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Toxicological information

Immunotoxicity

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Administrative data

Endpoint:
immunotoxicity
Remarks:
acute
Type of information:
experimental study
Adequacy of study:
other information
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: acceptable, well documented publication which meets basic scientific principles

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Comparison of the immunotoxicity of propanil and its metabolite, 3,4-dichloroaniline, in C57BI/6 mice
Author:
Barnett, JB et al.
Year:
1992
Bibliographic source:
Fundamental and applied toxicology, 18: 628-631
Reference Type:
publication
Title:
Effect of acute propanil exposure on the immunresponse of C57Bl/6 mice
Author:
Barnett, JB and Gandy, J
Year:
1989
Bibliographic source:
fundamental and applied toxicology 12: 757-764

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
7 to 12 weeks old female C57Bl/6 mice were treated with 0, 37, 75 or 150 mg/ kg bw 3,4-dichloroaniline via i.p. injection on day 0 and were subjected to a number of immunological assessments on day 7. Spleen cells were prepared and Leukocytes counted, T cell-dependent antibody response was determined after immunizing treated animals by i.v. injection with 200 Million sheep erythrocytes (day 3), T cell-independent antibody response after injection of 20 µg of DNP-Ficoll (Barnett and Gandy, 1989), Cytotoxic T-lymphocyte assay and the natural killer cell assay were performed.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
3,4-dichloroaniline
EC Number:
202-448-4
EC Name:
3,4-dichloroaniline
Cas Number:
95-76-1
Molecular formula:
C6H5Cl2N
IUPAC Name:
3,4-dichloroaniline
Details on test material:
- Name of test material (as cited in study report): 3,4-dichloroaniline
- Analytical purity: 97 %

Test animals

Species:
mouse
Strain:
C57BL
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Farms (Willmington, MA)
- Age at study initiation: 7 to 12 weeks
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days prior to experiment


Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
maize oil
Details on exposure:
i.p. exposure on day 0
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
no data
Duration of treatment / exposure:
single i.p. injection on day 0, mice sacrificed on day 7
Frequency of treatment:
single treatment
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0
Basis:
nominal conc.
mg /kg bw
Remarks:
Doses / Concentrations:
37
Basis:
nominal conc.
mg /kg bw
Remarks:
Doses / Concentrations:
75
Basis:
nominal conc.
mg /kg bw
Remarks:
Doses / Concentrations:
150
Basis:
nominal conc.
mg /kg bw
No. of animals per sex per dose:
4 to 6 animals per dose, all female
Control animals:
yes, concurrent vehicle

Examinations

Observations and clinical examinations performed and frequency:
no data
Sacrifice and pathology:
GROSS PATHOLOGY: No data
HISTOPATHOLOGY: No data
Cell viabilities:
SPLEEN: Yes
- Method: single-cell suspensions of spleen cells were prepared from individual animals as described by Barnett and Gandy (1989), counted electronically using a Coulter Model ZF cell counter, Viability was checked routinely by trypan blue exclusion

- Dose groups (including vehicle control): 3 (anti-DNP PFC) or 4 (NK activity)
- No. of animals (including vehicle control): 6 (anti-DNP PFC) or 4 (NK-activity)

Humoral immunity examinations:
ANTIBODY PLAQUE FORMING CELLS (PFC) ASSAY: Yes
Assay: T cell-dependent and T cell-independent, Immunisation of treated mice with either sheep erythrocytes or DNP-Ficoll, respectively
- Method: standard agarose hemolytic plaque assay (Barnett and Gandy, 1989)
- Dose groups: 0, 37 or 150 mg/ kg bw
- No. of animals: 6

Specific cell-mediated immunity:
CYTOTOXIC T-LYMPHOCYTE (CTL) ASSAY: Yes
- Method: Brunner et al. (1979), P815 tumor cells were utilized as the sensitizing and target call for these assays
- Dose groups: no data
- No. of animals: no data


Non-specific cell-mediated immunity:
NATURAL KILLER (NK) CELL ACTIVITY: Yes
- Method: spleen cell suspensions were prepared, YAC-1 lymphoma cells (target of NK-cells) were labeled with 51Cr. Effector cells (spleen cells) were added in triplicate at effector: target ratios of 25:1, 50:1 or 100:1 ; positive (0.1 % Triton X-100) and negative (medium) controls with only labeled YAc-1 cells were included.

- Dose groups: 0, 37, 75, 150 mg/ kg bw
- No. of animals: 4 per group

Other examinations:
no data
Positive control:
no data
Statistics:
no data

Results and discussion

Results of examinations

Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Gross pathological findings:
not specified
Details on results:
see remarks on results

Specific immunotoxic examinations

Cell viabilities:
effects observed, treatment-related
Humoral immunity examinations:
effects observed, treatment-related
Specific cell-mediated immunity:
effects observed, treatment-related
Non-specific cell-mediated immunity:
effects observed, treatment-related
Other functional activity assays:
not specified
Other findings:
not specified

Effect levels

Dose descriptor:
NOAEL
Sex:
female
Remarks on result:
not determinable
Remarks:
no NOAEL identified

Any other information on results incl. tables

Relative spleen weight and leukocyte count (=leukocytosis) increased significantly (compared to vehicle control) at highest dose (150 mg/ kg). On the other hand T cell-dependent and T cell-independent humoral immune response significantly decreased in the 150 mg/ kg bw group. A dose-dependent reduction in natural killer cell activiy was observed whereas an influence of DCA on the cytotoxic T lymphocytes was not evident.

Applicant's summary and conclusion

Executive summary:

Barnett, JB et al., (1992), Fundamental and applied toxicology, 18: 628-631

7 to 12 weeks old female C57Bl/6 mice were treated with 0, 37, 75 or 150 mg/ kg bw 3,4-dichloroaniline via i.p. injection on day 0 and were subjected to a number of immunological assessments on day 7. Spleen cells were prepared and Leukocytes counted, T cell-dependent antibody response was determined after immunizing treated animals by i.v. injection with 200 million sheep erythrocytes (day 3), T cell-independent antibody response after injection of 20 µg o DNP-Ficoll (Barnett and Gandy, 1989), Cytotoxic T-lymphocyte assay and the natural killer cell assay were performed. Relative spleen weight and leukocyte count (=leukocytosis) increased significantly (compared to vehicle control) at highest dose (150 mg/ kg). On the other hand T cell-dependent and T cell-independent humoral immune response significantly decreased in the 150 mg/ kg bw group. A dose-dependent reduction in natural killer cell activiy was observed whereas an influence of DCA on the cytotoxic T lymphocytes was not evident.