CHANGES IN THE VIRAL MESSENGER-RNA EXPRESSION PATTERN CORRELATE WITH A RAPID RATE OF CD4(-CELL NUMBER DECLINE IN HUMAN-IMMUNODEFICIENCY-VIRUS TYPE 1-INFECTED INDIVIDUALS() T)

The rate of disease progression varies considerably among human immuno deficiency virus type 1 (HIV-1)-infected individuals. Several cross-se ctional studies have shown an association between the stage of HIV-1 d isease and the viral burden or the relative levels of viral gene expre ssion. To study the extent of HIV-1 transcription and replication and its correlations with disease progression, we quantified serial, longi tudinal samples of blood cells from 10 HIV-1-infected individuals with markedly different rates of CD4(+) T-cell number decline following se roconversion. After normalization for the input nucleic acid content, multiply spliced viral mRNA and unspliced viral RNA were quantified by competitive reverse transcription-PCR using oligonucleotide primers w hich flank the major tat/rev/nef splice junction and span an internal region of the gag open reading frame, respectively. Coamplification of internal cRNA template controls was used to normalize for variation i n the efficiency of reverse transcription and in vitro enzymatic ampli fication. Similarly, proviral DNA was also quantified by competitive P CR performed within the linear range of amplification. Viral RNA was d etected in the blood cells of each individual from all time points reg ardless of the rate of CD4(+) T-cell decline. Unspliced genomic viral RNA rapidly increased in the blood cells from HIV-1-infected individua ls who had a precipitously declining CD4(+) T-cell number. In contrast , both unspliced and multiply spliced viral mRNAs remained relatively stable in the blood cells from HIV-1-infected individuals who have had a relatively benign clinical course. These data demonstrate that the extent of viral transcription and replication correlates with the rate of CD4(+) T-cell number decline and that quantifying intracellular vi ral RNA is of potential prognostic value.