Macrophages are a type of white blood cell found in horses that play a significant role in the immune system. They are involved in the detection, phagocytosis, and destruction of bacteria and other harmful organisms. Additionally, macrophages contribute to tissue homeostasis and repair by removing dead cells and stimulating tissue regeneration. In horses, macrophages are distributed throughout various tissues, including the liver, lungs, and lymph nodes, where they help orchestrate the body's response to infection and injury. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical implications of macrophages in equine health.
Threadgill DS, Steagall WK, Flaherty MT, Fuller FJ, Perry ST, Rushlow KE, Le Grice SF, Payne SL.The putative dUTPase domain was deleted from the polymerase (pol) gene of equine infectious anemia virus (EIAV) to produce a recombinant delta DUpol Escherichia coli expression cassette and a delta DU proviral clone. Expression of the recombinant delta DUpol polyprotein yielded a properly processed and enzymatically active reverse transcriptase, as determined by immunoblot analysis and DNA polymerase activity gels. Transfection of delta DU provirus into feline (FEA) cells resulted in production of virus that replicated to wild-type levels in both FEA cells and fetal equine kidney cells. In con...
Watson ED, Dixon CE.The distribution of T lymphocytes and of cells bearing MHC Class II antigens in the endometrium of the mare was studied using an avidin-biotin-peroxidase staining method. The cells within the endometrium which expressed MHC Class II were macrophages, lymphocytes, monocytes, dendritic cells, epithelial cells and endothelial cells. MHC Class II expression increased significantly (P < 0.05) in the luminal epithelium and tended (P = 0.0573) to increase in the subepithelial layers during oestrus. Numbers of T lymphocytes did not differ between oestrus and dioestrus. MHC Class II expression and T...
Atwal OS, Singh B, Staempfli H, Minhas K.The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monast...
Sellon DC, Perry ST, Coggins L, Fuller FJ.In situ hybridization of tissues from two horses infected with the wild-type Wyoming strain of equine infectious anemia virus (EIAV) identified the liver, spleen, lymph nodes, kidney, lung, and adrenal gland as the primary host tissue sites for viral transcription during acute infection. Combined immunohistochemistry, with a monoclonal antibody recognizing a cytoplasmic antigen of equine mononuclear phagocytes, and in situ hybridization for viral RNA identified most infected cells as mature tissue macrophages. In contrast, in situ hybridization of adherent peripheral blood mononuclear cells co...
Milam SB, Mackay RJ, Skelley LA.Secretion of tumor necrosis factor (TNF) by equine mammary exudate macrophages (MEM phi) exposed to bacterial lipopolysaccharide (LPS) was dose-dependent and was maximal (216.5 +/- 51.9 U/ml) at 100 micrograms LPS/ml, the highest concentration tested. All concentrations of dexamethasone tested (10(-8) to 10(-4) M) significantly (P less than or equal to 0.05) inhibited TNF production by MEM phi when the agent was added 1 hour before LPS. Pretreatment with pentoxifylline at concentrations greater than 3 micrograms/ml also significantly (P less than or equal to 0.05) reduced secretion of TNF by M...
Naylor JM, Clark EG, Clayton HM.We examined 18 horses with clinical signs of chronic obstructive pulmonary disease (COPD) using physical examination, cytological and bacteriological evaluation of bronchoalveolar fluid, and percutaneous lung biopsy. In 16 cases, histological examination of lung tissue confirmed the diagnosis of COPD. Two horses were excluded: one had uncomplicated bacterial pneumonia and in the other a satisfactory lung biopsy could not be obtained. In horses with COPD, the most common historical complaint was coughing, which was reported in 88%. The most frequently detected abnormal finding on physical exami...
Sweeney CR, Rossier Y, Ziemer EL, Lindborg S.Bronchoalveolar lavage (BAL) fluid was analyzed in healthy horses, using different lavage fluid volumes and lung sites. The only significant difference in the cellular composition of BAL fluid between the right and left lungs was the mast cell numbers, which were significantly higher in the left lung. Total cell count ranged from 34 to 330 cells/microliter for the right lung and 43 to 330 cells/microliter for the left lung. Percentage of neutrophils ranged from 1 to 7% in the right lung and 1 to 5% in the left lung. The small-volume (50 ml) lavage had a greater percentage of neutrophils and a ...
Morris DD, Crowe N, Moore JN, Moldawer LL.A study was performed to determine whether equine peritoneal macrophages produce interleukin 6 (IL-6) in vitro in response to endotoxin. Peritoneal fluid was collected from 14 clinically normal adult horses and was used as the source of peritoneal macrophages. Macrophages from each horse were isolated and cultured separately in vitro in the absence or presence of various concentrations (0.5, 5, 500 ng/ml) of endotoxin (lipopolysaccharide from Escherichia coli 055:B5). Culture medium supernatants were collected after 3, 6, 12, and 24 hours' incubation and were frozen at -70 C until assayed for ...
Crisman MV, Hodgson DR, Bayly WM, Liggitt HD.To determine whether road transport affected pulmonary phagocyte activity, 7 healthy Thoroughbred horses were shipped 1,160 kilometers over 36 hours. Fluid collected by bronchoalveolar lavage (BAL) 12 hours, and 7 and 14 days after transport was analyzed. Results were compared to those from the same horses pre-transport, and 7 non-transported control horses that had BAL performed at the same times as the transported horses. Of cells recovered with BAL the percentage of viable pulmonary alveolar macrophages (PAMs) declined from 90.0 +/- 0.9% pre-transport to 80.0 +/- 3.7% by 2 weeks post transp...
Ishino S, Kumagai K, Kuniyoshi S, Nakazawa M, Matsuda I, Oka M.An immunohistochemical analysis of Rhodococcus equi-induced pneumonia in 10 foals was performed by biotin-streptavidin system. The detection of R. equi was more sensitive in immuno-stain using anti-R. equi serum than in Gram's stain. This bacteria also reacted to anti-BCG serum. Lysozyme and alpha 1-antitrypsin were detectable in macrophages. A particularly intense staining was observed in association with intracellular bacteria. Though a degree of reaction for alpha 1-antichymotrypsin was very low in comparison with lysozyme and alpha 1-antitrypsin, it was also demonstrated in macrophages ing...
Kim CH, Casey JW.Equine infectious anemia virus (EIAV) is a lentivirus that infects and persists in the monocyte/macrophage populations of blood and tissues. We employed polymerase chain reaction to investigate the distribution and the level of genome variability of EIAV DNA in different tissues of a horse infected with a highly virulent variant of the Wyoming strain of the virus. Long terminal repeat, gag, and pol primer pairs were used to direct the amplification of EIAV DNA from the peripheral blood mononuclear cells and from cells, presumably the macrophage subtypes, of the kidney, spleen, liver, lymph nod...
Hondalus MK, Sweeney CR, Mosser DM.A Rhodococcus equi radiobinding assay has been developed using organisms labeled with 3H-uracil. These labeled organisms resemble their unlabeled counterparts with respect to colony morphology, viability, and buoyant density. Bacteria routinely incorporate between 5 x 10(-3) and 5 x 10(-2) counts per minute per colony forming unit (cfu) which in this assay allows the detection of fewer than 0.2 cfu per macrophage. Once incorporated, greater than 90% of the label remains bacterial associated for at least 4 h postlabeling. The majority of the label is trichloroacetic acid precipitable, partition...
Morris DD, Crowe N, Moore JN.The purpose of this study was to determine if a structurally novel dual inhibitor of arachidonic acid metabolism, SK & F 86002, would inhibit the endotoxin-induced production of tumor necrosis factor (TNF) activity by equine peritoneal macrophages. Equine peritoneal macrophages were variously pretreated for 0, 0.5 and 2 h with SK & F 86002 at 10(-9) to 10(-4) molar final concentrations or were left untreated. Then, the macrophages were cultured in vitro in the presence of endotoxin (5 ng/mL). Supernatant media were collected after 4 h and stored at -70 degrees C until assayed for TNF a...
Grünig G, Winder C, Hulliger C, Witschi U, Hermann M, Jungi TW, von Fellenberg R.Increases in procoagulant activities (PCA) in equine lung macrophages were induced by non-adherent blood lymphocytes which were prestimulated with phytohaemagglutinin for 48 to 72 hours or by supernatants harvested from prestimulated blood lymphocyte cultures. However, prestimulated lymphocyte suspensions themselves expressed PCA which was most probably derived from contaminating monocytes. Because non-adherent cells from lymphocyte suspensions may have attached to adherent macrophages, cells within lymphocyte suspensions might have contributed to the PCAs expressed by lymphocyte-stimulated lu...
Grünig G, Witschi U, Winder C, Hermann M, von Fellenberg R.Supernatants of equine respiratory secretions enhanced the migration of equine neutrophils into the lower compartments of Boyden chambers. Checkerboard analysis revealed that the neutrophil migration promoting activity (NMPA) of secretion specimens was in great part caused by chemokinesis, irrespective of the neutrophil score of the specimen. The NMPA of respiratory secretions was correlated neither with the neutrophil score of the secretion specimen nor with the severity of the chronic pulmonary disease. Respiratory secretions collected while horses were kept under low dust or under dusty hou...
MacKay RJ, King RR, Dankert JR, Reis KJ, Skelley LA.Blood monocytes and alveolar macrophages (AM) were harvested from foals (aged 46 days to 6 months) and cultured in either medium alone or medium containing 10 micrograms/ml bacterial lipopolysaccharide (LPS). After 24 h, culture supernates were collected and analyzed for cytotoxic activity on sensitized L929 cells. Both monocytes and AM that had been treated with LPS produced significantly more cytotoxic activity than the same cell type exposed to medium lacking LPS. LPS-treated macrophages secreted significantly more cytotoxic activity (120 +/- 17.8 U/ml) than did LPS-treated monocytes (47.3 ...
Arrighi S, Romanello MG, Domeneghini C.Following previous studies about the ultrastructure of male genital tract in parental species, a comparative study of epididymis of one of the possible hybrids, the mule, has been undertaken. Apart from small differences, general features of epididymal epithelium in the mule are similar to those of parental species. However, extension of our studies from the donkey to the horse to the hybrid permits a deeper insight into the morphology of this tract of excurrent duct. In the meantime, it is possible to evidence some features, sometimes shared with other species if taken separately, which in th...
Stitz L, Richt JA, Rott R.An overview of the pathogenesis of Borna disease (BD) in rats as a model for the naturally occurring infection in horses and sheep is presented. Our findings revealed a virus infection in which a virus-specific T cell-mediated immune response leads to disease. The immune cells capable of mediating this immunopathological reaction were defined as helper/inducer T cells. In all, the described observations indicate that CD4+ T cells and macrophages trigger a delayed-type hypersensitivity reaction and cause BD together with other cells of the immune system.
Morris DD.Endotoxemia remains the leading cause of death in horses, being intimately involved in the pathogenesis of gastrointestinal disorders that cause colic and neonatal foal septicemia. Endotoxins, normally present within the bowel, gain access to the blood across damaged intestinal mucosa, or endotoxemia occurs when gram negative organisms proliferate in tissues. Endotoxins are removed from the circulation by the mononuclear phagocyte system, and the response of mononuclear phagocytes to these lipopolysaccharides (LPS) play an important role in determining the severity of clinical disease. Macroph...
Morris DD, Henry MM, Moore JN, Fischer JK.A study was conducted to determine whether dietary supplements with alpha-linolenic acid altered the ability of equine peritoneal macrophages to produce tumor necrosis factor (TNF) in response to endotoxin. Peritoneal macrophages were harvested from 6 healthy adult horses before and after the horses were fed a nutritionally balanced ration that contained 8% linseed oil as a source of alpha-linolenic acid. The macrophages were cultured in media containing no additives (control), endotoxin (0.5 to 50 ng/ml), or the calcium ionophore, A23187. Macrophage supernatants were collected after 6 and 24 ...
Rossier Y, Sweeney CR, Ziemer EL.Bronchoalveolar lavage was performed in 22 horses with pneumonia or pleuropneumonia. All horses had clinical evidence of pneumonia or pleuropneumonia on the basis of physical, radiographic, ultrasonographic, tracheobronchial aspirate or post-mortem findings. Results of lavage fluid analysis were normal in 9 horses, equivocal in 3 horses, and abnormal in 10 horses. Abnormal lavage fluid had increased total cell count, increased relative and absolute neutrophil counts, degenerative neutrophils, and decreased relative and absolute macrophage and lymphocyte counts.
Winder NC, Pellegrini A, von Fellenberg R.Alpha 2-beta 1-glycoprotein may be found free in horse serum or complexed with alpha-1-proteinase inhibitor to form pre-alpha 2-elastase inhibitor. There has been little information published concerning alpha 2-beta 1-glycoprotein and its possible tissue sources in horses. A peroxidase-antiperoxidase technique was used to identify alpha 2-beta 1-glycoprotein in buffy coat and bone marrow neutrophils of healthy horses. Macrophages and neutrophils in bronchoalveolar lavage samples from clinically normal horses and from horses with chronic pulmonary disease also were positive for alpha 2-beta 1-g...
Fadok VA.Abstract- Urticarial eruptions, with or without pruritus, are common lesions in horses. The pathogenesis of these lesions can include immunological and other mechanisms. Research in the human field suggests that the mast cell co-ordinates the urticarial response by releasing a complex array of inflammatory mediators. Other cells, including the neutrophil, the eosinophil and the macrophage, may also play a role in the development of wheals. Elucidation of the role of many of these cells and mediators in the evolution of urticaria is only just beginning. Successful treatment of this dermatologic...
Grünig G, Hulliger C, Hermann M, Winder C, von Fellenberg R.Bronchopulmonary lavage was performed in 10 healthy horses and in 39 horses with chronic pulmonary disease. The predominant cell types were macrophages in healthy horses and neutrophils in severely diseased horses. Procoagulant activity (PCA) was detected in all 32 cell-free supernatants examined and in all 49 unpurified cell suspensions. Cells were separated by centrifugation on discontinuous gradients prepared either with Percoll or with Metrizamide. Macrophages were enriched in subpopulations of low density. Neutrophils could not be purified by density gradient centrifugation using either g...
Watson ED, Mair TS, Sweeney CR.Because of their capacity to produce prostanoids, alveolar macrophages may play a part in the pathogenesis of chronic obstructive pulmonary disease (COPD). Cultured equine alveolar macrophages and peripheral blood monocytes synthesised mainly prostaglandin (PG)F and PGE2. They also synthesised smaller quantities of PGI2, measured as the stable metabolite 6-keto-PGF1 alpha, and thromboxane B2. Concentrations of immunoreactive PGF and PGE2 were measured in bronchoalveolar lavage fluid collected from clinically normal horses (n = 3) and horses with COPD (n = 3). None of the normal horses had dete...
Winder NC, Grünig G, Hermann M, von Fellenberg R.The concentration of soluble fibrinogen derivatives (SFD) and protease and procoagulant activities were determined in cell-free supernatants of equine respiratory secretions obtained from horses with chronic pulmonary disease. The concentration of neutrophils was estimated from direct smears of the secretions. Lung specimens and smears of the secretions were evaluated for the presence of fibrin or fibrinogen by use of immunohistochemical methods. Thirty-five of 80 specimens tested contained SFD. Respiratory secretions from horses with moderate or severe chronic pulmonary disease contained SFD ...
Brumbaugh GW, Davis LE, Thurmon JC, Savage DC.Opsonized Rhodococcus equi activated the respiratory burst of resident alveolar macrophages (AM) from adult horses in a logarithmic-linear, mass-related manner. The effect of R equi was not significantly different from that of equal masses of opsonized zymosan A. Therefore, R equi does not appear to attenuate the respiratory burst of equine AM. The stimulatory effect of R equi was not reflected by increased production of superoxide anion (O2-), but increased activity of the hexose monophosphate shunt was observed. These results suggest a similarity between the respiratory burst of AM from hors...
Morris DD, Moore JN, Fischer K, Tarleton RL.A study was performed to determine whether equine macrophages produce tumor necrosis factor (TNF) activity in vitro in response to endotoxin and to study the effects of endotoxin concentration and incubation time on the amount of TNF produced. Equine peritoneal macrophages were isolated and cultured in vitro for 2, 6, 12, or 24 hr in tissue culture media containing 1) no additive (nonstimulated control), 2) endotoxin (0.5 ng/ml, 5 ng/ml, or 5 micrograms/ml), or 3) the calcium ionophore A23187 (0.95 microM). The supernatant media concentrations of TNF activity were determined by an in vitro cyt...
Wong CW, Thompson HL, Thong YH, Thornton JR.Bronchoalveolar lavage samples were collected using a fibreoptic endoscope from horses at specified times before and after single bouts of exercise. Lucigenin-dependent phagocytic chemiluminescence was used to assess the effect of exercise on the alveolar macrophage metabolic activity in response to stimulation by opsonised zymosan. A profound suppressive effect on the chemiluminescence production was present throughout the first three days after exercise. However, the cellular composition of lavage fluids was not altered by the exercise. It is suggested that strenuous exercise may jeopardize ...
McGuire TC, O'Rourke KI, Perryman LE.Virus replication and subsequent viremia are clearly correlated with clinical disease in EIAV infected horses. Termination of viremia is the result of specific immune responses. Recurrences of viremia are associated with antigenic variation of neutralization-sensitive epitopes. Immunosuppression experiments indicate that the eventual control of EIAV and development of carriers is mediated by the immune system. Even though the immune response to EIAV has a protective effect, immune responses also cause some of the lesions. At least one part of the anemia, erythrocyte destruction, is caused by t...
Singh B, Minhas KJ, Atwal OS.The pulmonary intravascular macrophages (PIMs) of the ponies possess all the characteristics of a fully differentiated resident macrophage, which forms adhesive plaques with the capillary-endothelial cells. In addition, it has a unique surface coat which does not conform to the concept of a carbohydrate rich membrane bound glycocalyx generally associated with macrophages. We studied the responses of these cells especially in the context of its globular surface coat to multiple doses of MB intravenously: 0.2 ml/kg body copper tracer substance. The ponies were treated with three doses of MB intr...
Del Piero F, Wilkins PA, Timoney PJ, Kadushin J, Vogelbacker H, Lee JW, Berkowitz SJ, La Perle KM.A case of fatal nonneurological equine herpesvirus 1 (EHV-1) infection in a yearling filly is described. Gross lesions included extensive pulmonary edema, prominent laryngeal lymphoid follicles, and congestion and edema of the dorsal third ventricle choroid plexus. Histologically, there was vasculitis, hemorrhage, and edema in the lungs and dorsal third ventricle choroid plexus as well as mild intestinal crypt necrosis with occasional intranuclear inclusion bodies. The perivascular and vascular inflammatory infiltrates were comprised mainly of T lymphocytes and macrophages. EHV-1 antigen was i...
Morris DD, Harmon BG, Moore JN.A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELI...
Liu Q, Wang XF, Du C, Lin YZ, Ma J, Wang YH, Zhou JH, Wang X.Integration is an important feature of retroviruses and retrovirus-based therapeutic transfection vectors. The non-primate lentivirus equine infectious anaemia virus (EIAV) primarily targets macrophages/monocytes . Investigation of the integration features of EIAV strains, which are adapted to donkey monocyte-derived macrophages (MDMs), is of great interest. In this study, we analysed the integration features of EIAV in equine MDMs during infection. Our previously published integration sites (IS) for EIAV in fetal equine dermal (FED) cells were also analysed in parallel as references. Sequenc...
Grünig G, Witschi U, Winder C, Hermann M, von Fellenberg R.Supernatants of equine respiratory secretions enhanced the migration of equine neutrophils into the lower compartments of Boyden chambers. Checkerboard analysis revealed that the neutrophil migration promoting activity (NMPA) of secretion specimens was in great part caused by chemokinesis, irrespective of the neutrophil score of the specimen. The NMPA of respiratory secretions was correlated neither with the neutrophil score of the secretion specimen nor with the severity of the chronic pulmonary disease. Respiratory secretions collected while horses were kept under low dust or under dusty hou...
Swardson CJ, Lichtenstein DL, Wang S, Montelaro RC, Kociba GJ.To characterize infection of bone marrow-derived macrophages (BMDM) with equine infectious anemia virus (EIAV) by determining virus production, effects on viability, and induction of cytokines. Methods: BMDM obtained from bone marrow of 6 clinically normal adult horses. Methods: BMDM were infected with EIAV at a multiplicity of infection of 8. Cell viability, percentage of cells with detectable viral protein, reverse transcriptase activity, and concentrations of infective virus (focus-forming units/ml), interleukin 6, and tumor necrosis factor-alpha were measured in culture supernatant samples...
Schneberger D, Sethi RS, Singh B.Endothelium plays an important role in maintaining the vascular barrier and physiological homeostasis. Endothelium also is fundamental to the initiation and regulation of inflammation. Endothelium demonstrates phenotypic and functional heterogeneity not only among various organs but also within an organ. One of the striking examples would be the pulmonary endothelium that participates in creating blood-air barrier. Endothelium in large pulmonary blood vessels is distinct in structure and function from that lining of the pulmonary capillaries. This chapter focuses on the comparative aspects of ...
Köhne M, Mönnig F, Papin J, Schöniger S, Tönissen A, Rohn K, Martinsson G, Schoon HA, Sieme H.The effects of standard uterine body and hysteroscopic insemination on endometrial health were investigated. For this purpose, 33 mares were assigned to five different protocols: control (no insemination; n = 7), sham AI (sham uterine body insemination; n = 6), sham HysAI (sham hysteroscopic insemination; n = 7), standard AI (standard uterine body insemination, 300 × 106 progressively motile sperms (PMS); n = 7) and HysAI (hysteroscopic insemination, 100 × 106 PMS; n = 6). Sampling included uterine swabbing for microbiological examination, cytology for determination of polymorphonucle...
Raidal SL, Bailey GD, Love DN.Flow cytometric techniques were developed for the evaluation of oxidative burst activity in equine peripheral blood neutrophils and lymphocytes, as well as bronchoalveolar lavage derived pulmonary alveolar macrophages and lymphocytes. The oxidation of dichlorofluorescin was measured by the increased fluorescence of cells stimulated with phorbol myristate acetate or a variety of other stimulants. Flow cytometry was a suitable method for the evaluation of the intracellular oxidation in all cell populations evaluated. Analysis was rapid and cell separation before analysis was not required. Hetero...
Atwal OS, McDonell W.The pulmonary intravascular macrophages (PIMs) of equines contain a unique electron-dense surface coat that is predominantly composed of lipoproteins. A single exposure of inhalatory halothane causes mobilization of the surface coat into the endocytotic system of the PIMs, followed by expansion of the Golgi apparatus and its enrichment with acid phosphatase. Simultaneously, the cells of the lymphocytic series show hyperplasia in the form of mitotic changes inside the microvascular compartment of the lung. Halothane is known to cause acute and chronic hepatotoxicity because of its biotransforma...
Rivolta AA, Bujold AR, Wilmarth PA, Phinney BS, Navelski JP, Horohov DW, Sanz MG.Neonates have different cellular composition in their bronchoalveolar lavage fluid (BALF) when compared to foals and adult horses; however, little is known about the non-cellular components of BALF. The objective of this study was to determine the proteomic composition of BALF in neonatal horses and to compare it to that of foals and adult horses. Bronchoalveolar lavage fluid samples of seven neonates (< 1 week age), four 5 to 7-week-old foals, and six adult horses were collected. Quantitative proteomics of the fluid was performed using tandem mass tag labeling followed by high resolution l...
MacKay RJ, King RR, Dankert JR, Reis KJ, Skelley LA.Blood monocytes and alveolar macrophages (AM) were harvested from foals (aged 46 days to 6 months) and cultured in either medium alone or medium containing 10 micrograms/ml bacterial lipopolysaccharide (LPS). After 24 h, culture supernates were collected and analyzed for cytotoxic activity on sensitized L929 cells. Both monocytes and AM that had been treated with LPS produced significantly more cytotoxic activity than the same cell type exposed to medium lacking LPS. LPS-treated macrophages secreted significantly more cytotoxic activity (120 +/- 17.8 U/ml) than did LPS-treated monocytes (47.3 ...
Dyer RM, Leid RW.Isolated equine alveolar macrophages obtained by bronchopulmonary lavage of four live ponies demonstrated surface receptors for equine IgG, equine IgM, and complement-coated sheep red blood cells, but not equine IgM or complement-coated erythrocytes alone. In addition, demonstration of IgG receptors was found to depend on the level of erythrocyte sensitization and could not be demonstrated by red blood cell rosetting techniques at low levels of sensitization. Demonstration of receptors for equine complement by red cell rosetting techniques required the presence of both IgM antibody and serum d...
Domańska D, Trela M, Pawliński B, Podeszewski B, Domino M.The somatic cell count in milk (SCC) and electrical conductivity of milk (ECM) are indicators of the health status of the mammary gland. Among somatic cells, mainly polymorphonuclear neutrophils (PMN), macrophages (MAC), and lymphocytes (LYM) are rated. This study aimed to determine the SCC, PMN, MAC, LYM, ECM, and bacteriological index (BII) in milk collected from mares with (CM) and without (NCM) clinical symptoms of mastitis concerning mares with (SM) and without (NSM) subclinical mastitis. Milk samples were collected from 27 mares divided into NCM (n = 12) and CM (n = 15) groups. In sample...
Sellon DC, Cullen JM, Whetter LE, Gebhard DH, Coggins L, Fuller FJ.An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The...
Lee GKC, Kang H, Beeler-Marfisi J, Sears W, Lillie BN, Bienzle D.Salivary scavenger and agglutinin (SALSA) is a secreted protein with various immunomodulatory roles. In humans, the protein agglutinates and inactivates microorganisms, and inhibits the release of pro-inflammatory cytokines. Saliva, which is rich in SALSA, accelerates bacterial phagocytosis, but SALSA's contribution is unclear. In horses, the functions of SALSA in inflammation remain undetermined, so they were investigated through phagocytosis and cytokine assays. Equine SALSA was purified from duodenal tissue, which contains abundant SALSA. To assess phagocytosis, fluorescently-labelled bacte...
Suzuki Y, Takai S, Kubota H, Hasegawa N, Ito S, Yabuuchi Y, Sasaki Y, van Duijkeren E, Kakuda T.Rhodococcus equiis the causative agent of pyogenic pneumonia in foals, and a virulence-associated protein A (VapA) encoded on the pVAPA virulence plasmid is important for its pathogenicity. In this study, we analyzed the virulence of R. equi strain U19, originally isolated in the Netherlands in 1997 and the genetic characteristics of the pVAPA_U19 plasmid. U19 expressed VapA that was regulated by temperature and pH and underwent significant intracellular proliferation in macrophages. The restriction fragment length polymorphism of pVAPA_U19 digested with EcoRI was similar to that of pREAT701 (...
Sellers RS, Toribio RE, Blomme EA.Idiopathic systemic granulomatous disease, which has been reported in horses, cattle and human beings, is characterized by perivascular granulomatous and lymphoplasmacytic inflammation in many organ systems. Diagnosis is based on the exclusion of possible viral, fungal or bacterial causes. The disease was identified in a miniature pony with widespread lymphoplasmacytic and granulomatous inflammation, special staining techniques having revealed no evidence of any aetiological agent. Skin lesions, which were severe, consisted of hyperkeratosis and serocellular crust formation, with inflammatory ...
Berghaus LJ, Cathcart J, Berghaus RD, Ryan C, Toribio RE, Hart KA.Rhodococcus equi (R. equi), a pneumonia-causing intracellular bacterium, results in significant morbidity and mortality in young foals, while healthy adult horses rarely develop disease. Survival and replication within alveolar macrophages (AMφ) are the hallmarks of R. equi's pathogenicity. The vitamin D receptor (VDR) and its ligand, the active vitamin D metabolite 1,25(OH)2D, are important in immune responses to intracellular bacteria. The vitamin D/VDR pathway regulates the downstream production of cytokines in infected human AMφ. The immunomodulatory role of the vitamin D/VDR pathway in ...
Tavanaeimanesh H, Alinia Z, Sadeghian Chaleshtori S, Moosavian H, Mohebi Z, Daneshi M.During transportation many horses develop post-transportation infection, which can be life-threatening and end their sport career. Preventing mucus accumulation and inflammation during transportation is vital, emphasizing the need for effective strategies to enhance overall horse health welfare. Objective: Assess the impact of N-acetylcysteine (NAC) on mucus accumulation and inflammation in horses subjected to 18 hours of head confinement. Methods: Six healthy crossbred horses, 5.3 ± 2.1 years of age and weighing 387 ± 30 kg. Methods: Prospective placebo-controlled cross-over d...
Barbazanges P, Richard EA, Lemonnier LC, Wouters CP, Toquet MP, Couroucé A.Cytology of airway samples is sensitive for diagnosis of exercise-induced pulmonary haemorrhage (EIPH), but the association between tracheal wash (TW) and bronchoalveolar lavage fluid (BALF) is unknown. The objective of this study was to determine whether diagnosis of EIPH, using haemosiderophages/macrophages (H/M) ratio, differs when based on TW or BALF. Methods: A prospective cross-sectional study was conducted on 102 standardbred horses in training. TW and BALF were collected concomitantly from all horses at rest (at least 24 hours after their last training or race), and their H/M ratios w...
Ammons DT, Chow L, Goodrich L, Bass L, Larson B, Williams ZJ, Stoneback JW, Dow S, Pezzanite LM.Osteoarthritis (OA) is a major source of pain and disability worldwide. Understanding of disease progression is evolving, but OA is increasingly thought to be a multifactorial disease in which the innate immune system plays a role in regulating and perpetuating low-grade inflammation. The aim of this study was to enhance our understanding of OA immunopathogenesis through characterization of the transcriptomic responses in OA joints, with the goal to facilitate the development of targeted therapies. Unassigned: Single-cell RNA sequencing (scRNA-seq) was completed on cells isolated from the syno...
Maloney SM, Shaw TM, Nennig KM, Larsen MS, Shah A, Kumar A, Marcotrigiano J, Grove J, Snijder EJ, Kirchdoerfer RN, Bailey AL.Arteriviruses are a family of single-stranded, positive-sense RNA (+ssRNA) viruses that infect diverse animal hosts. Many arteriviruses are macrophage-tropic, consistent with their utilization of the macrophage-specific molecule CD163 as a receptor. However, the horse arterivirus (equine arteritis virus, EAV), which infects additional cell types beyond macrophages, does not utilize CD163 in its entry mechanism. Here, we use a genome-wide CRISPR knockout screen to identify alternative receptors that could explain this discrepancy in arterivirus receptor utilization and tropism, identifying the ...
Kalsow CM, Albrecht TR, Steinberg NP, Lunn DP.Immunohistochemical evaluation of equine tissue necessitates the use of antibodies reactive with cells from a heterogeneous population. Lymphoid tissues from 12 horses were fixed in Bouin's fluid, ethanol or formalin and examined for immunohistochemical reactivity with anti-equine and anti-human monoclonal antibodies (MAbs) specific for MHC Class II antigens, T and B lymphocytes, and macrophages. Only a few of the anti-equine MAbs tested were reactive with fixed, paraffin wax-embedded tissue. Anti-human MAbs expanded the desired range of reactivity and increased the consistency in different an...
Winder NC, Pellegrini A, von Fellenberg R.A peroxidase antiperoxidase technique was used to identify alpha 2-macroglobulin in formalin-fixed sections of normal equine lung and liver and in tissue sections and bronchoalveolar lavage fluid from horses with various lung diseases. Equine peripheral blood leucocytes and bronchoalveolar lavage samples from clinically normal horses were negative for alpha 2-macroglobulin. It was concluded that liver and pulmonary macrophages may be potential sources of alpha 2-macroglobulin. Although alpha 2-macroglobulin may play a role in various chronic bronchointerstitial pneumonias of the horse, it is d...
Mérant C, Sheoran A, Timoney JF.Streptococcus equi (Se), the cause of equine strangles, is highly resistant to phagocytosis by neutrophils and is usually classified as an extracellular pathogen. Large numbers of the organism in tonsillar tissues during the acute phase of the disease are completely eliminated during convalescence by mechanisms not yet understood. In this study we demonstrate in an opsono-bactericidal assay and by cytometry and confocal microscopy that Se is interiorized and killed by equine blood monocytes. This finding supports the hypotheses that adaptive immune clearance is mediated by tonsillar macrophage...
Dyer RM, Liggitt HD, Leid RW.A device was constructed from an equine nasogastric tube, polyethylene tubing, and a 3-way stopcock and used to lavage the lungs of anesthetized ponies. The technique was safe and atraumatic in that 6.4 to 19.7 X 10(7) purified alveolar macrophages were removed from the lungs without harm to the ponies or contamination of the samples with blood. Studies of these highly purified cell suspensions revealed a mean viability of 85% as assessed by eosin dye exclusion with a mean recovery (+/- SD) of 12.5 +/- 4.8 X 10(7) pulmonary alveolar macrophages/pony.
Mariotti F, Cuteri V, Takai S, Renzoni G, Pascucci L, Vitellozzi G.Two horses with Rhodococcus equi infection were examined post mortem by an immunohistochemical method (peroxidase-antiperoxidase; PAP) with a monoclonal antibody (Mab 10G5) to the 15-17 kDa antigen of R. equi. One of the horses was also examined bacteriologically, R. equi being isolated in culture. Immunolabelling with this Mab was marked and widespread. On the other hand, the immunohistochemical reactivity of infected macrophages with a polyclonal antibody specific for lysozyme was slight. Thus, Mab 10G5 would appear to be a useful diagnostic reagent in R. equi infection, with or without cult...
van der Kolk JH, Grinwis GC.A three-year-old pregnant Dutch Warmblood mare was referred to the Department of Large Animal Medicine and Nutrition, Faculty of Veterinary Medicine, because of weight loss for 1 month. The main clinical features were (beside weight loss) moderate ventral oedema, enlarged mesenteric lymph nodes, and uniform thickening of the wall of the jejunum. Haematological evaluation revealed leukocytosis (15.9 G.l-1 with 18% lymphocytes and 1% eosinophils) and a decreased total serum protein and albumin concentration (35 g.l-1 and 36.3% albumin). At necropsy transmural eosinophilic enteritis was found in ...
Watson ED, Dixon CE.The distribution of T lymphocytes and of cells bearing MHC Class II antigens in the endometrium of the mare was studied using an avidin-biotin-peroxidase staining method. The cells within the endometrium which expressed MHC Class II were macrophages, lymphocytes, monocytes, dendritic cells, epithelial cells and endothelial cells. MHC Class II expression increased significantly (P < 0.05) in the luminal epithelium and tended (P = 0.0573) to increase in the subepithelial layers during oestrus. Numbers of T lymphocytes did not differ between oestrus and dioestrus. MHC Class II expression and T...
