Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 910-663-6 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Immunotoxicity
Administrative data
- Endpoint:
- immunotoxicity
- Remarks:
- other: in vitro human lymphocyte immunophenotype & NK function
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Meets generally acceptable scientific standards, well documented and acceptable for assessment.
Cross-referenceopen allclose all
- Reason / purpose for cross-reference:
- reference to same study
- Reason / purpose for cross-reference:
- reference to other study
Data source
Reference
- Reference Type:
- publication
- Title:
- The effect of nickel compounds on immunophenotype and natural killer cell function of normal human lymphocytes
- Author:
- Zeromski J, Jezewska E, Sikora J, Kasprzak KS
- Year:
- 1 995
- Bibliographic source:
- Toxicology. 97: 39-48
Materials and methods
Test guideline
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Peripheral blood mononuclear cells (PBMC) isolated from human blood (obtained from a blood bank) were exposed to Ni3S2 for 24 hrs. Immunophenotype and natural killer cell cytotoxicity were evaluated.
- GLP compliance:
- not specified
- Limit test:
- no
Test material
- Reference substance name:
- Trinickel disulphide
- EC Number:
- 234-829-6
- EC Name:
- Trinickel disulphide
- Cas Number:
- 12035-72-2
- Molecular formula:
- Ni3S2
- IUPAC Name:
- trinickel disulfide
- Details on test material:
- - Name of test material (as cited in study report): nickel subsulfide (Ni3S2)
- Physical state: particles of median size >= 30 um
Constituent 1
Test animals
- Species:
- other: human
- Strain:
- other: peripheral blood mononuclear cells (PBMCs) isolated from blood obtained from unidentified blood bank
- Sex:
- not specified
- Details on test animals or test system and environmental conditions:
- PBMC fractions (2 X 10^6 cells per 2 ml) were placed in 24-well tissue culture plates in RPMI-1640 medium for an 18-h pre-exposure incubation at 37°C in a 5% CO2 atmosphere.
Administration / exposure
- Route of administration:
- other: in vitro
- Vehicle:
- not specified
- Details on exposure:
- The PBMCs were cultured with three different concentrations, 0.01, 0.02, or 0.04 mM, of Ni3S2 alone, NiSO4 alone, and Mg(CH3COO)2 alone, or with three equimolar mixtures of a nickel and magnesium salt at the above concentrations, and the cells were incubated for an additional 24-h period. Control lymphocytes were cultured under the same conditions in the RPMI-1640 medium alone. After the incubation (all in triplicate), the cells were examined for immunophenotype expression and cytotoxic NK function.
- Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- Not applicable
- Duration of treatment / exposure:
- 24 hrs
- Frequency of treatment:
- Once
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0, 0.01, 0.02, and 0.04 mM Ni3S2
Basis:
other: Preliminary dose-finding study indicated that this dose range was nontoxic to cells
- No. of animals per sex per dose:
- Not applicable
- Details on study design:
- Dose Finding Study: Prior to the experiments, the toxicity of the individual metal salts for lymphocytes was tested in order to establish an optimal concentration which did not cause toxic effects on lymphocytes in short-term culture. The tests were carried out in plastic 96-well microplates kept for 24 h in a CO2 incubator. The cells (2 x 10^5 cells per 200 ul) were cultured in RPMI-1640 medium in the absence or presence of 0.01-1.0 mM concentrations of Ni3S2 and other metal salts. After a 24-h incubation, the percent of dead cells was determined by means of trypan blue exclusion using the cell culture without added metal salt as control.
Evaluation of immunophenotype: Indirect immunofluorescence was conducted to determine immunophenotype of control and treated PBMC cells. A cell suspension was overlayed on poly-L-lysine-coated slides. Cell sediments were fixed for 5 min in 1% buffered formalin and subsequently treated with mouse monoclonal antibodies (mAb) to lymphocyte or macrophage differentiation antigens (listed in Table 1 of publication). Following a 20-min incubation with the mAb and subsequent washing in phosphate buffered saline (pH 7.6), rabbit anti-mouse Ig(Fab)2 fluorescein-labelled reagent was used as a secondary antibody for another 20 min. Slides were then washed, mounted in buffered glycerin and evaluated using an Orthoplan fluorescence microscope with incident light illumination. Positive cells were counted in relation to total cells. At least 200 cells per sample were assessed.
Evaluation of NK cell cytotoxicity: PBMC were tested as effector cells versus K562 target cells labelled with 51Cr. Target cells (4 X 10^6 cells per sample) were incubated with 100 uCi of Na2·51CrO4 for 90 min at 37°C with gentle shaking. After labelling, the cells were washed five times with, and finally suspended in, RPMI-1640 medium. The test was carried out in plastic round-bottom microplates in triplicate. 100 ul aliquots of the effector cells were mixed with the same volume of target cells at effector-to-target cell ratios (E:T) of 12:1, 25:1, and 50:1. After 4-h incubation (37°C; 5% CO2), a 100 ul sample of the supernatant was collected from each well and radioactivity counted in a gamma counter. The result was calculated from the formula:
% Cytotoxicity = [(E - S)/(T - S)] x 100
Where E = activity of 51Cr released from target cells in the presence of effector cells. S = activity of 51Cr released spontaneously under identical conditions from target cells alone; this value did not exceed 10-15% of the maximal release (T). T = maximum activity of 51Cr released when all target cells are destroyed. T was determined by the addition of 10% Triton X-100 to the suspension of target cells.
Examinations
- Observations and clinical examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: No data
DETAILED CLINICAL OBSERVATIONS: No data
BODY WEIGHT: No data
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
OPHTHALMOSCOPIC EXAMINATION: No data
HAEMATOLOGY: No data
CLINICAL CHEMISTRY: No data
URINALYSIS: No data - Sacrifice and pathology:
- Not applicable
- Cell viabilities:
- SPLEEN: No data
THYMUS: No data
BONE MARROW: No data - Humoral immunity examinations:
- ANTIBODY PLAQUE FORMING CELLS (PFC) ASSAY: No data
ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA): No data - Specific cell-mediated immunity:
- ONE-WAY MIXED LYMPHOCYTE CULTURE (MLC) ASSAY: No data
DELAYED-TYPE HYPERSENSITIVITY (DTH) REACTION: No data
CYTOTOXIC T-LYMPHOCYTE (CTL) ASSAY: No data - Non-specific cell-mediated immunity:
- NATURAL KILLER (NK) CELL ACTIVITY: Yes
- Method: PBMC were tested as effector cells versus K562 target cells labelled with 51Cr. Target cells (4 X 10^6 cells per sample) were incubated with 100 uCi of Na2·51CrO4 for 90 min at 37°C with gentle shaking. After labelling, the cells were washed five times with, and finally suspended in, RPMI-1640 medium. The test was carried out in plastic round-bottom microplates in triplicate. 100 ul aliquots of the effector cells were mixed with the same volume of target cells at effector-to-target cell ratios (E:T) of 12:1, 25:1, and 50:1. After 4-h incubation (37°C; 5% CO2), a 100 ul sample of the supernatant was collected from each well and radioactivity counted in a gamma counter. The result was calculated from the formula:
% Cytotoxicity = [(E - S)/(T - S)] x 100
Where E = activity of 51Cr released from target cells in the presence of effector cells. S = activity of 51Cr released spontaneously under identical conditions from target cells alone; this value did not exceed 10-15% of the maximal release (T). T = maximum activity of 51Cr released when all target cells are destroyed. T was determined by the addition of 10% Triton X-100 to the suspension of target cells.
- Dose groups: each dose (0.01, 0.02, or 0.04 mM) or exposure condition (i.e., single metal salt alone, or equimolar mixture with magnesium salt) was run in triplicate.
- No. of animals: not applicable - Other functional activity assays:
- SPLEEN CELL PROLIFERATION ASSAY (ANTI-CD3 MEDIATED T CELL PROLIFERATION): No data
ENUMERATION TOTAL B CELLS, TOTAL T CELLS AND T CELL SUBPOPULATIONS: No data - Other examinations:
- Evaluation of immunophenotype: Indirect immunofluorescence was conducted to determine immunophenotype of control and treated PBMC cells. A cell suspension was overlayed on poly-L-lysine-coated slides. Cell sediments were fixed for 5 min in 1% buffered formalin and subsequently treated with mouse monoclonal antibodies (mAb) to lymphocyte or macrophage differentiation antigens (listed in Table 1 of publication). Following a 20-min incubation with the mAb and subsequent washing in phosphate buffered saline (pH 7.6), rabbit anti-mouse Ig(Fab)2 fluorescein-labelled reagent was used as a secondary antibody for another 20 min. Slides were then washed, mounted in buffered glycerin and evaluated using an Orthoplan fluorescence microscope with incident light illumination. Positive cells were counted in relation to total cells. At least 200 cells per sample were assessed.
- Positive control:
- Not reported
- Statistics:
- Significance levels of differences among the results were determined with the use of Student's t test.
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:
- NON-SPECIFIC CELL-MEDIATED IMMUNITY: Ni3S2 distinctly suppressed the NK cytotoxicity of cultured PBMC at all concentrations and E:T ratios tested. In contrast, Mg(CH3COO)2 alone tended to enhance NK cytotoxicity and to reverse the suppressive action of the nickel salts (see figures 4 & 5 in paper).
OTHER FINDINGS - Immunophenotyping: Ni3S2 had a marked inhibitory effect on the PBMCs. It consisted of a significant decrease in the number of CD4-positive cells at concentrations of 0.02 and 0.04 mM Ni3S2 and a decrease in NK (CD56-positive) cell number at all Ni3S2 concentrations tested. However, the percentages of CD3, CD8, CD20, and CD11a cells did not show major alterations. Mg(CH3COO)2 prevented the inhibitory effects of Ni3S2 on CD56 and CD4 cells.
Specific immunotoxic examinations
- Cell viabilities:
- not specified
- Humoral immunity examinations:
- not specified
- Specific cell-mediated immunity:
- not specified
- Non-specific cell-mediated immunity:
- effects observed, treatment-related
- Other functional activity assays:
- not specified
- Other findings:
- effects observed, treatment-related
Effect levels
open allclose all
- Dose descriptor:
- dose level:
- Effect level:
- >= 0.05 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: cell viability (from dose finding study): effect level = dose at which % cells alive below 95%.
- Dose descriptor:
- dose level:
- Effect level:
- > 0.04 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD3-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- >= 0.02 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD4-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- > 0.04 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD8-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- < 0.01 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD56-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- > 0.04 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD20-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- > 0.04 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects other: decrease in the number of CD11a-positive cells (relative to control)
- Dose descriptor:
- dose level:
- Effect level:
- < 0.01 other: mM
- Sex:
- not specified
- Basis for effect level:
- other: overall effects cell-mediated immunity (NK cell activity): suppression of the NK cytotoxicity of PBMC cells
Applicant's summary and conclusion
- Conclusions:
- The primary effects of Ni3S2 on human peripheral blood mononuclear cells in vitro consist of depletion of the CD4 and NK cell subsets and reduction of the NK cytotoxic function. The magnesium salt Mg(CH3COO)2 prevented these Ni3S2 effects.
- Executive summary:
Zeromski et al. (1995) examined a variety of immunotoxicity endpoints associated with Ni3S2 exposures in human lymphocytes (i.e., peripheral blood mononuclear cells – PBMCs) derived from samples of blood donors as obtained from an unidentified blood bank. Initially, a dose-finding study was conducted to identify doses of Ni3S2 that did not affect short-term cell viability in culture. Second, a series of assays was conducted to assess the natural killer activity of the PBMCs associated with Ni3S2 exposure in vitro.
The cell viability test was carried out in plastic 96-well microplates kept for 24 hr in a CO2 incubator. The cells (2 x 105 cells per 200 ul) were cultured in RPMI-1640 medium in the absence or presence of 0.01-1.0 mM concentrations of Ni3S2 and other metal salts. Trypan blue exclusion illustrated that 0.01, 0.02, and 0.04 mM Ni3S2 concentrations left greater than 95% of the cell populations alive and were thus considered “nontoxic” for the purposes of examining lymphocyte immunophenotype and function. PBMCs (2 X 106 cells per 2 ml) were cultured in 24-well tissue culture plates in RPMI-1640 medium for an 18-h pre-exposure incubation at 37°C in a 5% CO2 atmosphere, and then exposed to the three non-toxic concentrations of Ni3S2 alone or in an equimolar mixture with magnesium acetate for 24 hr. The immunophenotype of the cells was determined by indirect immunofluorescence, using monoclonal antibodies to major differentiation antigens of peripheral blood mononuclear cells. Exposure of cells to Ni3S2 resulted in the decline of CD4 (at 0.02 and 0.04 mM) and natural killer cell populations (i.e., CD56; at all concentrations). However, Ni3S2 did not affect CD3, CD8, CD20, or CD11a cell populations.
The natural killer activity of the PBMCs was studied by measuring the cytotoxic effect of Ni3S2 -exposed and control PBMCs toward K562 target cells. Target cells (4 X 106 cells per sample) were radiolabelled with 51Cr during a 90 min incubation at 37°C. The cells then were washed and suspended in, RPMI-1640 medium. 100 ul aliquots of the effector cells (i.e., PBMCs) were mixed with the same volume of target cells at effector-to-target cell ratios (E:T) of 12:1, 25:1, and 50:1. After 4-h incubation (37°C; 5% CO2), a 100 ul sample of the supernatant was collected from each well and radioactivity counted in a gamma counter. At all exposure concentrations, Ni3S2 suppressed the natural cytotoxic activity of PBMCs toward the target cells. The metal salt magnesium acetate, however, prevented all Ni3S2 -mediated immunotoxic effects studied. The authors broadly concluded that nickel subsulfide had deleterious effects on human peripheral blood mononuclear cells in short-term in vitro culture, but the presence and magnitude of these effects was dependent on the cell subsets. In this in vitro study, Ni3S2 selectively depleted helper/inducer T lymphocytes (i.e., CD4-presenting PBMCs) and natural killer cells (i.e., CD56-presenting PBMCs), while also reducing natural killer cell function. And since magnesium inhibited these effects, the authors further suggest that magnesium supplementation might be a useful prophylactic against immunotoxic effects of nickel exposure. STUDY RATED BY AN INDEPENDENT REVIEWER
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.