Limits...
Reduction of the HIV-1 reservoir in resting CD4+ T-lymphocytes by high dosage intravenous immunoglobulin treatment: a proof-of-concept study.

Lindkvist A, Edén A, Norström MM, Gonzalez VD, Nilsson S, Svennerholm B, Karlsson AC, Sandberg JK, Sönnerborg A, Gisslén M - AIDS Res Ther (2009)

Bottom Line: As yet, no approach to reduce this viral reservoir has proven effective.The latent HIV-1 pool decreased with in median 68% after IVIG was added to effective ART.We propose that IVIG should be further evaluated as an adjuvant to effective ART.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Laboratory Medicine, Division of Clinical Microbiology, Karolinska Institute, Stockholm, Sweden. annica.lindkvist@ki.se

ABSTRACT

Background: The latency of HIV-1 in resting CD4+ T-lymphocytes constitutes a major obstacle for the eradication of virus in patients on antiretroviral therapy (ART). As yet, no approach to reduce this viral reservoir has proven effective.

Methods: Nine subjects on effective ART were included in the study and treated with high dosage intravenous immunoglobulin (IVIG) for five consecutive days. Seven of those had detectable levels of replication-competent virus in the latent reservoir and were thus possible to evaluate. Highly purified resting memory CD4+ T-cells were activated and cells containing replication-competent HIV-1 were quantified. HIV-1 from plasma and activated memory CD4+ T-cells were compared with single genome sequencing (SGS) of the gag region. T-lymphocyte activation markers and serum interleukins were measured.

Results: The latent HIV-1 pool decreased with in median 68% after IVIG was added to effective ART. The reservoir decreased in five, whereas no decrease was found in two subjects with detectable virus. Plasma HIV-1 RNA >or= 2 copies/mL was detected in five of seven subjects at baseline, but in only one at follow-up after 8-12 weeks. The decrease of the latent HIV-1 pool and the residual plasma viremia was preceded by a transitory low-level increase in plasma HIV-1 RNA and serum interleukin 7 (IL-7) levels, and followed by an expansion of T regulatory cells. The magnitude of the viral increase in plasma correlated to the size of the latent HIV-1 pool and SGS of the gag region showed that viral clones from plasma clustered together with virus from activated memory T-cells, pointing to the latent reservoir as the source of HIV-1 RNA in plasma.

Conclusion: The findings from this uncontrolled proof-of-concept study suggest that the reservoir became accessible by IVIG treatment through activation of HIV-1 gene expression in latently-infected resting CD4+ T-cells. We propose that IVIG should be further evaluated as an adjuvant to effective ART.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic analysis of HIV-1 sequences from the latent reservoir and plasma. Phylogenetic trees of aligned sequences obtained by SGS from patient 2 (A) and 7 (B) were determined using the neighbour-joining distance method. From patient 2, a total of 15 SGS were obtained from the plasma sample 16 days after initiation of IVIG treatment (2.PLd16); 23 SGS from the supernatant at baseline (2.BL); and 38 SGS from the supernatant of activated T-cells 85 days after initiation of IVIG treatment (2.d85). From patient 7, 10 SGS were obtained from the plasma sample 15 days after initiation of IVIG (7.PLd15); 26 SGS from the supernatant at baseline (7.BL); and 23 SGS from the supernatant 57 days after initiation of IVIG treatment (7.d57). A close relationship was found between the HIV-1 RNA from plasma-activated T-cells and the SGS from the T-cell culture. This correlation falls within the cluster of plasma sequences, implying that activation of the latent reservoir can be the source of plasma HIV-1 RNA found during IVIG treatment. Bootstrap values > 70 are indicated in the trees.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2713257&req=5

Figure 3: Phylogenetic analysis of HIV-1 sequences from the latent reservoir and plasma. Phylogenetic trees of aligned sequences obtained by SGS from patient 2 (A) and 7 (B) were determined using the neighbour-joining distance method. From patient 2, a total of 15 SGS were obtained from the plasma sample 16 days after initiation of IVIG treatment (2.PLd16); 23 SGS from the supernatant at baseline (2.BL); and 38 SGS from the supernatant of activated T-cells 85 days after initiation of IVIG treatment (2.d85). From patient 7, 10 SGS were obtained from the plasma sample 15 days after initiation of IVIG (7.PLd15); 26 SGS from the supernatant at baseline (7.BL); and 23 SGS from the supernatant 57 days after initiation of IVIG treatment (7.d57). A close relationship was found between the HIV-1 RNA from plasma-activated T-cells and the SGS from the T-cell culture. This correlation falls within the cluster of plasma sequences, implying that activation of the latent reservoir can be the source of plasma HIV-1 RNA found during IVIG treatment. Bootstrap values > 70 are indicated in the trees.

Mentions: The latent HIV-1 pool decreased with a median of 68% after IVIG treatment (Table 2). When the individual subjects were scrutinized, a decrease in the latent reservoir was found in five (Figure 1a). Of the two subjects who experienced no decrease in the reservoir, one had a low pre-treatment viral load in resting cells, and in the other replication-competent virus went from undetectable to just detectable (0.5 IUPM) (Figure 1b). The five subjects with decrease of their reservoirs had a similar pattern of detectable HIV-1 RNA in plasma (6 to 27 copies/mL) two weeks after initiation of IVIG (Figure 1a). A close correlation was found between the maximal plasma viral load and levels of IUPM cells before IVIG treatment, rs = 0.86, p = 0.0045 (Figure 2). We also compared virus obtained from plasma and activated memory T-cells, using SGS of gag in subjects 2 and 7. Both had sufficiently high plasma viral loads to lead us to believe that sequencing would be possible. Plasma sequences were derived 15 days (subject 7) and 16 days (subject 2) after initiation of IVIG, when the plasma viral load was 19 and 27 copies/mL, respectively. In both, viruses from plasma and the T-cell reservoir were closely related, and clustered together in a distinct branch (bootstrap value > 90) in the phylogenetic trees (Figure 3). The SGS obtained from activated T-cells probably reflects an oligoclonal expansion in the culture of the most replication-competent HIV-1 in the resting T-cell population.


Reduction of the HIV-1 reservoir in resting CD4+ T-lymphocytes by high dosage intravenous immunoglobulin treatment: a proof-of-concept study.

Lindkvist A, Edén A, Norström MM, Gonzalez VD, Nilsson S, Svennerholm B, Karlsson AC, Sandberg JK, Sönnerborg A, Gisslén M - AIDS Res Ther (2009)

Phylogenetic analysis of HIV-1 sequences from the latent reservoir and plasma. Phylogenetic trees of aligned sequences obtained by SGS from patient 2 (A) and 7 (B) were determined using the neighbour-joining distance method. From patient 2, a total of 15 SGS were obtained from the plasma sample 16 days after initiation of IVIG treatment (2.PLd16); 23 SGS from the supernatant at baseline (2.BL); and 38 SGS from the supernatant of activated T-cells 85 days after initiation of IVIG treatment (2.d85). From patient 7, 10 SGS were obtained from the plasma sample 15 days after initiation of IVIG (7.PLd15); 26 SGS from the supernatant at baseline (7.BL); and 23 SGS from the supernatant 57 days after initiation of IVIG treatment (7.d57). A close relationship was found between the HIV-1 RNA from plasma-activated T-cells and the SGS from the T-cell culture. This correlation falls within the cluster of plasma sequences, implying that activation of the latent reservoir can be the source of plasma HIV-1 RNA found during IVIG treatment. Bootstrap values > 70 are indicated in the trees.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2713257&req=5

Figure 3: Phylogenetic analysis of HIV-1 sequences from the latent reservoir and plasma. Phylogenetic trees of aligned sequences obtained by SGS from patient 2 (A) and 7 (B) were determined using the neighbour-joining distance method. From patient 2, a total of 15 SGS were obtained from the plasma sample 16 days after initiation of IVIG treatment (2.PLd16); 23 SGS from the supernatant at baseline (2.BL); and 38 SGS from the supernatant of activated T-cells 85 days after initiation of IVIG treatment (2.d85). From patient 7, 10 SGS were obtained from the plasma sample 15 days after initiation of IVIG (7.PLd15); 26 SGS from the supernatant at baseline (7.BL); and 23 SGS from the supernatant 57 days after initiation of IVIG treatment (7.d57). A close relationship was found between the HIV-1 RNA from plasma-activated T-cells and the SGS from the T-cell culture. This correlation falls within the cluster of plasma sequences, implying that activation of the latent reservoir can be the source of plasma HIV-1 RNA found during IVIG treatment. Bootstrap values > 70 are indicated in the trees.
Mentions: The latent HIV-1 pool decreased with a median of 68% after IVIG treatment (Table 2). When the individual subjects were scrutinized, a decrease in the latent reservoir was found in five (Figure 1a). Of the two subjects who experienced no decrease in the reservoir, one had a low pre-treatment viral load in resting cells, and in the other replication-competent virus went from undetectable to just detectable (0.5 IUPM) (Figure 1b). The five subjects with decrease of their reservoirs had a similar pattern of detectable HIV-1 RNA in plasma (6 to 27 copies/mL) two weeks after initiation of IVIG (Figure 1a). A close correlation was found between the maximal plasma viral load and levels of IUPM cells before IVIG treatment, rs = 0.86, p = 0.0045 (Figure 2). We also compared virus obtained from plasma and activated memory T-cells, using SGS of gag in subjects 2 and 7. Both had sufficiently high plasma viral loads to lead us to believe that sequencing would be possible. Plasma sequences were derived 15 days (subject 7) and 16 days (subject 2) after initiation of IVIG, when the plasma viral load was 19 and 27 copies/mL, respectively. In both, viruses from plasma and the T-cell reservoir were closely related, and clustered together in a distinct branch (bootstrap value > 90) in the phylogenetic trees (Figure 3). The SGS obtained from activated T-cells probably reflects an oligoclonal expansion in the culture of the most replication-competent HIV-1 in the resting T-cell population.

Bottom Line: As yet, no approach to reduce this viral reservoir has proven effective.The latent HIV-1 pool decreased with in median 68% after IVIG was added to effective ART.We propose that IVIG should be further evaluated as an adjuvant to effective ART.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Laboratory Medicine, Division of Clinical Microbiology, Karolinska Institute, Stockholm, Sweden. annica.lindkvist@ki.se

ABSTRACT

Background: The latency of HIV-1 in resting CD4+ T-lymphocytes constitutes a major obstacle for the eradication of virus in patients on antiretroviral therapy (ART). As yet, no approach to reduce this viral reservoir has proven effective.

Methods: Nine subjects on effective ART were included in the study and treated with high dosage intravenous immunoglobulin (IVIG) for five consecutive days. Seven of those had detectable levels of replication-competent virus in the latent reservoir and were thus possible to evaluate. Highly purified resting memory CD4+ T-cells were activated and cells containing replication-competent HIV-1 were quantified. HIV-1 from plasma and activated memory CD4+ T-cells were compared with single genome sequencing (SGS) of the gag region. T-lymphocyte activation markers and serum interleukins were measured.

Results: The latent HIV-1 pool decreased with in median 68% after IVIG was added to effective ART. The reservoir decreased in five, whereas no decrease was found in two subjects with detectable virus. Plasma HIV-1 RNA >or= 2 copies/mL was detected in five of seven subjects at baseline, but in only one at follow-up after 8-12 weeks. The decrease of the latent HIV-1 pool and the residual plasma viremia was preceded by a transitory low-level increase in plasma HIV-1 RNA and serum interleukin 7 (IL-7) levels, and followed by an expansion of T regulatory cells. The magnitude of the viral increase in plasma correlated to the size of the latent HIV-1 pool and SGS of the gag region showed that viral clones from plasma clustered together with virus from activated memory T-cells, pointing to the latent reservoir as the source of HIV-1 RNA in plasma.

Conclusion: The findings from this uncontrolled proof-of-concept study suggest that the reservoir became accessible by IVIG treatment through activation of HIV-1 gene expression in latently-infected resting CD4+ T-cells. We propose that IVIG should be further evaluated as an adjuvant to effective ART.

No MeSH data available.


Related in: MedlinePlus