scholarly journals DEMONSTRATION BY IMMUNOFLUORESCENCE OF THE FIXATION OF PERFUSED ANTIBODY TO HUMAN COLLAGEN IN HUMAN KIDNEY

1967 ◽  
Vol 125 (4) ◽  
pp. 595-606 ◽  
Author(s):  
Sidney Rothbard ◽  
Robert F. Watson
Keyword(s):  
Serum Antibody ◽  
Human Kidney ◽  
Basement Membranes ◽  
Human Infant ◽  
Laboratory Animals ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Organs ◽  
Human Collagen ◽  
Testicular Hyaluronidase

Rabbit serum containing antibody to human collagen, perfused through human infant kidneys obtained at autopsy, gives an immunofluorescent reaction with an antigen in the basement membranes of the glomeruli and the tubules. This reaction was shown to be specific by the absence of reaction with normal rabbit serum, antibody to carp collagen, or anti-human collagen serum absorbed with human collagen. Slight cross-reactions were found in the human kidneys with antibody to chicken or rat collagen. There was no evidence that a collagen-like protein in human serum or an antigen common to human erythrocytes and renal glomeruli enters into this immunofluorescent reaction. Perfusion of the kidneys with purified collagenase before the introduction of the antibody to human collagen altered the antigen so that antibody could not be demonstrated in the basement membranes by immunofluorescence. Testicular hyaluronidase used in the same way did not affect the immunofluorescent reaction. This method of perfusion provides another means for studying antigens in human organs by immunofluorescence. These observations in human kidneys extend our earlier findings in laboratory animals and indicate that an antigen, collagen, is also present in human renal glomerular and tubular basement membranes.

scholarly journals RENAL GLOMERULAR LESIONS INDUCED BY RABBIT ANTI-RAT COLLAGEN SERUM IN RATS PREPARED WITH ADJUVANT

1959 ◽  
Vol 109 (6) ◽  
pp. 633-648 ◽  
Author(s):  
Sidney Rothbard ◽  
Robert F. Watson
Keyword(s):  
Cellular Proliferation ◽  
Basement Membranes ◽  
Rabbit Serum ◽  
Renal Lesion ◽  
Normal Rabbit Serum ◽  
Glomerular Injury ◽  
Glomerular Lesion ◽  
Renal Lesions ◽  
Homologous Antigen ◽  
Glomerular Lesions

Renal glomerular lesions were induced by rabbit serum containing antibody to rat collagen injected intravenously into rats prepared with subcutaneously administered Freund adjuvant. Neither the anti-collagen serum nor the adjuvant alone induced the lesion. The lesions were characterized by diffuse glomerular injury with swelling, shredding, and fusion of the basement membranes, crescent formation, cellular proliferation, numerous multinuclear giant cells, and capillary hyaline thrombi. Various rabbit antisera, including those against fish collagen or rat serum failed to induce the renal lesion when substituted for anti-rat collagen serum. Also, anti-rat collagen serum absorbed with its homologous antigen, native rat collagen, failed to induce the lesion. Although complete adjuvant, i.e. with mycobacteria, in which normal serum was incorporated enhanced the glomerular lesion which resulted from intravenous injection of anti-collagen serum, the incomplete adjuvant without serum was sufficient. Comparison of the renal lesions induced by anti-collagen serum with nephrotoxic nephritis induced in rats by rabbit anti-kidney serum showed that they differ histologically. Also the antisera used to produce these two renal lesions differ immunologically. Antibodies to normal rabbit serum developed in rats injected intravenously with rabbit anti-rat collagen serum after preparation with adjuvant, but not when adjuvant was omitted. The pathogenesis of the renal injury is discussed as a manifestation of an antigen-antibody reaction, with nephritis occurring only after the adjuvant-stimulated antibody to the rabbit globulin has been formed in the rat and has reacted with the rabbit anti-rat collagen already fixed by its homologous antigen in the kidney.

scholarly journals ANTIGENICITY OF RAT COLLAGEN

1961 ◽  
Vol 113 (6) ◽  
pp. 1041-1052 ◽  
Author(s):  
Sidney Rothbard ◽  
Robert F. Watson
Keyword(s):  
Basement Membranes ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Glomerular Injury ◽  
Green Fluorescence ◽  
Tissue Sections ◽  
Renal Glomeruli ◽  
Homologous Antigen ◽  
Fish Collagen ◽  
Immunologic Tests

Rabbit serum or globulin, containing antibody to rat collagen, injected intravenously or intracardially into normal or adjuvant-prepared rats becomes fixed in the basement membranes of renal glomeruli and, to a slight extent, of the tubules. When examined by ultraviolet light, this antibody can be identified in tissue sections by the yellow-green fluorescence occurring where the rabbit globulin, associated with the fixed collagen antibody, has reacted with fluorescein-conjugated anti-rabbit globulin from ducks. The reaction of the antibody to rat collagen with its antigen in the kidney is a primary factor in the production of the renal glomerular injury which occurs in rats prepared with adjuvant. Adjacent control sections failed to fluoresce when pretreated with unlabeled anti-rabbit globulin or when treated with heterologous conjugated anti-duck globulin from rabbits. The antibody to rat collagen remains in the kidney as long as 92 days and has been detected as early as 45 minutes after injection. When normal rabbit serum, rabbit anti-fish collagen serum, or rabbit anti-rat collagen serum absorbed with rat collagen was substituted for the rabbit anti-rat collagen serum, fixed antibody could not be demonstrated by fluorescence; but absorption of the anti-rat collagen serum with fish collagen did not affect the antibody fixation. This series of immunologic tests indicates that the anti-collagen serum reacts with its homologous antigen, presumably collagen, in the basement membranes of renal glomeruli and tubules, and that specific antibody can be used to identify collagen in other tissues of the animal body.

scholarly journals PREPARATION AND ANTIGENICITY OF M PROTEIN RELEASED FROM GROUP A, TYPE 1 STREPTOCOCCAL CELL WALLS BY PHAGE-ASSOCIATED LYSIN

1960 ◽  
Vol 112 (1) ◽  
pp. 77-96 ◽  
Author(s):  
Fred S. Kantor ◽  
Roger M. Cole
Keyword(s):  
Cell Walls ◽  
Serum Antibody ◽  
Intradermal Injection ◽  
Exchange Chromatography ◽  
M Protein ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Bactericidal Antibody ◽  
Group A

The lysis of Group A type 1 streptococcal cell walls by phage-associated lysin has been described. In the preparation of lysin, a new semisynthetic non-protein media to support growth of the propagating strain of Group C streptococci was employed. Following lysis of the cell walls, the resulting digest was partially purified by ion-exchange chromatography and ammonium sulfate precipitation. In addition to M protein, the resulting preparation (called lysin M protein) contained the group-specific carbohydrate and the T protein—but did not contain antigens, detectable by precipitin tests, which cross-reacted with absorbed heterologous group or type antisera. Capillary precipitin reactions between the lysin M protein and type-specific antiserum did not occur in the presence of high ionic strength buffers; these buffers did not similarly affect precipitin reactions of acid M protein. Type 1 lysin M protein is shown to be a good antigen. A total of 1.5 mg. injected intradermally in saline produced bactericidal antibody in eight of nine rabbits; when injected in adjuvant in one rabbit, protective serum antibodies developed. Streptococci grown in sera from seven of nine rabbits immunized with lysin M protein demonstrated significantly longer chains than when grown in normal rabbit serum. Antibody as demonstrated by each of these three tests was shown to be type-specific. No local or systemic toxicity was noted following intradermal injection in rabbits of lysin M protein.

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.

SEM localization of cell-surface-associated fibronectin in the cranium of chick embryos utilizing immunolatex microspheres

Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 175-195
Author(s):  
Stephen Meier ◽  
Christopher Drake
Keyword(s):  
Basement Membranes ◽  
Chick Embryos ◽  
Preimmune Serum ◽  
Fluorescent Staining ◽  
Cell Surfaces ◽  
Cell Soma ◽  
Rabbit Igg ◽  
Neural Ectoderm ◽  
Cranial Neural Crest Cells ◽  
Testicular Hyaluronidase

Fibronectin has been localized to basement membranes and cell surfaces with the light microscope by fluorescent staining of thick sections, and with the TEM by immunoperoxidase reaction. However, these methods are limited because it is difficult to appreciate the patterned distribution of fibronectin from sectioned material. We have developed a probe for fibronectin that facilitates its identification with the SEM. Our probe consists of two parts; the first component is a derivatized methacrylate microsphere 90 nm in diameter, linked to purified sheep anti-rabbit IgG. The second component is anti-fibronectin IgG raised in rabbits. Stage-3 to -12 chick embryos were fixed and the ectoderm covering the cranial mesoderm was removed. Embryos were treated with testicular hyaluronidase, exposed to rabbit antifibronectin IgG and finally to sheep anti-rabbit IgG conjugated microspheres. As expected, the basal lamina of surface and neural ectoderm as well as the remaining fibrous ECM were heavily decorated with microspheres, whereas control embryos treated with preimmune serum were beadless. Fibronectin was localized on the cell soma and processes of primary mesenchyme as early as stage 3. In addition, it was possible to decorate to various extents, populations of prosencephalic, mesencephalic, and rhombencephalic cranial neural crest cells. Our studies suggest that fibronectin is present in the cranium of chick embryos at earlier times than heretofore realized, and that fibronectin accumulates in a cranial to caudal gradient that reflects the sequential differentiation of the embryonic axis.

Central administration of alpha-MSH antiserum augments fever in the rabbit

1986 ◽  
Vol 250 (5) ◽  
pp. R803-R806 ◽  
Author(s):  
S. T. Shih ◽  
O. Khorram ◽  
J. M. Lipton ◽  
S. M. McCann
Keyword(s):  
Interleukin 1 ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Central Administration ◽  
Normal Body Temperature ◽  
Melanocyte Stimulating Hormone ◽  
Average Rise ◽  
The Brain ◽  
Antipyretic Action

alpha-Melanocyte-stimulating hormone (alpha-MSH) has a marked antipyretic action when given centrally or peripherally, and the concentration of this peptide within the septal region of the brain increases during fever. To assess the significance of endogenous central alpha-MSH in fever, antiserum was given to rabbits via a cannula implanted in the third cerebral ventricle. Each day for 3 days, the animals received 50 microliters of normal rabbit serum (NRS) or an equal volume of antiserum raised against alpha-MSH. Interleukin 1 (IL 1) was then injected intravenously to determine the effect of central immunoneutralization of alpha-MSH on the febrile response. Immunoneutralization markedly prolonged fever. The average rise in temperature and the area under the fever curve after IL 1 injection were also significantly increased. Antiserum treatment did not alter normal body temperature, and NRS had no effect on IL 1-induced fever. These results indicate that endogenous central alpha-MSH contributes to physiological limitation of fever and that the role of this peptide in temperature regulation is relevant to the febrile state but not to normothermia.

Sign in / Sign up

Export Citation Format

Share Document