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  • Author or Editor: Cynthia A. Lucidi x
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Abstract

OBJECTIVE To develop and characterize flow cytometric assays for detecting IgG bound to canine erythrocytes and bone marrow erythroid precursors.

SAMPLE Blood samples from 20 healthy and 61 sick dogs with (n = 33) or without (28) immune-mediated hemolytic anemia (IMHA) and bone marrow samples from 14 healthy dogs.

PROCEDURES A flow cytometric assay for measurement of IgG on RBCs was developed, and appropriate positive control cells were generated. Analytic and diagnostic performance were characterized. The RBC IgG assay was then combined with density-gradient fractionation of aspirated bone marrow cells and a 2-color process to yield an assay for detecting IgG on nucleated RBCs (nRBCs). Cell sorting and cytologic examination confirmed target cell populations, and anti–dog erythrocyte antigen 1 (DEA1) blood-typing serum was used to generate IgG-positive nRBCs.

RESULTS Within- and between-run coefficients of variation for the RBC IgG assay were 0.1% to 13.9%, and > 90% of spiked IgG-positive RBCs were detected. Diagnostic sensitivity and specificity of the assay for detection of IMHA were 88% and 93%, respectively. Cytologic findings for sorted bone marrow fractions rich in early-, mid-, and late-stage nRBCs from 3 healthy dogs indicated 89% to 98% nRBC purity. After IgG coating with anti-DEA1 blood-typing serum, IgG was detected on nRBCs from DEA1-positive, but not DEA1-negative, healthy dogs.

CONCLUSIONS AND CLINICAL RELEVANCE The developed RBC IgG assay had favorable analytic and diagnostic performance for detection of IMHA in dogs and was successfully adapted to detect IgG on canine nRBCs of various maturation stages. The findings supported the presence of DEA1 on canine nRBCs.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To characterize the clinical features of dogs with precursor-targeted immune-mediated anemia (PIMA).

ANIMALS

66 dogs with PIMA.

PROCEDURES

Electronic record databases of a teaching hospital were searched to identify dogs with a diagnosis of nonregenerative anemia between 2004 and 2013. Inclusion criteria included persistent nonregenerative anemia (Hct ≤ 30% and reticulocyte count < 76,000 reticulocytes/μL), cytologic findings supportive of ineffective bone marrow erythropoiesis, and absence of underlying disease. Information regarding clinical signs, clinicopathologic findings, treatment, and outcome was extracted from records of eligible dogs. A regenerative response was defined as a reticulocyte count > 76,000 reticulocytes/μL or sustained increase in Hct of > 5%. Remission was defined as a stable Hct ≥ 35%.

RESULTS

The median Hct was 13%, and reticulocyte count was 17,900 reticulocytes/μL. Rubriphagocytosis was identified in bone marrow aspirate samples from 61 of 66 dogs. Collagen myelofibrosis was detected in bone marrow biopsy specimens obtained from 31 of 63 dogs. Immune-mediated targeting of mature erythrocytes was uncommon. All dogs received immunosuppressive therapy. Fifty-five dogs developed a regenerative response at a median of 29 days, and 40 of those dogs went into remission at a median of 59 days after PIMA diagnosis. Thromboembolic events were confirmed for 9 dogs and were associated with a decreased survival time. Median survival time was 913 days for all dogs.

CONCLUSIONS AND CLINICAL RELEVANCE

Results indicated that most dogs with PIMA responded to prolonged immunosuppressive therapy. Studies to determine optimal immunosuppressive and thromboprophylactic protocols for dogs with PIMA are warranted.

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in Journal of the American Veterinary Medical Association
History and Clinical Findings

An 8-year-old 43.7-kg (96.1-lb) castrated male Labrador Retriever was referred for evaluation of progressive left thoracic limb lameness of 4 weeks’ duration. The lameness started suddenly following play with another dog, and the referring veterinarian localized pain to the lateral aspect of the left elbow joint upon palpation. Empirical treatment was not effective.

The referral physical examination revealed a grade 4/5 left thoracic limb lameness with offloading of the left paw when at rest. The dog's prostate was not palpable on rectal examination. Palpation of the left elbow region and flexion and extension of the left

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in Journal of the American Veterinary Medical Association