Antibodies to the CD4-binding site of HIV-1 gp120 suppress gp120-specific CD4 T cell response while enhancing antibody response

<p>Abstract</p> <p>Background</p> <p>The binding of Abs to the CD4-binding site (CD4bs) of HIV-1 envelope gp120 has been shown to obstruct the processing and generation of helper epitopes from this antigen, resulting in poor presentation of various gp120 epitopes by MHC class II to CD4 T cells. However, the physiologic significance of these inhibitory anti-CD4bs Abs <it>in vivo </it>has remained unclear. In this study, we evaluated the immunologic effects of anti-CD4bs Abs <it>in vivo </it>using a murine model.</p> <p>Results</p> <p>Animals were immunized with recombinant envelope proteins with or without CD4-binding activity (designated CD4bs⁺Env and CD4bs^–Env, respectively). As expected, anti-CD4bs Abs were generated only after immunization with CD4bs+ Env and not with CD4bs^–Env. The presence of anti-CD4bs Abs was associated with lower levels of envelope-specific lymphoproliferation in animals immunized with CD4bs+ Env. To further determine the specific role of the anti-CD4bs Abs, we immunized mice with gp120 in the presence of an inhibitory anti-CD4bs mAb or a non-inhibitory anti-gp120 mAb. The data show that the presence of anti-CD4bs mAb reduced CD4 T cell responses to gp120. However, we also detected significantly higher titers of anti-gp120 Abs following immunization with gp120 and the anti-CD4bs mAb.</p> <p>Conclusion</p> <p>Anti-CD4bs Abs can exert discordant effects on the gp120-specific CD4 T cell and Ab responses <it>in vivo</it>, indicating the importance of these particular Abs in influencing both the cellular and the humoral immune responses against HIV-1.</p

Small Tropical Mammals Can Take the Heat: High Upper Limits of Thermoneutrality in a Bornean Treeshrew

Tropical ectotherms are generally believed to be more vulnerable to global heating than temperate species. Currently, however, we have insufficient knowledge of the thermoregulatory physiology of equatorial tropical mammals, particularly of small diurnal mammals, to enable similar predictions. In this study, we measured the resting metabolic rates (via oxygen consumption) of wild-caught lesser treeshrews (Tupaia minor, order Scandentia) over a range of ambient temperatures. We predicted that, similar to other treeshrews, T. minor would exhibit more flexibility in body temperature regulation and a wider thermoneutral zone compared with other small mammals because these thermoregulatory traits provide both energy and water savings at high ambient temperatures. Basal metabolic rate was on average 1.03±0.10 mL O2 h−1 g−1, which is within the range predicted for a 65-g mammal. We calculated the lower critical temperature of the thermoneutral zone at 31.0°C (95% confidence interval: 29.3°–32.7°C), but using metabolic rates alone, we could not determine the upper critical temperature at ambient temperatures as high as 36°C. The thermoregulatory characteristics of lesser treeshrews provide a means of saving energy and water at temperatures well in excess of their current environmental temperatures. Our research highlights the knowledge gaps in our understanding of the energetics of mammals living in high-temperature environments, specifically in the equatorial tropics, and questions the purported lack of variance in the upper critical temperatures of the thermoneutral zone in mammals, emphasizing the importance of further research in the tropics

HIV Envelope gp120 Activates LFA-1 on CD4 T-Lymphocytes and Increases Cell Susceptibility to LFA-1-Targeting Leukotoxin (LtxA)

The cellular adhesion molecule LFA-1 and its ICAM-1 ligand play an important role in promoting HIV-1 infectivity and transmission. These molecules are present on the envelope of HIV-1 virions and are integral components of the HIV virological synapse. However, cellular activation is required to convert LFA-1 to the active conformation that has high affinity binding for ICAM-1. This study evaluates whether such activation can be induced by HIV itself. The data show that HIV-1 gp120 was sufficient to trigger LFA-1 activation in fully quiescent naïve CD4 T cells in a CD4-dependent manner, and these CD4 T cells became more susceptible to killing by LtxA, a bacterial leukotoxin that preferentially targets leukocytes expressing high levels of the active LFA-1. Moreover, virus p24-expressing CD4 T cells in the peripheral blood of HIV-infected subjects were found to have higher levels of surface LFA-1, and LtxA treatment led to significant reduction of the viral DNA burden. These results demonstrate for the first time the ability of HIV to directly induce LFA-1 activation on CD4 T cells. Although LFA-1 activation may enhance HIV infectivity and transmission, it also renders the cells more susceptible to an LFA-1-targeting bacterial toxin, which may be harnessed as a novel therapeutic strategy to deplete virus reservoir in HIV-infected individuals

Modulation of Antibody Responses to the V1V2 and V3 Regions of HIV-1 Envelope by Immune Complex Vaccines

Prophylactic HIV vaccines must elicit antibodies (Abs) against the virus envelope glycoproteins (Env) to effectively prevent HIV infection. We investigated a vaccine platform that utilizes immune complexes made of Env proteins gp120 and monoclonal Abs (mAbs) against different gp120 epitopes. We previously observed alterations in V3 antigenicity upon formation of certain gp120/mAb complexes and demonstrated the ability of these complexes to modulate the elicitation of V3 Ab responses. However, the effects on the V1V2 domain, an important target for Abs that correlate with vaccine-induced protection against HIV, have not been studied, nor have immune complex vaccines made with non-B subtype Env. This study compared subtypes B (JRFL) and CRF_01.AE (A244) Env gp120 proteins in complex with selected gp120-specific mAbs. Allosteric and antigenic changes were detected on these immune complexes, indicating that gp120/mAb interaction induces alterations on the Env surface that may modify the Env immunogenic properties. To evaluate this idea, mice were immunized with gp120/mAb complexes or their uncomplexed gp120 counterparts. The overall serum IgG titers elicited against gp120 were comparable, but a marked skewing toward V1V2 or V3 was evident and dependent on the gp120 strain and the specificity of the mAb used to form the complexes. Compared with uncomplexed gp120JRFL, gp120JRFL complexed with CD4bs or V1V2 mAbs, but not with C2 or V3 mAbs, elicited V3 Abs of greater titers and breadth, and Abs more capable of neutralizing tier 1 virus. Epitope mapping revealed a shift to a more conserved site in the V3 crown. However, the complexes did not enhance V1V2 Ab response, and the elicited V1V2 Abs were not cross-reactive. This profile contrasts with Ab responses to gp120A244/mAb complexes. Notably, gp120A244/mAb complexes induced higher levels of V1V2 Abs with some cross-reactivity, while also stimulating weak or strain-specific V3 Abs. Sera from gp120A244/mAb complex-immunized animals displayed no measurable virus neutralization but did mediate Ab-dependent cellular phagocytosis, albeit at levels similar to that induced by gp120A244 alone. These data indicate the potential utility of immune complexes as vaccines to shape Ab responses toward or away from Env sites of interest

Linking reactivity test outputs to properties of cementitious pastes made with supplementary cementitious materials

Recently developed reactivity tests for supplementary cementitious materials (SCMs) have shown the ability to differentiate between inert and reactive materials. Two promising SCM reactivity tests were used to study ten different SCMs. The first method involves quantifying the heat release, bound water, and calcium hydroxide consumption of SCMs in a simulated pore solution (modified R3 test), and the second method is a lime strength test that quantifies lime mortar strength development. Tests on cementitious pastes with a 20% SCM replacement level were conducted to determine heat release, calcium hydroxide content, bound water, compressive strength, and bulk resistivity for 35 days. Moderate correlations between the modified R3 heat release at 10 days and 1 day and between modified R3 heat release and bound water were shown, which suggests the possibility of reducing test duration and using bound water, instead of the heat release. The calcium hydroxide consumption measured from the modified R3 test correlates strongly with CaO content in the SCM and moderately to paste bulk resistivity at 35 days, demonstrating the fundamental importance of measuring calcium hydroxide consumption. Correlations between modified R3 heat release and bound water and most other paste properties were generally poor, presumably because of the high acceleration in the modified R3 tests. The compressive strength measured from the lime strength test showed moderate to strong correlation to heat release, calcium hydroxide content, bound water, and bulk resistivity of cementitious paste at 1–35 days. The fundamental importance of these reactivity tests is therefore clearly validated