Immunology.
Publisher:
Blackwell Scientific Publications
Frequency: Monthly
Country: England
Language: English
Author(s):
British Society for Immunology
Start Year:1958 -
ISSN:
0019-2805 (Print)
1365-2567 (Electronic)
0019-2805 (Linking)
1365-2567 (Electronic)
0019-2805 (Linking)
Impact Factor
6.4
2022
| NLM ID: | 0374672 |
| (DNLM): | I06200000(s) |
| (OCoLC): | 01752695 |
| Coden: | IMMUAM |
| Classification: | W1 IM53 |
Single-cell profiling of bronchoalveolar cells reveals a Th17 signature in neutrophilic severe equine asthma. Severe equine asthma (SEA) is a complex respiratory condition characterized by chronic airway inflammation. It shares many clinical and pathological features with human neutrophilic asthma, making it a valuable model for studying this condition. However, the immune mechanisms driving SEA have remained elusive. Although SEA has been primarily associated with a Th2 response, there have also been reports of Th1, Th17, or mixed-mediated responses. To uncover the elusive immune mechanisms driving SEA, we performed single-cell mRNA sequencing (scRNA-seq) on cryopreserved bronchoalveolar cells from 1...
Differential CD4+ T-cell responses of allergic and non-allergic subjects to the immunodominant epitope region of the horse major allergen Equ c 1. The responses of allergen-specific CD4(+) T cells of allergic and healthy individuals are still incompletely understood. Our objective was to investigate the functional and phenotypic properties of CD4(+) T cells of horse-allergic and healthy subjects specific to the immunodominant epitope region of the major horse allergen Equ c 1. Specific T-cell lines (TCLs) and clones were generated from peripheral blood mononuclear cells with Equ c 1(143-160), the peptide containing the immunodominant epitope region of Equ c 1. The frequency, proliferative response, cytokine production and HLA restriction...
Differentiation and activation of equine monocyte-derived dendritic cells are not correlated with CD206 or CD83 expression. Dendritic cells (DC) are the main immune mediators inducing primary immune responses. DC generated from monocytes (MoDC) are a model system to study the biology of DC in vitro, as they represent inflammatory DC in vivo. Previous studies on the generation of MoDC in horses indicated that there was no distinct difference between immature and mature DC and that the expression profile was distinctly different from humans, where CD206 is expressed on immature MoDC whereas CD83 is expressed on mature MoDC. Here we describe the kinetics of equine MoDC differentiation and activation, analysing both ph...
Equine CD4(+) CD25(high) T cells exhibit regulatory activity by close contact and cytokine-dependent mechanisms in vitro. Horses are particularly prone to allergic and autoimmune diseases, but little information about equine regulatory T cells (Treg) is currently available. The aim of this study therefore was to investigate the existence of CD4(+) Treg cells in horses, determine their suppressive function as well as their mechanism of action. Freshly isolated peripheral blood mononuclear cells (PBMC) from healthy horses were examined for CD4, CD25 and forkhead box P3 (FoxP3) expression. We show that equine FoxP3 is expressed constitutively by a population of CD4(+) CD25(+) T cells, mainly in the CD4(+) CD25(high)...
Monocyte-derived dendritic cells from horses differ from dendritic cells of humans and mice. Dendritic cells (DC) are the initiators of immune responses and are present in most tissues in vivo. To generate myeloid DC from monocytes (MoDC) in vitro the necessary cytokines are granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Using degenerated primers delineated from other species and rapid amplification of cDNA ends reverse transcription-polymerase chain reaction (RACE RT-PCR), the cDNA of equine (eq.) GM-CSF was cloned and found to have a point deletion at the 3'-end of eq.GM-CSF, resulting in a 24-nucleotide extended open reading frame not described ...
Identification of broadly recognized, T helper 1 lymphocyte epitopes in an equine lentivirus. Equine infectious anaemia virus (EIAV) is a horse lentivirus causing lifelong, persistent infection. During acute infection, CD8(+) cytotoxic T lymphocytes (CTL) are probably involved in terminating plasma viraemia. However, only a few EIAV CTL epitopes, restricted to fewer horse major histocompatibility complex (MHC) class I alleles, are known. As interferon-gamma (IFN-gamma)-secreting CD4(+), T helper 1 (Th1) lymphocytes promote CTL activity and help maintain memory CTL, identifying broadly recognized EIAV Th1 epitopes would contribute significantly to vaccine strategies seeking to promote s...
Characterization of equine E-selectin. Expression of E-selectin on activated endothelium is a critical initial step that leads to extravasation of leucocytes during inflammation, yet E-selectin is largely uncharacterized in several animal species including the horse. We have sequenced and compared E-selectin genes derived from activated cultures of purified equine (horse), cervid (black-tailed deer) and ovine (sheep) pulmonary artery endothelial cells (ECs). Phylogenetic and amino acid sequence comparisons indicate that bovine, cervid and ovine E-selectin are similar, whereas human and equine E-selectin are more closely related to ...
Transforming growth factor-beta induced by live or ultraviolet-inactivated equid herpes virus type-1 mediates immunosuppression in the horse. Up to 21 days after exposure to live or ultraviolet-inactivated equid herpesvirus type-1 (EHV-1) autologous serum from ponies caused an immunosuppressive effect if incorporated into T-cell proliferation assays to EHV-1. The suppressive factor in the sera of ponies also inhibited T-cell response to phytohaemagglutinin. Increased levels of circulating activated transforming growth factor-beta 1 (TGF-beta 1) were detected, and the suppressive activity of the serum could be reversed by antibody to TGF-beta 1. In a challenge experiment the ponies which exhibited circulating TGF-beta 1 activity succ...
Cross-antigenicity of horse serum albumin with dog and cat albumins: study of three short peptides with significant inhibitory activity towards specific human IgE and IgG antibodies. Horse serum albumin is present in the near vicinity of the animal, while dog and cat serum albumins are very common allergens present in house dust. Human patients clinically defined as allergic to horse could react with horse serum albumin by means of IgE or IgG antibodies. Studies regarding the specificities of these antibodies by inhibition enzyme-linked immunosorbent assay (ELISA) and depletion experiments have demonstrated that they are directed against dog serum albumin and cross-react not only with horse serum albumin but with other serum albumins from different origins. To investigate ...
Abnormal patterns of equine leucocyte differentiation antigen expression in severe combined immunodeficiency foals suggests the phenotype of normal equine natural killer cells. Severe combined immunodeficiency (SCID) is a fatal autosommal disease of Arabian horses that leads to failure of maturation of T- and B-lymphocyte populations, although natural killer (NK) cells are unaffected. Thymic and lymph node tissues from two foals suffering from SCID were examined in an immunohistological study using a panel of monoclonal antibodies recognising equine leucocyte differentiation antigens. In both foals, the majority of cells in lymphoid tissues had an EqCD3-EqCD4-EqCD8+ phenotype, although rare EqCD3+ cells were also detected. The EqCD3-EqCD4-EqCD8+ cells may represent a...
Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5. Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was ef...
Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets. The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-c...
Equine monoclonal antibodies recognize common epitopes on variants of equine infectious anaemia virus. Equine-murine xenohybridoma cells were produced using SP2/0 murine myeloma cells and splenic lymph node cells obtained from horses infected with 10(6) TCID50 of single cloned variants of equine infectious anaemia virus (EIAV). The xenohybridomas secreted equine IgG monoclonal antibodies reactive with EIAV in enzyme immunoassays employing purified virus. Seven antibodies were studied in detail. They bound to viral glycoproteins (gp90 or gp45) in radioimmunoprecipitation assays, and reacted with homologous EIAV as well as five other cloned variants of EIAV. When evaluated against a single cloned...
The characterization of equine herpes virus-1-infected cell polypeptides recognized by equine lymphocytes. Ponies, without evidence of previous exposure to Equine herpes virus-1 (EHV-1), were experimentally infected with EHV-1 subtype 2 and investigated for lymphocyte transformation to virus-infected cell polypeptides, as shown by separation with gel electrophoresis. Animals made significant responses to Western blot fractions that corresponded to molecular weights of approximately 30,000, 40,000-45,000, 60,000-65,000, 80,000-95,000 and 100,000-140,000 MW. These molecular weight ranges correlated with the positions of major EHV-1 subtype 2 glycoproteins that were found at migration distances approx...
Purification of the subunit Clq from the first component of equine complement. Initial separation and concentration of Clq from fresh, normal equine serum was accomplished by precipitation in 0.02 M acetate buffer, pH 5.5, at 4 degrees for 24 h. The re-dissolved precipitate was clarified by centrifugation at 80,000 g for 1 h and then dialysed against Tris-HCl buffer (0.05 M, pH 8.0) containing 10-3 M EDTA. The clarified dialysate remained biologically active at 5 degrees for at least 4 weeks. Biological activity of equine Clq was determined by assay of its ability to agglutinate sensitized sheep erythrocytes (EA). Following ammonium sulphate fractionation, Sepharose 4B g...
Homologous and cross-reactive precipitins in anti-pneumococcal sera raised in mules. Serial bleedings were obtained from two mules during prolonged immunization, one with type XXV the other with type VIII pneumococcal vaccine. IgGa, IgGb, IgGc, IgB, IgG(T) and IgM present among purified Pn anti-XXV and Pn anti-VIII immunoglobulin isolated from various bleedings were identified by use of rabbit anti-equine heavy chain specific reagents. Radioimmunodiffusion with 14C-labelled type XXV pneumococcal capsular polysaccharide and horse and donkey reagents with species specificity directed against donkey or horse IgGa respectively, demonstrated both parental horse and donkey IgGa heav...
Suppression of synthesis of an IgG subclass in a persistent viral infection. Comparison of immunoglobulin levels of nine horses before and after infection with equine infectious anaemia (EIA) virus demonstrated a significant depression of serum IgG(T) at 2 months (P less than 0-001) and at 1 year (P less than 0-01) after infection. In contrast, the levels of IgGa were significantly increased at both times after infection. Another sixteen horses with EIA for 1-4 months were examined and there was also significant depression (P less than 0-001) of IgG(T) when compared to pre-infection levels. No significant changes in IgG(T), IgGa and IgM were noted in fourteen normal ho...
Surface receptors on neutrophils and monocytes from immunodeficient and normal horses. Surface receptors on peripheral blood neutrophils and monocytes from normal and immunodeficient horses have been studied. Sheep erythrocytes (SRBC) coated with IgG, IgM, and complement but not IgG(T), readily bound to normal equine monocytes and neutrophils. More than 4000 molecules of IgG were required to sensitize each SRBC for adherence to monocytes, and more than 12,000 molecules were required for adherence to neutrophils. Young horses with a severe combined immunodeficiency had an almost total absence of lymphocytes, but normal numbers of monocytes and neutrophils. The number of receptors...
Studies on equine immunoglobulins. IV. Immunoglobulins of the donkey. Donkey IgGa was isolated in purified form from normal and immune donkey sera by column chromatography on DEAE-cellulose. Isolated donkey IgGa and mixtures of (IgGa+IgGb) were used as antigens to prepare rabbit reagents specific for equine IgGa or IgGb. Antibodies present in sera obtained from a single donkey at various times during the course of hyperimmunization with BSA were isolated by immuno-adsorption. The class or subclass of immunoglobulins present among isolated, donkey anti-BSA antibodies was determined by use of specific rabbit anti-equine immunoglobulin reagents. The homologues of h...
Passive cutaneous anaphylaxis and its enhancement by normal IgG. Rats were injected intradermally with rabbit anti-ovalbumin serum and 3 hours later were challenged intravenously with ovalbumin and Evans Blue dye. Inflammatory lesions were produced within 20 minutes and their size was markedly dose-dependent. Attempts were made to interfere with this passive cutaneous anaphylaxis (PCA) by admixture of normal IgG with the rabbit anti-ovalbumin to measure the relative tissue binding affinities of IgG from various species. It was found that normal IgG from any of the species tested had an enhancing effect on PCA in rats. These immunoglobulins serially arranged...
Studies on the IgA system of the horse. Equine serum and secretions were found to contain a protein which cross-reacted with an antiserum against human IgA, but not with antisera against any other human immunoglobulin. The physicochemical properties of equine IgA resembled those of human IgA. IgA was found to be the immunoglobulin having the highest secretion serum concentration ratio in equine lacteal and salivary secretions, and to be the protein produced by the majority of immunoglobulin-containing cells in the of the equine intestine.
In vitro synthesis of immunoglobulin-A by salivary glands from animals of different species. The synthesis of immunoglobulins by the salivary glands from eight different species was studied. It has been demonstrated that salivary glands from the cow, horse, sheep, pig, rat and guinea-pig preferentially synthesize a fast migrating immunoglobulin which seems to be analogous to IgA. In three of the species, the cow, sheep and pig, the IgA-like component cross-reacts with human IgA. The IgA synthesized by the salivary glands from the rat cross-reacts with the mouse IgA. When one compares the salivary IgA from the cow, horse, sheep, pig and rat with the IgA synthesized by the lymph nodes,...
The total protein and immunoglobulin profile of equine colostrum and milk. Thirty-six samples of colostrum and milk were collected from ponies at various intervals after parturition. Levels of total protein and immunoglobulins IgG and IgG(T) were determined. In samples collected in the first 3 hours , the total protein concentration was approximately twice that of serum protein and the immunoglobulins IgG and IgG(T) accounted for 30 per cent and 10 per cent respectively of this protein. The ratio of IgG to IgG(T) was similar to that in serum. After suckling, a marked decline in both total protein and immunoglobulin concentration occurred. In addition, the relative co...
Comparative studies on the haemolytic and Treponema pallidum immobilizing complement activity in the serum of different species. Complement activity in the serum of eight species has been studied in two ways: by immobilization of sensitized with human or rabbit antibody and by haemolysis of sheep red cells sensitized with rabbit antibody. Serum of the pig, monkey and man was actively haemolytic but contained a heatlabile factor that immobilized unsensitized in the presence of guinea-pig complement and precluded the detection of immune immobilizing activity. Sera of other species, although without action on unsensitized treponemes, even with added guinea-pig complement, differed in their relative haemolytic and immobil...
Bacteriostatic effects of horse sera and serum fractions on Clostridium welchii Type A, and the abolition of bacteriostasis by iron salts. Under a variety of conditions of concentration, Eh, and pH, horse anti- serum and normal horse serum exerted similar bacteriostatic effects against Type A. Ferric iron abolished the bacteriostatic effect when added during the first 2 hours of incubation at Eh+60 mV. Ferrous iron abolished the bacteriostatic effect when added after 3 hours. Ferric iron abolished the bacteriostatic effect at—140 mV. A mixture consisting of horse β- and γ-globulins together with human transferrin exerted a bacteriostatic effect similar to that of whole serum. This system responded in the same way as whole se...
The alexination and conglutination reactions. The reactions between sensitized erythrocytes and horse complement and between alexinated erythrocytes and conglutinin. A method is described for the quantitative measurement of the reactions between sensitized cells and horse complement and between alexinated cells and conglutinin. The method is laborious but its application has allowed the determination of the optimal times of the reactions at various temperatures. The results obtained in these experiments indicate that the alexinated configuration with which conglutinin and immuno-conglutinin react is not one of the recognized intermediates formed during the process of immune haemolysis.