ABSTRACT
OBJECTIVES: Canine allo- or autoantibodies are clinically important, but attachment of these immunoglobulin G (IgG) antibodies does not produce observable haemagglutination. Antibody to canine globulins is required to demonstrate sensitisation of red blood cells. Commercial reagents are available, but these often differ in sensitivity and specificity. Rabbit anticanine globulins (polyspecific) were produced for use in canine blood compatibility testing and in the investigation of immune-mediated haemolytic anaemia. METHODS: Canine sera was pooled, IgG was purified and subsequently used to immunise rabbits. A rising titre of anticanine IgG was demonstrated by indirect enzyme-linked immunosorbent assay. Rabbit anticanine complement was isolated and investigated by agglutination of complement-coated canine red blood cells. Both antibodies were purified and checked for crossreactivity before being combined to polyspecific anticanine globulins. The obtained reagent was used to indicate sensitised canine red blood cells and free antibodies in three different types of clinical samples, including blood for compatibility testing and that for investigation of immune-mediated haemolytic anaemia and screening for post-transfusion alloantibodies and was also compared to commercial Coombs' reagent. RESULTS: The product provided results in accordance with those from commercial Coombs' reagent. The sensitivity for canine crossmatching was 100% and specificity for diagnosing immune-mediated haemolytic anaemia was 87%. CLINICAL SIGNIFICANCE: This product is helpful for canine crossmatching purposes and in the investigation of immune-mediated haemolytic anaemia.
Subject(s)
Anemia, Hemolytic, Autoimmune/veterinary , Blood Grouping and Crossmatching/veterinary , Dog Diseases/diagnosis , Globulins , Anemia, Hemolytic, Autoimmune/blood , Anemia, Hemolytic, Autoimmune/diagnosis , Animals , Blood Grouping and Crossmatching/methods , Blood Transfusion/veterinary , Complement System Proteins/analysis , Coombs Test/veterinary , Dog Diseases/blood , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Globulins/immunology , Immunoglobulin G/immunology , Indicators and Reagents/therapeutic use , Rabbits , Reagent Kits, Diagnostic/veterinary , Sensitivity and SpecificityABSTRACT
We performed a placebo-controlled study to evaluate the effects of immunomodulatory treatment with thalidomide on HIV levels, TNF-alpha levels, and immune status of 31 HIV-infected individuals, after temporary suppression of viral replication with antiretroviral drugs. Treatment with a combination of zidovudine and lamivudine (ZDV/LMV) for 14 days resulted in a median decline in plasma viremia of 1.94 log10 RNA equivalents/ml. After discontinuation of ZDV/LMV, thalidomide therapy (200 mg/day for 4 weeks) did not retard the prompt return of HIV titers to the pretreatment levels, and had no effect on plasma levels of TNF-alpha. In contrast, thalidomide treatment resulted in significant immune stimulation. We observed increased levels of plasma soluble IL-2 receptor, soluble CD8 antigen, and IL-12 (p < 0.01 for all parameters), as well as increased cutaneous delayed-type hypersensitivity reactions to recall antigens (p < 0.01) in thalidomide-treated patients. These changes were associated with a median increase in HIV titer of 0.2 log10 RNA equivalents/ml in the thalidomide-treated group (p < 0.05), which resolved after stopping the drug. Further studies were performed in vitro to elucidate the mechanism of thalidomide-induced immune stimulation. When purified T cells from HIV-infected individuals were stimulated by immobilized anti-CD3 in the presence of thalidomide, a costimulatory effect of the drug was observed, resulting in increased production of IL-2 and IFN-gamma, and increased T cell-proliferative responses. Further experiments showed that thalidomide increased IL-12 production by antigen-presenting cells in a T cell-dependent manner. Our findings suggest a potential application for thalidomide as a novel immune adjuvant in HIV disease.