The following abstract presents results of a study conducted by a contract laboratory for the National Toxicology Program (NTP). The findings have not been peer reviewed and were not evaluated in accordance with the levels of evidence criteria established by the NTP on March 2009 (see http://ntp.niehs.nih.gov/ntp/htdocs/levels/09-3566%20NTP-ITOX-R1.pdf). The findings and conclusions for this study should not be construed to represent the views of the NTP or the US Government.
The Immunotoxicity of Para-nitrotoluene
99-99-0) in Female B6C3F1 Mice
NTP Report Number IMM87033
The mono-nitrotoluenes are synthesized by the nitration of toluene. Para-nitrotoluene (p-nitrotoluene) is a colorless solid at ambient temperature. The mono-nitrotoluenes are slightly soluble in aqueous solution but are readily soluble in organic solvents. The nitrotoluenes have been detected in air, water and soil.1, 2 Meta- and para-nitrotoluene have been identified by the National Toxicology Program (NTP) for detail study. P-nitrotoluene was the first mono-nitrotoluene to be investigated for its potential adverse actions on the immune system.The objective of the studies reported here was to determine the potential immunotoxicity of p-nitrotoluene in mice exposed by the oral route. The studies were divided into three areas: the more standard toxicology studies provided the base against which to compare the immunology studies; the immunology studies provided data on specific cellular targets of the xenobiotic; and the host resistance studies were used for a global assessment of the ability of the exposed animal to resist pathogenic challenge.
Table ES-1 shows a summary of the standard toxicology studies. Mice exposed to p-nitrotoluene by gastric gavage gained body weight over the 2 week experimental period in a manner similar to the control groups. Of the selected organs weighed, the spleen was the only organ to show an increase in weight. The liver of mice exposed to 400 and 600 mg/kg showed mild to moderate swelling of the hepatocytes adjacent to the central veins. The hepatocyte swelling appeared to be reversible and there was no evidence of necrosis. The selected hematological and serum chemistries were unaffected by p-nitrotoluene exposure. Bone marrow cellularity and the number of CFU/M and CFU/GM were unaffected by p-nitrotoluene exposure.
Table ES-2 summarizes the immunology studies of mice exposed to p-nitrotoluene. P-nitrotoluene suppressed the IgM response to sRBC and the DHR response to KLH. There was a slight (13%) decrease in the percentage of T lymphocytes in the spleen. All of the other immune parameters were unaffected by exposure to p-nitrotoluene. These parameters included macrophage numbers and function, natural killer cell activity, response to mitogens and allogeneic cells, and serum complement and interferon levels.
Table ES-3 summarizes the host resistance studies in mice exposed to p-nitrotoluene. Exposure of mice to p-nitrotoluene decreased resistance to Listeria monocytogenes. Resistance to Streptococcus pneumoniae and Plasmodium yoelii were unaffected by exposure as was the resistance to the two tumor models, the PYB6 tumor and the B16F10 melanoma.
The three phases of these studies indicate that the liver and, to an equal or greater extent, the immune system are a target for toxicity of p-nitrotoluene. The decreased host resistance to Listeria monocytogenes can be attributed to the decrease in T lymphocytes and, as seen in the number and activity as expressed, to the decreased delayed hypersensitivity response to Keyhole limpet hemocyanin. The decrease in IgM response did not correlate with a host resistance model, most likely because no effect was seen on the IgG response.
|Periodic Weights||Increase in |
|Histopathology||Moderate to mild |
swelling of hepatocytes
|Spleen||Increased||14%||No significance |
at high dose
|Retics||Increased||29%||No significance |
at high dose
|Albumin||Increased||25%||No significance |
at high dose
|Total Protein||Increased||17%||No significance |
at high dose
|Based on |
|DNA Synthesis||No effect|
|B Cells||No effect|
|IgM AFC to SRBC||Decreased||54%||Yes|
|IgG AFC to SRBC||No effect|
|DHR to KLH||Decreased||31%||Yes|
|Mitogens- Con A||No effect|
|- PHA||No effect|
|- LPS||No effect|
|- Medium||No effect|
|NK Cell Activity||No effect|
|Macrophage (RES)||No effect|
|Cell #||No effect|
|Phagocytosis Covaspheres||No effect|
|Phagocytosis CRBC||No effect|
|Adherence of CRBC||No effect|
|ß Glucuronidase||No effect|
|Listeria monocytogenes||Decreased resistance||41 % increase in |
|Streptococcus pneumoniae||No effect|
|Plasmodium yoelii||No effect|
1 Pellizzari, E.D. (1978): Quantitation of Chlorinated Hydrocarbons in Previously Collected Air Samples. EPA 450/3-78-412.
2 Howard, P.H., Santodonato, J., Saxena, J., Malling, J., and Greninger, D. (1976): Investigation of Selected Potential Environmental Contaminants: Nitroaromatics. U.S. EPA, RTP, NC, p.1407 EPA 560/2-76-010.
Report Date: November 1987