Warm red blood cell autoantibodies and clinical diagnoses in patients with or without autoimmune hemolysis.

OBJECTIVES: Red blood cell autoantibodies (RBC autoAbs) of IgG class are found in the majority of patients with warm autoimmune hemolytic anemia (wAIHA) but sometimes also during the pretransfusion testing of patients with different diagnoses but without hemolysis. The aim of the study was to identify the main differences between these two groups of patients according to age, gender, subclass and titer of IgG RBC autoAbs and diagnosis. MATERIAL AND METHODS: In the 9-year retrospective study, data were collected from records of 291 patients with IgG RBC autoAbs detected by gel technique, from which 111 with wAIHA. RESULTS: More than 85% of patients in both groups were over 40 years old, with male to female ratio 1:1.9 in wAIHA vs 1:1.3 in patients without hemolysis (P=0.0916). The main characteristics of patients with wAIHA vs patients without hemolysis were: IgG only 38% vs 70%, IgG+Complement 62% vs 30%, total IgG1 79% vs 55%, IgG1+IgG3 35% vs 11%, titer of 100 for IgG1+IgG3 17% vs 3% (P<0.0001), respectively, while titer of 100 for IgG1 18% vs 9% (P=0.0241). The underlying diagnosis in wAIHA vs patients without hemolysis: hematologic disorders 41% vs 22% (P=0.0006), autoimmune disorders 12% vs 13% (P=0.8033), solid tumors 5% vs 14% (P=0.0154) and surgery procedures 6% vs 26% (P<0.0001). CONCLUSION: We observed more wAIHA patients with high titer of IgG1 and high prevalence of IgG1+IgG3 and consider that patients without hemolysis having identical results might be interesting to find out how they are protected from damage by RBC autoAbs.

Thromboelastographic assessment of the contribution of platelets and clotting proteases to the hypercoagulable state of dogs with immune-mediated hemolytic anemia.

BACKGROUND: Hypercoagulability is a well-known feature of canine immune-mediated hemolytic anemia (IMHA) and is believed to increase the risk of thrombosis. This study was undertaken to differentiate the relative contribution of platelets and clotting proteases to this hypercoagulability using thromboelastography (TEG). STUDY DESIGN: Retrospective observational study. METHODS: Thromboelastograms from 27 dogs with IMHA were retrospectively evaluated. Standard TEG parameters (R, K, α, MA), the G value, and the novel parameter delta (Δ) were determined. Hypercoagulability was attributed to the platelet component of hemostasis when there was an increased G value with a normal Δ value. KEY FINDINGS: Nineteen of 27 dogs (70.4%) had ≥ 2 TEG variables suggestive of hypercoagulability, 18 (66.7%) had a hypercoagulable G value, and 11 (40.7%) had a hypercoagulable Δ value. Ten of 27 (37%) samples met the criteria for platelet hypercoagulability. SIGNIFICANCE: Our report documents the derivation and application of the Δ value to differentiate enzymatic from platelet hypercoagulability. Further studies are required to validate the use of these TEG variables in this manner. The hypercoagulable tendency in dogs with IMHA is complex and multifactorial, and in some dogs this hypercoagulability may be attributed primarily to platelet hyper reactivity. Our findings may support the use of anti-platelet drugs in some dogs with IMHA.

Serial assessment of the coagulation status of dogs with immune-mediated haemolytic anaemia using thromboelastography.

This study investigated the coagulation status of dogs with immune-mediated haemolytic anaemia (IMHA) over time. Thirty animals with primary IMHA were blood sampled on three occasions over a 5 day period and assays performed included prothrombin time, activated partial thromboplastin time, D-dimer and fibrinogen concentration, antithrombin activity and recalcified unactivated thromboelastography (TEG). Based on TEG, dogs with IMHA were significantly hypercoagulable vs. controls (P<0.001) and over the 5 day period, 3/4 of the TEG parameters reflected increased clotting kinetics (P ≤ 0.02). The 30 day survival of these patients was 80% and, at hospital admission, the TEG maximum amplitude (MA) was significantly higher in survivors than non-survivors (P=0.015). Each unit increase in MA was associated with an increased odds of 30 day survival of 1.13 (95%; CI 1.02-1.25). Based on TEG, most dogs with IMHA were hypercoagulable on admission and their clotting kinetics increased with time. Relative hypocoagulability identified by TEG at initial assessment was found to be a negative prognostic indicator.

Do patients with autoantibodies or clinically insignificant alloantibodies require an indirect antiglobulin test crossmatch?

BACKGROUND: Compatibility testing is the standard protocol that identifies suitable blood for patients requiring transfusion. If the antibody screen is negative or no clinically significant antibodies are detected, BCSH guidelines and AABB standards allow an immediate-spin crossmatch (IS XM) or even electronic issue. The testing requirement is less clear where autoantibodies or non-clinically significant alloantibodies compromise the indirect antiglobulin test crossmatch (IAT XM). Performing an IAT XM will give a mismatched result anyway, delays the supply of blood to the patient, and provides no additional benefit or safety. STUDY DESIGN AND METHODS: From January 2002 to April 2006, the provision of blood for autoimmune hemolytic anemia (AIHA) patients with autoantibodies and no alloantibodies as well as patients with alloantibodies that exhibited a "high-titer, low-avidity" (HTLA) mode of reactivity was reviewed. RESULTS: A total of 222 AIHA patients (428 samples) with autoantibodies had 1585 units of red cells supplied after IAT XM; 1308 (82.5%) were mismatched. In 50 patients (80 samples) with HTLA-like antibodies, 286 units of 328 (87.2%) were mismatched by IAT XM. CONCLUSION: No adverse reactions were reported for the study groups where "suitable" blood was provided after a serologically mismatched IAT XM. No additional benefit for these patients can be claimed by performing an IAT XM over an IS XM, as a check of ABO match. The IAT XM is both costly and time-consuming. It is proposed that for these study group patients, a reduction to an IS XM can be applied and can be beneficial.

Assessment of an EIA for measuring human serum erythropoietin as compared with RIA and an in-vitro bioassay.

A recently developed enzyme-linked immunosorbent assay (EIAZ, ELISA) using two murine monoclonal anti-erythropoietin antibodies was compared with a radioimmunoassay (RIA) and a commercial in-vitro bioassay, EPOS, for measuring serum erythropoietin (Epo) in humans. Specificity and validity for Epo-EIA and the other two assays were examined. The serum Epo in normal subjects was 18 +/- 12 mU/ml (mean +/- SD, n = 80) for EIA compared with 22.5 +/- 18.5 mU/ml (n = 20) for RIA and 136 +/- 132 mU/ml (n = 14) for the bioassay. The serum Epo concentrations in normals and patients were highly comparable between EIA and RIA for Epo (P less than 0.01, r = 0.95). Epo concentrations by the EIA for normal female and male subjects were 20.5 +/- 13 and 16.5 +/- 10 mU/ml, respectively. Epo levels in patients with secondary polycythaemia or autoimmune haemolytic anaemia were significantly higher than normal subjects by the three methods. Epo levels in patients with chronic renal failure were within the normal range. By the EPOS bioassay, the Epo concentrations of normals and patients with renal failure were significantly higher than expected (136 +/- 132 and 447 +/- 273, respectively). Due to its inherent design, the EPOS bioassay possibly measures bone marrow proliferative activity in response to other serum growth regulators besides erythropoietin and was found to be unsuitable for clinical assessment of Epo. We concluded that the new EIA and RIA were similarly sensitive, reliable and accurate for measurement of serum Epo. The EIA method has the advantage of being less time consuming, more convenient and avoids the use of a radioisotope.