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Immunologic cross-reactions among mammalian acute phase proteins.

Crystalline rabbit Cx-reactive protein has been compared immunologically with the analogous crystalline C-reactive protein of man. Immunologic cross-reactivity has been demonstrated between the acute phase proteins of man, rabbit, and monkey. Double-diffusion reactions in agar and passive cutaneous anaphylaxis reactions in vivo both indicate that these acute phase proteins are antigenically closely similar but not identical. Guinea pigs with delayed hypersensitivity to C-reactive protein exhibit delayed skin reactions when tested with Cx-reactive protein and vice versa.

Acute-Phase Proteins↗

Antigen-antibody reactions in allergic human tissues. I. Preparation and use of fluorescein-conjugated reagins for staining the reaction sites.

Skin sensitizing human antibodies were conjugated with various fluorescent dyes without significant loss in their ability to combine with specific antigen in vitro. A biopsy of the skin site challenged with egg albumin in a patient sensitive to this antigen could be stained specifically by the fluorescent reagins. The epithelial cells of the epidermis, sweat glands, hair follicles, and sebaceous glands in such a challenged site showed specific staining. In addition to the epithelial cells, the most intense staining was in macrophages and in pericapillary cells. The endothelium of the small blood vessels stained less intensely. Fibrous tissue bundles were specifically stained. The immunologic staining with the conjugated reagins was similar to but more intense than that obtained with conjugated rabbit anti-egg albumin globulins.

Animals↗

Biochemical study of cellular antigen-antibody reaction in tissue culture. I. Activation and release of a protease.

The correlation between morphological and biochemical changes produced by the antigen-antibody reaction was studied in cultures of tissue monocytes taken from sensitized animals. The cells were grown under conditions which allowed collection of samples from the culture fluid as well as microscopic observation. Introduction of the antigen into the culture medium causes rapid release of a protease characterized by its susceptibility to sulfhydryl block and its optimum pH in the neutral range. Protease activation occurs simultaneously with morphological changes in the cytoplasm of the cultured cells. Delayed changes affecting the mitochondria and Golgi bodies appear after the peak of the proteolytic reaction and may be secondary to it. The gradual inactivation of the protease observed in the course of the antigen-antibody reaction will be discussed in a separate paper.

Animals↗

Biochemical study of cellular antigen-antibody reaction in tissue culture. II. Release of a protease inhibitor.

Decrease in the protease activity of the culture fluid observed at later stages of the antigen-antibody reaction is believed to be due to the release of an inhibitor by the cells. The inhibitor was submitted to partial purification: it is heat-stable, non-precipitated by trichloracetic acid and non-dialyzable. It inhibits certain cellular and tissue proteases and papain but is inactive against trypsin. It is suggested that the balance between protease and anti-protease released may determine the intensity, extent, and duration of certain sensitization phenomena.

Antiemetics↗

Antigen-antibody reactions in allergic human tissues. II. Study by fluorescence technique of the localization of reagins in human skin and their relation to globulins.

The skin of atopic patients contains specific reaginic antibodies in all epidermal cells, in unchallenged as well as antigen-challenged areas. The reagins are also present in the epithelial cells of sebaceous glands, sweat glands, hair follicles and also in macrophages and pericytes. Judged by staining characteristics, their immunologic reaction in tissues with antigen is identical with that of rabbit antibodies. Comparison of tissues stained with conjugated reagins and conjugated anti-human globulin antibodies demonstrates the close relationship of reagins and globulins. In antigen-challenged tissues the macrophages and pericytes become enlarged and stain more intensely with toluidine blue. No such changes in morphology or staining are present in histamine-tested or unchallenged atopic tissues.

Antibodies, Anti-Idiotypic↗

Immunologic studies of heart tissue. IV. Serologic reactions with human heart tissue as revealed by immunofluorescent methods: isoimmune, Wassermann, and autoimmune reactions.

Reactions of normal and pathologic sera with heart tissue have been investigated by the immunofluorescent method, with particular reference to presumptive autoantibodies to heart and their differentiation from blood group isoantibodies and Wassermann antibody. In the heart, blood group substances A and B were found distributed in capillary walls, vascular endothelium, and interstitial connective tissue. In surveys of randomly selected sera, isoimmune reactions against tissue blood group substances A and B were noted infrequently. This finding was considered related to the limited sensitivity of fluorescent antibody methods. Heart tissue from blood group O individuals was used for screening of pathologic sera for presence of tissue-reactive factors. Wassermann antibody was found reactive with constituents of myocardial sarcoplasm, of which the major reactant was cardiolipin. Wassermann-positive sera absorbed with beef cardiolipin gave evidence of reaction with other constituents of myofiber sarcoplasm. Sera of patients with rheumatic fever, rheumatic heart disease, rheumatoid arthritis, disseminated lupus, and liver disease frequently showed a marked reactivity with constituents of myofiber sarcoplasm. These serum factors were differentiated from Wassermann antibody. At least three patterns of immunofluorescent staining could be differentiated by the distribution of reactants in the myofiber sarcoplasm. These reactants were extractable with ethanol and methanol but not by acetone. Sera found reactive by immunofluorescence frequently gave positive flocculation and complement-fixation tests with alcohol extracts of human heart. Immunofluorescent tests were best correlated with flocculation reactions. Sarcoplasmic-reactive factors were associated in some sera with 19S gamma globulin as demonstrated by the use of fluorescent anti-19S gamma globulin. Serologic reactions with homologous or autologous heart were observed particularly frequently with sera from rheumatic patients approximately 2 weeks following cardiac surgery, as well as in some non-rheumatic patients following cardiac or thoracic surgery or acute myocardial infarction. The pathogenetic significance of these presumptive autoantibodies to heart is unknown. As yet, no definite conclusions may be drawn regarding their relationship to bound gamma globulin in rheumatic hearts or to the postcardiotomy and post-infarction syndromes.

Antibodies↗

The analysis of streptococcal infections. VI. Immunoelectrophoretic observations on extracellular antigens detectable with human antibodies.

It has been found by immunoelectrophoresis, that Group A streptococci release at least 20 distinct extracellular antigens in human tissues, as judged by naturally occurring antibodies present in normal pooled human gamma globulin. Several of these precipitin arcs have been identified with streptococcal antigens previously purified by electrophoresis and chromatography. Human gamma globulin, as well as several rheumatic fever sera, were shown to be remarkably potent in antistreptococcal antibodies, when compared to four horse antibody concentrates obtained by hyperimmunization with several streptococcal filtrates. A Group C streptococcal culture concentrate revealed 8 or 9 antigens for which corresponding antibodies were present in human gamma globulin.

Animals↗

Immunologic studies with ethylene oxide-treated human serum.

The antigenicity of an ethylene oxide-treated human serum in humans has been studied. The immune response to the material had many of the characteristics of a delayed cellular skin reaction. Even after repeated immunizations by intradermal skin testing over a period of 1(1/2) years, no detectable antibody could be found in the sera. The antigenicity has been shown to be associated with drastic alteration of the homologous serum proteins as evidenced by (a) the formation of new proteins, and (b) the poor cross-reactions of the modified serum proteins with antisera against normal human serum albumin and normal human gamma globulin. The delayed hypersensitivity was transferable to normal recipients by either viable or killed leukocytes. The implications of these findings have been discussed with respect to the problem of sterilization of sera, the tuberculin reaction, and autoimmune phenomena.

Antiemetics↗

Antigenicity of rat collagen. Demonstration of antibody to rat collagen in the renal glomeruli of rats by fluorescence microscopy.

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.

Animals↗

The immune response of rabbits tolerant to bovine serum albumin to the injection of other heterologous serum albumins.

Immunological tolerance produced in rabbits by neonatal injections of BSA can be terminated by a series of injections of certain heterologous serum albumins which cross-react with BSA. Injections of albumins distantly related to BSA were more effective in terminating the tolerant state than injections of albumins closely related to BSA. It was concluded from results obtained with several heterologous albumins that immunological tolerance to BSA is directed to both the over-all antigenic or physical-chemical composition of the protein and the individual determinant groups present on the protein. Several possible mechanisms were given to explain the ability of cross-reacting albumins to terminate the tolerant state of BSA-tolerant rabbits. A possible relationship between the termination of tolerance in BSA-tolerant rabbits injected with cross-reacting albumins and autoimmunily was discussed. It was also suggested that the relative ease with which tolerance could be established to heterologous serum proteins in comparison to bacterial antigens is the result of the close serological and physical-chemical relationship of the heterologous serum proteins to the serum proteins of the rabbit. Rabbits injected with 500 mg of BSA during the first 5 days of life failed to form antibody capable of either eliciting an immune elimination of an injection of I* BSA given 3 to 4 months later or complexing with the circulating I* BSA.

Animals↗