Vaccine strategies for HIVAIDS

HIV/艾滋病疫苗策略

基本信息

批准号：
10926475
负责人：
JAY A BERZOFSKY
金额：
$ 107.36万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10926475
关键词：
ALVAC Acute Adjuvant Affect Affinity Agonist Animals Antibiotics Antibodies Antigens Avidity Back Binding Blood Platelets CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cancer Vaccines Cell Aggregation Cell Line Cells Citrobacter Clinical Trials Colitis Collaborations Colon Cross Presentation Cytoprotection Cytotoxic T-Lymphocytes DNA Data Dose Endosomes Epigenetic Process Epitopes Exclusion Fertilization Funding Genes Genetic Transcription Glycocalyx Goals Granulocyte-Macrophage Colony-Stimulating Factor Gut Mucosa HIV HIV Envelope Protein gp120 HIV Infections HIV vaccine HIV/SIV vaccine Helper-Inducer T-Lymphocyte Histocompatibility Home Homing Human IL17 gene Immune Immune checkpoint inhibitor Immune response Immunity Immunization Immunodominant Epitopes Immunoglobulin A Immunology Immunotherapy Infection Interferon Type II Interferons Interleukin-12 Interleukin-15 Large Intestinal Mucosa Large Intestine Ligands Macaca Malignant Neoplasms Memory Modeling Modification Mucosal Immunity Mucous Membrane Mus Myeloid-derived suppressor cells Natural Immunity OVA-8 Oral PF4 Gene PPBP gene Patients Phase Plasma Cells Plasmablast Predisposition Production Proteins Rectum Regulation Research Risk Role SIV SIV Vaccines STAT1 gene Safety Site Small Intestines Stomach Structure Surface T cell response T-Cell Depletion T-Lymphocyte TLR3 gene Testing Textbooks Time Toll-like receptors Training Transforming Growth Factor beta Transgenic Mice Translating Translations Tretinoin Vaccine Adjuvant Vaccine Design Vaccines Vaccinia virus Vagina Viral Viral Antigens Viral Load result Virus Virus Diseases Work aluminum sulfate anti-PD-L1 base cancer immunotherapy carboxypeptidase C chemokine receptor cytokine design gp160 gut microbiota immune activation imprint improved in vivo interleukin-21 invention knock-down lymphoid structures melanoma microbicide microbiome monocyte mucosal vaccine nanoparticle nanoparticle delivery novel novel strategies prevent receptor expression rectal response simian human immunodeficiency virus single-cell RNA sequencing synergism trafficking transcriptome sequencing transmission process vaccine delivery vaccine efficacy vaccine immunogenicity vaccine strategy vaginal mucosa viral detection viral transmission

项目摘要

The strategies above involve several steps that together comprise a push-pull approach, to optimize antigen structure, improve quantity and quality of the response, and remove regulatory barriers. Blocking negative regulation in HIV and vaccine strategies: We carried out epitope enhancement (sequence modification to improve MHC binding) to increase MHC binding of epitopes from several viral antigens including HIV, and showed these improved vaccine efficacy. We have pioneered the use of anti-TGFb as a novel checkpoint inhibitor, given that TGFb is one of the most immunosuppressive cytokines known. We even showed preliminary evidence of efficacy and safety in a clinical trial in melanoma patients. We have also compared combinations of checkpoint inhibitors and anti-TGF-b in enhancing vaccine efficacy in mice, and found preliminary data for synergy among anti-TGFb, anti-LAG3, anti-TIGIT and anti-PDL1. These strategies are applicable to both HIV and cancer. Cytokines as vaccine adjuvants for HIV and induction of high avidity T cells: Our earlier work first showed that high avidity T cells were more effective at clearing viral infections. We found ways to induce them with cytokines and TLR ligands, including expressing IL-15 in the vaccine, and inclusion of costimulatory molecules, in collaborations with Tom Waldmann and with Jeff Schlom, respectively. The quality of response proved more important than the quantity. We recently found, using a novel adjuvant, CAF09, that we could lower the vaccine dose sufficiently to induce higher avidity CD4 T cells to better clear HIV-gp160-expressing virus infection in mice. We also found that IL-1b induces Th17 helper cells that do not help Tc1 CD8 T cells that protect against vaccinia virus expressing HIV gp160. Rather, they skew the CD8 response to Tc17 cells that make IL-17 and do not protect. TGF-b blockade can prevent this problem, as TGF-b is involved in Th17 induction. We are examining the epigenetic and transcriptional mechanisms behind high avidity T cells by carrying out a single-cell RNAseq study of high vs low avidity T cell lines generated against the same antigen. T cells from OT-I TCR-transgenic mice were grown with high or low concentrations of antigen to produce low or high avidity lines, respectively, with the identical TCR, to exclude effects of intrinsic TCR affinity. These were stimulated with the same SIINFEKL peptide, and subjected to RNAseq. Also, T cells were generated against the same SIINFEKL peptide and subjected to concurrent TCR and RNAseq analysis, and high and low avidity lines with the same clonotype were compared. Promising genes are being tested in knock-down studies. We also found that IL-21 synergizes with IFN-g to induce IFN-stimulated genes and clear Citrobacter colitis through an effect on STAT1. We also examined combinations of cytokines and TLR ligands as vaccine adjuvants and found greatest efficacy of IL-15 + TLR3 and TLR9 agonists, but also some efficacy of IL-12 + GM-CSF. We are comparing these for qualitative differences in the immune response as well as quantity. Mucosal immunity, microbiome and HIV/SIV vaccines: About 85% of HIV transmission is mucosal. We found that a mucosal T cell vaccine can impact the initial mucosal nidus of infection. We are studying induction and trafficking of T cells, DCs, and MDSCs among mucosal compartments to optimize mucosal vaccine efficacy. In mice, we found that T cells could be directly primed in the vaginal mucosa, despite lack of organized lymphoid structures, contrary to textbook dogma. We also discovered that colonic DCs can imprint CD8 T cells to home back to the colon preferentially, based on differential retinoic acid expression vs. small intestine DCs. We are currently examining the mechanism of this effect in terms of chemokine receptor expression and homing activity, and by using an RNAseq comparison of clonal OT-1 T cells stimulated by DCs from the small vs large intestines. We discovered that altering a cathepsin S cleavage site could protect an immunodominant epitope of HIV gp120 from degradation in endosomes during cross-presentation, providing proof of concept for a novel mechanism of virus escape for HIV that infects mostly non-APCs. Using NHP models, we found that activated mucosal T cells determine susceptibility to SIV/HIV infection (transmission), eclipse time prior to systemic viral detection, and acute viral load. We found that even in naive animals, gut microbiota can strongly affect susceptibility to transmission, by immune activation, and also affect vaccine efficacy. We are exploring the mechanisms by testing the effect of antibiotics on multiple cell subsets in the large compared to the small intestine. Further, we found that vaccines can induce MDSCs that counteract vaccine protection, and also infection can affect trafficking of MDSCs. We also demonstrated for the first time that MDSCs could be infected by SHIV in vivo. As most HIV transmission is through mucosal surfaces and HIV homes to the gut mucosa, we have designed and invented a nanoparticle vaccine delivered orally in mice but coated to pass through the stomach intact and to be released selectively in the large intestine where it induces CTL responses in the large intestine and vaginal mucosa and protects against rectal or vaginal challenge with a virus. We translated our oral nanoparticle (NP) approach to macaque SIV vaccines, finding reduced risk against SHIV rectal acquisition in 2 studies. Surprisingly, we discovered that protection against SIV acquisition in 3 studies can occur without anti-envelope antibodies. While T cell immunity was induced, it did not correlate with protection, and protection was not abrogated by CD8 T cell depletion. Rather, protection correlated with trained innate immunity, involving monocyte "memory" for SIV in induction of cytokines maintained by epigenetic changes. We have carried out RNAseq to determine what changes occur in monocytes after immunization, leading to new mechanistic hypotheses including an unexpected role for platelets and their production of platelet factor 4 (PF4), also known as CXCL4. This is manifested by platelet-cell aggregates in which platelets actually coat cells. Also, we are combining an SIV vaccine and mucosal NP boost to increase mucosal immunity with a microbicide to reduce the viral inoculum in an OAR-funded study. Our hypothesis is that priming and boosting with mucosally delivered nanoparticles containing V2 loop antigens will both increase mucosal immunity to protect against intrarectal SIV challenge, and selectively expand immune responses against the V2 loop that have been shown to correlate with protection in the RV144 phase III human trial. Initial results show that priming and boosting orally with NPs containing a V2-loop pentamer can increase protection compared to the base vaccine alone consisting of SIV gag and env DNA, ALVAC expressing gp120, and deltaV1 gp120 protein in alum. Immune correlates are under study, including some evidence for mucosal IgA, ADCC, mucosal plasma cells and plasmablasts, and T cell responses.

上面的策略涉及多个步骤，这些步骤共同构成了推拉方法，以优化抗原结构，提高响应的数量和质量并消除调节屏障。阻止HIV和疫苗策略中的负调节：我们进行了表位增强（序列修饰以改善MHC结合），以增加来自包括HIV在内的几种病毒抗原的表位的MHC结合，并显示了这些提高的疫苗功效。鉴于TGFB是已知的最免疫抑制性细胞因子之一，我们已经开创了抗TGFB作为新型检查点抑制剂的使用。我们甚至在黑色素瘤患者的临床试验中显示了功效和安全性的初步证据。我们还比较了检查点抑制剂和抗TGF-B在增强小鼠疫苗功效中的组合，并发现了抗TGFB，抗LAG3，抗词素，抗词素和抗PDL1的协同作用的初步数据。这些策略适用于艾滋病毒和癌症。细胞因子作为HIV的疫苗佐剂和诱导高流行T细胞：我们先前的工作首先表明，高潮T细胞在清除病毒感染方面更有效。我们找到了用细胞因子和TLR配体诱导它们的方法，包括在疫苗中表达IL-15，并分别与汤姆·沃尔德曼（Tom Waldmann）和杰夫·施洛姆（Jeff Schlom）合作。响应的质量比数量更重要。我们最近使用一种新型辅助CAF09发现，我们可以降低足够的疫苗剂量，以诱导更高的亲发性CD4 T细胞，以更好地清除小鼠中表达HIV-GP160的病毒感染。我们还发现，IL-1B诱导Th17辅助细胞，这些细胞无助于预防表达HIV GP160的牛ac病毒的TC1 CD8 T细胞。相反，它们偏向于使IL-17且不能保护的TC17细胞的CD8响应。由于TGF-B参与TH17诱导，因此TGF-B阻滞可以防止此问题。我们正在研究高流经T细胞背后的表观遗传和转录机制，通过对针对相同抗原产生的高而低相位的T细胞系进行单细胞RNASEQ研究。来自OT-I TCR-转基因小鼠的T细胞用高或低浓度的抗原生长，分别与相同的TCR产生低或高发线，以排除固有的TCR亲和力的影响。用相同的siinfekl肽刺激这些，并受到RNASEQ。同样，针对相同的siinfekl肽产生T细胞，并进行并发的TCR和RNASEQ分析，并比较具有相同clonotype的高和低自发线。有希望的基因正在敲门研究中测试。我们还发现，IL-21与IFN-G协同诱导IFN刺激的基因并通过对STAT1的影响来诱导IFN刺激的基因。我们还研究了细胞因子和TLR配体的组合作为疫苗佐剂，发现IL-15 + TLR3和TLR9激动剂的功效最大，但也发现IL-12 + GM-CSF的功效。我们将这些与免疫反应和数量的质量差异进行比较。粘膜免疫，微生物组和HIV/SIV疫苗：大约85％的HIV传播是粘膜。我们发现，粘膜T细胞疫苗会影响最初的感染粘膜。我们正在研究粘膜室中T细胞，DC和MDSC的诱导和运输，以优化粘膜疫苗疗效。在小鼠中，我们发现，尽管缺乏有组织的淋巴结构，但与教科书教义相反，T细胞可以直接在阴道粘膜中引发。我们还发现，基于视黄酸的表达与小肠DC，结肠DC可以将CD8 T细胞重新归为归为结肠。我们目前正在研究这种作用在趋化因子受体表达和归巢活性方面，并使用rnaseq比较来自小型肠子的DC刺激的克隆OT-1 T细胞。我们发现，改变组织蛋白酶的裂解位点可以保护HIV GP120的免疫主导表位免受跨表达期间内体内降解的降解，从而为艾滋病毒的新型HIV机制提供了概念证明，该机制大多是非APC。使用NHP模型，我们发现活化的粘膜T细胞确定了对SIV/HIV感染（传播），全身病毒检测前的日食和急性病毒负荷的敏感性。我们发现，即使在天真的动物中，肠道菌群也会通过免疫激活强烈影响对传播的敏感性，并影响疫苗功效。与小肠相比，我们正在通过测试抗生素对大型细胞亚群的影响来探索机制。此外，我们发现疫苗可以诱导抵消疫苗保护的MDSC，并且感染也会影响MDSC的贩运。我们还首次证明了MDSC可以被Shiv In Vivo感染。 As most HIV transmission is through mucosal surfaces and HIV homes to the gut mucosa, we have designed and invented a nanoparticle vaccine delivered orally in mice but coated to pass through the stomach intact and to be released selectively in the large intestine where it induces CTL responses in the large intestine and vaginal mucosa and protects against rectal or vaginal challenge with a virus.我们将口服纳米颗粒（NP）方法转化为猕猴SIV疫苗，发现在2项研究中发现了针对SHIV直肠直肠采集的风险降低。出人意料的是，我们发现在没有抗Envelope抗体的情况下，可以在3项研究中进行SIV获取的保护。虽然诱导T细胞免疫，但与保护无关，并且CD8 T细胞耗竭不会消除保护。相反，保护与训练有素的先天免疫相关，涉及SIV的单核细胞“记忆”，以诱导通过表观遗传变化维持的细胞因子。我们已经进行了RNASEQ，以确定免疫后单核细胞中发生了什么变化，导致了新的机械假设，包括血小板的意外作用及其产生血小板因子4（PF4）（PF4），也称为CXCL4。这是由血小板聚集体表现出来的，该血小板实际上是覆盖细胞的。此外，我们正在结合SIV疫苗和粘膜NP的增强，以在一项由OAR资助的研究中降低菌心的粘膜免疫，以减少病毒接种。我们的假设是，含有V2环抗原的粘膜递送的纳米颗粒的启动和增强都将增加粘膜免疫，以防止直肠内SIV攻击，并选择性地扩展针对RV144阶段III阶段人类试验的V2环路的免疫反应。最初的结果表明，与仅由SIV GAG和ENV DNA，表达GP120的ALVAC和Deltav1 GP120蛋白组成的基本疫苗相比，用含有V2环五聚体的NP口服启动和增强可以增加保护。正在研究免疫相关性，包括一些证据表明粘膜IgA，ADCC，粘膜血浆细胞和浆质和T细胞反应。