Characterization of Anti-Dal Alloantibodies Following Sensitization of Two Dal-Negative DogsGoulet, S.; Blais, M. C.
doi: 10.1177/0300985816688746pmid: 28129096
Since its discovery, the immunogenicity of the Dal blood type has not been further investigated. The aim of this study was to better characterize anti-Dal alloantibodies produced following sensitization of Dal-negative dogs, notably their rate of appearance, the agglutination titer over time, and their immunoglobulin class. A secondary objective was to obtain polyclonal anti-Dal alloantibodies to increase the availability of Dal blood typing. Of 100 healthy laboratory Beagles tested, 2 Dal-negative dogs were identified as recipients. Ten healthy Dal-positive dogs were investigated as potential blood donors. All dogs were extensively blood typed for DEA 1, 3, 4, 5, and 7, as well as for Dal. Then, the recipients were transfused uneventfully with 10 ml/kg of Dal-positive but otherwise compatible packed red blood cells. Posttransfusion blood samples were collected routinely over a minimum of 1 year. Using a gel column technology, anti-Dal alloantibodies were detected as early as 4 days posttransfusion and remained detectable 2 years posttransfusion, with maximum agglutination titers reached at 1 and 2 months posttransfusion. The immunoglobulin class was IgG. The immunogenicity and clinical significance of the Dal blood type were confirmed. The results support the recommendations that previously transfused dogs be crossmatched starting 4 days posttransfusion and for the animal’s lifetime. The polyclonal anti-Dal antibodies produced will allow blood typing of a significant number of dogs, especially transfused dogs facing blood incompatibilities and canine blood donors.
Canine Neutrophil Extracellular Traps Enhance Clot Formation and Delay LysisJeffery, Unity; LeVine, Dana N.
doi: 10.1177/0300985817699860pmid: 28346125
Autoimmune diseases increase the risk of thrombosis. Neutrophil extracellular traps (NETs) are webs of DNA and protein that may mediate thrombosis in autoimmune diseases. Human and murine studies show NET-releasing neutrophils within a thrombus promote its growth, but it is unclear to what extent NET fragments released into circulation during inflammation are prothrombotic. This study hypothesized that canine NETs promote clot formation and impair lysis even in the absence of neutrophils. NETs were prepared from PMA-stimulated neutrophils and added to fibrinogen and thrombin or to recalcified pooled canine platelet-poor plasma, tissue factor, and tissue plasminogen activator. Clot formation and lysis were measured spectrophotometrically. NETs did not alter fibrin clot formation, but NETs increased maximum clot formation velocity (P = .001) and delayed lysis (P = .009) of plasma clots compared with supernatants from nonstimulated neutrophils. DNase digestion of NETs reduced their effect on clot lysis but not maximum clot formation velocity. This suggested impaired lysis was principally mediated by DNA within NETs but that NET proteins were principally responsible for increased speed of clot formation. Previous reports suggested elastase or histones might be responsible for the effect of NETs on clot formation. Elastase activity was greatly reduced by plasma, and addition of histones to plasma did not increase formation velocity, suggesting these proteins were not responsible for increasing maximum formation velocity. This study showed that NETs enhanced clot formation and impaired clot lysis in canine platelet-poor plasma. These in vitro findings suggest both NET proteins and DNA may contribute to thrombosis in inflammatory disease.
Analysis of Bacterial and Fungal Nucleic Acid in Canine Sterile Granulomatous and Pyogranulomatous Dermatitis and PanniculitisRosa, Fabio B.; Older, Caitlin E.; Meason-Smith, Courtney; Suchodolski, Jan S.; Lingsweiler, Sonia; Mansell, Joanne E.; Hoffmann, Aline Rodrigues
doi: 10.1177/0300985817738316pmid: 29145794
Next generation sequencing (NGS) studies are revealing a diverse microbiota on the skin of dogs. The skin microbiota of canine sterile granulomatous and pyogranulomatous dermatitis (SGPD) has yet to be investigated using NGS techniques. NGS targeting the 16S rRNA and ITS-1 region of bacterial and fungal DNA, respectively, were used to investigate if bacterial and fungal DNA were associated with skin lesions in cases of canine SGPD. The study included 20 formalin-fixed paraffin-embedded (FFPE) skin samples and 12 fresh samples from SGPD-affected dogs, and 10 FFPE and 10 fresh samples from healthy dogs. DNA was extracted from deep dermis and panniculus, and microbial DNA was amplified using primers targeting the bacterial 16S rRNA V1-V3 and fungal ITS-1 regions. The amplified DNA was utilized for NGS on an Illumina MiSeq instrument. The sequences were processed using QIIME. No differences in fungal or bacterial alpha diversity were observed between the SGPD and control samples. Beta diversity analysis demonstrated differences in the bacterial communities between SGPD and control, but not in the fungal communities. Compared to controls, the family Erysipelotrichaceae and genus Staphylococcus were significantly more abundant in the SGPD FFPE samples, and genus Corynebacterium were more abundant in fresh samples. The bacteria found to be more abundant in SGPD are common inhabitants of skin surfaces, and likely secondary contaminants in SGPD cases. This study provides additional evidence that SGPD lesions are likely sterile.
Horses With Pasture Asthma Have Airway Remodeling That Is Characteristic of Human AsthmaFerrari, Claudenir R.; Cooley, Jim; Mujahid, Nisma; Costa, Lais R.; Wills, Robert W.; Johnson, Melanie E.; Swiderski, Cyprianna E.
doi: 10.1177/0300985817741729pmid: 29254472
Severe equine asthma, formerly recurrent airway obstruction (RAO), is the horse counterpart of human asthma, affecting horses maintained indoors in continental climates. Equine pasture asthma, formerly summer pasture RAO, is clinically similar but affects grazing horses during hot, humid conditions in the southeastern United States and United Kingdom. To advance translational relevance of equine pasture asthma to human asthma, histologic features of airway remodeling in human asthma were scored in lung lobes from 15 pasture asthma-affected and 9 control horses of mixed breeds. All noncartilaginous airways were scored using a standardized grading rubric (0–3) in hematoxylin and eosin (HE) and Movat’s pentachrome-stained sections; 15 airways were chosen randomly from each lobe for analysis. Logistic regression identified disease, age, and lobe effects on probability of histologic outcomes. Airway smooth muscle (odds ratio [OR] = 2.5, P < .001), goblet cell hyperplasia/metaplasia (OR = 37.6, P < .0001), peribronchiolar elastic system fibers (OR = 4.2, P < .001), peribronchiolar fibrosis (OR = 3.8, P = .01), airway occlusion by mucus/inflammation (OR = 4.2, P = .04), and airway adventitial inflammation (OR = 3.0, P = .01) were significantly greater in diseased airways. A novel complex tissue disorganization, designated terminal bronchiolar remodeling, was overrepresented in diseased airways (OR = 3.7, P < .0001). Distribution of terminal bronchiolar remodeling corresponded to putative sites of air trapping in human asthma, at secondary pulmonary lobules. Age (>15 years) was an independent risk factor for increased peribronchiolar fibrosis, elastic system fibers, and terminal bronchiolar remodeling. Remodeling differed significantly between lung lobes, congruent with nonhomogeneous remodeling in human asthma. Equine pasture asthma recapitulates airway remodeling in human asthma in a manner not achieved in induced animal asthma models, endorsing its translational relevance for human asthma investigation.
Acute Leukemia in HorsesCooper, Carina J.; Keller, Stefan M.; Arroyo, Luis G.; Hewson, Joanne; Kenney, Daniel; Bienzle, Dorothee
doi: 10.1177/0300985817720983pmid: 28812528
Leukemia is broadly divided into acute and chronic lymphocytic and myeloid types based on the proportion of blasts, morphology of cells, and expression of specific antigens on neoplastic cells. Classifying leukemia in horses can be challenging if blasts predominate and since few antibodies to identify cell types are available. The objective of this study was to describe in detail the clinical and pathologic features of acute leukemia in horses. Twelve horses ranging from 0.2 to 25.9 years of age were diagnosed with acute leukemia. Six cases were classified as acute lymphocytic leukemia (ALL) based on predominance of blasts, lack of granulocytic or monocytic differentiation, and detection of CD3, CD20, and/or CD79a antigens by immunohistochemistry. Six other cases were classified as acute myeloid leukemia (AML) with myelomonocytic (n = 4), basophilic (n = 1), and eosinophilic (n = 1) differentiation based on > 20% bone marrow blasts and partial leukocytic differentiation. Reactivity with antibodies to Iba-1/AIF-1, CD172a, and CD163 was determined for all cases of AML. Eleven horses had thrombocytopenia, 10 had neutropenia, 8 had anemia, all had blasts on blood films, and none had leukocytosis. Ten horses had increased serum acute phase proteins. Bone marrow cellularity ranged from 30% to 100%, and the proportion of blasts ranged from 80% to 100% and 30% to 60% in ALL and AML, respectively. Horses were severely ill at diagnosis and euthanized within days or weeks. Unique features of acute leukemia in horses compared to other species were variable lymphocyte antigen expression (ALL) and frequent inflammation (ALL and AML).
Relationship Between Immunoglobulin Deposition and Early Lesions of Progressive Glomerulonephropathy in Young Common MarmosetsYamada, Naoaki; Hashimoto, Nanako; Kamiie, Junichi; Doi, Takuya; Sato, Junko; Inoue, Takeshi; Shirota, Kinji; Tsuchitani, Minoru
doi: 10.1177/0300985817712556pmid: 28578628
The authors previously investigated progressive glomerulonephropathy in 2- to 11-year-old common marmosets and characterized age-related changes of the renal glomeruli and development of tubulointerstitial lesions. In this study, immunoglobulin deposition and ultrastructural changes of the glomeruli were investigated in 5 young marmosets from 6 months to 3 years of age with pre-onset or early glomerulonephropathy. In all animals, the foot processes of podocytes were effaced, and IgM was deposited into the glomeruli. In glomeruli without glomerular basement membrane (GBM) alteration, IgM was the only immunoglobulin type deposited in the glomeruli. In cases with more advanced lesions of reticulation and thickening of GBM, IgA and IgG deposits were also observed. Therefore, the authors conclude that IgM may be the primary or earliest immunoglobulin deposited in this nephropathy, whereas IgA and IgG deposition may be connected to the progression of the glomerular lesions. IgM deposition and foot process effacement of podocytes occur early in the life of affected marmosets.
Lymphocyte Subsets in the Adrenal Glands of Dogs With Primary HypoadrenocorticismFriedenberg, S. G.; Brown, D. L.; Meurs, K. M.; Law, J. McHugh
doi: 10.1177/0300985816684914pmid: 28005496
Primary hypoadrenocorticism, or Addison’s disease, is an autoimmune condition common in certain dog breeds that leads to the destruction of the adrenal cortex and a clinical syndrome involving anorexia, gastrointestinal upset, and electrolyte imbalances. Previous studies have demonstrated that this destruction is strongly associated with lymphocytic-plasmacytic inflammation and that the lymphocytes are primarily T cells. In this study, we used both immunohistochemistry and in situ hybridization to characterize the T-cell subtypes involved. We collected postmortem specimens of 5 dogs with primary hypoadrenocorticism and 2 control dogs and, using the aforementioned techniques, showed that the lymphocytes are primarily CD4+ rather than CD8+. These findings have important implications for improving our understanding of the pathogenesis and in searching for the underlying causative genetic polymorphisms.
Hepatic Mastocytosis in Japanese Black CattleOhfuji, Susumu
doi: 10.1177/0300985817736115pmid: 29050542
In 5 Japanese Black steers (2-2.4 years old) that originated from 5 different feedlots, the livers were found at slaughter to have multiple nodular or cordlike lesions (5 steers) and an extensive fibrotic area (1 steer). Microscopic changes included extensive fibroplasia in the portal tracts and chronic proliferative endophlebitis-like lesions confined to the portal vein branches. Fibroplasia was much more prominent in the macroscopic fibrotic lesion of 1 steer. Portal vein branches presented irregular variciform dilation of the vascular lumen and fibroplastic changes in the subendothelial areas that showed occasional hemorrhage and were simultaneously infiltrated with large numbers of mast cells and moderate to large numbers of eosinophils. Within these subendothelial regions, not only did mast cells exhibit cytologically atypical features, but they also formed multifocal nodules. The venous lesions may represent a variant of mastocytosis with specific involvement of the hepatic portal vein branches in cattle.