Isolation frequency and growth properties of HIV‐variants: Multiple simultaneous variants in a patient demonstrated by molecular cloning

Briesen, Hagen von; Becker, Walter; Henco, Karsten; Helm, E. B.; Gelderblom, Hans R.; Brede, H. D.; Rübsamen‐Waigmann, Helga

doi:10.1002/jmv.1890230107

Cited by 95 publications

(43 citation statements)

References 28 publications

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“…This suggests that at least two qualitatively different types of R5 virus (or host responses to R5 virus) can occur in vivo. This idea parallels early observations (1,13,65) and a recent study (49) suggesting that NSI virus late-growth characteristics are different from those of the NSI virus that tends to initiate infection. A persistent R5 population may have evolved more efficient binding to the CCR5 receptor, as has been shown to occur with in vitro-passaged virus under pressure from a small-molecule CCR5 inhibitor (60), making it better able to exploit diminishing resources.…”

Section: Discussionsupporting

confidence: 88%

Improved Coreceptor Usage Prediction and GenotypicMonitoring of R5-to-X4 Transition by Motif Analysis of HumanImmunodeficiency Virus Type 1 env V3 LoopSequences

Jensen

Wout

et al. 2003

J Virol

391

423

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Early in infection, human immunodeficiency virus type 1 (HIV-1) generally uses the CCR5 chemokine receptor (along with CD4) for cellular entry. In many HIV-1-infected individuals, viral genotypic changes arise that allow the virus to use CXCR4 (either in addition to CCR5 or alone) as an entry coreceptor. This switch has been associated with an acceleration of both CD3؉ T-cell decline and progression to AIDS. While it is well known that the V3 loop of gp120 largely determines coreceptor usage and that positively charged residues in V3 play an important role, the process of genetic change in V3 leading to altered coreceptor usage is not well understood. Further, the methods for biological phenotyping of virus for research or clinical purposes are laborious, depend on sample availability, and present biosafety concerns, so reliable methods for sequencebased "virtual phenotyping" are desirable. We introduce a simple bioinformatic method of scoring V3 amino acid sequences that reliably predicts CXCR4 usage (sensitivity, 84%; specificity, 96%). This score (as determined on the basis of position-specific scoring matrices [PSSM]) can be interpreted as revealing a propensity to use CXCR4 as follows: known R5 viruses had low scores, R5X4 viruses had intermediate scores, and X4 viruses had high scores. Application of the PSSM scoring method to reconstructed virus phylogenies of 11 longitudinally sampled individuals revealed that the development of X4 viruses was generally gradual and involved the accumulation of multiple amino acid changes in V3. We found that X4 viruses were lost in two ways: by the dying off of an established X4 lineage or by mutation back to low-scoring V3 loops.Early studies of the biological properties of human immunodeficiency virus type 1 (HIV-1) found that virus isolates could be placed into as few as two phenotypic categories (defined in vitro as either non-syncytium-inducing [NSI] or syncytium-inducing [SI]) in certain CD4 ϩ T-cell lines. These phenotypes were often found to be associated with differences in growth properties and cytopathicity on peripheral blood mononuclear cells (PBMC) (1,14,46) and in cellular host range (3,48). Ultimately, the difference between the NSI and SI phenotypes was shown to be due largely to the differential use of chemokine receptors as coreceptors for viral entry: NSI viruses predominantly use CCR5, while SI viruses can use CCR5 and CXCR4 or CXCR4 exclusively (2, 29, 31, 52, 54). Results determined on the basis of SI phenotype and/or coreceptor usage typing showed that although HIV-1 present at primary infections used the CCR5 coreceptor (R5 virus) ϳ90% of the time (63, 67, 68), a substantial proportion of individuals eventually developed virus that used the CXCR4 coreceptor (X4 virus). These X4/SI viruses are associated with accelerated CD4 decline and more rapid progression of HIV-1 disease (8,28,33,43,47). Little is known about the mechanisms by which these viruses come to predominate among the HIV-1 strains present in an infected person. For example, it is no...

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Section: Discussionsupporting

confidence: 88%

Improved Coreceptor Usage Prediction and GenotypicMonitoring of R5-to-X4 Transition by Motif Analysis of HumanImmunodeficiency Virus Type 1 env V3 LoopSequences

Jensen

Wout

et al. 2003

J Virol

391

423

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“…We performed repeated isolations from patients in each category at 3-month intervals, each cytopathic pattern (NS, SI, CI) and NC pattern was reproducible for a particular patient. The heterogeneity among HIV isolates with regard to cytopathic effect that we observed has been reported from several laboratories (9,11,14,16,24). Dahl et al have used a lymphoblastoid cell line immortalized by Epstein-Barr virus to distinguish the capacity of various HIV isolates to induce cytolysis (9).…”

Section: Mechanism Of Cytolysis Of P4 Cells In the Presence Of Hiv-1 supporting

confidence: 61%

“…The physiopathology of HIV infection is not yet well understood but differences in the pathogenicity of HIV strains or the development of a more virulent strain in the course of infection may play a role in the outcome of HIV infection (1,2,6,9,11,21,24). Isolates of HIV may differ in biological properties such as replication rate, host range, and syncytium-inducing (SI) capacity (2,6,11,21,24). Fenyo et al reported that efficiently replicating HIV isolates causes syncytium formation in vitro, and poorly replicating HIV isolates show syncytium formation and single-cell killing simultaneously or cell killing only (13).…”

mentioning

confidence: 99%

A Microassay for Determination of the Cytopathogenicity of Human Immunodeficiency Virus Type‐1 Isolates

Benyoucef

Hober

Shen

et al. 1996

Microbiology and Immunology

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Correlations between the in vitro biological properties of HIV strains isolated from patients and the prognosis of their disease have been reported. We developed a technique to study the phenotype of HIV strains isolated from patients. We used the P4 cell line, derived from HeLa cells, which has been transfected with receptor CD4 gene. HIV laboratory strain (HIVLAI) and peripheral blood leukocytes (PBLs) from donors infected with HIVLAI induce syncytium in P4 cell cultures in vitro. The presence of reporter gene (LacZ gene) under the control of the HIV‐1 long terminal repeat (LTR) in these cells allows colorimetric visualization of syncytia in the cytoplasm using a β‐galactosidase (βgal) assay in the presence of X‐gal. We cocultivated 1 × 106 patient PBLs with 2 × 106 normal PHA‐activated normal PBLs for 4 days in the presence of IL‐2 in 24‐well plates. Half of the medium was replaced twice a week and PHA‐activated normal PBLs were added every 7 days. HIV‐1 was isolated from cocultured PBLs of 18 patients with advanced‐stage HIV infection as assessed by the production of HIV p24 detected with a commercially available HIV‐1 p24 ELISA. Supernatant and 105 cells were collected twice a week from cocultured PBLs and were added to P4 cells in 96‐well microtiter plates. The cultures were observed every day for 3 days and then the βgal assay was performed. We did not observe any effect with cells and supernatant from 8 patients, harvested from cultures incubated for as long as 28 days. The phenotype of these isolates was called NC (noncytopathic). In cells from 2 patients, we obtained blue multinucleated giant cells; the phenotype of these strains was called SI (syncytium inducing). In cultures from 8 other patients, we obtained the death of P4 cells without syncytium formation, and the phenotype of these strains was called CI (cell‐killing inducing). In every case, the cytopathic effect of HIV‐1 isolates could be detected with cocultured PBLs collected as early as day 4 of culture. Cocultured PBLs from 13 healthy controls did not alter the P4 cells. We displayed the replication of CI strains of HIV‐1, but not the one of NC strains in P4 cell line. Our micromethod allowed the detection of cytopathic effects of HIV isolates. Further investigations should define the clinical applications of this method.

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“…Changes in the biological behaviour of HIV-1 during the course of infection have been reported by a number of workers (Asjo et al, 1986;von Briesen et al, 1987;Evans et al, 1987;Cheng-Mayer et al, 1988;Tersmette et aI., 1988). These include the onset of syncytium-inducing (SI) capacity, enhancement of the replication level, and alteration in cytotropism from growth in monocytes to the ability to grow in lymphocytes.…”

Section: Other Factors In Hiv Disease Progressionmentioning

confidence: 76%

Herpesviruses as Co-Factors of HIV Disease: Case not Proven

Jeffries

1995

Antivir Chem Chemother

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SummaryFactors other than human immunodeficiency virus (HIV) may be important in determining disease progression in AIDS. The concept that viruses may act as co-factors in HIV pathogenesis has been based on in vitro studies of potential molecular or biological interactions between HIV and other viruses, some epidemiological evidence for an adverse effect of cytomegalovirus on progression to AIDS, and the apparent survival advantage conferred by aciclovir prophylaxis in people with AIDS.It is possible that molecular or cellular interaction between herpesviruses or other infectious agents and HIV are necessary to encourage the virus into maximal production or to amplify pre-existing replication. However, there is no evidence as yet that this occurs, and studies that claim to show such an effect cannot be convincing unless they take account of the significant associations of progression with virus phenotype, host response, age and genetic factors. This paper was presented as the case against in a debate on 'Herpesviruses as co-factors of HIV disease'. The case for was made by Paul Griffiths (see page 17).

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Isolation frequency and growth properties of HIV‐variants: Multiple simultaneous variants in a patient demonstrated by molecular cloning

Cited by 95 publications

References 28 publications

Improved Coreceptor Usage Prediction and GenotypicMonitoring of R5-to-X4 Transition by Motif Analysis of HumanImmunodeficiency Virus Type 1 env V3 LoopSequences

Improved Coreceptor Usage Prediction and GenotypicMonitoring of R5-to-X4 Transition by Motif Analysis of HumanImmunodeficiency Virus Type 1 env V3 LoopSequences

A Microassay for Determination of the Cytopathogenicity of Human Immunodeficiency Virus Type‐1 Isolates

Herpesviruses as Co-Factors of HIV Disease: Case not Proven

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