A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay.

We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.     

Duck lymphocytes. VI. Requirement for phagocytic and adherent cells in lymphocyte transformation.

Preparations of duck (Anas platyrhynchos) spleen and blood lymphocytes depleted of cells capable of phagocytosing carbonyl iron gave lower transformation responses to the mitogens phytohaemagglutinin (PHA), concanavalin A (Con A), Bandeiraea simplicifolia seed lectin (BSS), wheat germ agglutinin (WGA), lentil lectin (LL) and phorbol ester (PMA) than intact cell preparations. When cell populations were fractionated on the basis of their adherence to plastic, it was found that the adherent cells were responsive to PHA, Con A, BSS, WGA and PMA, while the non-adherent cells responded to LL. These observations confirm the expected requirement for phagocytic accessory cells in the induction of in vitro mitogen-driven duck lymphocyte responses. The responses of plastic-adherent populations of cells to most mitogens are believed to reflect the generally close physical relationship between the adherent accessory cells and the lymphocytes, although it remains possible that duck monocytes respond to some of the mitogens employed. The data also suggest that LL stimulates a population of cells different to those responding to other mitogens.     

Immunocytochemical Study of the Ultimobranchial Tubule in Wistar Rats

A systematic immunohistochemical study of the ultimobranchial tubule (UBT) has been carried out in 45 Wistar rats of different ages (0, 5, 10, 15, 20, 25, 30, 60 and 120 days). The existence of calcitonin immunoreactive cells in the UBT wall has been demonstrated in a 5-days old rat. In addition, immunohistochemical studies for thyroglobulin revealed positive staining in follicular cells connected to the UBT and, occasionally, in isolated cells lying within solid clusters from the UBT. These last results together with the continued and repeated existence of numerous mitosis and PAS (+) microfollicles, apparently rising from the UBT, support the hypothesis that the ultimobranchial body (UBB) may contribute partially to the formation of a part of the follicular component.     

A comparison of the gland of Harder response and head-associated lymphoid tissue (HALT) morphology in chickens and turkeys.

The immunofunctional response of the gland of Harder (GH) was compared in chickens and turkeys using an in vivo assay previously developed for use in chickens. The GH were surgically removed (GHx) from leghorn chicks at 1 day of age and from poults at 2 days of age. Intact birds of each species served as controls. During the fourth week of age, both GH-intact and GHx chicks were exposed to killed Brucella abortus antigen by the ocular or intraperitoneal route. One week later, serum and tears were collected and assayed for antibodies to B. abortus. In addition, all birds were killed at the end of the trial period, and the heads were fixed and processed for histologic examination. Various components of the head-associated lymphoid tissue (HALT) including the GH, nasal glands, lacrimal glands, lacrimal ducts, eyelid conjunctiva, and nasal cavity mucosa/submucosa, were evaluated microscopically using a scoring system to estimate quantity and degree of development of immune tissue in those sites. Results of all analyses indicate that functional response and morphology of the HALT are comparable in turkeys and chickens.     

Maturation of functional antibody affinity in animals immunised with synthetic foot-and-mouth disease virus.

A good correlation exists between specific neutralising antibody titre and protection against challenge with foot-and-mouth disease virus (FMDV) in infected or virus-vaccinated cattle, but not in the case of animals immunised with synthetic FMDV peptides. Therefore, mechanisms other than simple neutralisation are likely to be important in vivo. Antibody affinity may influence the protective capacity of sera from immunised animals and experiments were carried out to measure the functional affinity for synthetic FMDV peptide of sera from guinea pigs and cattle given various synthetic vaccines. In guinea pigs given a single dose of synthetic vaccine, antibody affinity increased with time after immunisation. In cattle, however, administration of a second dose of peptide 21 days after the first markedly retarded the process of affinity maturation. For guinea pig sera of equivalent neutralising activity, those of higher functional affinity had higher protective indices than those of lower functional affinity. Knowledge of the importance of antibody affinity in protection against FMD is important for an improved understanding of the mechanisms of protection and for the design of novel vaccines.     

Use of an indirect enzyme-linked immunosorbent assay (elisa) in the detection of channel catfish, Ictalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri

Abstract. The use of an indirect enzyme-linked immunosorbent assay ( elisa ) for the detection of channel catfish antibody to Edwardsiella ictaluri is described. Changes in agglutination titre in fish immunized with Edwardsiella ictaluri heat killed whole bacterins or lipopolysaccharides were reflected by corresponding changes in elisa readings. Relatively high correlations were observed among elisa OD readings, computed elisa titres and corresponding agglutination titres.     

Serum amyloid A protein (SAA) in horses: objective measurement of the acute phase response

A sensitive and precise immunoassay for equine serum amyloid A protein (SAA) was established and used to determine, for the first time, the circulating concentration of this protein in health and disease. As in other species, equine SAA was present only at trace levels in healthy animals but behaved as an extremely sensitive and rapidly responding acute phase reactant following most forms of tissue injury, infection and inflammation, objectively reflecting the extent and activity of disease. Measurements of SAA should make a significant contribution to diagnosis and management of viral and bacterial infection in horses, and routine serial assays could provide an objective criterion for monitoring prospectively the general health of horses in training and racing.     

Incidence of microscopically detectable degenerative characteristics in skeletal muscle of turkey

1. The incidence of microscopically detectable degenerative characteristics in 5 skeletal muscles (m. pectoralis thoracicus, m. supracoracoideus, m. biceps femoris, m. semitendinosus, m. femorotibialis medius) of turkeys was investigated. 2. Samples were obtained from 30 Large White turkey males 14, 16 and 18 weeks old. Hyaline degeneration, infiltration of mononuclear cells and necrotic fibres were observed. 3. Individual fibres varied greatly in size and muscle fibre nuclei were often shrunken and pyknotic. 4. Weak and/or uniform reaction for Ca++-ATPase and SDH in all types of muscle fibres and loss of alkaline phosphatase activity in cell membranes were noted. A positive reaction for acid phosphatase occurred in regions of perivascular infiltration and in necrotic muscle fibres. The majority of muscle fibres possessed high activity for phosphorylase a and b. 5. Based on the use of fluorescein alpha-bungarotoxin conjugate, motor end-plates appeared to be morphologically intact. Direct immunofluorescence with anti-chicken IgG showed positive reaction in muscle fibres undergoing necrosis and in the involved connective tissue. 6. Degenerative changes varied with age and were most marked in the oldest birds. 7. Because gross degenerative symptoms were absent from both the birds and the meat from them, the condition appears to be either different from or a precursor to the degenerative myopathy characterised by other authors.