Clinical Immunology

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The Clinical Immunology laboratory of KSVDL provides high quality services to practicing veterinarians, the Veterinary Health Center, researchers and industry. This is an AAVLD accredited laboratory.

Tests

If you do not find a specific test listed, please contact Client Services at (785) 532-5650, toll free at 1-866-512-5650 or clientcare@vet.k-state.edu for special testing submission and fees.

RBC- surface associated immunoglobin test for dogs and horses with immune mediated anemia

The erythrocyte surface antibody test detects antibodies on the surface of erythrocytes. For dogs, we use fluorescently- labeled polyclonal antisera to canine IgG, IgM, IgA, and C3 and flow cytometry to quantitate the percentage of antibody coated cells in the sample. For horses, we use fluorescently- labeled polyclonal antisera to equine IgG, IgM and IgA. This test is more sensitive that the Coombs anti-globulin agglutination test, because it is not dependent on agglutination as an endpoint and the false negatives that occur at low end (prozone) or high end (post zone) of the dilution of Coombs reagent. The sensitivity is 100% and the specificity is 87.5% determined by population of dogs with immune mediated hemolytic anemia (J. Vet. Intern. Med. 2000, 14:190-196). The test does not differentiate primary (idiopathic or autoimmune) immune mediated hemolytic anemia (IMHA) from secondary causes for IMHA.

We modified this assay to detect penicillin specific antibodies in equine serum in cases of penicillin induced IMHA.

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Anti-Nuclear Antibody (ANA)

Dilutions of patient serum are tested for antibodies to nuclear substances 1:80 through 1:640. We have noted ANA titers of 1:80 and 1:160 in healthy dogs and in dogs with inflammatory conditions. Positive ANA titers should be considered when the titer is >1:160. Interpretation of a positive ANA as diagnostic of SLE should always accompany specific clinical criteria for SLE because some dogs can have positive ANA titers without disease or with inflammatory conditions (Compendium, 1999:21:135 and 402).

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Rheumatoid Factor Assay (RF)

We use a latex agglutination slide test to detect canine rheumatoid factor in serum (antibody to canine IgG). A definitive diagnosis of canine rheumatoid arthritis should be made by meeting 5/11 criteria established by the American Rheumatoid Association. (Berkow et al, Merck Manual of Diagnosis and Therapy, thirteenth edition, Rahaw, N.J.).

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Immunophenotyping

Lymphocyte phenotyping is requested in cases of immunodeficiency, chronic inflammatory conditions (lymphocytosis), and determining the cell lineage of lymphomas or leukemias. Lymphocyte phenotyping is requested in cases of immunodeficiency, chronic inflammatory conditions (lymphocytosis), and lymphoma or leukemia typing.

Monoclonal antibodies (VMRD, Pullman, WA) Surface Determinants % positive lymphocytes*
HB19A + 1.9/3.2 CD5 + IgM+ B 92.0 ± 5.8
HB19A CD5 91.8 ± 2.5
HB61A CD4 66.2 ± 12.3
HT14A CD8 16.6 ± 2.8
1.9/3.2 IgM+B 12.6 ± 3.2
TH14B MHC II (DR) 70.7 ± 8.3
H42A MHC II (DP) 81.1 ± 5.6

Equine lymphocyte subsets determined by immunophenotyping and flow cytometry
*percentages determined on gated lymphocytes excluding monocytes
(values based on 10 adult horses)

Flow Cytometry PowerPoint Presentation

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