The extent to which virus-specific CD8 T cell responses contribute to control of viral load and prevention of disease progression in HIV infection has long been a subject of conflicting data and controversy. Early tests of the ability of CD8 T cells to kill target cells expressing HIV antigens were misleading because they relied on extended culture with IL-2, a process that turned out to have restorative properties on otherwise dysfunctional cells. The advent of class I MHC tetramers, which facilitate the counting of CD8 T cells specific for individual HIV epitopes, was an advance but also initially caused confusion: an early paper using tetramers reported an inverse correlation between CD8 T cells specific for epitopes from HIV Gag and Pol and viral load, but a subsequent more comprehensive analysis of CD8 T cell responses to all viral proteins found the opposite: a positive correlation with viral load.
To some extent, these disparate findings have been reconciled by data indicating that CD8 T cell targeting of certain epitopes, especially those in Gag, may be particularly important in controlling HIV. But it has also become clear that cell function is critical, and a number of research groups have developed assays designed to directly measure the ability of CD8 T cells to kill HIV-infected CD4 T cells in vitro. For example, the laboratory of Mark Connors at NIAID has pioneered this approach and demonstrated that elite controllers display functionally superior HIV-specific CD8 T cells compared to individuals with progressive infection.
In new a study published in the current issue of the Journal of Infectious Diseases, Lucy Dorrell and colleagues use an assay developed in their lab to investigate the relationship between the killing function of HIV-specific CD8 T cells and the rate of decline in peripheral blood CD4 T cell counts. The researchers first established that the CD8 T cell antiviral activity they were measuring was specific: at a CD8:CD4 ratio of 2:1 in the in vitro assay, the median inhibition was 78% in HIV-positive individuals compared to 0% in HIV-negative controls. The activity was only seen when the CD4 T cell targets shared a class I HLA allele with the study participant from whom the CD8 T cells were obtained.
In an initial retrospective analysis, the slopes of CD4 T cell decline were determined in a group of 30 chronically infected individuals using a median of nine counts obtained over the course of 4.5 years. For individuals with a CD8 T cell inhibition value of 90%, the mean rate of CD4 T cell loss was 20 cells/mL per year (assuming a baseline count of 500 cells), whereas for an inhibition value of 50% the mean rate of loss was 85 cells/mL per year. The relationship between CD8 T cell antiviral activity and CD4 T cell decline was highly statistically significant (p < 0.0001). Notably, there was no correlation between the inhibitory capacity of the CD8 T cells and the magnitude of interferon gamma-producing HIV-specific T cell responses targeting Gag, other proteins, or the entire virus proteome.
The researchers also studied a cohort of 20 individuals recruited during acute HIV infection in China (in the Beijing PRIMO study) followed prospectively. CD4 T cell slopes were calculated from a median of 13.5 measurements over an average of around 2.5 years of follow-up. The results showed that the antiviral activity of CD8 T cells in early infection was predictive of the rate of CD4 T cell loss over time. The relationship was strongest when reduced ratios of CD8:CD4 T cells were used (either 1:10 or 1:100), leading the authors to state that: “titration of CD8 T cells to sub-optimal levels improved the capacity of the assay to distinguish fast and slow progressors.” The percentage of CD8 T cell inhibition was calculated to explain 73% of the variance in CD4 slopes at a CD8:CD4 ratio of 1:10. The researchers found that adding HIV viral load set point into a model together with percentage of CD8 T cell inhibition improved their ability to predict CD4 T decline: at a CD8:CD4 ratio of 1:10, the combined measures explained a remarkable 91% of the variance in the slope of CD4 T cell loss.
These findings make an important contribution to the understanding of differences in individual rates of CD4 T cell loss and disease progression among HIV-positive individuals. The researchers note, however, that their study has some limitations: their model assumes constant CD8 T cell antiviral activity, when in reality it is likely that this function becomes impaired over the course of infection. The retrospective nature of the analysis of the chronic infection cohort also makes discerning cause and effect difficult, and the prospective cohort only had limited samples from early infection available. To address these limitations, they recommend the conduct of a longitudinal study assessing CD8 T cell antiviral activity repeatedly from the earliest stages of infection onwards.
J Infect Dis. 2012 Jul 3. [Epub ahead of print]
Yang H, Wu H, Hancock G, Clutton G, Sande N, Xu X, Yan H, Huang X, Angus B, Kuldanek K, Fidler S, Denny TN, Birks J, McMichael A, Dorrell L.
Oxford NIHR Biomedical Research Centre.
Background. Rare human immunodeficiency virus type 1 (HIV-1)-infected individuals who maintain control of viremia without therapy show potent CD8+ T-cell-mediated suppression of viral replication in vitro. Whether this is a determinant of the rate of disease progression in viremic individuals is unknown.
Methods. We measured CD8+ T-cell-mediated inhibition of a heterologous HIV-1 isolate in 50 HIV-1-seropositive adults with diverse progression rates. Linear mixed models were used to determine whether CD8+ T-cell function could explain variation in the rate of CD4+ T-cell decline.
Results. There was a significant interaction between CD8+ T-cell antiviral activity in vitro and the rate of CD4+ T-cell decline in chronically infected individuals (P < .0001). In a second prospective analysis of recently infected subjects followed for up to 3 years, CD8+ T-cell antiviral activity strongly predicted subsequent CD4+ T-cell decline (P < .0001) and explained up to 73% of the interindividual variation in the CD4+ T-cell slope. In addition, it was inversely associated with viral load set point (r = -0.68 and P = .002).
Conclusions. The antiviral inhibitory capacity of CD8+ T cells is highly predictive of CD4+ T-cell loss in early HIV-1 infection. It has potential as a benchmark of effective immunity in vaccine evaluation.