About: Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles

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About: Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles
Creator	Jones, R Longmoor, B Offutt, D
Source	Elsevier; Medline; PMC
abstract	Abstract The purpose of this subacute 22-day study was to evaluate methods for canine circulating immunoglobulins (IgM, IgG, and IgE) and select B- and T-lymphocyte populations (CD4-helpers, CD8-suppressors, pan-T and pan-B) for immunotoxicity testing using an organ system (concordance) approach. The challenge substance for immunoglobulin testing was repeated immunization with six-way distemper vaccination (DHLAPP), while the challenge substance for leukocyte subpopulations was treatment with cyclophosphamide. Immunoglobulin measurements were made by capture enzyme-linked immunosorbent assay (ELISA), and leukocyte immunophenotyping by fluorescein isothiocyanate/phycoerythrin conjugation (flow cytometry). A battery of parameters that would be used in a typical regulatory study were taken to aid interpretation of the data generated by these methods. Body weights, food consumption, clinical observations, complete clinical chemistry and urinalysis measurements were taken. Gross pathology and micropathology of sternal bone marrow, spleen, mesenteric and retropharyngeal lymph nodes, thymus, liver and kidney were completed. The ELISA method demonstrated acceptable intra-assay reproducibility for IgM, IgG and IgE, with values in good agreement as reported for radial immunodiffusion. The immunologic challenge demonstrated a biological trend of an increase in IgM that preceded an increase in IgG with no discernible trend in IgE response, and no abnormalities in lymphocyte subpopulations. Principle flow cytometry findings related to cyclophosphamide were that the relative percent of B cells decreased dramatically and progressively after compound administration; being statistically decreased in males on day 22 compared with day −5. The relative percent CD4 and CD8 contribution increased, but the CD4/CD8 ratio remained relatively unchanged as total white blood cells decreased progressively. The increase in relative percent CD4 (males only) was statistically significant according to a two-sample t-test on days 17, 20 and 22 when compared with the pre-treatment day −5. There was a relative percent increase in CD5-panT, but absolute numbers were dramatically decreased. We conclude that an organ system approach to assessment of the immune system which incorporates humoral antibody, enumeration of lymphocyte populations and pathologic evaluation of the lymphoreticular organs assists in the interpretation of an adverse toxicological response. The ELISA method for measurement of Igs detected the expected levels of IgG, IgM and IgE due to repeated vaccinations and to cyclophosphamide treatment. The flow cytometry method was acceptable for measuring select canine lymphocyte populations and detecting the expected decrease in B cells due to cyclophosphamide treatment. Both methods may be added to a testing battery for assessing immunotoxicity in canine regulatory studies.
has issue date	2000-04-10(xsd:dateTime)
bibo:doi	10.1016/s0378-4274(00)00173-9
bibo:pmid	10817629
has license	els-covid
sha1sum (hex)	29350aee2c60ec77463b4d59a60574becf0a2e68
schema:url	https://doi.org/10.1016/s0378-4274%2800%2900173-9
resource representing a document's title	Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles
has PubMed Central identifier	PMC7127553
has PubMed identifier	10817629
schema:publication	Toxicology Letters
resource representing a document's body	covid:29350aee2c60ec77463b4d59a60574becf0a2e68#body_text
is schema:about of	named entity 'aid' named entity 'liver' named entity 'subpopulations' named entity 'sternal' named entity 'canine' named entity 'canine' named entity 'Both' named entity 'IgE' named entity 'compound' named entity 'substance' named entity 'evaluate' named entity 'organ system' named entity 'immunologic' named entity 'expected' named entity 'CD8' named entity 'OBSERVATIONS' named entity 'LEUKOCYTE' named entity 'LYMPHORETICULAR' named entity 'IMMUNOPHENOTYPING' named entity 'INTRA' named entity 'ENUMERATION' named entity 'IGG' named entity 'PAN' named entity 'NO ABNORMALITIES' named entity 'CONTRIBUTION' named entity 'COMPLETED' named entity 'JONES' named entity 'WWW' named entity 'B CELLS' named entity 'TOTAL' named entity 'REGULATORY' named entity 'COMPLETE' named entity 'REPORTED' named entity 'ACCEPTABLE' named entity 'IMMUNOGLOBULIN' named entity 'ENZYME-LINKED IMMUNOSORBENT ASSAY' named entity 'BEING' named entity 'T-TEST' named entity 'DUE TO' named entity 'RELATIVE' named entity 'SUBACUTE' named entity 'METHODS' named entity 'CONJUGATION' named entity 'RESPONSE' named entity 'IMMUNE SYSTEM' named entity 'URINALYSIS' named entity 'SPLEEN' named entity 'USED' named entity 'WHITE BLOOD CELLS' named entity 'TOXICOLOGY' covid:arg/29350aee2c60ec77463b4d59a60574becf0a2e68 named entity 'BEAGLES' named entity 'CAPTURE' named entity 'FOLLOWING' named entity 'CHALLENGES' named entity 'IMMUNIZATION' named entity 'ASSESSMENT' named entity 'COMPARED' named entity 'CAPTURE' named entity 'PRINCIPLE' named entity 'KIDNEY' named entity 'FLUORESCEIN ISOTHIOCYANATE' named entity 'TESTING' named entity 'LYMPHOCYTE' named entity 'RETROPHARYNGEAL LYMPH NODES' named entity 'FLOW CYTOMETRY' named entity 'T-LYMPHOCYTE' named entity '115' named entity 'ELISA' named entity 'METHODS' named entity 'CYCLOPHOSPHAMIDE' named entity 'BIOLOGICAL' named entity 'GENERATED' named entity 'ANTIBODY' named entity 'DAY' named entity 'ADMINISTRATION' named entity 'PATHOLOGIC' named entity 'MALES' named entity 'DISTEMPER'

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