Genomic variation of human immunodeficiency virus type 1 (HIV-1): molecular analyses of HIV-1 in sequential blood samples and various organs obtained at autopsy

Ball, Jonathan K.; Holmes, Edward C.; Whitwell, Helen; Desselberger, Ulrich

doi:10.1099/0022-1317-75-4-867

Cited by 63 publications

(63 citation statements)

References 59 publications

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“…DNA was extracted from Ficol~Hypaque purified PBMCs or organ samples as described previously (Ball et al, 1994) using either the sarcosyl-proteinase K phenol method of Simmonds et al (1990a) or a commercially available extraction kit (Stratagene). RNA was extracted from plasma derived from the cardiac blood and used as template in a reverse transcription procedure to produce cDNA, as described previously (Ball et al, 1994).…”

Section: Methodsmentioning

confidence: 99%

“…The following env (e) and pol (p) primers described by Simmonds et al (1990a, b), Ball et al (1994) and Sheehy & Desselberger (1993) were used in nested primer reactions. The env andpolcoordinates correspond to those of H IV-1 HXB2 (Myers et al, 1994): 401 (e, sense, 6539, 5' GAG GAT ATA ATC AGT TTA TGG 3'); 306 (e, sense, 7009, 5' TGG CAG TCT AGC AGA AGA AG 3'); 307 (e, antisense, 7331, 5' CTG GGT CCC CTC CTG AGG 3'); 502 (e, sense, 7360, 5' GGG GAA TTT TTC TAC TGT AAT 3'); 503 (e, antisense, 7665, 5' CTT CTC CAA TTG TCC CTC ATA 3'); 504 (e, antisense, 7814, 5' CCA TAG TGC TTC CTG CTG CT 3'); RVL8 (p, sense, 1877, 5' GTT GAC TCA GAT TGG TTG CAC 3'); WHO1 (p, sense, 1947, 5' CCA GGA ATG GAT GGC CCA 3'); WHO2 (p, antisense, 3578, 5' CCT GAT TCC AGC ACT GAC 3'); NS12 (p, antisense, 3877, 5' GTT TCC TGC CCT GTC TCT GCT 3').…”

Section: Pcrmentioning

confidence: 99%

“…The amplification of single DNA/cDNA molecules was carried out using methods described previously (Ball et al, 1994). Amplification of a portion of the gpl20 coding region encoding all the hypervariable regions of env was achieved using the primer pair 401 and 504 in a first round reaction.…”

Section: Pcrmentioning

confidence: 99%

“…Positive single molecule PCR amplification products were identified using ethidium bromide stained agarose gel electrophoresis and purified using the commercially available Isogene kit (Perkin Elmer Cetus). Purified V3-and RT-specific PCR products were used as templates in sequencing reactions using components of the Sequenase version 2 kit (USB) in the presence of E. coli single-stranded DNA binding protein (Ball & Desselberger, 1992) and only two labelling nucleotides (Tsang & Bentley, 1989;Ball et al, 1994).…”

Section: Pcrmentioning

confidence: 99%

“…Extensive temporal and topographical genetic variations are observed in human immunodeficiency virus type 1 (HIV-1) populations in different individuals and also in those present within the same individual Epstein et al, 1991 ;Delassus et al, 1992;Ball et al, 1994). The structural genes of HIV-1 differ in their levels of genetic variation, with the gag and pol genes being more conserved than the env gene (Myers et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Concurrent evolution of regions of the envelope and polymerase genes of human immunodeficiency virus type 1 observed during zidovudine (AZT) therapy

Sheehy

Desselberger²,

Whitwell³

et al. 1996

Journal of General Virology

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Nucleotide sequences of regions of the envelope (env) and polymerase (po/) genes of human immunodeficiency virus type 1 (HIV-1) proviral DNA were obtained from sequential blood and autopsy samples from an AIDS patient who had been treated with zidovudine for 9 months, Phylogenetic analyses showed that a reduction in genetic heterogeneity of the env regions of viruses present in the proviral blood population occurred during therapy, and this coincided with an increased pol gene heterogeneity. Differences were observed in different organs obtained post mortem for both the env and pol coding regions. The cardiac blood proviral population consisted mainly of variants which possessed sequences containing mutations at position 215 of the pol gene, associated with drug resistance. By contrast, the brain population consisted entirely of zidovudine sensitive genotypes, and this organ also harboured variants with genetically distinct env sequences. The lymph tissues obtained after death held more diverse proviral env and pol populations, containing both zidovudine sensitive and resistant genotypes.

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Section: Methodsmentioning

confidence: 99%

Section: Pcrmentioning

confidence: 99%

Section: Pcrmentioning

confidence: 99%

Section: Pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Concurrent evolution of regions of the envelope and polymerase genes of human immunodeficiency virus type 1 observed during zidovudine (AZT) therapy

Sheehy

Desselberger²,

Whitwell³

et al. 1996

Journal of General Virology

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GBV-C/HGV coinfection in HIV-1-positive men: Frequent detection of viral RNA in blood plasma but absence from seminal fluid plasma

Hollingsworth¹,

Jameson²,

Minton³

et al. 1998

J. Med. Virol.

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Sequential paired samples of blood and seminal fluid were obtained from a cohort of 54 HIV-1-infected homosexual males. The prevalence of GBV-C/HGV RNA in the cell-free fractions of some of these patients was determined using reverse-transcription polymerase chain reaction (RT-PCR). To assess the effects of HIV-1 and HCV infection upon GBV-C/HGV RNA status, blood CD4 cell counts, HCV RNA status, and HIV-1 proviral DNA and viral RNA titres were also determined. GBV-C/HGV RNA was detected in 8/30 (27%) of the blood plasma samples obtained at the start of the study, and was present at a frequency of 14/64 (22%) in all the blood plasma samples tested. By contrast, GBV-C/HGV RNA was not detected in the 26 seminal fluid samples obtained at the start of the study, including 8 samples obtained from patients for which GBV-C/HGV RNA was detected in the corresponding blood sample. Of the samples tested for the presence of both GBV-C/HGV and HCV RNA, there was no evidence of coinfection. Although GBV-C/HGV RNA detection rates were significantly higher in individuals with blood CD4 cell counts greater than 200 cells per microlitre, there were no significant differences in the median blood CD4 cell counts or HIV-1 proviral DNA or viral RNA titres observed between the GBV-C/HGV-positive and -negative individuals. The failure to detect GBV-C/HGV RNA in seminal fluid samples obtained from this cohort would suggest that further studies need to be carried out to determine the roles of sexual transmission and of seminal fluid in GBV-C/HGV infection.

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Concordance between semen‐derived HIV‐1 proviral DNA and viral RNA hypervariable region 3 (V3) envelope sequences in cases where semen populations are distinct from those present in blood

Curran

Ball

2002

Journal of Medical Virology

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Sequence analysis of the third hypervariable region (V3) of the envelope gene of the HIV-1 was carried out on HIV proviral and viral populations present in blood and semen. Phylogenetically distinct populations of virus were observed in three of the 10 patients analysed. Although the majority of the viruses were predicted to have an R5 phenotype, amino acid differences between blood and semen-derived virus and provirus sequences were observed at sites previously shown to affect cell tropism. Importantly, the semen proviral population was representative of that observed for cell-free virus. This indicates that seminal fluid mononuclear cells are possible sources for the cell-free virus in found in semen.

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Genomic variation of human immunodeficiency virus type 1 (HIV-1): molecular analyses of HIV-1 in sequential blood samples and various organs obtained at autopsy

Cited by 63 publications

References 59 publications

Concurrent evolution of regions of the envelope and polymerase genes of human immunodeficiency virus type 1 observed during zidovudine (AZT) therapy

Concurrent evolution of regions of the envelope and polymerase genes of human immunodeficiency virus type 1 observed during zidovudine (AZT) therapy

GBV-C/HGV coinfection in HIV-1-positive men: Frequent detection of viral RNA in blood plasma but absence from seminal fluid plasma

Concordance between semen‐derived HIV‐1 proviral DNA and viral RNA hypervariable region 3 (V3) envelope sequences in cases where semen populations are distinct from those present in blood

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