Possible incorporation of an immunotoxicological functional assay for assessing humoral immunity for hazard identification purposes in rats on standard toxicology study.
The objective of this study was to examine the feasibility of conducting an immunotoxicological assay for assessing humoral immunity in rats on standard toxicology study. Male CD rats were untreated or dosed intraperitoneally daily for 30 or 90 days, excluding weekends, with vehicle or 2 mg/kg cyclophosphamide (CY). Six days prior to sacrifice, selected rats were injected intravenously with sheep red blood cells (SRBC). One day prior to necropsy, blood samples for hematological and clinical chemical measurements were collected from each rat. On the day of necropsy standard protocol tissues were collected, weighed, processed to slides, and examined microscopically. One-half of each spleen was used to prepare a single cell suspension in order to assess spleen cell numbers. Serum was analyzed for anti-SRBC IgM antibody using an enzyme-linked immunosorbent assay. A second set of studies was performed to examine further the effect of SRBC administration on lymphoid organ weights using 30- and 90-day study age-equivalent naive male CD rats. Exposure of animals to 2 mg/kg CY for 30 or 90 days resulted in a 28% and 61% decrease, respectively, in SRBC-specific serum IgM levels. CY treatment also caused mild alterations in some leukocytic parameters, with significant decreases of 35% and 33% in white blood cell and lymphocyte counts, respectively, observed in 30-day CY-treated animals receiving SRBC. Injection of SRBC alone did not alter hematological or clinical chemistry parameters. With the expected exception of the spleen (increased number and size of germinal centers), administration of SRBC did not significantly alter the weights or morphology of routine protocol tissues. Furthermore, administration of SRBC did not mask the immunosuppressive effects of CY treatment under the conditions of this study. Based on our preliminary findings, a functional assay for assessing humoral immunity may be conducted in animals on standard toxicology study.
Ladics
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GS
E.I. du Pont de Nemours and Co., Haskell Laboratory for Toxicology and Industrial Medicine, Newark, DE 19714.
Smith
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C
Heaps
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K
Elliott
G S
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Slone
T W
TW
Loveless
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D000818:Animals; D000917:Antibody Formation / Q000187:drug effects* / Q000276:immunology; D003520:Cyclophosphamide / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D017707:Erythrocyte Transfusion; D004912:Erythrocytes / Q000276:immunology; D005240:Feasibility Studies; D007075:Immunoglobulin M / Q000097:blood; D007274:Injections, Intraperitoneal; D007958:Leukocyte Count; D008099:Liver / Q000187:drug effects / Q000473:pathology; D008221:Lymphoid Tissue / Q000187:drug effects / Q000473:pathology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012756:Sheep; D013154:Spleen / Q000166:cytology / Q000187:drug effects* / Q000473:pathology
D007075:Immunoglobulin M; D003520:Cyclophosphamide