Registration Dossier

Administrative data

Endpoint:
immunotoxicity, other
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
other information
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature

Data source

Reference
Reference Type:
secondary source
Title:
Immunotoxicological evaluation of environmental chemicals utilizing the mouse lymphocyte mitogenesis test
Author:
Sakazaki, H. et al.
Year:
2001
Bibliographic source:
Journal of Health Sciences, 47(3), pp. 258-271; cited in OECD SIDS "Isobutanol", Final September 2004

Materials and methods

Principles of method if other than guideline:
Immunotoxicity - Lymphocyte Mitogenesis Test
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
mouse
Strain:
Balb/c
Remarks:
and C3H/He
Sex:
male

Administration / exposure

Vehicle:
water
Remarks:
or DMSO (max. 0.3 %)
Duration of treatment / exposure:
96 h
Frequency of treatment:
continous
Doses / concentrations
Remarks:
Doses / Concentrations:
10e-9 to 10e-3 mol/L
Control animals:
yes, concurrent vehicle
Details on study design:
B cells were isolated from the spleen of male C3H/He mice following injection of 10 ml of RPMI-1640 medium into the spleen. T cells were isolated from the spleen of male BALB/C mice in a similar manner. The cells were flushed out of the spleen using a syringe and the suspended cells were removed from the connective tissue, washed, and counted with a hematocytometer. The cells were dispensed into 96-well microplates at 10e-5 cells/well in 200 µL RPMI-1640 medium containing 5 mM HEPES, 50 PM 2-mercaptoethanol, 100 IU/mI penicillin, 50 µg/ml streptomycin, 0.18% NaHC03, and 10% fetal calf serum. Mitogenic stimuli for the B and T cells were provided by addition of 100 µg/ml lipopolysaccharide or 200 µg/ml concanavalin A, respectively. The test chemical was added to stimulated cells and incubations proceeded for 96 hours at 37' C in a 5% C02 atmosphere. At the end of the exposure period, the total amount of DNA in the grown cells was determined by the ethidium bromide fluorescence method. The control cells were treated in the same manner as the treated cells with the exception that no test chemical was added (only vehicle) . A cell growth curve was plotted against test chemical concentration and a concentration for 50% growth inhibition (IC50) was determined .

Results and discussion

Effect levels

Key result
Dose descriptor:
other: inhibition of mitogenic actitivity
Basis for effect level:
other: Isobutanol showed no inhibition of mitogenic activity in stimulated B and T cells in the concentration range tested.
Remarks on result:
not measured/tested

Applicant's summary and conclusion