A predictive F344 rat immunotoxicology model: cellular parameters combined with humoral response to NP-CgammaG and KLH.
The purpose of this study was to examine the predictive value of humoral and cellular immune parameters in determining the immunotoxic effects of the oral administration of azathioprine (AZA), cyclophosphamide (CY), or cyclosporin A (CsA) at doses of 25/17, 10, or 25 mg/kg per day, respectively, for 30 days in F344 female rats. The effect of these known immunosuppressive compounds on the immune response was assessed in a humoral model that consisted of the administration of nitrophenyl-chicken gamma globulin (NP-CgammaG) and keyhole limpet hemocyanin (KLH) antigens during immunosuppressive treatment and the measurement of resulting rat antigen-specific IgG and IgM, as well as total IgG, levels. Cellular assessment parameters were collected from the same groups of animals as the humoral parameters and included organ weights and cellularity, hematology, lymphocyte phenotype characteristics, spleen cell mitogen stimulation (T and B cell-dependent), splenic natural killer (NK) cell cytotoxicity, and bone marrow cellularity and lymphocyte phenotype differential. Although decreases in several of the cellular assay parameters were observed, the only functional assays to demonstrate a statistically significant immunosuppressive effect by all three immunosuppressive agents were the antigen-specific serum IgG levels. The primary (day 10; 15 days post-immunization) and secondary (day 25; 5 days post-rechallenge) nitrophenyl (NP) responses were significantly suppressed by > or =60%. The use of NP hapten provided consistent responses when analyzed with a sensitive, well developed, ELISA methodology. Absolute lymphocyte phenotyping and lymphocyte hematology were also predictive of T cell immunosuppression for all three compounds. The data presented herein suggests that these two parameters, NP-IgG humoral response and lymphocyte phenotyping, are sufficient for identifying immunosuppressive compounds.
Smith
Holly W
HW
Eli Lilly and Company, 2001 W Main Street, Greenfield, IN 46140, USA. smith_holly_w@lilly.com
Winstead
Colleen J
CJ
Stank
Kristen K
KK
Halstead
Bartley W
BW
Wierda
Daniel
D
D000818:Animals; D000917:Antibody Formation / Q000187:drug effects* / Q000276:immunology; D001379:Azathioprine / Q000633:toxicity; D001854:Bone Marrow Cells / Q000187:drug effects / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D003520:Cyclophosphamide / Q000633:toxicity; D016572:Cyclosporine / Q000633:toxicity; D005260:Female; D006241:Haptens / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology*; D007111:Immunity, Cellular / Q000187:drug effects* / Q000276:immunology; D007166:Immunosuppressive Agents / Q000633:toxicity*; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology; D023421:Models, Animal; D009596:Nitrophenols / Q000276:immunology*; D051381:Rats; D011916:Rats, Inbred F344; D013154:Spleen / Q000187:drug effects / Q000276:immunology; D013950:Thymus Gland / Q000187:drug effects / Q000276:immunology; D018675:Toxicity Tests / Q000379:methods
D006241:Haptens; D007166:Immunosuppressive Agents; D009596:Nitrophenols; D016572:Cyclosporine; D003520:Cyclophosphamide; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D001379:Azathioprine