Department of Health and Human Services Public Health Service, Grant Application

Princit al Investigator/Program DirectorLast, first, middle):

Duesber, Peter H.

"P11""V. I Invi tr /Pro... am. Director...ast... first middle): Duesber 'Peter H.
Animal tests of the AIDS risks of nitrite inhalants
1. Specific Aims
It is proposed to test the hypothesis that characteristic AIDS diseases such as Kaposi's sarcoma
and pneumonia are caused by nitrite inhalants (poppers) which are used as psychoactive drugs and
sexual stimulants, primarily by male homosexuals. This drug-AIDS hypothesis was originally
advanced as "lifestyle" hypothesis, but was abandoned by most investigators in 1984 in favor of the
HIV-AIDS hypothesis. The drug hypothesis is based on epidemiological and toxicological evidence.
The epidemiological evidence includes pre- and post-1984 statistics demonstrating consistent
correlations between nitrite use and AIDS. Kaposi's sarcoma, for example, occurs almost exclusively
(>95%) in male homosexual AIDS patients (Beral et al., 1990) and male homosexuals are almost the
exclusive (>95%) users of nitrite inhalants (see 2c). Moreover HIV-free AIDS has been documented
in users of nitrites (see 2d). The toxicity of nitrites includes a) oxidation of hemoglobin to
methemoglobin, b) mutagenesis of DNA by deamination and c) carcinogenicity.
Following and extending unfinished studies by others from the mid-1980s, it is proposed here
to determine the effects of the long-term inhalation of nitrites on mice (e.g. 6 hrs/5 days/week for 6 to
12 months). To distinguish between the roles of nitrites as autonomous pathogens and as co-factors of
HIV, murine retrovirus-infected and un-infected mice will be studied in parallel. The parameters to be
monitored include a) weight loss, b) anemia and lymphocytopenia, c) clinical manifestations of
immunodeficiency, particularly Pneumocystis pneumonia, d) evidence for Kaposi's sarcoma.
In addition we propose to study the effects of nitrites on the growth rate of human T-cell lines,
H9 and CEM, in culture (Rasheed et al., 1986) as a model for the immunotoxicity of nitrites in
humans.
Finally we plan to examine the Kaposi sarcomagenic potential of nitrites on rodent cells in
culture. Avoiding the toxicity to vital organs, like the lung and the bone marrow, the cell culture
system will permit us to test nitrites at higher-than-in-vivo concentrations. This is critical to detect
sarcomagenic potential, because Kaposi sarcomas are typically restricted to humans with the highest
cumulative lifetime doses of nitrites (see 2c). If confirmed the nitrite hypothesis would be directly
relevant for the prevention and therapy of AIDS.
2. Background and significance
a)The drug-AIDS hypothesis. The epidemiology of AIDS is very peculiar,'and hard to
reconcile with the hypothesis that AIDS is a sexually transmitted, viral disease. About 90% of all
American AIDS patients are males (Centers for Disease Control, 1993) and male homosexuals have a
20-fold higher risk of developing Kaposi's sarcoma than other HIV-infected risk groups, including
intravenous drug users, hemophiliacs and recipients of transfusions (Selik et al., 1987; Beral et al.,
1990). Typically, infectious diseases spread equally between the sexes and cause the same symptoms
or collection of symptoms in all risk groups (Duesberg, 1992a).
A consistent alternative explanation suggests that AIDS is caused in part or entirely by toxins,
e.g., recreational drugs and AZT (Duesberg, 1992a). This drug-AIDS hypothesis was originally
advanced as "lifestyle" hypothesis, but has been abandoned by most scientists in 1984 in favor of
Gallo's HIV-AIDS hypothesis (Centers for Disease Control, 1986; Oppenheimer, 1992). Indeed,
recreational drugs are consumed primarily by males: males consume about 80% of injected
recreational drugs (Duesberg, 1992a), and homosexual males are almost exclusive users of oral
aphrodisiacs, including above all nitrite inhalants, who are used to facilitate anal intercourse (Mirvish
and Haverkos, 1987; Haverkos and Dougherty, 1988b). The almost exclusive use of nitrite inhalants

("poppers") and other chemical stimulants by male homosexuals also offers a plausible explanation for
the almost exclusive occurrence of Kaposi's sarcoma in this AIDS risk group (Guinan et al., 1984;
PHS 398 (Rev.9/91) Page 24
Number pages consecutively at the bottom throughout the application. Do not use suffixes such as 3a, 3b.