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Topic:Immunology

The equine immune system is a complex network of cells, tissues, and organs that work collaboratively to defend against pathogens and maintain homeostasis. It consists of innate and adaptive components, each with distinct functions and mechanisms. The innate immune system provides the first line of defense through physical barriers, phagocytic cells, and the complement system. The adaptive immune system involves lymphocytes, such as B cells and T cells, which generate specific responses to antigens and provide immunological memory. Research in equine immunology explores the interactions between these components, the impact of genetic and environmental factors on immune function, and the development of vaccines and therapeutics. This page gathers peer-reviewed studies and scholarly articles focusing on the mechanisms, regulation, and clinical applications of the equine immune system in health and disease.
Generic immunoassay of corticosteroids with minimum pre-treatment of urine samples.
The Analyst    December 1, 1994   Volume 119, Issue 12 2631-2634 doi: 10.1039/an9941902631
Rodriguez ML, McConnell I, Lamont J, Campbell J, FitzGerald SP.A generic, rapid and sensitive enzyme linked immunosorbent assay (ELISA) test has been developed which allows large-scale simultaneous testing of synthetic corticosteroids viz., flumethasone, dexamethasone and betamethasone. This assay can be directly applied to diluted urine samples (1 + 9) without hydrolysis of glucuronide or sulfate conjugates or any other treatment of samples. The polyclonal antibody was obtained by immunizing sheep with a flumethasone derivative linked to human serum albumin. This polyclonal antibody displayed high-reactivity with several synthetic corticosteroids whilst ...
Treatment of aplastic anemia with an investigational antilymphocyte serum prepared in rabbits.
The American journal of the medical sciences    December 1, 1994   Volume 308, Issue 6 338-343 doi: 10.1097/00000441-199412000-00005
Stein RS, Means RT, Krantz SB, Flexner JM, Greer JP.The authors evaluated antilymphocyte serum prepared in rabbits (ALS-R) as an alternative to antilymphocyte serum prepared in horses (ALG-H) in the therapy of aplastic anemia. Between 1980 and 1993, 57 evaluable patients received ALS-R and prednisone +/- cyclosporine +/- androgens. Standard response criteria were used and patients were evaluated at 3 months from the start of therapy. Median age was 43 years. Disease was present for up to 2 months in 24 patients, 2-5 months in 14 patients, and 6 months or more in 19 patients. Disease was severe in 30 patients and moderate in 27. Responses occurr...
Monoclonal antibodies to the equine CD2 T lymphocyte marker, to a pan-granulocyte/monocyte marker and to a unique pan-B lymphocyte marker.
Immunobiology    December 1, 1994   Volume 192, Issue 1-2 48-64 doi: 10.1016/S0171-2985(11)80407-9
Tumas DB, Brassfield AL, Travenor AS, Hines MT, Davis WC, McGuire TC.Murine monoclonal antibodies, HB88A, B29A and DH59B separately identify the CD2 T lymphocyte molecule, a unique pan-B lymphocyte surface marker and a pan-granulocyte/monocyte surface molecule, respectively, in the horse. Specificity was shown by two-color immunofluorescent flow cytometry and immunofluorescent microscopy. MAb HB88A reacted with a 52 kDa pan-T lymphocyte molecule present on 75% +/- 7 of peripheral blood lymphocytes (PBL) (n = 15 horses). It also reacted with lymphocytes restricted to T lymphocyte dependent areas of lymph node and spleen. Specificity of mAb HB88A to CD2 was demon...
Receptor specificity in human, avian, and equine H2 and H3 influenza virus isolates.
Virology    November 15, 1994   Volume 205, Issue 1 17-23 doi: 10.1006/viro.1994.1615
Connor RJ, Kawaoka Y, Webster RG, Paulson JC.The receptor specificity of 56 H2 and H3 influenza virus isolates from various animal species has been determined to test the relevance of receptor specificity to the ecology of influenza virus. The results show that the receptor specificity of both H2 and H3 isolates evaluated for sialic acid linkage specificity and inhibition of hemagglutination by horse serum correlates with the species of origin, as postulated earlier for H3 strains based on a limited survey of five human, three avian, and one equine strain. Elucidation of the amino acid sequence of several human H2 receptor variants and a...
Distribution of equid herpesvirus-1 (EHV-1) in respiratory tract associated lymphoid tissue: implications for cellular immunity.
Equine veterinary journal    November 1, 1994   Volume 26, Issue 6 470-473 doi: 10.1111/j.2042-3306.1994.tb04052.x
Kydd JH, Smith KC, Hannant D, Livesay GJ, Mumford JA.Twelve adult ponies and 2 conventional foals were exposed intranasally to EHV-1, strain Ab4 (TCID50 10(-6.6) and samples of respiratory tract associated lymphoid tissues were recovered between 12 h and 13 days after infection. Infectious virus was detected in tissue homogenates using susceptible cell monolayers and expression of viral antigens was monitored using indirect immunoperoxidase histochemistry on paraffin sections. The results showed both infectious EHV-1 and viral antigens in respiratory tract associated lymph nodes 12 h after exposure. Infected leucocytes were identified morphologi...
Resistance of castrated male horses to attempted establishment of the carrier state with equine arteritis virus.
Journal of comparative pathology    November 1, 1994   Volume 111, Issue 4 383-388 doi: 10.1016/s0021-9975(05)80096-9
McCollum WH, Little TV, Timoney PJ, Swerczek TW.Twelve geldings all became infected when inoculated intranasally with the KY-84 strain of equine arteritis virus (EAV), a strain previously shown to be capable of establishing the carrier state in the stallion. With the exception of one animal that showed no effects other than pyrexia, all of the geldings developed clinical signs characteristic of equine viral arteritis (EVA). The geldings were febrile for varying periods within the range of 2-10 days after inoculation. Viraemia occurred from day 2 onwards, for periods varying from 9 to at least 19 days. Nasal shedding of virus began 2-4 days ...
Distribution of equid herpesvirus-1 (EHV-1) in the respiratory tract of ponies: implications for vaccination strategies.
Equine veterinary journal    November 1, 1994   Volume 26, Issue 6 466-469 doi: 10.1111/j.2042-3306.1994.tb04051.x
Kydd JH, Smith KC, Hannant D, Livesay GJ, Mumford JA.Twelve adult ponies and 2 conventional foals were exposed to 10(6.6) TCID50 of Equid herpesvirus-1 (EHV-1), strain Ab4 and samples of respiratory tract tissues were recovered. Infectious virus in tissue homogenates was detected using susceptible cell monolayers and expression of viral antigens was monitored using indirect immunoperoxidase histochemistry of paraffin sections. The results illustrated the rapid dissemination of EHV-1 throughout the respiratory tract, with early replication in the lungs one day after exposure. Endothelial cell infection was prominent in all areas of the nasopharyn...
Comparative analysis of phagocytosis of fungal cells by insect hemocytes versus horse neutrophils.
Developmental and comparative immunology    November 1, 1994   Volume 18, Issue 6 455-466 doi: 10.1016/s0145-305x(06)80001-7
Mazet I, Pendland J, Boucias D.In this study, the phagocytic ability of Spodoptera exigua hemocytes was compared to horse neutrophils. In vitro assays showed that the insect granulocytes and horse neutrophils actively phagocytose FITC-labeled Paecilomyces farinosus blastospores opsonized with S. exigua hemolymph lectin or horse serum, respectively. Killing of fungal cells by the neutrophils and hemocytes was analyzed under in vitro conditions. Neutrophils reduced the growth of P. farinosus up to 65% whereas no fungicidal activity was observed with hemocyte monolayers. The production of oxygen metabolites by both phagocytic ...
Distribution of SP- and CGRP-like immunoreactive nerve fibers in the lower respiratory tract of neonatal foals: evidence for loss during development.
Anatomy and embryology    November 1, 1994   Volume 190, Issue 5 469-477 doi: 10.1007/BF00235494
Sonea IM, Bowker RM, Robinson NE, Holland RE.The lungs of neonatal foals contain many nerves immunoreactive for substance P and calcitonin gene-related peptide. These nerves are closely associated with the epithelium, bronchial and pulmonary vessels and the airway smooth muscle of all intrathoracic airways, including non-cartilaginous bronchioles. Activation of sensory nerves in the respiratory epithelium could thus potentially affect, via local axon reflexes, vascular and respiratory smooth muscle in neonatal equine airways. Nerves immunoreactive for these peptides are much more widely distributed within the lung than in adult horses; t...
An immunohistochemical study of an equine B-cell lymphoma.
Journal of comparative pathology    November 1, 1994   Volume 111, Issue 4 445-451 doi: 10.1016/s0021-9975(05)80102-1
Asahina M, Murakami K, Ajito T, Goryo M, Okada K.The tissues of an 8-year-old thoroughbred castrated male horse with equine lymphoma were examined immunohistochemically. Neoplastic masses were observed in the mediastinum, mesenteric lymph nodes, gastric mucosa and serosa, liver capsule, and spleen capsule with associated lymph nodes. Histopathologically, the neoplastic cells were seen to consist predominantly of a mixture of well differentiated small and large types. Immunohistochemically, the small lymphoid cells were MHC class IIlow+ and PanT- and the large lymphoid cells were MHC class IIhigh+ and PanT-. These findings revealed that the n...
Genetic and antigenic analysis of the influenza virus responsible for the 1992 Hong Kong equine influenza epizootic.
Virology    November 1, 1994   Volume 204, Issue 2 673-679 doi: 10.1006/viro.1994.1583
Lai AC, Lin YP, Powell DG, Shortridge KF, Webster RG, Daly J, Chambers TM.An outbreak of influenza occurred among thoroughbred racehorses in Hong Kong in November-December 1992, with morbidity of 37%. All horses involved had been vaccinated against equine-1 and equine-2 influenza viruses but not against the virus responsible for the 1989 equine influenza outbreak in northern China (influenza A/equine/Jilin/89, subtype H3N8). Therefore the source and nature of the virus causing the Hong Kong outbreak was investigated. Virus isolated from a horse infected during the outbreak was used for genetic analysis. All the viral gene segments were similar to those of equine-2 (...
Structural protein relationships among eastern equine encephalitis viruses.
The Journal of general virology    November 1, 1994   Volume 75 ( Pt 11) 2897-2909 doi: 10.1099/0022-1317-75-11-2897
Strizki JM, Repik PM.We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins. Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the E1, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidly-migrating E2-associated protein and a high M(r) (HMW) protein. In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype. The pro...
Genetic Bit Analysis: a solid phase method for typing single nucleotide polymorphisms.
Nucleic acids research    October 11, 1994   Volume 22, Issue 20 4167-4175 doi: 10.1093/nar/22.20.4167
Nikiforov TT, Rendle RB, Goelet P, Rogers YH, Kotewicz ML, Anderson S, Trainor GL, Knapp MR.A new method for typing single nucleotide polymorphisms in DNA is described. In this method, specific fragments of genomic DNA containing the polymorphic site(s) are first amplified by the polymerase chain reaction (PCR) using one regular and one phosphorothioate-modified primer. The double-stranded PCR product is rendered single-stranded by treatment with the enzyme T7 gene 6 exonuclease, and captured onto individual wells of a 96 well polystyrene plate by hybridization to an immobilized oligonucleotide primer. This primer is designed to hybridize to the single-stranded target DNA immediately...
Monocyte maturation controls expression of equine infectious anemia virus.
Journal of virology    October 1, 1994   Volume 68, Issue 10 6270-6279 doi: 10.1128/JVI.68.10.6270-6279.1994
Maury W.In vivo, equine infectious anemia virus (EIAV) replicates in tissues rich in macrophages, and it is widely believed that the tissue macrophage is the principal, if not sole, cell within the host that replicates virus. No viral replication has been detected in circulating peripheral blood monocytes. However, proviral DNA can be detected in these cells, and monocytes may serve as a reservoir for the virus. In this study, an in vitro model was developed to clarify the role of monocyte maturation in regulating EIAV expression. Freshly isolated, nonadherent equine peripheral blood monocytes were in...
Characterization of a novel streptokinase produced by Streptococcus equisimilis of non-human origin.
Thrombosis and haemostasis    October 1, 1994   Volume 72, Issue 4 595-603 
Nowicki ST, Minning-Wenz D, Johnston KH, Lottenberg R.Streptokinases are proteins with plasminogen activator activity produced by certain hemolytic streptococci. We previously identified equine streptococcal isolates which produced streptokinases (ESKs) that bound both human and equine plasminogen but only readily activated equine plasminogen (14). This property was exploited to purify a representative ESK produced by Streptococcus equisimilis strain 87-542-W. Affinity chromatography with human plasminogen resulted in the isolation of a M(r) approximately 49,000 molecule with two isoforms. This ESK was subsequently compared to well characterized ...
Survival and replication of Rhodococcus equi in macrophages.
Infection and immunity    October 1, 1994   Volume 62, Issue 10 4167-4175 doi: 10.1128/iai.62.10.4167-4175.1994
Hondalus MK, Mosser DM.Rhodococcus equi is a facultative intracellular bacterium of macrophages that can cause serious pneumonia in both young horses and immunocompromised people. Essential to understanding rhodococcus pathogenesis is a quantitative documentation of the intracellular events that follow macrophage phagocytosis of the organism. By using a bacterial immunofluorescence staining assay, we verified the intracellular survival and replicative potential of R. equi in both murine peritoneal macrophages and equine alveolar macrophages in vitro. Following an initial lag period of 6 to 12 h, the intracellular nu...
Rapid diagnosis of equine influenza by the Directigen FLU-A enzyme immunoassay.
The Veterinary record    September 17, 1994   Volume 135, Issue 12 275-279 doi: 10.1136/vr.135.12.275
Chambers TM, Shortridge KF, Li PH, Powell DG, Watkins KL.The Directigen FLU-A enzyme immunoassay was tested for its ability to detect equine-2 influenza viruses in nasopharyngeal fluids from horses and ponies. A total of 125 swabs from experimental infections and from different sources of natural infection in the USA and Hong Kong were examined. The assay results were compared with the results of standard virus culture in embryonated chicken eggs or Madin-Darby canine kidney cells, and with the serology of the horses sampled. In comparison with virus culture the enzyme immunoassay exhibited 83 per cent sensitivity, 78 per cent specificity, 70 per ce...
A new type of staphylococcal exfoliative toxin from a Staphylococcus aureus strain isolated from a horse with phlegmon.
Infection and immunity    September 1, 1994   Volume 62, Issue 9 3780-3785 doi: 10.1128/iai.62.9.3780-3785.1994
Sato H, Matsumori Y, Tanabe T, Saito H, Shimizu A, Kawano J.A new type of staphylococcal exfoliative toxin (sET) was isolated from the culture filtrate of a Staphylococcus aureus strain isolated from a horse with skin infection including phlegmon. The new sET was purified by precipitation with 80% saturated ammonium sulfate, column chromatography on DEAE-cellulofine A-500, gel filtration on a Sephadex G-75 column, and polyacrylamide gel electrophoresis (7.5% polyacrylamide). The new sET elicited general exfoliation of the epidermis with the so-called Nikolsky sign when inoculated into both 3-day-old mice and 1-day-old chicks, whereas sETA and sETB from...
Proteins induced by recombinant equine interferon-beta 1 within equine peripheral blood mononuclear cells and polymorphonuclear neutrophilic granulocytes.
Veterinary immunology and immunopathology    September 1, 1994   Volume 42, Issue 3-4 221-235 doi: 10.1016/0165-2427(94)90069-8
Heinz H, Marquardt J, Schuberth HJ, Adolf GR, Leibold W.Peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophilic granulocytes (PMN) as well as embryonic equine dermal fibroblasts and the equine fibroblast line E. Derm which were used as controls, were treated with recombinant equine interferon-beta 1 (rEqIFN-beta 1) in vitro which induced the expression of different proteins in these cells. A 74 kDa protein was induced in PBMC and an 82 kDa protein was additionally found in the equine fibroblast E. Derm cell line following treatment with rEqFN-beta 1. Both proteins reacted with anti-mouse and anti-human Mx protein antisera in im...
Monoclonal antibodies to equine arteritis virus proteins identify the GL protein as a target for virus neutralization.
The Journal of general virology    September 1, 1994   Volume 75 ( Pt 9) 2439-2444 doi: 10.1099/0022-1317-75-9-2439
Deregt D, de Vries AA, Raamsman MJ, Elmgren LD, Rottier PJ.Monoclonal antibodies (MAbs) to equine arteritis virus (EAV) proteins were produced and characterized. The protein specificities of eight MAbs were determined definitively by immunoprecipitation of EAV proteins expressed from vaccinia virus recombinants (VVRs). Included were two new VVRs produced for this study, expressing the M and the GL proteins, respectively. Three MAbs were determined to be N-specific and five MAbs recognized the GL protein. One GL-specific MAb, 17F5, of the IgA class, efficiently neutralized EAV infectivity. In competitive binding assays (CBAs), the N-specific MAbs defin...
Proteolytic processing of the replicase ORF1a protein of equine arteritis virus.
Journal of virology    September 1, 1994   Volume 68, Issue 9 5755-5764 doi: 10.1128/JVI.68.9.5755-5764.1994
Snijder EJ, Wassenaar AL, Spaan WJ.To study the proteolytic processing of the equine arteritis virus (EAV) replicase open reading frame 1a (ORF1a) protein, specific antisera were raised in rabbits, with six synthetic peptides and a bacterial fusion protein as antigens. The processing of the EAV ORF1a product in infected cells was analyzed with Western blot (immunoblot) and immunoprecipitation techniques. Additional information was obtained from transient expression of ORF1a cDNA constructs. The 187-kDa ORF1a protein was found to be subject to at least five proteolytic cleavages. The processing scheme, which covers the entire OR...
Differential distribution of immunoreactive S-100 protein in mammalian testis.
Histochemistry    September 1, 1994   Volume 102, Issue 3 241-245 doi: 10.1007/BF00268901
Amselgruber WM, Sinowatz F, Erhard M.The present study deals with the immunohistochemical localization of S-100 protein in the testes of seven mammalian species including rat, cat, dog, pig, sheep, cattle and horse. Significant differences are demonstrated in the cellular distribution and intensity of immunoreaction for the protein. In bull, ram, boar and cat testes S-100 protein was localized in the cytoplasm and nuclei of Sertoli cells. A particularly intense staining was seen in the modified Sertoli cells of the terminal tubular segment. With the exception of the cat and horse S-100 protein immunoreactivity was additionally fo...
Potency of partially purified malleo-proteins for mallein test in the diagnosis of glanders in equines.
Veterinary microbiology    August 15, 1994   Volume 41, Issue 4 391-397 doi: 10.1016/0378-1135(94)90035-3
Verma RD, Venkateswaran KS, Sharma JK, Agarwal GS.Malleo-proteins from synthetic broth mallein of six strains of Pseudomonas mallei were separated by trichloroacetic acid precipitation, ammonium sulfate precipitation and Ultrogel AcA 34 gel filtration chromatography. When tested comparatively with Dutch PPD mallein as standard on P. mallei-sensitized and normal horses all the strains were found to be malleinogenic, trichloroacetic acid precipitated proteins were comparable to Dutch PPD mallein in potency and innocuity whereas ammonium sulfate-precipitated proteins elicited non-specific reactions. Ultrogel AcA 34 chromatographed high molecular...
Evaluation of a monoclonal antibody-based colony blot test for rapid identification of virulent Rhodococcus equi.
The Journal of veterinary medical science    August 1, 1994   Volume 56, Issue 4 681-684 doi: 10.1292/jvms.56.681
Takai S, Morishita T, Nishio Y, Sasaki Y, Tsubaki S, Higuchi T, Hagiwara S, Senba H, Kato M, Seno N.We recently generated a monoclonal antibody immunoglobulin G1 (MAb 10G5), which can recognize 15- to 17-kDa antigens, virulence-associated antigens of Rhodococcus equi, and developed a colony blot enzyme-linked immunosorbent assay with MAb 10G5 for the rapid identification of virulent R. equi. In this epidemiologic study, we evaluated the results of the colony blot test in the identification of virulent isolates of R. equi from feces of horses and soil and compared them with those from a conventional procedure (plasmid profiles of isolates by agarose gel electrophoresis). Environmental isolate...
The immunology of companion animals: reagents and therapeutic strategies with potential veterinary and human clinical applications.
Immunology today    August 1, 1994   Volume 15, Issue 8 347-353 doi: 10.1016/0167-5699(94)90171-6
Cobbold S, Holmes M, Willett B.There is now a wide range of immunological reagents that can be used in the diagnosis and treatment of diseases in the companion animals (dogs, cats and horses). Many of these diseases are the veterinary equivalents of human conditions, and may therefore provide good models to study basic pathogenic mechanisms.
African horsesickness: pathogenesis and immunity.
Comparative immunology, microbiology and infectious diseases    August 1, 1994   Volume 17, Issue 3-4 275-285 doi: 10.1016/0147-9571(94)90047-7
Burrage TG, Laegreid WW.African horsesickness (AHS) is a serious, non-contagious disease of horses and other solipeds caused by an arthropod-borne orbivirus of the family Reoviridae. In horses, AHS causes three distinct clinicopathologic syndromes, the pulmonary, cardiac and fever forms of the disease. Recent work has shown that the primary determinant of the form of disease expressed by naive horses is the virulence of the virus inoculum. Horses which recover from AHS exhibit solid humoral immunity against homologous challenge. Protective antibodies appear to be directed towards neutralizing epitopes on AHS virus VP...
Molecular weight alterations of alpha-1 proteinase inhibitor in equine bronchoalveolar lavage fluid.
Electrophoresis    August 1, 1994   Volume 15, Issue 8-9 1193-1197 doi: 10.1002/elps.11501501180
Milne EM, Pemberton AD, McGorum BC, Dixon PM, Miller HR.The equine alpha-1 proteinase inhibitor (alpha 1PI) system differs from that of man in that the equine system consists of four closely-linked genes (Spi1-Spi4) whereas in man, a single gene encodes for alpha 1PI. We have previously found differences in the proportion of the Spi proteins in equine serum and bronchoalveolar lavage fluid (BALF). We therefore wished to determine whether, as reported in man, there was any molecular weight difference between the Spi proteins in serum and BALF. alpha 1PI and albumin from equine BALF migrated further towards the anode compared with serum alpha 1PI on ...
Substance P and calcitonin gene-related peptide-like immunoreactive nerve fibers in lungs from adult equids.
American journal of veterinary research    August 1, 1994   Volume 55, Issue 8 1066-1074 
Sonea IM, Bowker RM, Robinson NE, Broadstone RV.Distribution of pulmonary nerves immunoreactive for either substance P or calcitonin gene-related peptide was determined, using immunohistochemical methods on healthy lungs from adult equids. The overall patterns of distribution of substance P and calcitonin gene-related peptide-like immunoreactivity were similar. Distribution of immunoreactive nerves was not uniform throughout the lungs; nerve fibers immunoreactive for these peptides were more frequently observed near the hilus of the lung than in the caudal lobes or in the periphery of the lung. Nerve fibers immunoreactive for substance P or...
Light chain isotype regulation in the horse. Characterization of Ig kappa genes.
Journal of immunology (Baltimore, Md. : 1950)    August 1, 1994   Volume 153, Issue 3 1099-1111 
Ford JE, Home WA, Gibson DM.Horse Ig kappa genes have been characterized to determine whether there may be a structural basis for the low level of kappa expression in this species. The overall organization of the J kappa-C kappa locus is remarkably similar to that of the mouse and human loci. A single C kappa exon is separated by 2.9 kb from five J kappa segments, four of which seem functional and three of which are associated with canonical recombination signal sequences. A highly conserved intron enhancer was identified upstream of the C kappa exon and a single restriction fragment in horse genomic DNA hybridized stron...
The trigeminal ganglion is a location for equine herpesvirus 1 latency and reactivation in the horse.
The Journal of general virology    August 1, 1994   Volume 75 ( Pt 8) 2007-2016 doi: 10.1099/0022-1317-75-8-2007
Slater JD, Borchers K, Thackray AM, Field HJ.Four specific pathogen-free ponies were infected intranasally with equine herpesvirus 1 (EHV-1) and two were similarly infected with an EHV-1 thymidine kinase deletion mutant. The primary infections were characterized by a transient fever accompanied by virus shedding into nasal mucus and viraemia. No virus was detected in clinical specimens after 15 days post-infection. Two months later a reactivation stimulus was administered to all six ponies and only the four that had been previously inoculated with wild-type EHV-1 shed virus into nasal mucus (for 10 days), proving the presence of a latent...