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1.
Vet J ; 192(2): 246-8, 2012 May.
Article in English | MEDLINE | ID: mdl-21704542

ABSTRACT

An outbreak of tuberculosis (TB), caused by Mycobacterium bovis, was investigated in a small herd of llamas (Lama glama). Based on three ante-mortem diagnostic methods (clinical signs, tuberculin skin test reactions, and 'Rapid Test' serology), 12 llamas were selected for examination post-mortem. Grossly visible lesions suspicious of TB were observed in eight animals, four of which had exhibited clinical signs, one was a skin test 'reactor', and three had been seropositive. M. bovis was isolated from seven of these eight animals. Clinical signs combined with serology were found to be useful in identifying infected animals, but tuberculin skin testing had limited negative predictive value as four llamas that were subsequently confirmed as infected were not detected using this assay.


Subject(s)
Camelids, New World , Disease Outbreaks/veterinary , Tuberculin Test/veterinary , Tuberculosis/veterinary , Animals , Autopsy/veterinary , Disease Outbreaks/prevention & control , Mycobacterium bovis/isolation & purification , Predictive Value of Tests , Respiration Disorders/veterinary , Serologic Tests/veterinary , Tuberculosis/diagnosis , Tuberculosis/epidemiology , Tuberculosis/prevention & control , United Kingdom/epidemiology , Weight Loss
2.
Vet Rec ; 167(13): 475-80, 2010 Sep 25.
Article in English | MEDLINE | ID: mdl-20871080

ABSTRACT

An outbreak of tuberculosis (TB) caused by Mycobacterium bovis in a llama herd is described. Over a 25-month period, a total of 70 llamas were selected for postmortem examination using four distinct criteria: clinical suspicion of disease (15 animals), positive tuberculin skin test result (three animals), antibody positive using a novel serological test (Rapid Test, 54 animals) and elective cull (five animals). Some animals qualified on more than one criterion. Gross lesions of TB were detected in 15 animals, with lung and lymph node lesions consistently observed. Samples were collected from 14 of 15 animals with visible lesions as well as those with no visible lesions, for histopathology and mycobacterial culture. All 14 llamas with visible lesions had caseonecrotic granulomatous lesions associated with acid-fast bacteria and variable mineralisation, and M bovis was isolated from 13. There were no histopathological lesions of TB in llamas with no grossly visible lesions, and M bovis was not isolated from any of these. The predictive value of suspicious gross lesions at postmortem examination was therefore high in the herd. Molecular typing results indicated that the outbreak was caused by a single strain likely to have originated from a local reservoir, probably cattle or wildlife. Antemortem indicators of infection assisted control of the outbreak, but no single test accurately identified all TB cases. Visible lesions were detected in nine of 15 llamas with clinical suspicion of disease, in two of three that had positive tuberculin skin test results and in 10 of 54 that were antibody positive; there was none (zero out of five) in llamas that were electively culled.


Subject(s)
Camelids, New World/microbiology , Disease Outbreaks/veterinary , Mycobacterium bovis , Tuberculin Test/veterinary , Tuberculosis/veterinary , Animals , Antibodies, Bacterial/blood , Disease Outbreaks/prevention & control , Female , Immunohistochemistry/veterinary , Male , Mycobacterium bovis/immunology , Seroepidemiologic Studies , Tuberculosis/epidemiology , Tuberculosis/pathology , Tuberculosis/prevention & control , United Kingdom/epidemiology
4.
Tuberculosis (Edinb) ; 88(6): 586-94, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18450515

ABSTRACT

Cattle infected with Mycobacterium bovis spoligotype 9 were treated with Isoniazid (INH) from three to 14 weeks post infection, rested for fourweeks to allow INH depletion and then challenged with M. bovis spoligotype 35. Post mortem examination (PME) 35 weeks after the initial infection showed partial protection against infectious challenge following INH-attenuated infection compared with the spoligotype 35 challenge controls. Antigen-specific IFN-gamma responses decreased over time with INH therapy, following a similar pattern to that observed in the treatment of M. tuberculosis infection in humans. Following cessation of therapy, specific IFN-gamma responses increased more strongly in those calves that were visibly lesioned at PME. IFN-gamma responses were also used to identify two antigens, TB10.4 and Acr2, that induced anamnestic responses in INH-treated, re-challenged calves, suggesting a role for both antigens in protective immunity. Specific IL-10 responses were observed in all calves following treatment with INH suggesting a role for IL-10 in the resolution of infection.


Subject(s)
Antitubercular Agents/therapeutic use , Isoniazid/therapeutic use , Mycobacterium bovis/drug effects , Tuberculosis, Bovine/drug therapy , Tuberculosis, Bovine/immunology , Animals , Antigens, Bacterial/immunology , Cattle , Cattle Diseases/immunology , Cytokines , Disease Models, Animal , Female , Immunohistochemistry , Interferon-gamma/immunology , Interleukin-10/immunology , Male , Minisatellite Repeats , Mycobacterium bovis/classification , Mycobacterium bovis/genetics , Mycobacterium bovis/immunology , Random Allocation , Tuberculin Test , Tuberculosis, Bovine/pathology
5.
Lupus ; 11(8): 493-500, 2002.
Article in English | MEDLINE | ID: mdl-12220103

ABSTRACT

Systemic lupus erythematosus (SLE) is characterized by B cell hyperactivity and the production of autoantibodies, some of which (antibodies to dsDNA) are thought to be pathogenic. T helper cells drive the production of autoantibodies and the aim of this study is to characterize phenotypically a subpopulation of T cells (the CD3+ CD4- CD8-, double negative (DN) T cells) previously identified as helping to enhance anti-DNA antibodies in patients with SLE. Data were obtained using FACS staining of DN T cells that had been purified from PBMCs by magnetic bead separation. The percentage of TCR alphabeta+ DN T cells was found to be significantly higher in patients with SLE as compared with controls (P = 0.02), although there was no significant increase in total percentage of DN T cells, which includes TCR gammadelta+ cells. Activation markers HLA-DR and CD69, the costimulatory molecule CD28 and CTLA-4 were all expressed on the surface of a higher percentage of DN T cells in patients with SLE than in patients with rheumatoid arthritis (RA) or healthy controls (HC). More DN T cells from patients with SLE were of CD45RA phenotype than was found in controls, while CD45RO-expressing cells were reduced. In addition, DN T cells from patients with SLE expressed significantly higher levels of HLA-DR (P = 0.006), CD28 (P = 0.05), CTLA4 (P = 0.03) and CD45RA (P = 0.05) on the cell surface than those from the CD4/8 population. Correlation of expression of the markers measured with various parameters of disease activity and severity showed that high levels of HLA-DR expression correlated with high circulating serum C3 (> 0.9 IU/ml), indicating that an activated phenotype is consistent with severe disease.


Subject(s)
Biomarkers/analysis , CD3 Complex/analysis , Immunoconjugates , Lupus Erythematosus, Systemic/diagnosis , Lupus Erythematosus, Systemic/immunology , T-Lymphocytes/chemistry , Abatacept , Adolescent , Adult , Aged , Antigens, CD/analysis , Antigens, Differentiation/analysis , Antigens, Differentiation, T-Lymphocyte/analysis , Antigens, Surface/analysis , CD28 Antigens/analysis , CD4 Antigens/analysis , CD8 Antigens/analysis , CTLA-4 Antigen , Female , HLA-DR Antigens/analysis , Humans , Lectins, C-Type , Leukocyte Common Antigens/analysis , Lymphocyte Activation , Male , Middle Aged
6.
Lupus ; 11(8): 501-7, 2002.
Article in English | MEDLINE | ID: mdl-12220104

ABSTRACT

Systemic lupus erythematosus (SLE) is a chronic autoimmune rheumatic disease that may affect every organ or system in the body. We have shown previously that the TCR alphabeta+ subpopulation of CD3+ CD4- CD8-, DN T cells is expanded in patients with SLE and that double negative T cells express increased levels of activation markers compared both with healthy people and with patients with rheumatoid arthritis, (RA) as autoimmune controls. The aim of this study was to characterize these cells in terms of their ability to produce IL4, a Th2 cytokine, both spontaneously and after mitogen stimulation. It was found that a higher percentage of TCR alphabeta+ double negative T cells from patients with SLE contained IL4 constitutively than did the same population of cells from healthy people or from those with RA. After mitogen stimulation, there was no significant difference in the amount of IL4 produced by each of the three groups. Further study of patients producing high levels of IL4 (about one third of the patients) indicated that they had a lower percentage of alphabeta+ T cells in the double negative compartment than did patients with fewer IL4 containing cells.


Subject(s)
CD3 Complex/analysis , Interleukin-4/biosynthesis , Lupus Erythematosus, Systemic/diagnosis , Lupus Erythematosus, Systemic/immunology , T-Lymphocytes/chemistry , Adolescent , Adult , Aged , Antigens, Surface/analysis , Autoantibodies/blood , Biomarkers/analysis , CD4 Antigens/analysis , CD8 Antigens/analysis , Complement C3/metabolism , DNA/immunology , Female , Histocompatibility Testing , Humans , Immunosuppressive Agents/therapeutic use , Lupus Erythematosus, Systemic/drug therapy , Lupus Erythematosus, Systemic/ethnology , Male , Middle Aged , Phytohemagglutinins/pharmacology , Racial Groups , Receptors, Antigen, T-Cell, alpha-beta/analysis , T-Lymphocytes/drug effects , T-Lymphocytes/metabolism
8.
Clin Exp Rheumatol ; 19(3): 251-7, 2001.
Article in English | MEDLINE | ID: mdl-11407076

ABSTRACT

OBJECTIVE: We have attempted to devise a method for measuring the levels of anticardiolipin antibodies produced in in vitro culture of peripheral blood mononuclear cells isolated from patients with systemic lupus erythematosus (SLE), with or without circulating anticardiolipin antibodies, patients with the primary antiphospholipid syndrome (PAPS) and normal controls. METHODS: Peripheral blood mononuclear cells were isolated and cultured for up to six days, in the presence or absence of added cytokines in the culture medium. Supernatants harvested after culture were tested by ELISA for the presence of anticardiolipin antibodies. RESULTS AND CONCLUSION: Despite variation of the culture conditions and of the cell numbers and populations used, it was found that accurately measurable levels of anticardiolipin antibodies could not be detected reliably in any of the culture supernatants. It is concluded that alternative methods of measurement of antibody production need to be explored.


Subject(s)
Antibodies, Anticardiolipin/analysis , Leukocytes, Mononuclear/immunology , Lupus Erythematosus, Systemic/diagnosis , Lupus Erythematosus, Systemic/immunology , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Humans , Immunoglobulin G/analysis , Immunoglobulin M/analysis , In Vitro Techniques
9.
Clin Orthop Relat Res ; (386): 106-13, 2001 May.
Article in English | MEDLINE | ID: mdl-11347822

ABSTRACT

The purpose of the current study was to review the demographics and etiologies of symptomatic femoral head osteonecrosis in the pediatric and adolescent population and to assess the results of treatment using free vascularized fibular grafting. A group of patients with femoral head osteonecrosis who were treated with free vascularized fibular grafting was reviewed. Patients who were studied were 18 years of age or younger at the time of surgery. Records were examined for demographic data, etiology of osteonecrosis, stage of the disease at time of surgery, and results of treatment including preoperative and postoperative Harris hip scores. Eighty-two pediatric and adolescent patients with osteonecrosis of the femoral head underwent 90 free vascularized fibular grafting procedures. Fifty patients (54 hips) who have been followed up at least 2 or more years (average, 4.3 years) constituted the study group. At the last followup, total hip arthroplasty was performed in seven hips (seven patients) and hip fusion was performed in one hip (one patient). The average Harris hip scores in patients who did not undergo total hip arthroplasty improved from a preoperative average of 55.3 points to 90.2 points at the latest followup. Treatment of patients with osteonecrosis with free vascularized fibular grafting resulted in a lower rate of conversion to total hip arthroplasty or fusion (16%) in pediatric and adolescent patients when compared with conversion to total hip arthroplasty in adults (25%). The quality of life as evidenced by the increased Harris hip scores was improved significantly in this group of pediatric and adolescent patients.


Subject(s)
Bone Transplantation/methods , Femur Head Necrosis/diagnostic imaging , Femur Head Necrosis/surgery , Fibula/blood supply , Fibula/transplantation , Adolescent , Child , Child, Preschool , Female , Follow-Up Studies , Graft Rejection , Graft Survival , Humans , Male , Radiography , Retrospective Studies , Treatment Outcome
10.
Orthopedics ; 23(6): 573-8, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10875418

ABSTRACT

This study examined the mechanisms of anterior cruciate ligament (ACL) injury. In the first part of the study, using a comprehensive, standardized questionnaire, 89 athletes (100 knees) were interviewed about the events surrounding their ACL injury. A noncontact mechanism was reported in 71 (72%) knees and a contact injury in 28 (28%) knees; one patient was unsure if there was any contact. Most of the injuries were sustained at footstrike with the knee close to full extension. Noncontact mechanisms were classified as sudden deceleration prior to a change of direction or landing motion, while contact injuries occurred as a result of valgus collapse of the knee. Hamstring flexibility parameters revealed a statistically higher level of laxity in the injured athletes compared with a matched group of 28 controls. In the second part of the study, videotapes of 27 separate ACL disruptions were reviewed and confirmed that most noncontact injuries occur with the knee close to extension during a sharp deceleration or landing maneuver. Because the knee is in a position to allow the extensor mechanism to strain the ACL and maximum, eccentric muscle force conditions usually apply, the quadriceps may play an important role in ACL disruption. Passive protection of the ACL by the hamstring muscles may be reduced in patients with above-average flexibility.


Subject(s)
Anterior Cruciate Ligament Injuries , Athletic Injuries/etiology , Anthropometry , Athletic Injuries/physiopathology , Biomechanical Phenomena , Electromyography , Humans , Surveys and Questionnaires , Video Recording
12.
Immunol Lett ; 68(2-3): 333-7, 1999 Jun 01.
Article in English | MEDLINE | ID: mdl-10424440

ABSTRACT

The aim of this work was to raise allotype specific monoclonal antibodies to human CD45, with the long-term objective of producing a reagent which could be used to prolong graft survival in renal transplantation through removal of passenger leukocytes from the graft. At present there are no anti-CD45 monoclonal antibodies able to distinguish between host and donor leukocytes. An in vitro immunisation technique has been developed through which donated human leukocytes are sensitised to CD45 prior to fusion with a myeloma cell line. IgM was produced by all the anti-CD45-positive clones. Flow cytometric analysis using these antibodies showed their ability to differentiate between blood from individual donors, indicating the existence of allotypic forms of human CD45, in conformity with the findings in rats and pigs. Therefore, a reagent which could be used in renal transplantation is a technical possibility.


Subject(s)
Antibodies, Monoclonal , Leukocyte Common Antigens/immunology , Flow Cytometry , Humans , Hybridomas , Kidney Transplantation/immunology
13.
Int J Cancer ; 82(4): 525-31, 1999 Aug 12.
Article in English | MEDLINE | ID: mdl-10404066

ABSTRACT

Different epitopes on the extracellular domain of the HER-2 receptor can serve as distinct targets for immunotoxins. To determine the optimal epitope target for immunotoxin therapy, 7 anti-HER-2 ricin A chain murine monoclonal immunotoxins, each reactive with different epitopes of HER-2 receptor, were tested for cytotoxic activity. The immunotoxins produced 1.2-4.6 logs of cytotoxicity in limiting dilution clonogenic assays with 2 breast cancer cell lines that overexpressed HER-2. Cytotoxicity did not correlate with immunoglobulin isotype, binding affinity, relative position of epitopes or internalization of the anti-HER-2 immunotoxins. Interestingly, the most and least effective immunotoxins bound to epitopes in very close proximity. Competitive binding assays with unconjugated antibodies have previously indicated that our antibodies recognized epitopes that are arranged in a linear array. To orient this relative epitope map, deletions were prepared in the HER-2/neu gene and these mutant constructs were expressed in NIH3T3 cells. Epitope expression was determined by antibody binding and radioimmunoassay. Epitopes targeted by the PB3, 454C11 and NB3 antibodies are localized N-terminal to the epitopes recognized by ID5, BD5, 741F8 and 520C9 antibodies. The 2 non-conformational epitopes PB3 and NB3 were localized to regions corresponding to amino acides 78-242 of the p185(HER-2) protein.


Subject(s)
Antibodies, Monoclonal/therapeutic use , Epitopes/immunology , Immunotoxins/therapeutic use , Neoplasm Proteins/immunology , Receptor, ErbB-2/immunology , 3T3 Cells/immunology , 3T3 Cells/metabolism , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/metabolism , Breast Neoplasms/immunology , Breast Neoplasms/metabolism , Breast Neoplasms/therapy , Epitopes/chemistry , Epitopes/metabolism , Female , Humans , Immunoglobulin G/immunology , Immunoglobulin G/metabolism , Immunotoxins/immunology , Immunotoxins/metabolism , Mice , Neoplasm Proteins/chemistry , Neoplasm Proteins/metabolism , Receptor, ErbB-2/chemistry , Receptor, ErbB-2/metabolism , Sequence Deletion , Transfection , Tumor Cells, Cultured
14.
Clin Cancer Res ; 4(10): 2545-50, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9796989

ABSTRACT

Approximately 30% of ovarian and breast cancers overexpress p185(c-erbB-2) with as many as 10(6) receptors/cell. Normal cells have as few as 10(4) receptors/cell. We have examined the susceptibility of SKOv3 human ovarian cancer cells to anti-c-erbB2 antibodies and immunotoxins as a function of c-erbB-2 density on the cell surface. A panel of SKOv3 clones that expressed different densities of p185(c-erbB-2) receptor were generated through transfection with the c-erbB-2 gene. A significant correlation was found between p185(c-erbB-2) density and susceptibility to killing by anti-p185(c-erbB-2)-ricin A chain (anti-p185(c-erbB-2)-RTA) immunotoxins. With 10(5) copies/cell of p185(c-erbB-2), <10% of clonogenic ovarian cancer cells could be eliminated, whereas in clones that expressed 10(6) copies/cell of p185(c-erbB-2), 99.9% of clonogenic tumor cells were killed. In cell lines that overexpressed p185(c-erbB-2) and also expressed p170(EGFR), anti-p185(cerbB-2)-RTA and anti-p170(EGFR)-RTA immunotoxins exerted synergistic cytotoxicity. Treatment with the two immunotoxins could eliminate 99.99% of clonogenic cells. Importantly, tumor cells that had survived first treatment with anti-p185(c-erbB2)-RTA alone still retained sensitivity to repeat treatment with the same immunotoxin and also proved susceptible to the synergistic cytotoxicity of anti-p185(cerbB-2)-RTA in combination with anti-p170(EGFR)-RTA. Growth characteristics of the clones expressing various levels of p185(c-erbB-2) were also studied. No correlation was found between p185(c-erbB-2) expression levels and the rate of anchorage-dependent growth, anchorage-independent growth, or in vivo growth in nude mice.


Subject(s)
ErbB Receptors/analysis , Immunotoxins/therapeutic use , Ovarian Neoplasms/therapy , Receptor, ErbB-2/analysis , Ricin/therapeutic use , Animals , Drug Synergism , ErbB Receptors/immunology , Female , Humans , Mice , Receptor, ErbB-2/immunology , Tumor Cells, Cultured
15.
Clin Orthop Relat Res ; (341): 12-22, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9269149

ABSTRACT

Transplantation of total elbow allografts has been used as a salvage procedure in an attempt to provide patients with functional, painless range of motion of the elbow. This procedure is appropriate for patients with massive bone loss after trauma, tumor resection, or revision elbow arthroplasty. During the past 20 years, 23 patients have undergone elbow allograft reconstruction with variable results and a high complication rate. Ten of 14 patients with elbow allografts observed for an average of 7.5 years report satisfactory results. Allograft removal was required in six patients: for infection (two), instability (three), and nonunion and resorption (one). Three patients with instability have since undergone successful total elbow arthroplasty. Two patients have been observed less than 1 year and another patient died during the study period. Complications occurred in 16 of 23 patients. This operation is not recommended for routine use and is viewed as a salvage procedure. The use of allografts in elbow reconstruction does not preclude subsequent reconstruction with another allograft or fusion. In patients with deficient bone stock, the allograft reestablishes bone mass to permit an arthrodesis or reconstructive arthroplasty.


Subject(s)
Bone Diseases/surgery , Bone Transplantation , Elbow/surgery , Adult , Aged , Aged, 80 and over , Female , Humans , Joint Instability/etiology , Joint Instability/surgery , Joint Prosthesis , Male , Middle Aged , Orthopedics/methods , Postoperative Complications , Transplantation, Homologous , Treatment Outcome , Elbow Injuries
16.
Clin Exp Immunol ; 52(2): 293-6, 1983 May.
Article in English | MEDLINE | ID: mdl-6345037

ABSTRACT

H-10407 (078, H11) an enteropathogenic strain of Escherichia coli carries a fimbriate plasmid-mediated adherence antigen on the cell surface (CFA/I) which facilitates colonization of human small intestine. This shows strong similarities to the K88 antigen of porcine enteropathogenic E. coli. K88 expression may be suppressed by antibody both in vivo and in vitro. Expression of CFA/I, detected by agglutination of human erythrocytes was progressively lost during in vitro culture with antisera containing antibodies specific for CFA/I but, unlike K88, CFA/I was re-expressed during further culture in the absence of antibody. Antibody to CFA/I seemed to exert a switching effect on expression of the adherence antigen.


Subject(s)
Antibodies, Bacterial/immunology , Antigens, Bacterial/immunology , Escherichia coli/immunology , Fimbriae Proteins , Antibody Specificity , Antigen-Antibody Reactions , Hemagglutination Tests
17.
Parasite Immunol ; 5(2): 113-22, 1983 Mar.
Article in English | MEDLINE | ID: mdl-6302626

ABSTRACT

Cats infected with Brugia pahangi were immunosuppressed and the development of lymphatic pathology, as measured xeroradiographically by dilatation of the vessels, in these animals was compared with that of normally infected cats. Individual cats showed wide variations in response. Niridazole and antilymphocyte serum both reduced dilatation, niridazole being particularly effective in the early stages of infection. When silica, used as an inflammatory agent, was injected subcutaneously in conjunction with worm infection lymphatic dilatation developed, this being potentiated in degree and rate of development as compared with control animals. The possible role of a non-filarial inflammatory response in the development of lymphatic dilatation is discussed.


Subject(s)
Filariasis/immunology , Immunosuppressive Agents/pharmacology , Lymphatic System/pathology , Silicon Dioxide/pharmacology , Animals , Antilymphocyte Serum , Brugia , Cats , Female , Filariasis/pathology , Inflammation , Lymphography , Male , Niridazole/pharmacology , Xeroradiography
18.
Am J Clin Pathol ; 58(5): 603, 1972 Nov.
Article in English | MEDLINE | ID: mdl-4565893
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