◽  
Macrophage Migration ◽  
Neutralizing Capacity

Macrophage migration inhibitory factor (MIF) has recently been forwarded as a critical regulator of inflammatory conditions, and it has been hypothesized that MIF may have a role in the pathogenesis of asthma and chronic obstructive pulmonary disease (COPD). Hence, we examined effects of MIF immunoneutralization on the development of allergen-induced eosinophilic inflammation as well as on lipopolysaccaride (LPS)-induced neutrophilic inflammation in lungs of mice. Anti-MIF serum validated with respect to MIF neutralizing capacity or normal rabbit serum (NRS) was administered i.p. repeatedly during allergen aerosol exposure of ovalbumin (OVA)-immunized mice in an established model of allergic asthma, or once before instillation of a minimal dose of LPS into the airways of mice, a tentative model of COPD. Anti-MIF treatment did not affect the induced lung tissue eosinophilia or the cellular composition of bronchoalveolar lavage fluid (BALF) in the asthma model. Likewise, anti-MIF treatment did not affect the LPS-induced neutrophilia in lung tissue, BALF, or blood, nor did it reduce BALF levels of tumor necrosis factor-α (TNF-α) and macrophage inflammatory protein–1 α (MIP–1 α). The present data suggest that MIF is not critically important for allergen-induced eosinophilic, and LPS-induced neutrophilic responses in lungs of mice. These findings do not support a role of MIF inhibition in the treatment of inflammatory respiratory diseases.

scholarly journals Fibronectin in artery subendothelium is important for platelet adhesion

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 598-604
Author(s):  
WP Houdijk ◽  
JJ Sixma
Keyword(s):  
Shear Rate ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Fluorescein Isothiocyanate ◽  
Wall Shear Rate ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Rabbit Igg ◽  
Before And After ◽  
Wall Shear

The role of subendothelial fibronectin in platelet interaction with subendothelium was studied. Human umbilical artery subendothelium was exposed to flowing blood containing 111In-labeled platelets in an annular perfusion chamber. Platelet adhesion was determined from the 111In radioactivity on the vessel wall. When perfusions were performed for five minutes at a wall shear rate of 1,800 s-1, platelet adhesion was the same whether normal plasma or fibronectin-free plasma was used. Preincubation of subendothelium with rabbit anti-human fibronectin serum, however, resulted in a marked inhibition of platelet adhesion. Preincubation with normal rabbit serum had no effect. Platelet adhesion was also diminished when the vessel wall was preincubated with anti- fibronectin IgG fraction or F(ab')2 fragment. After the latter preincubations, frozen sections of 4 micron were incubated with fluorescein isothiocyanate-conjugated goat anti-rabbit IgG, F(ab')2 fragment specific. Fluorescence was seen throughout the subendothelium both before and after perfusion. No fluorescence was seen when subendothelium was preincubated with normal rabbit IgG or F(ab')2 or with anti-fibronectin IgG that had been absorbed with purified fibronectin. After absorption of anti-fibronectin IgG with purified fibronectin, the inhibiting effect on platelet adhesion was also no longer present. Preincubation of the vessel wall with anti-fibronectin IgG reduced platelet adhesion significantly at a wall shear rate of 800 s-1. This effect was even greater at 1,800 s-1. At low shear rate (400 s-1), there was no inhibition.

scholarly journals THE EFFECTS OF VARIOUS HORSE SERUM FRACTIONS IN PRODUCING CARDIOVASCULAR AND RENAL LESIONS IN RABBITS

1949 ◽  
Vol 90 (6) ◽  
pp. 577-594 ◽  
Author(s):  
Robert W. Wissler ◽  
Katharine Smull ◽  
James B. Lesh
Keyword(s):  
Sedimentation Rate ◽  
Heart Valves ◽  
Horse Serum ◽  
Rabbit Serum ◽  
Pathological Findings ◽  
Serum Fractions ◽  
Before And After ◽  
Circulating Antigen ◽  
Acute Necrotizing ◽  
Tissue Sensitivity

Five groups of 10 rabbits each were injected intravenously 2 times at 15 day intervals with either whole horse serum or one of its cold alcohol-precipitated fractions. Suitable serological and general observations were made at appropriate intervals before and after each injection. All animals were sacrificed on the 22nd day of the experiment. A study of the antemortem and pathological findings led to the following conclusions. 1. Allergie arteritis, valvulitis, and to a lesser degree, focal pericarditis, Aschoff-like nodules, and glomerulitis can be produced by several of the cold alcohol-precipitated fractions of horse serum as well as by whole serum. 2. Most of the acute arteritis was seen in rabbits receiving fraction V (albumin). These rabbits showed the largest amounts of circulating antigen, low antibody titers, low tissue sensitivity, and slight elevation in sedimentation rate and temperature. 3. There was a high incidence of chronic arteritis in the rabbits receiving fraction III which is almost devoid of albumin, suggesting that the alpha and beta globulins in addition to albumin may produce arteritis. 4. A state most nearly resembling that of acute rheumatic fever was produced by either fractions III or IV-3,4 (alpha and beta globulins). Pancarditis (pericarditis, Aschoff-like lesions, and valvulitis) was found relatively frequently. Many of the rabbits developed a high sedimentation rate, elevated temperature, and high tissue sensitivity, but little acute arteritis was found in this group. 5. Gamma globulin (fraction II) produced little reaction either in the antemortem determinations or histopathologically. 6. Glomerulitis of an acute necrotizing type was seen in a few rabbits without particular correlation to the fraction injected. 7. The frequency of involvement of heart valves in rabbit serum disease follows a pattern very similar to that of rheumatic heart disesae. 8. Attempts to correlate antemortem observations and pathological findings either on a group basis or for individual animals failed.

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).

Localization of oxytocin and neurophysin in baboon (Papio anubis) corpus luteum by immunocytochemistry

1986 ◽  
Vol 113 (4) ◽  
pp. 570-575 ◽  
Author(s):  
Firyal S. Khan-Dawood
Keyword(s):  
Corpus Luteum ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Positive Staining ◽  
Normal Rabbit Serum ◽  
Fallopian Tubes ◽  
Papio Anubis ◽  
Luteal Cells ◽  
Luteal Tissue ◽  
Immunocytochemical Reaction

Abstract. Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and preoxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.

scholarly journals Increased cytotoxicity of normal rabbit serum for lectin-resistant mutants of animal cells.

1984 ◽  
Vol 160 (4) ◽  
pp. 1241-1246
Author(s):  
C Jones
Keyword(s):  
Structural Changes ◽  
Chinese Hamster ◽  
Rabbit Serum ◽  
Cytotoxic Action ◽  
Normal Rabbit Serum ◽  
Animal Cells ◽  
Naturally Occurring ◽  
Alternate Complement Pathway ◽  
Surface Carbohydrates ◽  
Naturally Occurring Antibodies

Plant lectins are cytotoxic and can be used to select for mutants of animal cells that exhibit structural changes in cell surface carbohydrates reflecting glycosylation defects. We isolated eight lectin mutants of Chinese hamster ovary (CHO) cells that appear to represent three different phenotype classes. These lectin mutants were much more sensitive to the cytotoxic action of normal rabbit serum (NRS) than were the parental cells. This increased cytotoxicity was heat sensitive, specifically absorbed, and inhibited by simple and complex carbohydrates. No killing was observed under conditions in which only the alternate complement pathway was active. An NRS-resistant subclone that was isolated from one lectin mutant was shown to have also regained wild type behavior when tested with the lectins. The possibility that naturally occurring antibodies in rabbit serum are reacting with incomplete carbohydrate chains on the surface of the lectin mutants is discussed.

scholarly journals THE EFFECT OF SALICYLATES ON THE PRECIPITATION OF ANTIGEN WITH ANTIBODY

1943 ◽  
Vol 77 (2) ◽  
pp. 173-183 ◽  
Author(s):  
Alvin F. Coburn ◽  
Eleanor M. Kapp
Keyword(s):  
Immune System ◽  
Sodium Salicylate ◽  
Sodium Tungstate ◽  
Horse Serum ◽  
Rabbit Antiserum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Egg Albumin ◽  
Immune Precipitation ◽  
Lyotropic Series

1. Sodium salicylate modifies the precipitation of normal rabbit serum protein by sodium tungstate, and partially inhibits the precipitation of horse serum euglobulin by rabbit antiserum. Sodium salicylate added to a system containing crystalline egg albumin and its antibody partly prevents the formation of precipitate, the degree of inhibition being related to the concentration of salicylate. 2. Precipitation in the equivalence zone is more readily prevented by salicylate than precipitation in the region of antibody excess, the immune system becoming progressively less sensitive to the action of salicylate as the excess of antibody becomes larger. 3. Formed precipitates were partly dissolved following resuspension in the presence of salicylate. 4. The salicylate effect on immune precipitation is reversible, and appears to be due to inactivation of antibody. 5. Salicylate was more effective in preventing specific precipitation than other anions of a lyotropic series tested.

scholarly journals STUDIES ON THE MECHANISM OF THE FORMATION OF THE PENICILLIN ANTIGEN

1961 ◽  
Vol 114 (6) ◽  
pp. 875-940 ◽  
Author(s):  
Bernard B. Levine ◽  
Zoltan Ovary
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Penicillin G ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Beings ◽  
Mixed Disulfide ◽  
Diastereomeric Mixture ◽  
Potassium Penicillin

An excess of D-benzylpenicillenic acid (BPE) was reacted with human γ-globulin, human serum albumin, gelatin, and poly-L-lysine in aqueous solution buffered at pH 7.5–8.0. Under these conditions, BPE reacted predominantly with lysine ϵ-amino groups of the proteins to form the mixture of diastereomers of ϵ-N-(D-α-benzylpenicilloyl)-lysine groups (Di-BPO-Lys). BPE reacted also, but to a considerably smaller extent, with cystine disulfide linkages of human γ-globulin and human serum albumin to form D-benzylpenicillenic acid-cysteine mixed disulfide groups (BPE-SS-Cys). Conjugates containing large numbers of BPE or D-penicillamine mixed disulfide groups were prepared by reaction of BPE or D-penicillamine with thiolated human γ-globulin under mild oxidizing conditions. Anti-penicillin antibodies were produced in rabbits by immunization with either potassium penicillin G (PG) or a preincubated mixture of PG with normal rabbit serum (PG-NRS) in complete Freund's adjuvant. Specific precipitation analyses in aqueous and gel media (Ouchterlony), PCA analyses, and specific inhibition of these reactions with haptens were carried out on the rabbit anti-PG and anti-(PG-NRS) sera, using the above conjugates as antigens. The anti-penicillin antibodies were found to be directed against the diastereomeric mixture of N-(D-α-benzylpenicilloyl) groups, predominantly the Di-BPO-Lys groups. By these techniques, no antibodies directed against the BPE-mixed disulfide or the D-penicillamine mixed disulfide groups were detected. Three out of six patients with histories of allergic reactions to PG responded with wheal-and-erythema reactions to the N-(D-α-benzylpenicilloyl) (BPO) groups contained in BPE-human gamma globulin conjugate. Another such patient exhibited serum antibodies specific for the BPO group. One patient being treated with 25 gm per day of PG showed the presence of non-dialyzable antigenic BPO-conjugates in his serum. These results demonstrate that the diastereomeric BPO groups (predominantly Di-BPO-Lys groups) are major antigenic determinant groups responsible for PG hypersensitivity in rabbits and human beings. The possible clinical usefulness of multivalent Di-BPO conjugates and univalent Di-BPO haptens is discussed.

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