Document 2876
 DOCN  M94A2876
 TI    Use of N-(4-[[(2,4-diamino-6-pteridinyl)methyl]methylamino]-1- glutamic
       acid (methotrexate) (MT) in inhibiting replication of human
       immunodeficiency virus-1(HIV-1) in vitro.
 DT    9412
 AU    Kumar S; Clinical Regional Laboratory, Flossmoor, IL 60422.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):204 (abstract no. PB0246). Unique
       Identifier : AIDSLINE ICA10/94369699
 AB    OBJECTIVE: To study the possible effects of different drugs on the
       activity and growth and on controlling, delaying, or preventing the
       replication of the AIDS virus (HIV-1) in humans, we have used peripheral
       blood lymphocytes from untreated AIDS-positive patients and studied the
       effect of MT, a drug used for the treatment of various types of
       carcinomas, acute lymphocytic leukemia, meningeal leukemia, etc. on the
       growth of HIV in culture. METHODS: Phytohaemagglutin (PHA)-stimulated
       peripheral blood lymphocytes (PHABL) were infected with isolates of
       HIV-1 from five untreated patients and grown in a serum-free, synthetic,
       completely chemically defined tissue culture medium containing varying
       concentrations of MT. The cells were cultured at an initial
       concentration of 1 x 10(6) per milliliter in 5 ml of culture media.
       Appropriate concentrations of MT (0.01, 0.02, 0.04, 0.08 and 0.12
       microM) were added to the cultures with no MT added to control cultures
       containing cells from AIDS-positive blood samples. Cells were passaged
       every 3 days and MT added so that the original drug level was
       maintained. The cultures were monitored for 30 days. Fresh PHABL from
       uninfected donors were added every week to replenish aged cultures.
       RESULTS: Activity of supernate viral reverse transcriptase (RT)
       (measured as an indicator of HIV replication) increased 2 to 5 times in
       cultures that contained no MT or 0.01 uM/L of MT. No viral cells, viral
       enzyme (RT) activity or HIV-1 p24 antigen production was detected in
       cultures containing 0.12 uM/L of MT by day 15. No cell toxicity at
       concentrations used in our experiments was observed. DISCUSSIONS AND
       CONCLUSIONS: Results of our study indicate that MT inhibits the
       replication of HIV-1 in culture. Virus breakthrough suppression and
       complete inhibition of HIV-1 RT by MT, should prevent emergence of the
       virus, once inactivated, in humans. Complete inhibition of HIV-1
       replication indicates that MT therapy may be beneficial in the treatment
       of HIV-1 infections and possibly, in post-exposure prophylaxis. To our
       knowledge, this is the first time that a drug has been shown to inhibit
       HIV-1 replication and inactivate the virus completely. While our results
       strongly suggest use of MT as a drug to control replication of HIV-1,
       extrapolation of in vitro experiments to clinical therapy will require
       further work to determine a direct relationship on the effect of MT and
       the level of HIV-1 in patients with HIV-induced persistent generalized
       lymphadenopathy. Experiments are presently being performed to determine
       if MT will affect the progression of viral infection, in vivo, in
       animals infected with HIV.
 DE    Antiviral Agents/*PHARMACOLOGY  Cells, Cultured  Culture Media,
       Conditioned/CHEMISTRY  Human  HIV Core Protein p24/ANALYSIS  HIV-1/*DRUG
       EFFECTS  Lymphocytes/MICROBIOLOGY  Methotrexate/*PHARMACOLOGY  Reverse
       Transcriptase/ANALYSIS  Virus Replication/*DRUG EFFECTS  MEETING
       ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).