Conserved positive selection signals in gp41 across multiple subtypes and difference in selection signals detectable in gp41 sequences sampled during acute and chronic HIV-1 subtype C infection

<p>Abstract</p> <p>Background</p> <p>The high diversity of HIV variants driving the global AIDS epidemic has caused many to doubt whether an effective vaccine against the virus is possible. However, by identifying the selective forces that are driving the ongoing diversification of HIV and characterising their genetic consequences, it may be possible to design vaccines that pre-empt some of the virus' more common evasion tactics. One component of such vaccines might be the envelope protein, gp41. Besides being targeted by both the humoral and cellular arms of the immune system this protein mediates fusion between viral and target cell membranes and is likely to be a primary determinant of HIV transmissibility.</p> <p>Results</p> <p>Using recombination aware analysis tools we compared site specific signals of selection in gp41 sequences from different HIV-1 M subtypes and circulating recombinant forms and identified twelve sites evolving under positive selection across multiple major HIV-1 lineages. To identify evidence of selection operating during transmission our analysis included two matched datasets sampled from patients with acute or chronic subtype C infections. We identified six gp41 sites apparently evolving under different selection pressures during acute and chronic HIV-1 infections. These sites mostly fell within functional gp41 domains, with one site located within the epitope recognised by the broadly neutralizing antibody, 4E10.</p> <p>Conclusion</p> <p>Whereas these six sites are potentially determinants of fitness and are therefore good candidate targets for subtype-C specific vaccines, the twelve sites evolving under diversifying selection across multiple subtypes might make good candidate targets for broadly protective vaccines.</p

Features of recently transmitted HIV-1 clade C viruses that impact antibody recognition : implications for active and passive immunization.

CAPRISA, 2016.Abstract available in PDF file

Differences in HIV-1 neutralization breadth in two geographically distinct cohorts in Africa.

CAPRISA, 2015.Abstract available in pdf

Maximum likelihood phylogenetic analysis of southern African clade C acute/early envelope nucleotide sequences (n = 200) (Table 1).

<p>Branches are colored according to country/region. Bootstrap values > 80% of 100 resampled replicates are illustrated as filled circles on nodes. South African samples from Soweto/Johannesburg (Gauteng province), Cape Town (Western Cape Province), and KwaZulu-Natal are highlighted in light green, red, and blue respectively, sequences from other locations in South Africa are shown in grey. Clades that were from the same geographic region are highlighted. Only one of these regional grouping (>2 sequence clusters) had strong bootstrap support (4 sequences from Tanzania, highlighted in orange).</p

Comparison of neutralization susceptibility and viral characteristics of pre- and post-seroconversion clade C panel viruses.

<p>Pseudotyped clade C viruses were partitioned into three infection stage groups according to their sequential gain of HIV-1 specific antibody responses as a marker of time from infection; pre-seroconversion (Ab-) (n = 58) indicated in blue, indeterminate (Ab-/+) (n = 26) indicated in grey and post-seroconversion (Ab+) (n = 55) indicated in red. Each point represents the geometric mean of viruses tested for neutralization sensitivity using a panel of 30 South African sera. (A) Serum neutralization potency as measured by GMT, over the 30 sera/plasma included in this study; sera below the threshold of detection (dilutions of 1:20 was the limit tested) were given the value of 10. The two subgroups evident among the pre-seroconversion viruses did not cluster phylogenetically. One-sided Wilcoxon rank sum test employed with median values shown in red and interquartile ranges in black (B) Neutralization breadth per infection stage against 30 clade C serum samples measured as the percentage of viruses neutralized at ID₅₀ > 1:20. Three very sensitive viruses, two tier 1A (SO032_A2.8–1, CH0505.w4.3) and one tier 1B, (6644.v2.c33), were excluded from the analysis. (C), (D) V1V2 loop amino acid length variation and glycan density per infection stage. Mann-Whitney two-sided tests used in panel B, C and D, with median values shown in red and interquartile ranges in black. Uncorrected p-values <0.05 are provided, however as four comparisons were made, only p-values <0.015 should be considered significant.</p

K-means (n = 4) neutralization clustering of clade C Env pseudoviruses for tier classification (n = 200).

<p>Neutralization sensitivity was assessed by assaying pseudoviruses against 30 sera from chronically infected individuals. Individual viruses are indicated to the right of the heatmap with individual serum listed at the bottom. Viruses are ranked according to respective sensitivities and categorized into tiers as determined by k-means clustering. Neutralization sensitivity (log₁₀ ID₅₀ titer) is denoted by color key provided in the inset. Viral isolates clustering, together with >95% probability, was performed to assess the stability of the Env pseudovirus clusters based on a random-with replacement resampling of the sera (rows), or to assess the stability of the serological clusters based on resampling of the viruses (columns), and were boxed on the heatmap indicating categorization into their respective tiers. Bootstrap support for clusters was determined by resampling the data sets from the individual serum and re-evaluating the k-means clusters 10,000 times giving an indication of how many times each virus was a member of the originally assigned tier classification in the resampled datasets. Env pseudoviruses that clustered with the assigned tier are shown in the column on the left as blue for tier 1A (very sensitive), yellow for tier 1B, magenta for tier 2 (intermediate), and black for tier 3 (very resistant). This follows the tier classification of Seaman et al (2010) [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005742#ppat.1005742.ref016" target="_blank">16</a>]. Similarly sera potential is illustrated at the top with the most uniformly potent sera indicated as blue, potent sera as yellow, modestly potent as magenta, and least potent as black. Color intensity indicates the probability of falling within a cluster and the degree of blending the frequency of falling into each of the respective tiers, with strong unblended colors, associated with 95% bootstrap support, boxed. The poor resolution of the k- means bootstrapping emphasizes that the sensitivity of Envs is a continuum, and will be dependent on the specific sera or antibodies used for the evaluation.</p

bnAb VRC01 coverage against clade C pseudoviruses viruses.

<p>Potency-breadth curves are presented for both IC₅₀ in red and IC₈₀ titers in blue. Dashed vertical lines indicate concentrations of 50 ug/ml, 10 ug/ml and 1 ug/ml tested. Assays using starting concentrations of 50μg/ml were used for VRC01 data reported here.</p

Characteristics of the acute/early clade C panel virus donors by infection stage and country/region of origin.

<p>Characteristics of the acute/early clade C panel virus donors by infection stage and country/region of origin.</p

Geometric mean titer and tier 1A and 1B classification (n = 200).

<p>(A) Viruses are rank ordered according to neutralization sensitivity to 30 clade C chronic infection serum samples, from the least sensitive to the most sensitive along the x-axis by average log₁₀ GMTs. Two viruses were classified as highly sensitive tier 1A and an additional 17 as above-average sensitive tier 1B. A previously determined cut off ID₅₀ = 200, was used to distinguish between tier 1A and tier 1B is indicated on the graph, with tier 1B classified viruses above this cut off colored in red and those below in orange [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005742#ppat.1005742.ref016" target="_blank">16</a>]. Twelve pseudoviruses classified by both Seaman et al., (2010) and this study were found to be discrepant (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005742#ppat.1005742.s009" target="_blank">S1 Table</a>): Du156.12 consistently falls near the boundary of the tier 2 and tier 1B, was classified as tier 1B here however was previously classified as tier 2; and ZM197M and SM109F were classified as tier 2 here, however were previously classified as 1B. (B) Maximum likelihood phylogenetic analysis of southern African clade C acute/early envelope nucleotide sequences (n = 200) with branches colored according to tier. Bootstrap values > 80% of 100 resampled replicates are illustrated as filled circles on nodes.</p