3,4-dichloroaniline

Immunotoxicity

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

Reference

Reference 1

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 2

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

Test material information

Constituent 1

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 / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0
Basis:
nominal conc.
mg /kg bw

Doses / concentrations 2

Remarks:

Doses / Concentrations:
37
Basis:
nominal conc.
mg /kg bw

Doses / concentrations 3

Remarks:

Doses / Concentrations:
75
Basis:
nominal conc.
mg /kg bw

Doses / concentrations 4

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.