Inhibition of HIV-1 infection by monoclonal antibodies to carbohydrates of Schistosoma mansoni

Patients infected with HIV-1 develop a potent humoral immune response against the virus, but HIV-1 primary isolates are remarkably resistant to neutralizing antibodies. Considering that the envelope glycoprotein of HIV-1 (gp120/41) is heavily glycosylated, we investigated whether anti-carbohydrate antibodies could inhibit HIV-1 infection in vitro. We studied the neutralizing activity of three monoclonal antibodies (mAbs) raised to carbohydrates of Schistosoma mansoni, against seven primary isolates of HIV-1. Assays were performed infecting peripheral blood mononuclear cells from normal donors with viral isolates previously treated with mAbs. Viral strains used were tropic for the coreceptors CCR5, CXCR4, and dual-tropic ones. We found that the anti-glycan mAbs vigorously inhibited HIV-1 infection, regardless of the preferential coreceptor usage of the isolate, in a dose-response manner. Importantly, five isolates were resistant to neutralization by two HIV-1 antibody-positive human sera endowed with potent anti-HIV-1 inhibitory activity. Our findings suggest that carbohydrates of the HIV-1 viral envelope may be a target of an effective humoral immune response elicited by vaccination.The first two authors contributed equally to this work     

Role of Humoral Immunity in Host Defense Against HIV

Individuals infected with HIV-1 and nearly everyone vaccinated with HIV-1 vaccines will, in time, generate antibodies against viral proteins. These antibodies do not resolve natural infection, and vaccine candidates that successfully stimulate the production of high titers of neutralizing antibodies have failed to protect against infection. In spite of this, antibodies continue to be a focus of vaccine research. One reason for the continued interest in antibodies is the failure of a vaccine engineered to generate cell-mediated immunity against HIV. Successful protective immunity against most intracellular pathogens involves several arms of the immune response. A successful vaccine should also stimulate both protective cell-mediated immunity and specific antibody. Efforts should be directed toward making a vaccine that will stimulate the production of 1) more antibody, 2) more broadly cross-reactive neutralizing antibody (broadly neutralizing antibodies), and 3) antibody with a particular functional activity (antibody-dependent cell-mediated cytotoxicity; catalytic antibodies).     

Neutralizing antibodies mechanism of neutralization and protective activity against HIV-1

The role of the humoral immune response in prevention against HIV-1 infection is still incompletely understood. However, neutralizing antibodies to certain epitopes on HIV-1 envelope glycoproteins inhibit HIV-1 infection in vitro and in vivo. Passive administration of these antibodies by themselvesor in combination completely protected hu-PBL-SCID mice or macaques from intravenous, vaginal, as well as maternal-fetal mucosal transmission. All these studies provide direct experimental evidence that neutralizing antibodies are potentenough to prevent HIV infection, and strongly suggest that neutralizing-antibody-based vaccines could provide effective protection against HIV-1, despite the potent action of CTLs. Some neutralizing epitopes have been defined in vitro and in vivo. Unfortunately, none of the neutralizing-antibody-based candidate vaccines has been demonstrated to induce enough protective activity. Weak antigenicity and immunogenicity of neutralizing epitopes on native or recombinant proteins and other factors made it difficult to induce neutralizing-epitope-specific antibody responses in vivo enough to prevent against primary isolates. Recent studies indicated that HIV-1 variations resulted in escape from neutralization or the CTL responses, which may be the principal challenge for HIV-1 prevention. Epitope vaccine as a new strategy activating both arms of the immune system, namely, using the “principal neutralizing epitopes” and the CTL epitopes in combination, should provide new hope for developing an effective vaccine to halt the HIV-1 epidemic.     

Comparison of human immunodeficiency virus (HIV)-specific infection-enhancing and -inhibiting antibodies in AIDS patients

The humoral immune response of the human host against the human immunodeficiency virus (HIV) type 1 (HIV-1) envelope glycoproteins comprises virus-neutralizing antibodies (NAs), antibody-dependent cellular cytotoxicity-mediating (ADCC) antibodies, and infection-enhancing antibodies (IEAs). Because of their potential significance for the outcome of infection with this virus, we have studied the relative prevalence of NAs, ADCC antibodies, and IEAs in the sera of patients infected with HIV. Our results demonstrate that while >or=60% of serum samples are positive for NAs or ADCC antibodies, 72% of these serum samples mediate the enhancement of infection in the presence of complement. In patients with low CD4 counts, NA and ADCC antibody levels tend to decrease, while IEA levels increase. A significant positive correlation was found only between the presence of ADCC antibodies and the presence of antibodies that neutralized HIV-1 in the presence of complement. These results show that the anti-HIV-1 humoral immune response consists of a mixture of antibodies that may inhibit or enhance HIV infection and whose ratios may vary in different stages of the infection.     

Anti-HIV-1 humoral immune response in Brazilian patients

Data on humoral immune response against Brazilian human immunodeficiency virus type 1 (HIV-1) isolates have been collected and reviewed in reference to data published in the international literature. The results obtained up to now indicate that the Brazilian humoral immune response can be divided into at least three antibody binding serotypes: anti-Bbr, anti -B/F and anti-C/F. No neutralization serotypes have been distinguished up to now, just different degrees of resistance to neutralizing antibody (NAb), similar to data obtained by neutralization analyses of European/North American HIV-1isolates. No correlation between V3 binding antibodies, genotype and HIV-1 NAb could be observed.     

Virological,immunological and host factors in HIV-1 discordant couples in Thailand

The potential factors of resistance to HIV-1 infection were investigated in 23 HIV discordantly infected couples, of each, one partner had HIV infection and the matched spouse was not infected. Both partners of the HIV discordant couples possessed comparable number of CD4+ cells expressing CCR5. Our study demonstrated that resistance to HIV-1 infection was not due to low level of HIV viral load in their infected-matched spouses. In addition, selective biological phenotype of HIV clinical isolates, which is indicative for risk of transmission, could not be determined in this study. However, we have demonstrated that the unknown genetic factor(s), and neutralizing antibody of broad and high activity could be taken into an account for resistance to HIV infection in the HIV discordant couples.     

Predominance of Detrimental Humoral Immune Responses to HIV-1 in AIDS Patients with CD4 Lymphocyte Counts Less Than 400/mm3

The humoral immune response to human immunodeficiency virus type 1 (HIV-1) was studied in 25 AIDS patients with CD4 lymphocyte counts of less than 400/mm³. Humoral immune responses against tissue culture adapted strains of HIV-1, and two limited-passage patient isolates were investigated. Total anti-HIV antibody levels were not significantly different between different individuals. Neutralizing titres against HIV_LAI and HIV_SF2 were 10- to 100-fold higher than against clinical isolates. The complement-mediated, antibody-dependent enhancement of HIV-1 infection titre was high (mean 1:14,000). Antibody-complement mediated cytotoxicity of both HIV_LAI and HIV_SF2 was ineffective using human complement as a complement source. The antibody-dependent, cell-mediated cytotoxicity (ADCC) activity varied against the four isolates with tissue culture-adapted strains being more susceptible than clinical isolates. Finally, an ADCC effector cell function, natural killer or NK activity, was measured for all 25 patients, and NK activity of patients was decreased by nearly 75% compared to uninfected individuals. In summary, beneficial humoral immune responses are low in HIV-1 infected individuals with CD4 counts of less than 400/mm³ if the in vitro assay system is constructed to best mimic the in vivo situation. These results suggest that the lack of functional antibody responses to HIV may play an important role in viral pathogenesis.     

Site-directed primary in vitro immunization: production of HIV-1 neutralizing human monoclonal antibodies from lymphocytes obtained from seronegative donors.

The design of an in vitro immunization system based on a synthetic heterotope immunogen, which was a peptide containing both T- and B-cell epitopes, that elicited a neutralizing, primary human humoral immune response against the human immunodeficiency virus (HIV-1) is reported here. This heterotope construct contained the major neutralizing B-cell epitope, within the V3 region of glycoprotein 120 (gp120), linked to a promiscuous helper T-cell epitope of tetanus toxin. The peptide was used to induce a human humoral in vitro immune response against the V3 region, using lymphocytes obtained from healthy, sero-negative blood donors. The in vitro immunized peripheral blood lymphocytes were Epstein-Barr virus infected and the antibody response to the synthetic peptide was evaluated using a solid-phase ELISA with the recombinant C-terminal fragment of gp120 (pB1, amino acid residues 287-467, derived from the HIV-1 LAI isolate). The heterotope construct yielded a significantly frequency of specifically immunized B cells, in contrast to the control immunizations with individual T and B epitopes, mixtures of these epitopes or no immunogen at all. This approach allowed us to generate human monoclonal antibodies, using lymphocytes derived from sero-negative donors, that cross-neutralized several HIV-1 strains, inhibited syncytia formation as well as prevented spreading of the viral infection from cell to cell. Thus, site-directed in vitro immunization using synthetic heterotopes might prove valuable in the dissection and induction of a protective humoral immune response.     

Mother to child transmission of HIV‐1 in a Thai population: Role of virus characteristics and maternal humoral immune response

The objective of this study was to investigate factors influencing mother to child transmission of HIV‐1 in Thailand, where HIV‐1 CRF01_AE, the major subtype in Southeast Asia, predominates. Samples from 84 HIV‐1 infected, anti‐retroviral treatment‐naïve, non‐breast feeding mothers, 28 who transmitted HIV‐1 to their babies (transmitters) and 56 who did not (non‐transmitters), were studied for maternal humoral immune response and virus characteristics. Maternal humoral immune response was measured by lymphocyte phenotyping; neutralizing antibodies to laboratory HIV‐1 MN strain and two clinical isolates; peptide binding antibody to gp41 and V3 from strains CRF01_AE, B, and MN; autologous antibodies; and quasispecies diversity. Virus characteristics studied were viral load, co‐receptor usage, and viral replication capacity. No significant difference between transmitters and non‐transmitters was found for any parameter of maternal humoral immune response. However, viral load and viral replication capacity were significantly higher in transmitters versus non‐transmitters and were not correlated with each other. This suggests that viral replication capacity may be a transmission factor independent of viral load, which is already well established as a risk factor for transmission of HIV‐1. All except four viral isolates used the CCR5 co‐receptor. This is one of few studies of vertical transmission in a population where HIV‐1 CRF01_AE predominates. The data suggest that in this population the maternal humoral immune response was not important in preventing transmission at parturition, but that virus characteristics were key factors, and that viral replication capacity may contribute to birth‐associated mother to child transmission of HIV‐1. J. Med. Virol. 81:768–778, 2009. © 2009 Wiley‐Liss, Inc.     

A computer-based, image-analysis method to quantify HIV-1 infection in a single-cycle infectious center assay

Rapid and precise quantitation of the infectivity of HIV is important for molecular virologic studies as well as for measuring the activities of antiviral drugs and neutralizing antibodies. In the present study, an indicator cell line and image-analysis software were used to quantify HIV infectivity. Cells of the P4R5 line, which express the receptors for HIV infection as well as beta-galactosidase under the control of the HIV-1 long terminal repeat, were infected either with CXCR4- or CCR5-using viruses, including primary isolates, then stained 2 days later with X-gal to turn infected cells blue. Digital images of monolayers of the infected cells were captured using a high resolution CCD video camera and a macro video zoom lens. A software program was developed to process the images and to count the blue-stained foci of infection. The assay was applied to assess the infectivity of site-directed viral mutants, and to measure the activity of antiviral drugs and neutralizing antibody. The results indicate that the described method allows for the rapid quantitation of infected cells over a wide range of viral inocula with reproducibility, accuracy and relatively low cost.     

中国HIV-1 B'/C亚型感染者对异体病毒中和作用与疾病进展关系研究

目的 探讨中国HIV-1 B'/C亚型感染者对异体病毒中和作用与疾病进展的关系.方法 根据CD4 T淋巴细胞数量和有尤临床症状将HIV-1 B'/C亚型感染者分为HIV慢性感染组和AIDS组.将HIV-1感染者血清稀释(1/10～1/320)后,与在基因结构特点上同源性很低的3株HIV-1作用,以检测其中和作用.同时以正常人血清加病毒悬液为对照孔,能够抑制对照孔50%病毒复制的血清为中和作用阳性.将某个HIV-1感染者血浆能够中和异体病毒的个数占3个异体病毒的百分率定义为HIV-1感染者中和异体病毒的宽度;将某个HIV-1感染者血浆中和3个异体病毒抗体滴度的几何平均滴度定义为HIV-1感染者中和异体病毒的强度.结果 HIV-1慢性感染组与AIDS组之间中和异体病毒的宽度和强度差异有统计学意义,HIV-1慢性感染组显著高于AIDS组.HIV-1慢性感染组中和异体病毒的宽度和强度与病毒载量呈正相关,而AIDS组巾和异体病毒的宽度和强度与病毒载量没有显著的相关性.HIV-1慢性感染组和AIDS组中和异体病毒的宽度和强度与CD4 T淋巴细胞数均没有显著的相关性.结论 中国HIV-1B'/C亚型感染者不同疾病进展阶段针对异体病毒中和作用能力不同,HIV慢性感染组显著高于AIDS组,当疾病进展到AIDS期时,失去对异体病毒的中和作用,提示针对异体病毒的中和抗体与疾病进程有关.     

Progress on the Induction of Neutralizing Antibodies Against HIV Type 1 (HIV-1)

Infection with HIV type 1 (HIV-1), the causative agent of AIDS, is one of the most catastrophic pandemics to affect human healthcare in the latter 20th century. The best hope of controlling this pandemic is the development of a successful prophylactic vaccine. However, to date, this goal has proven to be exceptionally elusive. The recent failure of an experimental vaccine in a phase IIb study, named the STEP trial, intended solely to elicit cell-mediated immune responses against HIV-1, has highlighted the need for a balanced immune response consisting of not only cellular immunity but also a broad and potent humoral antibody response that can prevent infection with HIV-1. This article reviews the efforts made up to this point to elicit such antibody responses, especially with regard to the use of a DNA prime-protein boost regimen, which has been proven to be a highly effective platform for the induction of neutralizing antibodies in both animal and early-phase human studies.     

Epitope-vaccine strategy against HIV-1: today and tomorrow

Vaccines play important roles in preventing infectious diseases caused by different pathogens. However, some pathogens such as HIV-1 challenge current vaccine strategy. Poor immunogenicity and the high mutation rate of HIV-1 make great difficulties in inducing potent immune responses strong enough to prevent infection via vaccination. Epitope-vaccine, which could intensively enhance predefined epitope-specific immune responses, was suggested as a new strategy against HIV-1 and HIV-1 mutation. Epitope-vaccines afford powerful approaches to elicit potent, broad and complete immune protection against not only primary homologous viral isolates but also heterologous viral mutants. Although most studies are still preliminary now, epitope-vaccine as a novel strategy against the AIDS epidemic has great developmental potential. To trigger T-cell-dependent IgG antibody responses and improve affinities of the epitope-specific antibodies, approaches such as recombinant multi-epitope-vaccination and prime-boosting vaccination were suggested. Cellular immune responses, especially CTL responses, could also be elicited and enhanced in addition to humoral immune responses. Developed epitope-vaccines activating both arms of the immune system would benefit prevention and immunotherapy not only against HIV but also other chronic infections.     

HIV-1 and the self-nonself connection: how to sleep with the enemy and be much better off

Envelope-based immunogens capable of generating high titers of neutralizing antibodies have until now been difficult to generate, or failed to act as useful vaccines to prevent HIV-1 infection and disease progression. On the other hand, humoral immune responses to self and allogeneic cellular antigens involved in HIV-1 docking and entry are present both in infected patients and in subjects with natural resistance to HIV-1 infection, where they share similarities but also display definite differences. By dissecting these subtle differences, crucial cellular and molecular markers, possibly correlated with natural resistance to HIV-1 and with the modulation of clinical progression in stably infected patients, have been identified. Here, state-of-the art knowledge on anti-self immune responses following infection or exposure to HIV will be reviewed. The possible implications of these mechanisms in the design of unconventional therapies aimed to counteract the peculiar HIV-1 capability to circumvent the immune system will be discussed.     

Characterization of the HIV-1 specific humoral immune response during highly active antiretroviral therapy (HAART).

Plasma samples from 19 patients were analyzed for HIV-1 directed humoral immune responses prior to and 1 year after initiation of HAART. Eight of the subjects were classified as virologic successes, defined by a >100-fold decrease in viral load (VL) over the 1-year study period and a final VL <500 copies/ml. The eleven HAART failures were defined as subjects with <10-fold decrease in VL. At study entry (before HAART), VL and CD4 counts were similar between the two groups. Humoral immune responses before therapy and after 1 year of therapy were measured by V3 peptide antibody binding titers and neutralization of HIV-1 MN and four subtype B clinical isolates. Before HAART, neutralizing antibody titers to the clinical isolates and HIV(MN), as well as HIV V3 envelope binding titers to several V3 peptides, were significantly higher among treatment successes compared with treatment failures. After 1 year on HAART, neutralization declined in titer and narrowed in specificity among the HAART successes. In contrast, a significant increase in both neutralizing titer and breadth was seen among HAART failures.     

Neutralizing antibodies have limited effects on the control of established HIV-1 infection in vivo

Neutralizing antibodies can protect against challenge with HIV-1 in vivo if present at appropriate concentrations at the time of viral challenge, but any role in the control of established infection is unclear. Here, we show that high serum concentrations of neutralizing monoclonal antibodies, either singly or as a cocktail, have little sustained effect on viral load in established HIV-1 infection in hu-PBL-SCID mice. In some instances, virus replication of neutralization-sensitive virus continues even in the presence of high levels of neutralizing antibody. In most instances, neutralization escape occurs in a few days, even from a cocktail of three antibodies that recognize distinct epitopes. The results imply that humoral immunity is unlikely to play a significant role in the control of established HIV-1 infection in humans.     

Combination DNA plus protein HIV vaccines

A major challenge in developing an HIV vaccine is to identify immunogens and delivery methods that will elicit balanced humoral and cell mediate immunities against primary isolates of HIV with diverse sequence variations. Since the discovery of using protein coding nucleic acids (mainly DNA but also possible RNA) as a means of immunization in the early 1990s, there has been rapid progress in the creative use of this novel approach for the development of HIV vaccines. Although the initial impetus of using DNA immunization was for the induction of strong cell-mediated immunity, recent studies have greatly expanded our understanding on the potential role of DNA immunization to elicit improved quality of antibody responses. This function is particularly important to the development of HIV vaccines due to the inability of almost every previous attempt to develop broadly reactive neutralizing antibodies against primary HIV-1 isolates. Similar to the efforts of developing cell mediated immunity by using a DNA prime plus viral vector boost approach, the best antibody responses with DNA immunization were achieved when a protein boost component was included as part of the immunization schedule. Current experience has suggested that a combination DNA plus protein vaccination strategy is able to utilize the benefits of DNA and protein vaccines to effectively induce both cell-mediated immunity and antibody responses against invading organisms.     

Live attenuated measles vaccine expressing HIV-1 Gag virus like particles covered with gp160ΔV1V2 is strongly immunogenic

Although a live attenuated HIV vaccine is not currently considered for safety reasons, a strategy inducing both T cells and neutralizing antibodies to native assembled HIV-1 particles expressed by a replicating virus might mimic the advantageous characteristics of live attenuated vaccine. To this aim, we generated a live attenuated recombinant measles vaccine expressing HIV-1 Gag virus-like particles (VLPs) covered with gp160ΔV1V2 Env protein. The measles-HIV virus replicated efficiently in cell culture and induced the intense budding of HIV particles covered with Env. In mice sensitive to MV infection, this recombinant vaccine stimulated high levels of cellular and humoral immunity to both MV and HIV with neutralizing activity. The measles-HIV virus infected human professional antigen-presenting cells, such as dendritic cells and B cells, and induced efficient presentation of HIV-1 epitopes and subsequent activation of human HIV-1 Gag-specific T cell clones. This candidate vaccine will be next tested in non-human primates. As a pediatric vaccine, it might protect children and adolescents simultaneously from measles and HIV.     

Nonneutralizing Functional Antibodies: a New “Old” Paradigm for HIV Vaccines

Animal and human data from various viral infections and vaccine studies suggest that nonneutralizing antibodies (nNAb) without neutralizing activity in vitro may play an important role in protection against viral infection in vivo. This was illustrated by the recent human immunodeficiency virus (HIV) RV144 vaccine efficacy trial, which demonstrated that HIV-specific IgG-mediated nNAb directed against the V2 loop of HIV type 1 envelope (Env) were inversely correlated with risk for HIV acquisition, while Env-specific plasma IgA-mediated antibodies were directly correlated with risk. However, tier 1 NAb in the subset of responders with a low level of plasma Env-specific IgA correlated with decreased risk. Nonhuman primate simian immunodeficiency virus (SIV) and simian-human immunodeficiency virus (SHIV) challenge studies suggest that Env-mediated antibodies are essential and sufficient for protection. A comparison of immune responses generated in human efficacy trials reveals subtle differences in the fine specificities of the antibody responses, in particular in HIV-specific IgG subclasses. The underlying mechanisms that may have contributed to protection against HIV acquisition in humans, although not fully understood, are possibly mediated by antibody-dependent cell-mediated cytotoxicity (ADCC) and/or other nonneutralizing humoral effector functions, such as antibody-mediated phagocytosis. The presence of such functional nNAb in mucosal tissues and cervico-vaginal and rectal secretions challenges the paradigm that NAb are the predominant immune response conferring protection, although this does not negate the desirability of evoking neutralizing antibodies through vaccination. Instead, NAb and nNAb should be looked upon as complementary or synergistic humoral effector functions. Several HIV vaccine clinical trials to study these antibody responses in various prime-boost modalities in the systemic and mucosal compartments are ongoing. The induction of high-frequency HIV-specific functional nNAb at high titers may represent an attractive hypothesis-testing strategy in future HIV vaccine efficacy trials.