Skip to Main Navigation
Skip to Page Content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.


The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Share This:

Abstract for IMM90012

Immunotoxicity of Oxymetholone in Male B6C3F1 Mice

CASRN: 434-07-1
Chemical Formula: C21H32O3
Molecular Weight: 332.4808
Report Date: December 1990


The following abstract presents results of a study conducted by a contract laboratory for the National Toxicology Program. The findings have not been peer reviewed and were not evaluated in accordance with the levels of evidence criteria established by NTP in March 2009. The findings and conclusions for this study should not be construed to represent the views of the NTP or the U.S. Government.


Oxymetholone is a synthetic androgen, structurally related to testosterone. Originally, it was used in the treatment of pituitary dwarfism and adjunctive therapy in osteoporosis. Its current use is limited to treatment of certain anemias. Anabolic steroids have been taken to improve strength and orally active anabolic steroids, such as oxymetholone, represent a class of abused drugs. Since sex steroids, including estrogen and testosterone, are well known to influence immune function at pharmacological or suprapharmacological levels the potential effects of oxymetholone on the immune system of mice were examined.


Oxymetholone was obtained from Sigma Chemical Corp. (Lot #037F0705) and was determined to be greater than 97% pure by HPLC. The material was prepared in saline containing 0.5% methylcellulose, and was administered daily for 14 consecutive days at doses of 50, 150, or 300 mg/kg by oral gavage in volumes of 0.2ml/10g body weight. Control mice received the vehicle and the positive control group received cyclophosphamide (Sigma Chemical Corp. Lot #67F-0155) by i.p. injection at daily doses of 25 mg/kg on days 11 through 14 of the study except in the host resistance tests where cyclophosphamide was administered at a single dose of 200mg/kg on day 14. Since oxymetholone is a synthetic androgen, male B6C3F1 mice, rather than female mice were used. On day 15, one day after the last exposure, the animals were evaluated for the indicated immunological and toxicological endpoints. Unless indicated, 7-8 mice were examined per treatment group for each endpoints.


General toxicological and immunological data are summarized in Table 1. There were no treatment-related effects on mortality, body weights or selected organ weights including spleen, thymus or liver (latter not shown). Hematological parameters were not remarkably affected, nor were serum chemistries (SGPT, glucose, total protein, albumin, globulin) although BUN levels were slightly elevated at the two higher doses. While bone marrow cell DNA synthesis in the 300 mg/kg treatment group was decreased, no effects were seen on cellularity or CFU-C2 progenitor cell formation (data not shown). There were no oxymetholone-related effects on the antibody response to sheep erythrocytes, natural killer cell activity, mitogenic responses to PHA, Con A or LPS, or the numbers of splenic T and B lymphocytes or T lymphcytes in subsets. The mixed leukocyte response was suppressed in the high- dose group and there was a slight decrease in the cytotoxic T lymphocyte response for all doses of oxymetholone. Macrophages, obtained from the peritoneal cavity of oxymetholone-treated mice, were equally responsive to activation by interferon-gamma as those from control mice when assessed by in vitro tumor cell killing (data not shown).

The ability of animals to resist infection with Listeria monocytogenes was not significantly compromised by oxymetholone treatment (Table 2).


Under these experimental conditions, the results suggest that treatment with oxymetholone can result in a mild decrease in cell-mediated immunity. However, when contrasted to the degree of suppression induced by cyclosphosphamide, and the limited alterations in other immune functions associated with oxymetholone exposure, the changes are considered minimal. The chemical treatment did not affect the ability of the animals to resist infection in the host resistance tests.


Table 1. Summary of Oxymetholone Immune Studies
Parameter Dose (mg/kg) CTX
  0 50 150 300 25 mg/kg
Body Weight (g) 28.6 28.2 27.8 28.6 28.1
Spleen (mg) 134 117 139 154 74#
Spleen:Body wt. (%) 0.48 0.41 0.50 0.55 0.26#
Thymus (mg) 55 44 43 49 45
Thymus:Body wt. (%) 0.19 0.15 0.16 0.17 0.16
Spleen cell no. (x 10E7) 15.4 13.2 14.9 14.1 8.3#
Peripheral Leukocyte Count
(10E3/mm3) 4.6 6.7 4.5 5.6 3.3
Thy 1.2+ (%) 39.2 41.1 47.3 32.8 57.4*
CD4+ (%) 22.6 25.4 26.0 22.0 32.0#
CD8+ (%) 10.8 13.4 13.4 10.6 16.6#
sIg+ (%) 41.7 40.9 41.3 39.3 36.4
Cytotoxic T Lymphocytes
(% cytotoxicity) 25:1 99.6 88.6* 87.5# 84.3# 83.0#
Natural Killer Cell Activity
(% cytotoxicity) 100:1 7.8 ND 9.3 9.2 ND
IgM PFC/10E6 2143 2253 2347 2183 33#
IgM PFC/spleen (x 10E3) 366 432 434 422 4#
Lymphoproliferation: 3H-TdR Incorporation (cpm x 10E3)
Concanavalin A 107 128 113 87 66
Phytohemagglutinin 94 128 123 76 110
Lipopolysaccharide 11 13 10 9 5*
Mixed Leukocyte Response 47 34 45 29* 12#

Abbreviation: CTX = cyclophosphamide;
* different from vehicle control at P less than 0.05
# different from vehicle control at P less than 0.01
ND = no data

Table 2: Summary of Oxymetholone Host Resistance Studies
L. Monocytogenes Dose (mg/kg/day) CTX
Host Resistance Test   200mg/kg
(12 Per Group) 0 50 100 300
Percent Morbidity 17 8 8 25 100
Low Infectivity challenge
Percent Morbidity
High Infectivity Challenge 100 100 100 100 100