Immunized Dogs Research Articles

Immunological Properties of a High Molecular Weight Component from Yeast Cell Autolysate in Dogs and Evaluation of Its Potential Role in Human Dextran Reactions

A high molecular weight component (HMC) of autolysate from Saccharomyces cerevisiae yeast cells was prepared. HMC was found to be immunogenic in dogs, inducing hemagglutinating antibody formation. Upon HMC challenge of immunized dogs, systemic anaphylactoid reactions were observed in 4/5 animals. The most prominent symptom was decreased cardiac output. Decrease in mean arterial pressure and increase in pulmonary arterial pressure were also observed. Consumption of total serum complement activity amounted to 22% of initial values. HMC also exhibited mitogenic activity in lymphocyte cultures from nonimmunized and immunized dogs. Since yeast autolysate is used as nitrogen source for Leuconostoc mesenteroides in the production of clinical B 512 dextran it is a theoretically possible trace contaminant of such solutions. Therefore, dogs hyperimmunized with HMC were also challenged with clinical dextran. No anaphylactoid signs were observed. These data suggest a negligible causal role of macromolecular contaminants derived from yeast cell autolysate in rare human anaphylactoid reactions following infusion of clinical dextran.

Experimental gingivitis in immunized dogs.

The present experiment was performed to study the clinical and histopathological features of developing gingivitis in eight dogs, of which four were repeatedly immunized with plaque antigens prior to the start of a defined plaque accumulating period. The dogs were, after weaning, fed a soft diet but also twice daily subjected to meticulous tooth cleaning. When they were 10 months old and their gingiva were in excellent health, all tooth cleaning procedures were terminated and dental plaque was allowed to accumulate for 4 weeks. Clinical examinations and gingival biopsy sampling were performed during this period. The biopsies were analyzed morphometrically.In the injected dogs the levels of specific serum antibodies, as determined by an immunoradiometric assay (IRMA), were raised 3–4 fold. No antibody increase was observed in the controls. After the termination of the tooth‐cleaning period, the immunized dogs seemed to accumulate dental plaque at the same rate as non‐immunized controls. The gingiva of the immunized animals reacted to initial plaque formation with symptoms such as redness, swelling and bleeding in a manner very similar to the controls. The degree of gingival exudation and the size of the gingival connective tissue lesion was, however, less pronounced in the immunized dogs compared to the non‐immunized controls.The results indicate that raised immunity to plaque antigens may have a protective function in the dento‐gingival tissues during the development of plaque induced gingivitis.

Functional role of antibody against "core" glycolipid of Enterobacteriaceae.

Antibodies against the "core" glycolipid of Enterobacteriaceae (2-keto, 3-deoxyoctonate-Lipid A) have been associated with protection against the sequelae of gram-negative rod bacteremia. To investigate the nature of this protection, dogs and rabbits were immunized with purified glycolipid prepared by phenol-chloroform-petroleum ether extraction of the "Re" mutant of Salmonella minnesota 595 and opsonophagocytic and bactericidal tests were carried out using lapine peritoneal granulocytes and serum factors. Whereas 1-4 mug/kg of glycolipid i.v. produced hypotension in dogs and 8 mug/kg i.v. was lethal, a rising dosage schedule of immunization with an average total dose of 1 mg/kg produced striking protection against shock and death against challenge with heterologous organisms. 20 control dogs, given approximately 10(10) live, serum-resistant Escherichia coli 0.85:H9 or Serratia marcescens 03 during a continuous intra-arterial pressure transducer recording, showed a drop in mean systolic pressure from 186 (+/- 6 SE) to 101 (+/- 12 SE) MM Hg and a fall in mean diastolic pressure from 118 (+/- 3 SE) to 64 (+/- 8 SE) mm Hg within 60-120 min. Minor pressure changes (average less than 12% of prechallenge levels) were seen in the same number of immunized dogs. In contrast, no significant difference was noted in the bloodstream clearance of these serum-resistant organisms over a period of 4-6 h between immunized and control dogs. Intravascular clearance was greater in animals immunized with the challenged strain or in glycolipid-immunized animals challenged with highly serum-sensitive E. coli 0.14:K7. Antibody against core glycolipid protected against the hemodynamic sequelae of bacillemia, augmented intravascular clearance of serum-sensitive organisms, and abrogated the pyrogenic response to enteric bacilli, but did not enhance clearance of serum-resistant organisms. Although canine and lapine antiserum against core glycolipid passively protected mice against a heterologous challenge, opsonophagocytic and bactericidal activity was at least 100-fold less than type-specific antiserum.

Immunization of dogs against Echinococcus granulosus using worm secretory antigens

A new approach to the immunization of dogs against Echinococcus granulosus, using secretory antigens derived from adult tapeworms cultivated in vitro, resulted in a highly significant suppression of egg production by Echinococcus granulosus in immunized dogs. There was also a significant reduction in the number of proglottids per worm in immunized dogs. It appeared that the suppression of egg production was due to an immune effect on both somatic growth and germinal development. The implications of this work with respect to hydatidosis control programmes are discussed.

Immunoradiometric assay for quantification of serum antibodies to dental plaque antigen in immunized dogs.

An immunoradiometric assay (IRMA) was used for quantifying dog serum antibodies to antigens from dental plaque collected from full‐grown dogs. The antigens were adsorbed onto the inner surface of plastic tubes and then incubated with dog‐anti‐plaque serum. 125I‐labelled anti‐dog plasma‐immunoglobulin was used for quantification of the specific antibodies.Four 10 months old Beagle dogs in excellent gingival health were immunized for 10 weeks with ultrasonicated dog dental plaque. The anibody levels in antisera sampled 6, 8, 10 and 11 weeks after the first antigen injection were 2 to 5 times as high as those recorded before the immunizing period.The variability of the assay as judged from the difference between duplicate samples was found to be 18 %± 4 (p < 0.01) of the mean value and the variability between the same serum ran on different test occasions 13 %± 7 (p < 0.01). The specificity of the antigen‐antibody reaction in the immuno assay was tested by inhibition experiments, Preincubation of the antisera with dental plaque antigen significantly inhibited the antigen‐antibody reaction in the IRMA, while bovine serum albumin did not.

Evaluation of protective immunity against experimental Dirofilaria immitis infection in Beagle dogs

Prevention of heartworm is a key goal of pet wellness. An experimental trial was carried out to induce the immunologic protection against Dirofilaria immitis infection in dogs during the period from July 2005 to November 2006. Â To evaluate the three protocols of immunization, dogs were separately immunized with gamma-irradiated infective larvae; with chemical abbreviation of infection; with chemical abbreviation plus Freund's complete adjuvant respectively. Each trial consisted of immunized and control groups and each group composed of two dogs. All dogs used for this study were subcutaneously challenged with 100 intact third-stage larvae at designated times after the last immunization following the above three protocols. The dogs were euthanized and necropsized between 120 to 175 days after challenge infection for the worms in the right ventricle of the heart and pulmonary arteries. Number of worms and sexes were determined. A mean of 36 worms from the immunized groups with irradiated L3; 33 worms from the chemically-abbreviated group whereas 13 worms from the chemical abbreviation plus Freund's complete adjuvant group were recovered. The percentages of the average protection in three groups were 45.80%, 55.05% and 77.90% respectively. The adjuvant enhanced the protective immunity against L3 challenge infection. The ELISA values do not explain the intensity of the protection, but shown adequately the immunized dogs responding to the immunization performed in this experiment. Key words: Heartworm, protective immunity, chemical-abbreviation, gamma-irradiated larvae DOI = 10.3329/bjvm.v5i1.1323 Bangl. J. Vet. Med. (2007). 5 (1 & 2): 93-98

Aldosterone Secretion in Dogs Immunized with Hog Renin

Confirming previously reported observations, the plasma of dogs treated for prolonged periods with hog renin was found to neutralize the pressor and aldosterone-stimulating activities of hog and dog renin. The neutralizing activity was presumably due to antirenin antibodies, since the activity was found in the 7S γ-globulin fraction of the plasma. Sodium and potassium excretion and aldosterone secretion were studied in dogs with high circulating antirenin titers. These immunized dogs conserved sodium normally and increased their aldosterone secretion rate when fed a low sodium diet. However, aldosterone secretion was also increased when these dogs were hypophysectomized and subjected to constriction of the aorta just above the openings of the renal arteries. The immunized dogs were found to have high renal renin contents, the amount of renin paralleling the antirenin titer in the blood. Since there is good evidence that aortic constriction increases renin secretion, the data suggest that dogs immunized wi...

A LABILE SERUM FACTOR IN EXPERIMENTAL ENDOTOXIN SHOCK: CROSS-TRANSFUSION STUDIES IN DOGS

Peripheral vascular failure caused by endotoxin in the dog has an initial stage of vasoconstriction. Preliminary studies in vitro demonstrated that the constriction was due to the interaction of endotoxin with a heat-labile serum or plasma factor and platelets, resulting in the liberation of histamine. Further studies on the intact dog support and extend this concept. A standardized dose of Escherichia coli endotoxin produced fatal shock in control adult mongrel dogs within 28 hours. The characteristic pattern of changes included progressive hypotension, oliguria and anuria, hemoconcentration, and acidosis. Normal dogs were protected against endotoxin by transfusions of blood in which the essential serum factor was depleted in one of two ways. First, plasma separated from the blood of normal animals was heated at 56 degrees C for 30 minutes, and the infused reconstituted whole blood protected normal dogs. Protection was not afforded by unheated reconstituted blood. Second, blood from immune dogs obtained within 24 hours after a second lethal dose of endotoxin protected recipient dogs. However, protection was not demonstrated with blood collected 72 hours after a second injection of endotoxin. The nature of the serum factor essential for endotoxin activity is not known. It is postulated that an enzyme or enzyme system is involved, and the possible role of complement is discussed.

ジステンパーの中和抗体に関する実験

Distemper neutralizing antibodies in sera of various immunized dogs and guinea pigs were investigated by means of neutralization tests using the chorioallantoic membrane of the chick embryo and an egg-adapted strain of distemper virus as indicative virus. The results were as follows.1. The presence of neutralizing antibodies was hardly indicated in sera collected from three-to four-month-old puppies, but was indicated with remarkably high titers.2. Very variable titers of neutralizing antibodies were shown in sera collected from 18 dogs clinically diagnosed as distemper.3. In the case of guinea pigs inoculated intraperitoneally and of dogs inoculated subcutaneously with the egg-adapted distemper virus or a formolized tissue vaccine, neutralizing antibodies in sera of the guinea pigs indicated the highest level on the 21st day after the inoculation and those in sera of the dogs the highest level between 14 and 42 days. In addition, the production of neutralizing antibodies in sera collected from guinea pigs was in proportion to the inoculated dose of virus. The neutralizing antibody levels were exceedingly low in both dogs and guineapigs receiving formolized tissue vaccines.4. Neutralizing antibodies were found in abundance in commercial immune sera or gammaglobulin products.

On the participation of the renal tubule in the mechanism of excretion of dysentery antigens by the kidney

In 56% of cases primary introduction of dysentery antigen to unimmunized dogs was accompanted by slight excretion of antigens. Excretion of antigens by kidneys after immunization was accelerated by 5–6 times. Increased diuresis in immunized dogs increased antigen excretion correspondingly. Prior to immunization, accelerated diuresis decreased the concentration index, the antigen excretion not being pronounced. Glucose loading, instituted to maximal reabsorption, increased dysentery antigen excretion. They dysentery antigen depressed the maximal reabsorption of glucose in immunized dogs. A certain relation in connection with increased dysentery antigen excretion by kidneys in the process of immunization may be attributed to functional changes of cells of the glomerular apparatus.

Relationship of duration of phases of cardiac cycle length as determined in humans by the electrokymograph

407 (and by invitation) Philip Y. Paterson, M.D. and Elizabeth Smithwick, M.D. New Orleans, La. (From the Division of Infectious Disease, Department of Medicine, Tulane University School of Medicine.) Allergic encephalomyelitis has been produced in dogs by immunization with homologous brain extracts mixed with adjuvants. The histopathology of the disease resembles that described by others in monkeys but a longer time is required for the development of lesions and the incidence is lower. An antibody has been demonstrated in the serum of immunized dogs which reacts with an antigenic component of normal dog brain and peripheral nerve in complement fixation tests. The relationship between this antigen-antibody system and the experimental disease is not known; all dogs showing demyelination have had antibody but others without lesions have also had antibody. The antigen is demonstrable in aqueous and lipid-solvent extracts of brain. It is present in brain and nerve tissue from other species but not in other organs. It is not demonstrable in newborn dog brain tissue. It is resistant to boiling and to treatment with 10 per cent formalin. It is a component of the acetone-soluble, unsaponifiable fraction of white matter lipids and is separable from cholesterol. Specific immunologic activity may be demonstrated with less than 1 microgram of the partially purified material. Further studies bearing on the nature of this material will be presented. EFFECTS OF POSTURE AND OF COMPRESSION (OF THE NECK ON GLOMERULAR FILTRA:TION. William Viar, M.D. and Thomas Lombardo, M.D. (introduced by T. R. Harrison, M.D.). Dallas, Tex. (From the Department of Medicine, Southwestern Medical College.) The rise in sodium excretion produced by the recumbent position or by compression of the neck in the sitting position is not attended by significant change in glomerular filtration. Alterations in tubular activity must therefore be held responsible. RELATIONSHIP UF DURATION OF PHASES w CARDIAC CYCI.E LENGTH AS DETERMINED IN HUMANS BY THE ELECTROKYMOGRAPH. kathryn Willis, M.D. (introduced by Tin&y R. Harrison, M.D.). Dallas, Tex. (From the Department of Internal Medicine, Southwestern Medical College.) There is a paucity of data in the literature

Acute disseminated encephalomyelitis following immunization with homologous brain extracts; studies on the role of a circulating antibody in the production of the condition in dogs.

1. A severe demyelinating condition characterized by ataxia and paralysis, in some instances leading to death, was produced in thirty-five of a total of fifty-five dogs following immunization with homologous brain tissue combined with Freund's adjuvants. In more than 30 per cent of instances paralysis did not occur until immunization was continued for 6 or more months. Only eight dogs became paralyzed after a single injection of antigen. The condition appeared between 6 and 15 days after the last injection in all animals, irrespective of the total number of injections or the duration of immunization. 2. An antibody which reacted in complement fixation tests with aqueous and alcoholic extracts of homologous brain tissue was demonstrable in the majority of immunized dogs, whether or not the animals became paralyzed. It appeared during or after the 3rd week of immunization, and its occurrence or titer could not be correlated with the incidence of the encephalomyelitis. In general, there were fewer dogs with demonstrable antibody in the paralyzed group than in the non-paralyzed group. 3. A flocculation reaction with alcohol extracts of homologous brain was demonstrated in the serum of immunized dogs. The antigen and antibody involved were apparently identical with those responsible for the complement fixation reactions. 4. The brain tissue component which reacted as antigen in the complement fixation test was present in adult brain from several mammalian species, and peripheral nerve. It was not present in the brain of newborn dogs nor in other unrelated organs. It was demonstrable in brain tissue which had been allowed to autolyze, or treated with 10 per cent formalin. It was not impaired by boiling, or by acid hydrolysis, and was contained in the unsaponifiable fraction of brain lipids. It was separable from cholesterol by digitonin precipitation of the latter. 5. Immunization of dogs with the unsaponifiable fraction of homologous brain, in adjuvants, caused the appearance of antibrain antibody similar to that in animals injected with whole brain. Encephalomyelitis was not observed during a 2 month period of immunization with this fraction. 6. In guinea pigs, an injection of the unsaponifiable fraction of brain, in adjuvants, was followed by fatal meningoencephalitis, but the identity of the state with that caused by whole brain antigens was not established.

The Quantitative Estimation of the Removal of Bacteria from the Blood by the Various Organs of the Immunized Animal

Summary By a method of constant intravenous infusion of bacteria combined with the determination of the numbers of bacteria in the circulation at various sites by venous catheterization, further quantitative data have been obtained concerning factors influencing the removal of bacteria from the blood stream. Total canine splanchnic removal of Klebsiella pneumoniae type B averaged 42 ± 15% (S.D.). There was no significant removal of organisms in the pulmonary circulation. During a second infusion of K. pneumoniae type B given after a two to four week interval, the average splanchnic removal rose to 77 ± 11% (S.D.). The degree of removal in the pulmonary area may have been significant. The splanchnic removal of K. pneumoniae type B rose abruptly to 83 ± 16% (S.D.) following intravenous administration of homologous plasma obtained from immunized dogs. Normal dog plasma (given 1.5–3 hours prior to the immune plasma) failed to affect significantly the splanchnic removal during this period. In dogs given a K. pneumoniae type B infusion two weeks after having received plasma obtained from an immunized dog, splanchnic removal averaged 70 ± 10% (S.D.) and was not altered by administration of additional immune dog plasma.

ANTISTREPTOLYSIN CONTENT OF CEREBROSPINAL FLUID

INVESTIGATIONS on the concentration of antibodies in the cerebrospinal fluid in man and in animals have been made previously with varied results. Hektoen and Carlsson 1 found opsonins but no agglutinins or lysins in the cerebrospinal fluid of actively or passively immunized dogs. Becht and Greer 2 also failed to demonstrate lysins or agglutinins in the spinal fluid of these animals. Kafka, 3 however, found traces of hemolysins and bacterial agglutinins in the cerebrospinal fluid of immunized dogs. Starkenstein and Zitterbart 4 reported that only undiluted spinal fluid agglutinated typhoid bacilli in rabbits, although the serum titer was as high as 1: 10,000. Ransom 5 compared the titer for tetanus antitoxin in the serum with that in cerebrospinal fluid of a strongly immunized horse. He observed that the ratio in this animal was 100: 0.4. Freund 6 recorded that in 16 rabbits which he had immunized with dead typhoid bacteria

Agglutination of Antigens from Distemper-Infected Dogs and Ferrets by Anticanine-Distemper Immune Sera

Summary Organs and body fluids of dogs and ferrets experimentally infected with the virus of canine distemper contain an antigen that reacts specifically with an antibody present in the serum of immunized and hyperimmunized dogs. The antibody-antigen reaction is demonstrated by the agglutination of heat-killed suspensions of B. prodigiosus that had been in contact with the organ-extracts or the body-fluids of infected animals.

Study on reagin content of spinal fluid

A CCORDING to Merritt and Fremont-Smith,l normal cerebrospinal fluid contains none of the antibodies normally present in the blood; nor do any of the antibodies acquired as a result of vaccination or immunization by disease enter the spinal fluid through the intact choroid plexus or meninges. Kolme? likewise says that cerebrospinal fluid is free of normal or natural antibodies. However, there is evidence to show that even in the uninjured state the choroid apparatus is not entirely impermeable. according to Kolmer,2 in typhoid fever and other infections accompanied by a large amount of agglutinins in the blood, agglutinins in a lesser quantity Will be found in the cerebraspinal fluid. Nelis,3 after injecting Paratyphoid B into the blood stream of rabbits, demonstrated antibodies in the spinal fluid in low titer, when high titer was found in the blood stream. Sohier and his coworker9 recovered tetanus antitoxin in the spinal fluid of human beings passively immunized with tetanus antitoxin. These investigator9 also demonstrated antitoxin in the spinal fluid of human beings actively immunized with tetanus and diphtheria toxins. Becht and Gree9 found traces of bacterial agglutinins in spinal fluid of immunized dogs. Hektoen and Carlson injected dogs with goat blood and found tra,ces of lysins and agglutinins at the height of immunization in the spinal fluid. This combined experience clearly demonstrates that the above mentioned antibodies may traverse the choroid barrier. So far we have been dealing with antibodies which show a eomparatively ready diffusibility. The atopic reagin, however, has a strong affinity for fixed tissue and evinces a refractoriness toward passage through an anatomic membrane. Coca and Grove* have shown that local passive sensitization of the skin endures for at least four weeks. Precipitins and antitoxins readily pass the placental barrier; yet Bell and Erikssong were unable to find reagins in cord blood from atopic women showing high titers of reagins in their blood. ZohnlO made a similar investigation in artificially sensitized women. He injected prepartum women with extract of Ascaris lunzbricoides until reagins appeared in the blood. Cord blood failed to show Ascaris reagins. Spain and NewellI and Parlato12 demonstrated reagins in blister fluid

A Comparative Study of Phagocytosis and Digestion of Pneumococci by Macrophages and Polymorphonuclear Leucocytes in Normal and Immune Dogs

Summary A comparative study was made of the phagocytosis and digestion of pneumococci by macrophages and polymorphonuclear leucocytes in normal and immune dogs. Cellular exudates of the two types of cell were produced in dogs by the intrapleural injection of gum arabic on the one hand, and aleuronat on the other. Observations were carried on both in vivo and in vitro employing a highly virulent Pneumococcus type I. It was found that while the macrophages showed a somewhat greater phagocytic activity than the polymorphonuclears their ability to digest pneumococci was much more pronounced than that of the latter cells. This difference was most marked in the presence of opsonic fluids of relatively low concentration. The use of suspensions of macrophages which had been concentrated by differential sedimentation brought out the superior antipneumococcal activity of these cells most clearly. Macrophages secured from the pleural exudates of dogs highly immune to pneumococcus as well as from the lungs of dogs recovering from experimental pneumonia were found to be just as dependent on opsonins for the phagocytic activity as were cells of normal animals.

Relation of Protein Denaturization Rate to Specific Antibodies.

Horse proteins injected in routine doses intravenously into normal dogs are so completely denatured by the end of about 4 days that they no longer call forth recognizable anaphylactic reactions on massive blood transfusion into partially exsanguinated horse-serum-hypersensitive dogs.1 Representing this normal denaturization rate by the coefficient 1.0, the following approximate coefficients summarize our present data:Horse protein denaturization rate in:The rapid protein denaturization in hypersensitive dogs, therefore, is apparently due to some specific antibody which is absent or inoperative in immune dogs. This is best explained by the assumption that anaphylactic antibodies and immune antibodies are of different chemical compositions and of different physiological functions, both conceivably defensive in character.

Parenteral Denaturization of Foreign Proteins

Summary and Conclusions The rate of parenteral denaturization of horse proteins in horse serum immune dogs, therefore, is distinctly slower than the rate in normal dogs. There is not the rapid denaturization previously observed in hypersensitive dogs, all tests with hypersensitive donors being negative after the eighteenth hour (1). Nor is there the still more rapid denaturization one would predict from the unitarian conception of the relation of anaphylaxis to immunity. The biochemical properties or relationships responsible for rapid protein denaturization in hypersensitive dogs are apparently wholly absent or inoperative in immune dogs. This finding is consistent with our previous conclusion of the non-identity of specific antibodies in hypersensitive and immune dogs (3).

Relation of Anaphylaxis to Immunity

Summary and Conclusions Urinary bladders of goat serum immune dogs, therefore, show no suggestion of an acquired insusceptibility to the histamine-like substances explosively formed or liberated by the hypersensitive liver during the initial stages of horse serum shock. Horse serum immune bladders show no suggestion of an acquired insusceptibility to the hepatic anaphylatoxins of goat serum shock. This apparent specificity of the acquired fixed-tissue antianaphylatoxic insusceptibility, is in harmony with our conception that the hepatic anaphylatoxin is a specific secondary antigen, presumably of protein nature, conceivably a denaturization product of the primary protein used in sensitization or immunization.