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Biomedical subjects

L H Arp

Publications and source records attributed to L H Arp.

At least 19 recordsLinked to original sources

Reduced incidence and severity of collagen-induced arthritis in interleukin-12-deficient mice.

Collagen-induced arthritis (CIA) is an animal model for rheumatoid arthritis. The disease is elicited by immunization of genetically susceptible DBA/1 mice with type II collagen, resulting in a debilitating arthritis characterized by inflammation and involvement of multiple joints. We investigated the role of endogenous interleukin (IL)-12 in the pathogenesis of this disease by undertaking an analysis of IL-12-deficient mice on the DBA/1 genetic background after immunization with type II collagen. Both the incidence and severity of disease were significantly reduced in mice unable to produce biologically active IL-12. Concomitant decreases were observed in serum levels of pathogenic, collagen-specific IgG2a antibodies and collagen-induced secretion of interferon-gamma by immune splenocytes in vitro, consistent with an impaired T helper-1 response. There were, however, a few animals which developed severe disease in a single paw in spite of this highly diminished Th1 response. Taken together, these results demonstrate an important role for IL-12 in the pathogenesis of CIA, although it is not absolutely required for disease development.

Animals↗

Effect of passively administered immunoglobulin G on the colonization and clearance of Bordetella avium in turkeys.

This study was conducted to detect the effect of parenterally administered immunoglobulin isotype G (IgG) on the colonization and clearance of Bordetella avium at the tracheal surface in young turkeys. In two separate experiments, 3-week-old turkeys were infected with B. avium either after or before IgG administration. Comparisons were made between a control group which received an irrelevant IgG (specific for keyhole limpet hemocyanin [KLH]) and the experimental group which received a B. avium-specific IgG. When given before the bacteria, IgG reduced the numbers of colony-forming units (CFUs) in the trachea. As a supplement to non-specific respiratory defense mechanisms, B. avium-specific IgG also appears to inhibit colonization of the tracheal mucosa. In a second experiment designed to study the role of IgG in bacterial clearance, administration of B. avium-specific IgG after bacterial inoculation significantly reduced the number of CFUs on the tracheal surface. These studies support the role of B. avium-specific IgG in resistance to and recovery from B. avium infection.

Animals↗

Uptake of ferritin and Bordetella avium in bronchus-associated lymphoid tissue of turkeys.

The uptake of macromolecular and particulate materials in bronchus-associated lymphoid tissue (BALT) in turkeys was examined using transmission electron microscopy. Tracer materials used were live and ultraviolet-killed (UV-killed) Bordetella avium and ferritin. Suspensions of bacteria and ferritin were instilled via intratracheal catheterization and allowed to remain in contact with the respiratory surfaces for 0, 10, 30, 60, 90, and 120 min. Ferritin and B. avium were taken up by both ciliated and non-ciliated cells of the epithelium overlying BALT (BALT epithelium). Ferritin was found in organelles associated with endocytosis (i.e. apical vesicles, endosomes, cytoplasmic vacuoles) and was apparently transported across epithelial cells, since it was also found in intercellular spaces. Bacteria were found in vacuoles within BALT epithelial cells, but not free in intercellular spaces. Some macrophages in BALT epithelium also contained bacteria. No differences were observed between uptake of live and UV-killed bacteria. We conclude that both ciliated and non-ciliated cells of BALT epithelium in turkeys are able to take up macromolecular and particulate materials. Bacteria are also accessible to intraepithelial macrophages, although whether they are taken up directly from the bronchial surface or whether they pass through epithelial cells first could not be determined. This evidence suggests that antigens, including respiratory pathogens, could gain access to cells of the avian immune system by transepithelial passage in BALT.

Animals↗

Canine distemper epizootic in lions, tigers, and leopards in North America.

Canine distemper virus (CDV) infection occurred in captive leopards (Panthera pardus), tigers (Panthera tigris), lions (Panthera leo), and a jaguar (Panthera onca) in 1991 and 1992. An epizootic affected all 4 types of cats at the Wildlife Waystation, San Fernando, California, with 17 mortalities. CDV-infected raccoons were thought to be the source of infection in these cats. Two black leopards died at the Naibi Zoo, Coal Valley, Illinois, and 2 tigers died at the Shambala Preserve, Acton, California. Initial clinical signs were anorexia with gastrointestinal and/or respiratory disease followed by seizures. Canine distemper virus was isolated from 3 leopards, 3 tigers, and 3 lions that died or were euthanized when moribund. Monoclonal antibody testing identified the virus isolates as CDV. Gross and histopathologic findings were similar to those found in canids with distemper with a few exceptions. There were fewer lesions in the brain, and there was a pronounced type 2 cell proliferation in the lung, with inclusion bodies and CDV antigen demonstrated by immunohistology. Neutralizing antibody to CDV was found in high titers in serum from most animals but was absent or was found only in low titers in some cats that succumbed after CDV infection. There was a marked difference in neutralizing antibody titers when tests were done with different strains of CDV.

Animals↗

Glycoconjugates as components of receptors for Bordetella avium on the tracheal mucosa of turkeys.

Bordetella avium is an important respiratory tract pathogen of turkeys. In common with other pathogenic bordetellae, B avium manifests a tissue tropism for cilia of the respiratory tract epithelium. To determine the molecular characteristics of the host cell receptors for B avium, we used hemagglutination and in vivo adherence assays. Carbohydrates, mucus, sialic acid-specific lectin, and other glycoconjugates were evaluated for their ability to competitively inhibit binding of B avium to host cells. The gangliosides, GD1a and GT1b, completely inhibited hemagglutination, whereas N-acetylneuraminic acid (sialic acid) partially inhibited hemagglutination. Adherence to turkey tracheal mucosa in vivo was significantly (P < 0.01) inhibited by GD1a and GT1b gangliosides, N-acetylneuraminic acid, bovine submaxillary mucin, and horseshoe crab (Limulus polyphemus) lectin. Treatment of the tracheal mucosa with neuraminidase also inhibited adherence of B avium. We conclude that N-acetylneuraminic acid and the gangliosides, GD1a and GT1b, may be important components of the tracheal mucosa receptor for B avium in turkeys.

Animals↗

Colonization of the small intestine of weaned pigs by enterotoxigenic Escherichia coli that lack known colonization factors.

Intestinal colonization of 3-week-old weaned pigs by enterotoxigenic Escherichia coli (ETEC) strains that were originally isolated from weaned pigs with fatal diarrhea and that lacked K88, K99, F41, and 987P adhesins (4P- ETEC) was studied by histologic, immunofluorescent, and electron microscopic techniques. In the first experiment, 16 principal pigs were inoculated orogastrically with ETEC strain 2134 (serogroup O157: H19) or 2171 (serogroup 0141:H4), and eight control pigs were not inoculated. In the second experiment, 24 principals were inoculated with ETEC strain 2134, and 12 controls were inoculated with a nonenterotoxigenic strain of E. coli. Principal and control pigs were necropsied at intervals from 24 to 72 hours after inoculation of principals to provide the tissues used for this report. Results from the two experiments and with both ETEC strains were similar and therefore were combined. Adhesion by 4P- ETEC was demonstrated in ileum but not in cecum or colon in 22/40 principal pigs sampled at 24 to 72 hours after orogastric inoculation. Adherent bacteria were most apparent on the intestinal villi covering Peyer's patches. Only occasional adherent bacteria were detected in ileal sections from a few (4/20) of the control pigs. Adherence by 4P- ETEC was characterized by "patches" of bacteria closely associated with the lateral surfaces and less frequently with the tips and the bases of intact villi. In most cases, the adherent bacteria were separated from epithelial cell microvilli and other bacterial cells by a 50-400-nm space. Filamentous bacterial appendages bridged this space and formed a network among adjacent bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

An experimental model for subclinical edema disease (Escherichia coli enterotoxemia) manifest as vascular necrosis in pigs.

An experimental model for subclinical edema disease was developed in weanling pigs. In multiple experiments, 3-week-old pigs were weaned, then inoculated intragastrically with 10(10) colony-forming units of an SLT-IIv-positive strain of Escherichia coli originally isolated from a pig with edema disease (principals). Control pigs were inoculated with a nonpathogenic E coli strain. Of 39 principals, 8 developed clinical edema disease within 14 days after inoculation. However, 20 of 21 principals that did not develop clinical signs of edema disease, but were submitted for necropsy examination at 14 days after inoculation, had characteristic vascular lesions of edema disease. Vascular lesions, found principally in ileum and brain, consisted of segmental necrosis of myocytes in the tunica media of small arteries and arterioles. None of the pigs inoculated with a nonpathogenic strain of E coli developed edema disease or vascular lesions. None of the principals necropsied at 2 days after inoculation had vascular lesions. Development of vascular lesions by 14 days after inoculation was used as the end point for detecting subclinical edema disease in the model.

Animals↗

Morphologic characterization of conjunctiva-associated lymphoid tissue in chickens.

Conjunctiva-associated lymphoid tissue (CALT) in the eyelids of chickens was studied by gross, histologic, and electron microscopic techniques. Structural features were characterized at 1 day of age and at posthatching week (PHW) 1, 2, 3, 4, 6, 8, 12, and 16. Beginning at PHW 1, prominent lymphoid nodules containing a heterogenous population of lymphocytes, lymphoblasts, and macrophages were first observed within conjunctival folds and fissures of the lower eyelid. Nodules contained germinal centers by PHW 2 and plasma cells by PHW 4. The epithelium associated with these nodules was flat, had short, irregular microvilli, contained intraepithelial lymphocytes, and lacked goblet cells. High endothelial venules were located at the base of lymphoid nodules and contained lymphocytes within and below the cuboidal endothelium. In the upper eyelid, CALT was morphologically similar to lymphoid tissue in the lower eyelid, but nodules were smaller and more random, lacked association with epithelial folds and fissures, and were clustered around the opening of the nasolacrimal duct. By PHW 12, CALT was characterized by basal germinal centers outlined by collagenous stroma, suprafollicular plasma cells, columnar epithelium with goblet cells, and fewer intraepithelial lymphocytes. On the basis of these features, CALT in chickens has morphologic characteristics similar to other components of the mucosal immune system and, therefore, may have a role in mucosal immunity.

Animals↗

A morphologic study of bronchus-associated lymphoid tissue in turkeys.

Bronchus-associated lymphoid tissue (BALT) in normal turkeys of ages 1 day and 1, 2, 3, 4, 8, and 18 weeks was examined by light microscopy and by scanning and transmission electron microscopy. Turkey BALT resembled other mucosa-associated lymphoid tissues; it was made up of a population of lymphocytes covered by a specialized epithelium different from typical pseudostratified ciliated columnar bronchial epithelium. There were distinct age-related differences in BALT structure. Bronchus-associated lymphoid nodules were larger and more numerous in older turkeys. In 1-day- to 2-week-old turkeys, the primary cell type of BALT epithelium was nonciliated cuboidal; in 2-week old turkeys it was squamous; and in turkeys older than 4-weeks of age, the epithelium was primarily ciliated columnar. In 1- to 4-week old turkeys, large numbers of intraepithelial lymphocytes disrupted the normal organization of the epithelium. In older turkeys, epithelial and lymphoid cells were in discrete compartments separated by connective tissue. Lymphocytes in 1-day-old turkeys were found in loose aggregates around venules and within the epithelium. In 1-week old turkeys, lymphocytes were organized into compartments of morphologically similar cells. By 3-weeks of age, lymphocytes were present in distinct germinal centers. Epithelial cells of BALT did not have large numbers of apical vesicles and thus were not structurally specialized for antigen uptake by endocytosis. However, the epithelial barrier appeared to be disrupted over lymphoid nodules, suggesting that antigen would be readily available to lymphocytes and phagocytes in BALT. Age-related differences in turkey BALT structure may have functional consequences with respect to the respiratory immune response.

Animals↗

Orally administered clonidine as a secretagogue of growth hormone and as a thymotrophic agent in dogs of various ages.

The growth hormone (GH) secretagogue activity of variable dosages of clonidine (16.5, 50, 150, and 450 micrograms/kg of body weight), given orally mixed with the daily food ration, was evaluated in young and old dogs. Significant (P less than 0.05) increase in plasma GH concentration was detected at all dosages tested in young dogs and in response to all but the lowest dose tested in the old dogs fed the clonidine-containing diet. Old dogs had plasma GH concentration that exceeded that of young dogs when higher doses of clonidine were used. A clonidine (100 micrograms/kg)-supplemented diet was fed to middle-aged dogs twice daily for 30 days. Significant (P less than 0.01) increase of plasma GH concentration was observed on the first day of the feeding trial, but was undetectable by day 30. After feeding the clonidine-enhanced diet for 30 days, the effects on thymic morphology were variable, and there was no effect on plasma thymulin titer. Clonidine-fed dogs had significantly increased lymphocyte blastogenic responsiveness to mitogens, compared with that of control dogs, when evaluated as stimulation index.

Administration, Oral↗

Identification of Bordetella avium antigens recognized after experimental inoculation in turkeys.

Sera and tracheal washings (TW) were used to identify antigens of Bordetella avium recognized during experimentally induced bordetellosis in young turkeys. Pooled sera and TW were examined for antibody by a microtitration agglutination test and by western immunoblotting. In addition, comparable samples collected from 1-day-old turkeys and uninoculated control turkeys also were examined. At least 8 outer membrane proteins of B avium were recognized in immunoblots of sera and TW from infected turkeys. Reactivity of TW in immunoblots was qualitatively similar but less intense, compared with reactivity of corresponding sera collected on postinoculation (PI) weeks 2, 3, and 4. Molecular weights of the major outer membrane proteins of B avium recognized by sera and TW at PI week 4 were 100,000, 97,000, 36,000, 31,000, 21,000, 18,000, 14,000, and less than 14,000. A protein with a molecular weight of 55,000 reacted nonspecifically in all samples tested. Antibody, detectable by microtitration agglutination, was in sera of 1-day-old turkeys and in sera and TW of B avium-infected turkeys during PI weeks 2 to 4.

Agglutination Tests↗

Conjunctiva-associated lymphoid tissue (CALT) in normal and Bordetella avium-infected turkeys.

Conjunctiva-associated lymphoid tissue (CALT) was characterized in normal and Bordetella avium-infected turkey poults during the first 5 weeks of life. At 1, 5, 12, 19, 25, and 33 days post-hatching (DPH), upper and lower eyelids were examined by gross, histologic, and electron microscopic techniques. CALT was confined to the proximal part of the lower eyelid near the conjunctival fornix; it appeared by 5 DPH as individual lymphoid nodules and as dense masses by 19 DPH. In the upper eyelid, CALT was present only as isolated nodules. Histologically, CALT was composed of dense lymphocyte infiltrates within subepithelial connective tissue, intraepithelial lymphocytes, and flattened lymphoid-associated epithelium that lacked goblet cells. Germinal centers were in CALT by 19 DPH. By scanning electron microscopy, epithelial cells over lymphoid areas were flat and had short, irregular microvilli; non-lymphoid areas were covered by cells with tall, regular microvilli. Transmission electron microscopy revealed that with increasing age of birds, the epithelium over conjunctival lymphoid infiltrates became progressively flattened and infiltrated by lymphocytes. Some blood vessels in CALT had high endothelial cells; lymphocytes were in the lumen and between or beneath endothelial cells. In B. avium-infected poults, CALT was increased, developed earlier, and contained more germinal centers than in normal poults. We conclude that CALT of turkeys closely resembles other mucosal lymphoid tissues and may serve as a site for local antigen uptake.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of complement activity in turkeys: evidence for classical and alternative complement pathways.

Complement activity in turkey serum was examined by using inhibitors or activators of mammalian complement. Hemolytic test systems using sheep red blood cells sensitized with specific antibody (SSRBC) and horse red blood cells (HRBC) were developed to measure residual complement activity of turkey sera treated with the various inhibitors or activators. Lysis of SSRBC was blocked by treatment with 6-mM EDTA, 10-mM ethylene glycol-bis-beta aminoethylether N,N,N',N' tetraacetic acid (EGTA), and carrageenan. In contrast, lysis of HRBC was blocked by 6-mM EDTA, but not by 10-mM EGTA or carrageenan. Addition of magnesium to EGTA-chelated serum facilitated the lysis of HRBC but not the lysis of SSRBC. Treatment of serum with zymosan at 1 mg/mL and inulin at 5 mg/mL depleted hemolytic activity against both SSRBC and HRBC, suggesting depletion of components common to both pathways. Differences in the hemolytic activities of sera against SSRBC and HRBC after treatment with the various complement inhibitors and activators demonstrate the presence of two complement pathways in turkeys.

Animals↗

Interaction of turkey complement with Escherichia coli isolated from turkeys.

The role of turkey complement in a serum bactericidal reaction was determined using serum-sensitive and serum-resistant Escherichia coli isolated from turkeys. Inactivation of complement by heating serum (56 C for 40 minutes) or by treating serum with 10 mM EDTA eliminated bactericidal activity. Serum-sensitive E coli organisms were killed by turkey serum treated with 10 mM ethylene glycol-bis-beta-(aminoethyl ether)-N,N,N',N'-tetraacetic acid and 5 mM MgCl2. Exposure of normal turkey serum to serum-sensitive or serum-resistant E coli resulted in equivalent reductions in hemolytic activity of serum. Treatment of serum-resistant E coli with antibody rendered the bacteria sensitive to bactericidal effects of normal turkey serum. Serum-sensitive E coli organisms were readily killed by an alternative complement pathway, serum-sensitive and serum-resistant E coli activated the complement system equally well, and antibody was required for complement-mediated killing of certain serum-resistant E coli organisms from turkeys.

Animals↗

Transposon mutagenesis in Bordetella avium.

Transposon mutagenesis was used to introduce specific mutations in the chromosome of Bordetella avium. The transposon Tn5 was transferred by conjugation to B avium from Escherichia coli strain SM10 by the suicide plasmid vector pSUP1011. A defined minimal medium for growth of B avium was developed for scoring transposon-containing mutants for specific nutritional requirements. Approximately 1% of all mutants tested were auxotrophic in some form, and these organisms were grouped into 13 classes. Also, 2 classes of hemagglutination-negative mutants were isolated. Stability of Tn5 insertion was examined, and frequencies of reversion less than 10(-7) were observed, indicating stable mutations.

Bordetella↗

Serum resistance and virulence of Escherichia coli isolated from turkeys.

Twenty-five strains of Escherichia coli isolated from turkeys were characterized for their serum resistance and virulence. An in vitro bactericidal assay was used to determine the serum resistance of E coli. Virulence was determined by survival time after IV inoculation of each strain into 3-week-old turkeys. Serum-resistant E coli strains were generally found to be virulent for turkeys, whereas serum-sensitive E coli strains were avirulent. Of the 25 strains, 18 strains were placed in the 2 categories of serum-resistant/virulent and serum-sensitive/avirulent. Five strains were serum-resistant and avirulent, and 2 strains were serum-sensitive and virulent. Serum resistance appears to be an important determinant of virulence for E coli in turkeys; however, the requirement for other virulence factors, in addition to serum resistance, was suggested by the finding that 5 serum-resistant strains were avirulent in turkeys.

Animals↗

Adherence of Bordetella avium to tracheal mucosa of turkeys: correlation with hemagglutination.

Adherence to turkey tracheal mucosa and agglutination of guinea pig erythrocytes were determined for 20 strains of Bordetella avium. Ten type-I strains, 7 type-II strains, 2 transposon-induced mutants, and 1 revertant were evaluated. All type-I strains adhered readily to tracheal mucosa and agglutinated erythrocytes, whereas no type-II strains adhered to trachea or caused hemagglutination (HA). Two mutants selected for loss of HA activity were less adherent to tracheal mucosa, compared with the parent strain. Reversion of one mutant to HA-positive status was accompanied by reconstitution of much of its adherence capacity. Results of this study provide preliminary evidence that tracheal adherence and HA of B avium are closely related.

Animals↗

Histologic evaluation of lung and bronchus-associated lymphoid tissue in young turkeys infected with Bordetella avium.

One-day-old turkeys were inoculated intranasally with Bordetella avium. Noninoculated hatchmates were housed separately. At postinoculation weeks 1, 2, 3, 4, and 5, B avium-infected (BA+) and B avium-free (BA-) turkeys were necropsied; specimens of tracheas, intrapulmonary primary bronchi, and lung adjacent to primary bronchi were bacteriologically cultured. Lung tissue was collected for histologic examination. Lungs perfused with acetic acid were collected for evaluation to determine the size, number, and distribution of lymphoid nodules associated with primary bronchi. Bordetella avium was isolated from trachea and primary bronchi of all BA+ turkeys, but was never isolated from lung parenchyma. Acute purulent bronchitis was associated with colonization of the primary bronchi by B avium from postinoculation weeks 1 to 3. Macrophages and lymphocytes persisted in the peribronchial connective tissue for 5 weeks after inoculation. Bronchus-associated lymphoid tissue consisted of discrete lymphoid nodules protruding into the lumens of primary bronchi. Lymphoid nodules, morphologically similar in BA+ and BA- turkeys, were composed of nonciliated, cuboidal epithelium covering a zone of loosely arranged lymphocytes and macrophages and a deeper, sharply demarcated lymphoid follicle. Compared with bronchus-associated lymphoid tissue of BA- turkeys, lymphoid nodules of bronchus-associated lymphoid tissue in BA+ turkeys were more numerous and widely distributed along primary bronchi. In both BA- and BA+ turkeys, the mean diameter of lymphoid nodules doubled between 1 and 5 weeks of age.

Animals↗