Petroleum kerosene fraction, co-processed with renewable hydrocarbons of plant and/or animal origin

Immunotoxicity

Description of key information

The NOAEL for jet fuel immunotoxicity is > 495 mg/kg/day administered as a 60% solution in mineral oil over at least 10% of the body surface.

Key value for chemical safety assessment

Effect on immunotoxicity: via oral route

Endpoint conclusion

Endpoint conclusion:

no study available

Effect on immunotoxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Effect on immunotoxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

immunotoxicity: short-term dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2003-01-14 to 2003-02-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study is classified as reliable without restrictions because it was conducted according to EPA OPPTS 870.7800.

Justification for type of information:

Read across justification included in Section 13

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.7800

GLP compliance:

yes

Species:

rat

Strain:

other: Crl:CD(SD)IGS BR

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Stone Ridge, New York
- Age at study initiation: 7 to 8 weeks old
- Weight at study initiation: 146 to 209 grams
- Housing: Individually in suspended stainless steel cages with wire mesh
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:15 to 22 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 25
- Humidity (%): 11 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: 2003-01-14 To: 2003-02-18

Route of administration:

dermal

Vehicle:

other: mineral oil

Details on exposure:

TEST SITE
- Area of exposure: Dorsal surface from the shoulder region to the lumbar region
- % coverage: Approximately 10%
- Type of wrap if used: None
- Time intervals for shavings or clippings: At least once a week except where there was severe dermal irritation


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Removed by gently wiping the exposed area with dry gauze
- Time after start of exposure: 6 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1 ml/kg/day
- Concentration (if solution):20%, 40%, or 60% w/v in mineral oil
- Constant volume or concentration used: yes


VEHICLE
- Justification for use and choice of vehicle (if other than water): Not reported
- Amount(s) applied (volume or weight with unit): 1 ml/kg/day
- Lot/batch no. (if required): 1F46205

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Solutions were analysed for uniformity and stability. The test compound was found to have a uniform distribution in the mineral oil. The relative standard deviation was 0.608% for the 20% samples and 4.85% for the 60% samples. The solutions were stable for 10 days. Concentration verification indicated that the concentrations were within 13.5%, 10.3%, and 18.8% of the nominal concentrations of 20%, 40%, and 60% samples, respectively.

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
165, 330, 495 mg/kg/day
Basis:
nominal conc.

No. of animals per sex per dose:

20 females per dose

Control animals:

other: sham exposed and concurrent vehicle

Details on study design:

- Dose selection rationale: Not reported
- Other: The study was conducted in two phases, each with 10 females per treatment group

Observations and clinical examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once or twice a day
- Cage side observations for mortality and viability.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily, included dermal observations at the treatment site on days 0, 1, 3 or 4, 7, 10, 14, 17, 21, and 24


BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 1, 3 or 4, 7, 10, 14, 17, 21, and 24


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: Study termination
- Anaesthetic used for blood collection: No
- Animals fasted: No data
- How many animals: All animals from Phase 2 of the study
- Parameters checked: Leukocyte counts (total and differential)


CLINICAL CHEMISTRY: No



URINALYSIS: No



OTHER: Immunotoxicity parameters were measured.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, a full postmortem exam was performed on all animals
HISTOPATHOLOGY: Yes, but only on treated and untreated skin

Cell viabilities:

SPLEEN: Yes
- Method: Not reported
- Dose groups: Untreated control, mineral oil control, 165 mg/kg/day, 330 mg/kg/day, 495 mg/kg/day, and positive control
- No. of animals: 10 animals per group


THYMUS: No


BONE MARROW: No

Humoral immunity examinations:

ANTIBODY PLAQUE FORMING CELLS (PFC) ASSAY: Yes
- Method: The plaque-forming cell assay to T-dependent antigen sheep erythrocytes, on day 24 (4 days prior to sacrifice), animals were injected via the tail vein with 200,000,000 sheep red blood cells in Earle's Balanced Salt Solution with HEPES.
- Dose groups: Untreated control, mineral oil control, 165 mg/kg/day, 330 mg/kg/day, 495 mg/kg/day, and positive control
- No. of animals: 10 animals per group


ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA): No

Non-specific cell-mediated immunity:

NATURAL KILLER (NK) CELL ACTIVITY: Yes
- Method: Not reported
- Dose groups: Untreated control, mineral oil control, 165 mg/kg/day, 330 mg/kg/day, 495 mg/kg/day, and positive control
- No. of animals: 10 animals per group


MACROPHAGE NUMBER AND FUNCTION: No

Other functional activity assays:

SPLEEN CELL PROLIFERATION ASSAY (ANTI-CD3 MEDIATED T CELL PROLIFERATION)
- Method: Not reported
- Dose groups: Untreated control, mineral oil control, 165 mg/kg/day, 330 mg/kg/day, 495 mg/kg/day, and positive control
- No. of animals: 10 animals per group

ENUMERATION TOTAL B CELLS, TOTAL T CELLS AND T CELL SUBPOPULATIONS
- Method: Not reported
- Dose groups: Untreated control, mineral oil control, 165 mg/kg/day, 330 mg/kg/day, 495 mg/kg/day, and positive control
- No. of animals: 10 animals per group

Other examinations:

The spleen, thymus, kidneys, and liver were weighed.

Positive control:

Cyclophosphamide was administered by intraperitoneal injection on the 4 days prior to sacrifice in both Phase 1 and Phase 2. Additionally, in Phase 2 Anti-asialo GM1 was used as a positive control and was injected via intraperitoneal injection 24 hours prior to sacrifice.

Statistics:

A Bartlett's test was performed to determine if the data had equal variances. A standard one-way ANOVA with F distribution followed by Dunnett's test was used for data with equal variances. A Kruskal-Wallis test followed by Dunn's Summed Rank Test was used for data with unequal variances. For the immunotoxicology data a Bartlett's Chi Square Test was used to test for homogeneity of the variances. A one-way analysis of variance was used for homogeneous data. When significant differences occurred, the treatment groups were compared to the vehicle control using Dunnett's t Test. A non-parametric analysis of variance was used on the non-homogeneous data. When a significant difference occurred, the treatment groups were compared to the vehicle control using a Gehan-Wilcoxon Test. Positive controls were compared to the corresponding vehicle control using a Student's T-test. A Jonckheere's test was used to test for trends.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not specified

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Gross pathological findings:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: Although there were no clinical signs of toxicity or treatment-related mortality, dermal irritation occurred in the vehicle control and treated rats. Eight vehicle control animals displayed transient dermal irritation. In treated animals there was a dose-dependent increase in dermal irritation with the 495 mg/kg/day dose causing considerable irritation. Irritation was noted by the presence of erythema, oedema, and desquamation. While dermal irritation was not noted until day 10 in the low- and mid-dose groups, it was evident by day 4 in the high-dose group.


BODY WEIGHT AND WEIGHT GAIN: There were no treatment-related effects on body weight.


FOOD CONSUMPTION: There were no treatment-related effects on food consumption.


HAEMATOLOGY: Several of the haematology samples were lost due to shipping problems. Therefore, the results could not be statistically analyzed.



GROSS PATHOLOGY: The only findings at necropsy were the observations of skin irritation.


CELL VIABILITIES: There were no treatment-related effects.


HUMORAL IMMUNITY EXAMINATIONS: There were no treatment-related effects.

NON-SPECIFIC CELL-MEDIATED IMMUNITY: There were no treatment-related effects.


OTHER FUNCTIONAL ACTIVITY ASSAYS: There were no treatment-related effects.


OTHER FINDINGS: There was a statistically significant decrease in thymus weights in the 330 and 495 mg/kg/day groups in Phase 1, but since the decrease did not occur in Phase 2 animals it was not considered treatment-related and was likely due to the fact that the thymus weights in Phase 1 vehicle controls were substantially larger than Phase 2 controls (sham and vehicle). Histopathology of the skin found treatment-related changes.

Cell viabilities:

no effects observed

Humoral immunity examinations:

no effects observed

Specific cell-mediated immunity:

not examined

Non-specific cell-mediated immunity:

no effects observed

Other functional activity assays:

no effects observed

Other findings:

no effects observed

Dose descriptor:

NOAEL

Effect level:

> 495 mg/kg bw/day (nominal)

Sex:

female

Basis for effect level:

other: based on overall effects; no adverse effect on immune responses of female rats at any dose level

The test compound did not affect any of the immunotoxicity parameters measured.

Conclusions:

The NOAEL for jet fuel immunotoxicity is > 495 mg/kg/day administered as a 60% solution in mineral oil over at least 10% of the body surface. However, dermal irritation occurred in a dose-dependent manner with irritation noted even in the vehicle controls.

Executive summary:

In an immunotoxicity study, 20 female Crl:CD(SD)IGS BR rats were dermally exposed to jet fuel at doses of 0 (a sham control and a mineral oil vehicle control), 165, 330, or 495 mg/kg/day (administered as 20%, 40%, or 60% solutions in mineral oil) for at least 6 hours a day for 28 days. Site was unoccluded and the rats wore Elizabethan collars to prevent ingestion. The experiment was separated into two phases. In Phase 1 there were 10 animals per group that were evaluated for the antibody plaque-forming (PFC) cell assay. Endpoints examined in Phase 1 included spleen cell viability, number of IgM PFC per spleen, number of IgM PFC per 1,000,000 spleen cells, spleen cell number, spleen weight, and body weight. Cyclophosphamide was used as the positive control. Phase 2 also used 10 animals per group and evaluated splenocyte phenotyping, the subpopulation of the cell types in the spleen (including total B and T cell populations and T-cell subsets), cell-mediated immunity as measured by proliferation of T-cells following stimulation with anti-CD3 antibody, and Natural Killer cell function. The positive controls were cyclophosphamide and anti-asialo GM1.

Vehicle control and treated rats exhibited dermal irritation characterized by erythema, oedema, and desquamation in a dose-dependent manner. Besides the dermal irritation noted, there were no treatment-related effects on clinical signs, mortality, body weight, food consumption, any of the immunotoxicity parameters measured, or macroscopic and microscopic findings. There was a significant decrease in thymus weight in the mid- and high-dose groups in Phase 1, but since this was not replicated in Phase 2 and the vehicle control rats in Phase 1 has substantially higher thymus weights than either sham or vehicle controls from Phase 2 this is not considered to be related to treatment. The positive controls had the expected results.

The NOAEL for jet fuel immunotoxicity is > 495 mg/kg/day administered as a 60% solution in mineral oil over at least 10% of the body surface. No LOAEL could be determined in this study.

This study received a Klimisch score of 1 and is classified as reliable without restrictions because it was conducted according to EPA OPPTS 870.7800.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

495 mg/kg bw/day

Study duration:

subacute

Species:

rat

Additional information

Data on related substances have been used to 'read-across' and predict the hazard properties. A 'read-across' justification document can be found in section 13.

In an immunotoxicity study, 20 female Crl:CD(SD)IGS BR rats were dermally exposed to jet fuel at doses of 0 (a sham control and a mineral oil vehicle control), 165, 330, or 495 mg/kg/day (administered as 20%, 40%, or 60% solutions in mineral oil) for at least 6 hours a day for 28 days. Site was unoccluded and the rats wore Elizabethan collars to prevent ingestion. The experiment was separated into two phases. In Phase 1 there were 10 animals per group that were evaluated for the antibody plaque-forming (PFC) cell assay. Endpoints examined in Phase 1 included spleen cell viability, number of IgM PFC per spleen, number of IgM PFC per 1,000,000 spleen cells, spleen cell number, spleen weight, and body weight. Cyclophosphamide was used as the positive control. Phase 2 also used 10 animals per group and evaluated splenocyte phenotyping, the subpopulation of the cell types in the spleen (including total B and T cell populations and T-cell subsets), cell-mediated immunity as measured by proliferation of T-cells following stimulation with anti-CD3 antibody, and Natural Killer cell function. The positive controls were cyclophosphamide and anti-asialo GM1.

Vehicle control and treated rats exhibited dermal irritation characterized by erythema, oedema, and desquamation in a dose-dependent manner. Besides the dermal irritation noted, there were no treatment-related effects on clinical signs, mortality, body weight, food consumption, any of the immunotoxicity parameters measured, or macroscopic and microscopic findings. There was a significant decrease in thymus weight in the mid- and high-dose groups in Phase 1, but since this was not replicated in Phase 2 and the vehicle control rats in Phase 1 has substantially higher thymus weights than either sham or vehicle controls from Phase 2 this is not considered to be related to treatment. The positive controls had the expected results.

The NOAEL for jet fuel immunotoxicity is > 495 mg/kg/day administered as a 60% solution in mineral oil over at least 10% of the body surface.

Justification for classification or non-classification

This endpoint is not a REACH requirement.