A new multiplex PCR strategy for the simultaneous determination of four genetic polymorphisms affecting HIV-1 disease progression

Kristiansen, Thomas Birk; Knudsen, Troels Bygum; Ohlendorff, Stine; Eugen‐Olsen, Jesper

doi:10.1016/s0022-1759(01)00349-0

Cited by 34 publications

(27 citation statements)

References 13 publications

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“…CD4 ϩ T-cell counts were determined by the University of Washington (UW) Laboratory Medicine clinical laboratory by combining percent CD4 ϩ T cells from flow cytometry and lymphocyte counts from hemocytometry. Before July 2003, HIV-1 viral-load testing employed the Amplicor HIV-1 Monitor Test (Roche Molecular Systems, Inc.) with a lower limit of detection of 50 copies per ml; thereafter, an in-house real-time reverse transcription (RT)-PCR method was used, with a lower limit of detection of 30 copies per ml (17). The HIV-1 plasma viral load was measured at time point 1 for viral-load analyses.…”

Section: Methodsmentioning

confidence: 99%

Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Dinges

Richardt

Friedrich

et al. 2010

J Virol

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؉ T-cell counts (4 to 8 times longer), while responses to variable epitopes presented by B35 alleles (DL9 and IL9) resulted in more rapid progression. The plasma viral load was higher in B57/58 ؉ and B27 ؉ subjects lacking the conserved B57/58-and B27-restricted responses. The presence of certain B57/58-, B27-, and B35-restricted HIV-specific CD8 ؉ T-cell responses after primary HIV-1 infection better defined disease progression than the HLA genotype alone, suggesting that it is the HIV-specific CD8 ؉ T cells and not the presence of a particular HLA allele that determine disease progression. Further, the most effective host CD8 ؉ T-cell responses to HIV-1 were prevalent within an HLA allele, represented a high total allele fraction of the host CD8 ؉ T-cell response, and targeted conserved regions of HIV-1. These data suggest that vaccine immunogens should contain only conserved regions of HIV-1.

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Section: Methodsmentioning

confidence: 99%

Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Dinges

Richardt

Friedrich

et al. 2010

J Virol

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“…HLA class I and class II typing for HLA-A, -B, -C, -DRB and -DQB1 was carried out using a polymerase chain reaction (PCR) technique with sequence-specific primers as reported previously [30,31]. The D32 CCR5 polymorphism was determined by modified PCR with sequence specific primers, with the primer 5′-TCA TCA TCC TCC TGA CAA TCG-3′ and the anti-sense primer 5′-CCA GCC CCA AGT TGA CTA TC-3′ [32]. The PCR products were separated on a 2% agarose gel to maximize the separation between the wildtype (262 bp) and deletion (230 bp) products.…”

Section: Hla Typing and Ccr5 Polymorphismmentioning

confidence: 99%

Immunogenic and tolerogenic signatures in human immunodeficiency virus (HIV)-infected controllers compared with progressors and a conversion strategy of virus control

Whittall

Peters²,

Rahman³

et al. 2011

Clinical and Experimental Immunology

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“…The extraction was performed following to the manufacturer's protocol, DNA was re-suspended in 200 mL of AE buffer (QIAgen, Hilden, Germany) and stored at 4°C for further analyses. To assess the distribution of single nucleotide polymorphisms, PCR-RFLP protocols were used except for the D32 CCR5 variation, which was screened by a previously described PCR technique (Umehara et al 2004;Kristiansen et al 2001;Rector et al 2001) .…”

Section: Dna Extraction and Genotypingmentioning

confidence: 99%

Sequence variants of chemokine receptor genes and susceptibility to HIV-1 infection

et al. 2009

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Genetic susceptibility to HIV infection was previously proven to be influenced by some chemokine receptor polymorphisms clustering on chromosome 3p21. Here the influence of 5 genetic variants was studied: Delta32 CCR5, G(-2459)A CCR5, G190A CCR2, G744A CX3CR1 and C838T CX3CR1. They were screened in a cohort of 168 HIV-1 positive adults [HIV(+) group] and 151 newborns [control group] from northwestern Poland. PCR-RFLP was performed to screen for the variants (except for Delta32 CCR5 polymorphism, where PCR fragment size was sufficient to identify the alleles) and then electrophoresed on agarose gel to determine fragment size. Distribution of genotypes and alleles was not significantly different between the groups except for the CCR5 polymorphisms, with the Delta32 allele and the (-2459)A CCR5 allele more frequent among neonates than in the HIV(+) group. No Delta32/Delta32 homozygotes were found in the HIV(+) group, but 16.1 percent were Delta32/wt heterozygotes. In the control group, 1.3 percent; were Delta32/Delta32 homozygotes and 26.0percent were Delta32/wt heterozygotes. Linkage between the chemokine polymorphisms was calculated using the most informative loci for haplotype reconstruction. Haplotypes containing Delta32 CCR5, 190G CCR2 and 744A CX3CR1 were found to be significantly more common in the control group. This suggests an association between these haplotypes and resistance to HIV-1 infection.

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A new multiplex PCR strategy for the simultaneous determination of four genetic polymorphisms affecting HIV-1 disease progression

Cited by 34 publications

References 13 publications

Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Immunogenic and tolerogenic signatures in human immunodeficiency virus (HIV)-infected controllers compared with progressors and a conversion strategy of virus control

Sequence variants of chemokine receptor genes and susceptibility to HIV-1 infection

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A new multiplex PCR strategy for the simultaneous determination of four genetic polymorphisms affecting HIV-1 disease progression

Cited by 34 publications

References 13 publications

Virus-Specific CD8+T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Virus-Specific CD8+T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Immunogenic and tolerogenic signatures in human immunodeficiency virus (HIV)-infected controllers compared with progressors and a conversion strategy of virus control

Sequence variants of chemokine receptor genes and susceptibility to HIV-1 infection

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Product

Resources

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Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype

Virus-Specific CD8⁺T-Cell Responses Better Define HIV Disease Progression than HLA Genotype