Dynamics of viral replication in infants with vertically acquired human immunodeficiency virus type 1 infection.

Abstract

About one-third of vertically HIV-1 infected infants develop AIDS within the first months of life; the remainder show slower disease progression. We investigated the relationship between the pattern of HIV-1 replication early in life and disease outcome in eleven infected infants sequentially studied from birth. Viral load in cells and plasma was measured by highly sensitive competitive PCR-based methods.Although all infants showed an increase in the indices of viral replication within their first weeks of life, three distinct patterns emerged: ( a ) a rapid increase in plasma viral RNA and cell-associated proviral DNA during the first 4-6 wk, reaching high steady state levels ( Ͼ 1,000 HIV-1 copies/10 5 PBMC and Ͼ 1,000,000 RNA copies/ml plasma) within 2-3 mo of age; ( b ) a similar initial rapid increase in viral load, followed by a 2.5-50-fold decline in viral levels; ( c ) a significantly lower ( Ͼ 10-fold) viral increase during the first 4-6 wk of age. All infants displaying the first pattern developed early AIDS, while infants with slower clinical progression exhibited the second or third pattern.These findings demonstrate that the pattern of viral replication and clearance in the first 2-3 mo of life is strictly correlated with, and predictive of disease evolution in vertically infected infants. ( J. Clin. Invest. 1996. 97:323-330.)

Dynamics of HIV-1 mRNA expression in patients with long-term nonprogressive HIV-1 infection.

“…The use of internal standards for the quantification of total extracted DNA ( ␤ -globin) and RNA ( ␤ -actin) appears very important for these quantitative evaluations, since the total amount of extracted nucleic acids from clinical samples is often very variable. We have already used this procedure to study the regulation of HIV-1 gene expression in in vitro-infected experimental models (37) and to monitor viral load in HIV-1-infected patients (38,40,41).…”

“…DNA was extracted from frozen cellular pellets according to standard procedures (54). Viral RNA was extracted from the virion pellet obtained after ultracentrifugation (40,000 rpm on an SW41t rotor for 60 min; Beckman Instruments, Fullerton, CA) of 5 ml plasma (38,40), according to the guanidinium thiocyanate procedure (55). Cellular RNA was extracted from frozen cellular pellets by cell lysis in solution D according to the guanidinium thiocyanate procedure (55).…”

“…Plasmid pSPLI-II was used as a competitor for the competitive PCR determination of the copy number of HIV-1 proviral DNA and of the reference single-copy ␤ -globin gene in DNA samples, while the in vitro transcription product from plasmid pSPLI-II was used for the RT-PCR quantification of HIV-1 genomic RNA in serum, of HIV-1 viral mRNAs, and of the cellular ␤ -actin mRNA in cellular RNA samples. The construction (37) and the use (38,40) of this multicompetitor plasmid for the quantitation of HIV-1 nucleic acids have already been described; a schematic representation of the insert of this plasmid is shown in Fig. 1 B .…”

“…In recent years, we have developed and extensively applied a competitive PCR technology for the exact quantitation of low abundance nucleic acids in clinical samples (30)(31)(32)(33)(34)(35), including HIV DNA and RNA in different clinical settings (36)(37)(38)(39)(40)(41). Competitive PCR and RT-PCR (using internal DNA and RNA competitors, respectively) couple the sensitivity of conventional PCR to the accuracy that is necessary for the exact quantification of the target molecules, irrespective of sample nucleic acid purity, primer pair efficiency and specificity, or amplification kinetics profile.…”

Functional state of CD4+ and CD8+ T lymphocytes and their role in the slow progression of HIV infection in pediatric patients

“…Foi identificado um comportamento clínico bimodal nas crianças infectadas por transmissão vertical. Assim, 10 a 30% dos pacientes, chamados de progressores rápidos (PRs), começam a apresentar os sintomas da AIDS nos primeiros meses devida (no período de 1 ano), enquanto que um grupo minoritário de pacientes, denominados progressores lentos (PLs), permanece assintomático por vários anos e pode inclusive chegar à idade escolar [3][4][5] . Embora alguns fatores imunológicos e virológicos 6,7 associados à progressão para a doença tenham sido identificados, tais como sintomas clínicos (hepatomegalia, esplenomegalia e linfadenopatia manifestadas nos primeiros anos de vida) 6,8 , os mecanismos celulares e moleculares subjacentes à não progressão em crianças infectadas pelo HIV-1 ainda não foram bem compreendidos.…”