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Biomedical subjects

D M Haines

Publications and source records attributed to D M Haines.

At least 91 records · Page 5Linked to original sources
PubMed1991

The role of passive immunity in bovine respiratory syncytial virus-infected calves.

The role of passive immunity in bovine respiratory syncytial virus (BRSV) infections in neonatal calves was evaluated. Calves were divided into groups as follows: colostrum-deprived, sham-inoculated; colostrum-deprived, BRSV-inoculated; and colostrum-fed, BRSV-inoculated. Calves were inoculated with a low-passage field isolate of BRSV for 4 consecutive days by a combined respiratory tract route and were euthanized 6 days after receiving the last inoculation. Arterial oxygen tension (Pao2) decreased significantly over time in colostrum-deprived, BRSV-inoculated calves (P less than .01) and was significantly different among treatment groups (P less than .05). A significant decrease in arterial oxygen saturation was observed in this same group over time (P less than .01). Mean percentage of pneumonic lung volume (determined by computer data digitalization) was significantly greater in infected, colostrum-deprived calves compared with the other groups (P less than .01), and BRSV antigen was detected in these calves by avidin-biotin immunoperoxidase staining. Thus, passive immunity derived from colostrum feeding decreased the severity of BRSV infections in calves.

Analysis of Variance↗
PubMed1991

The relationship between autoantibodies to triiodothyronine (T3) and thyroglobulin (Tg) in the dog.

The relationship between T3 autoantibodies (T3AA) and thyroglobulin autoantibodies (TgAA) in dogs was investigated by determining the inhibitory effect of triiodothyronine (T3), thyroxine (T4) and thyroglobulin (Tg) on T3AA and TgAA binding activity and by determining the pattern of occurrence of the two activities in canine serum samples. Strong similarity in binding characteristics between the two activities, as one might expect if T3AA activity were merely a cross-reactivity of TgAA, was not observed. Canine T3AA activity exhibited a cross-reactivity to purified canine Tg that was intermediate between that of T3 and T4, indicating an antigenic relationship to an epitope of Tg. Average affinity constants of canine T3AA (N = 11) for T3, Tg and T4 were 1.76 x 10(10) M-1, 2.29 x 10(9) M-1, and 1.02 x 10(8) M-1, respectively. Canine TgAA activity, however, did not cross-react significantly with T3 or T4. Canine TgAA (N = 21) binding to canine Tg was not inhibited by T4 or T3 at concentrations up to 2 x 10(-4) M. Each of 23 canine serum samples containing T3AA also exhibited TgAA activity, although there was poor correlation between the magnitudes of the two activities. Neither T3AA nor TgAA activity was observed in serum samples from 16 euthyroid dogs; however, 46.7% of the samples from 15 hypothyroid dogs had detectable TgAA activity. T3AA is so rare that is was not observed in this small population of samples from hypothyroid dogs. The [125I] T3 binding in serum from hypothyroid dogs was elevated compared to that in euthyroid dogs, but was considerably lower than in samples generally designated as containing T3AA. These results suggest that T3AA found in occasional canine serum samples are due to the presence of autoantibodies recognizing a T3 containing epitope of Tg that is different from the epitopes involved in eliciting the predominant population of canine Tg autoantibodies.

Animals↗
PubMed1990

Systemic lupus erythematosus in a filly.

Systemic lupus erythematosus (SLE) was diagnosed in a 2-year-old Standardbred filly. Clinical signs of SLE included weight loss, bilateral symmetric alopecia, seborrhea, oral ulceration, and lymphadenopathy. Abnormal laboratory findings included a Coombs test-positive hemolytic anemia and positive antinuclear antibody test result. Histologic evaluation of multiple skin biopsy specimens revealed interface dermatitis with linear deposition of IgG at the basement membrane zones of the epidermis and hair follicles. The filly did not respond to glucocorticoid treatment and was euthanatized. Necropsy findings included membranous glomerulonephritis and fibrous synovitis. On the basis of these findings, SLE should be considered in the differential diagnosis of immune-mediated skin disease in horses. Definitive diagnosis of SLE relies on recognition of multisystemic disease and confirmatory histopathologic and immunopathologic findings.

Animals↗
PubMed1990

Idiopathic granulomatous disease involving the skin in a horse.

Clinical signs of generalized granulomatous disease in a horse included depression, generalized lymphadenopathy, scaly skin, and dependent edema. Diagnosis was confirmed histopathologically by diffuse granulomas in more than one organ system (lymph nodes and skin), and by ruling out etiologic agents. Response to treatment with corticosteroids was favorable. The clinical features and response to treatment in this horse were different from classic textbook descriptions of equine generalized granulomatous disease. Equine granulomatous disease appears to be comparable to human sarcoidosis. To avoid confusion between human medical and veterinary medical terminology, the authors suggest that the equine disease be referred to as equine idiopathic granulomatous disease and that a complete diagnostic description should include a reference to the organ systems involved.

Animals↗
PubMed1990

Immunohistochemical detection of intermediate filament proteins in formalin fixed normal and neoplastic canine tissues.

Normal and well differentiated neoplastic canine tissues were immunohistochemically stained for keratin, vimentin and desmin intermediate filament proteins using commercially available monoclonal antibodies. Keratin was detected in 56 of 57 carcinomas, vimentin in 59 of 62 sarcomas and desmin in three of four muscle cell tumors. Most normal and neoplastic tissues expressed only one type of intermediate filament; exceptions were one hemangiosarcoma and one pulmonary carcinoma in which there was coexpression of vimentin and keratin proteins. Since immunohistochemical detection of intermediate filaments has tissue-specific distribution in the majority of well differentiated canine neoplasms, these stains may be useful in the differential diagnosis of anaplastic canine tumors. However, the monoclonal antibodies to cytokeratin which were tested in this study failed to detect intermediate filaments in liver, pancreas and salivary glands which suggests that these antibodies may also be unable to detect epithelial tumors derived from these tissues. In addition, in nine neoplasms, the normal tissues adjacent to neoplastic cells failed to stain for the intermediate filament normally expressed. When this occurs, evaluation of intermediate filament expression is invalid for the determination of tissue of origin of the neoplastic cells.

Animals↗
PubMed1990

Immunohistochemical demonstration of Trypanosoma evansi in tissues of experimentally infected rats and a naturally infected water buffalo (Bubalus bubalis).

Trypanosoma evansi was demonstrated by an immunohistochemical technique in formalin-fixed paraffin-embedded tissues of experimentally infected rats. Trypanosoma evansi was visible readily, nuclei were stained darkly, the cytoplasm was stained moderately, and the cell membranes were delineated clearly. The parasites were present in small- to large-sized blood vessels of all organs, in extravascular spaces of ventricles and neuropil of the brain, and in interstitial tissues of the lung and testes. This method also stained nuclei but not cytoplasm or cell membranes of Trypanosoma congolense, and did not stain Trypanosoma theileri. In a water buffalo (Bubalus bubalis) with nonsuppurative meningoencephalitis, the presence of T. evansi could not be demonstrated by conventional histological stains. However, the trypanosomes were recognized readily in the Virchow-Robin spaces and neuropil of the brain by the immunohistochemical method.

Animals↗
PubMed1989

A novel marker for basal (stem) cells of mammalian stratified squamous epithelia and squamous cell carcinomas.

We have developed a monoclonal antibody (174H.64) which selectively recognizes antigens shared by the basal cells of mammalian stratified squamous epithelium and squamous cell carcinoma (SCC). Histopathological studies of the frozen tissue sections demonstrated selective binding of this antibody to SCCs of human, bovine, canine, feline, and murine origin. Tumors of other histological types did not show reactivity with the antibody. In well-differentiated SCCs the peripheral layer of the tumor showed preferential binding of the antibody, suggesting that the antigens are associated with the proliferative compartment of the tumor. Studies on normal human tissues showed selective binding of the antibody to the basal layer of stratified squamous epithelia, thymic epithelial cells, and myoepithelial cells around breast ducts, while no antibody binding was observed for the suprabasal layers of stratified epithelia, simple epithelia, or tissues of nonepithelial origin. A similar pattern of antibody binding was also observed for bovine and murine skin with staining of the basal layer. The antigens detected by monoclonal antibody 174H.64 were characterized from cytoskeletal protein extracts of normal human keratinocytes as well as human and bovine SCC tissues by using an immunoblotting technique. The antigens detected in normal human keratinocytes consisted of two major protein bands of approximate molecular weights of 48,000-50,000 and 57,000. In bovine SCC tumor the antigen detected was the Mr 48,000-50,000 band and in the human SCC tumor it was the Mr 57,000 band. A murine lung SCC model was developed with a murine SCC cell line KLN-205. The lung tumor obtained was reactive against the antibody and showed selective staining of the peripheral layer of the tumor containing the stem cell population. The antigens described by monoclonal antibody 174H.64 appear to be molecules associated with the stem cell populations of normal stratified epithelium and squamous cell carcinoma.

Animals↗
PubMed1989

Monoclonal antibodies to bovine coronavirus glycoproteins E2 and E3: demonstration of in vivo virus-neutralizing activity.

Six monoclonal antibodies (MAbs) to bovine coronavirus (BCV, Quebec isolate) E2 and E3 glycoproteins which were found previously to be neutralizing in vitro were examined for virus-neutralizing activity in vivo. Surgically ligated intestinal loops of newborn colostrum-deprived calves were virus-inoculated, mock-infected or inoculated with a mixture of virus and antibody. Of the six BCV-specific MAbs, four were found to be protective against a virulent field isolate of BCV, as indicated by a reduction in villous atrophy. These MAbs were specific to antigenic domain A and antigenic domains A1 and A2 on the E2 and E3 glycoproteins respectively. MAbs to antigenic domains B and C on the E2 and E3 glycoproteins, respectively, were not protective.

Animals↗
PubMed1989

Immunohistochemical staining and radionuclide imaging of canine tumors, using a monoclonal antibody recognizing a synthetic carbohydrate antigen.

The in vitro and in vivo binding of a monoclonal antibody (MAB) that recognizes a tumor-associated carbohydrate antigen was studied in dogs. Monoclonal antibody 155H.7 was raised in response to innoculation of mice with beta-galactose(1-3)beta N-acetylgalactosamine conjugated to human serum albumin. Avidin-biotin-complex immunohistochemical staining of cryostat sections of normal and neoplastic canine tissue specimens revealed heterogenous binding of MAB 155H.7 to the cells of many canine mammary and lung carcinomas and homogenous staining of many sarcomas, including osteogenic sarcoma. In addition, there was variable staining of a variety of normal tissues including some ductual epithelium, peripheral nerve fibers, and some endothelial cells and fibroblasts. Immunoscintigraphy with 131I-labeled MAB 155H.7 was used to study the in vivo distribution of the antibody. The 131I-labeled MAB 155H.7 was administered to 1 clinically normal dog, 7 dogs with osteogenic sarcoma, 1 dog with undifferentiated sarcoma, and 2 dogs with mammary tumor. Scintigraphy revealed concentration of radioactivity in 8 of 10 tumor sites within 24 hours after MAB administration. The ratio of 131I in tumor sites to 131I in the surrounding normal tissues, compared with the similar ratio of 99mTc-labeled erythrocytes ranged from 1.1 to 4.3, in tumor vs normal tissue with a mean value of 2, confirming tumor localization of the radiolabeled MAB in excess of that associated with enhanced tumor vascularization.

Animals↗
PubMed1989

The detection of bovine respiratory syncytial virus in formalin fixed bovine lung with commercially available monoclonal antibodies and avidin biotin complex immunohistochemistry.

Eight commercially available monoclonal antibodies directed against respiratory syncytial virus antigens were tested for ability to detect bovine respiratory syncytial virus (BRSV) antigen in formalin fixed, paraffin embedded bovine lung using avidin-biotin complex immunohistochemical staining. Monoclonal antibodies from clone 18B2 purchased from Biosoft, Paris, France and those from clone 8G12 purchased from the Department of Veterinary Sciences, University of Nebraska, Lincoln, Nebraska stained BRSV antigen in infected bovine lung with acceptable background staining of uninfected tissues. This method offers advantages over other techniques for BRSV diagnosis in that fresh tissue is not required and all reagents may be purchased commercially.

Animals↗
PubMed1988

Immunocytochemical identification of bovine Langerhans cells by use of a monoclonal antibody directed against class II MHC antigens.

We undertook a study to develop a reliable light microscopic technique for identifying Langerhans cells (LC) in bovine epidermis. Monoclonal antibodies (MCA) detecting bovine class II MHC antigens were used in conjunction with an avidin-biotin-peroxidase complex (ABC) immunocytochemical staining method. The specificity of the MCA for LC was confirmed ultrastructurally by use of gold-labeled second antibody. Epidermal sheets and epidermal single-cell suspensions examined by light microscopy confirm that bovine epidermal LC express class II antigens. Anti-bovine class II MCA is a dependable reagent for identification of LC in normal bovine epidermis.

Animals↗
PubMed1987

A molecular approach to immunoscintigraphy: a study of the T-antigen conformation on the surface of tumors.

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both alpha and beta configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the beta configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

Animals↗
PubMed1987

Avidin-biotin-peroxidase complex immunohistochemistry to detect immunoglobulin in formalin fixed skin biopsies in canine autoimmune skin disease.

Avidin-biotin-peroxidase complex immunohistochemistry was used on formalin fixed, paraffin embedded, trypsin digested, skin biopsies to detect immunoglobulin deposition in dogs with autoimmune skin disease. Immunostaining by the avidin-biotin-peroxidase complex technique revealed intercellular and/or basement membrane immunoglobulin deposits in 27 of 28 dogs considered to have autoimmune skin disease by clinical and histological evaluation and in six of 19 dogs considered to have autoimmune skin disease by clinical evaluation but without histological confirmation. Similar immunostaining was not evident in five biopsies of normal skin or in biopsies from four dogs with noninflammatory dermatoses, but was present in biopsies from one of ten dogs considered by clinical and histological criteria to have an inflammatory dermatosis other than autoimmune skin disease. Detection of immunoglobulin deposits in skin biopsies by avidin-biotin-peroxidase complex immunohistochemistry offers numerous advantages over conventional immunofluorescence methods including the opportunity to precisely compare histological and immunological findings.

Animals↗
PubMed1986

A re-examination of islet cell cytoplasmic antibodies in diabetic dogs.

Serum samples were obtained from 48 dogs with recently diagnosed untreated diabetes mellitus. Serums were tested for cytoplasmic autoantibodies to normal canine pancreatic islet antigens by indirect immunofluorescence, peroxidase-anti-peroxidase, and avidin-biotin complex, immunohistochemistry. Autoantibodies were not detectable in any of the samples. Serums were also examined from 20 diabetic dogs maintained on exogenous insulin therapy for periods of one month to five years. Positive reactions were seen in 11 dogs. These positive responses were completely absorbed by preincubation of serums with commercial insulin preparations or with purified pork or beef insulin. Newly diagnosed diabetic dogs do not have readily detectable autoantibodies to islet cytoplasmic antigens. Our previous report (Haines and Penhale, 1985) of islet antibody in diabetic dogs with unknown clinical histories was likely demonstrating antibody to insulin in patients treated with exogenous insulin. Antibodies to insulin were detected in approximately half of the insulin treated dogs tested. These antibodies were induced by commercial beef and pork insulin preparations and were found to be broadly cross-reactive recognizing epitopes on canine, bovine and porcine insulins.

Animals↗
PubMed1985

Experimental thyroid autoimmunity in the dog.

Thyroid autoimmunity was induced in 6 crossbred dogs by a single injection of thyroid extract in complete Freund's adjuvant alone, or followed by reimmunization with thyroid extract and incomplete Freund's adjuvant or peanut oil. Total thyroxine levels, autoantibodies to thyroglobulin and thyroid microsomal antigen and thyroid histopathological changes were sequentially monitored for up to 150 days. All dogs developed fluxuating levels of thyroid autoantibodies detectable by enzyme-linked immunosorbent-assay (ELISA). Histopathological damage was observed in one or more thyroid gland biopsies in 4 of the dogs, in 3 cases the lesions were of lymphocytic thyroiditis, in the fourth dog the lesion was a granulomatous reaction. Histopathological thyroid damage was observed in biopsies taken during periods of maximal autoantibody titer. Fixed immunoglobulin was not demonstrable in thyroid gland biopsies by direct immunofluorescence. In 5 dogs total thyroxine levels fell to below the normal range at some period during the study. However in only one dog did this correlate with a time when biopsy findings revealed histopathological thyroid damage. High levels of thyroid autoantibody may indicate histopathological thyroid damage in dogs, however lower levels of thyroid antibody may be found without evidence of histological damage or alteration in thyroid function.

Adjuvants, Immunologic↗
PubMed1985

Autoantibodies to pancreatic islet cells in canine diabetes mellitus.

Indirect immunofluorescence on normal canine pancreatic tissue fixed in Bouin's solution was used to detect islet cell antibodies in dogs with diabetes mellitus, other endocrine diseases, and pancreatitis. 18 of 25 dogs with diabetes mellitus alone, 2 of 8 dogs with diabetes mellitus and concurrent pancreatitis, and 2 of 2 dogs with diabetes mellitus and concurrent pancreatic exocrine insufficiency were positive for autoantibody. 2 of 12 dogs with hypoadrenocorticism, 3 of 6 dogs with hyperadrenocorticism, 6 of 28 dogs with hypothyroidism and one of 19 dogs with pancreatitis alone were also antibody positive. None of 20 healthy dogs or 20 dogs with disorders other than those of the pancreas or endocrine organs were antibody positive. Islet cell antibodies were demonstrated in dogs with diabetes mellitus and other endocrine disorders. The possibility of autoimmune involvement in the development of diabetes mellitus in the dog should be considered.

Animals↗
PubMed1984

The detection of canine autoantibodies to thyroid antigens by enzyme-linked immunosorbent assay, hemagglutination and indirect immunofluorescence.

Autoantibodies to thyroid antigens in serums from 34 clinically hypothyroid dogs were detected by various methods. Antibodies to thyroglobulin were detectable by enzyme-linked immunosorbent assay (ELISA) in 59%, by chromic chloride hemagglutination in 53% and by indirect immunofluorescence on formalin-fixed, paraffin-embedded, trypsin-digested thyroid tissue in 73% of samples. Antibody to thyroid microsomal antigen was detectable by ELISA in 29% of serums. Indirect immunofluorescence showed cytoplasmic fluorescence of thyroid follicular cells in several serums, however, this could not be confirmed as specific for microsomal antigen by absorption trials. Hemagglutination tests using commercially available tanned red cells coated with human antigens and indirect immunofluorescence assays on formalin-fixed tissue without trypsin digestion, on Bouin's fixed tissue, or on cryostat, methanol-fixed sections, were insensitive. Cryostat sections without methanol fixation were unsuitable due to tissue fragility. No method was recommended for routine diagnostic use. The ELISA test, because of its convenience, may be useful as a screening aid or adjunct to the diagnosis of thyroid disease.

Animals↗