Chemoattractant-based immunotherapeutics to modulate immune and cancer cells

基于化学引诱剂的免疫疗法可调节免疫细胞和癌细胞

• 批准号: 7732323
• 负责人: Arya Biragyn
• 金额: $ 21.45万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 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/7732323
• 关键词: Adjuvant; Animal Model; Antigen Targeting; Antigen-Presenting Cells; Antigens; Apoptosis; Autoantigens; Autoimmunity; B lymphoid malignancy; Binding; Biological; Blood; Breast; CCR6 gene; CD4 Positive T Lymphocytes; CD8B1 gene; Cancer Vaccine Related Development; Cancer Vaccines; Cell Death; Cell Nucleus; Cells; Cessation of life; Characteristics; Chemotactic Factors; Chronic Disease; Colon; Data; Death Domain; Defensins; Development; Effector Cell; Epitopes; Exhibits; Family; Genus Cola; HLA-A2 Antigen; Hand; Human; Immune; Immune response; Immunity; Immunization; Immunodominant Epitopes; Immunotherapeutic agent; Laminin Receptor; Lead; Long-Term Effects; Lymphoma; MAPK14 gene; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of testis; Mediating; Membrane; Memory; Modeling; Molecular; Mus; NF-kappa B; Necrosis; Numbers; Ovarian; Paper; Pathway interactions; Pattern; Peptides; Process; Proteins; Publications; Publishing; Reporting; Research; Signal Transduction; Staging; Surface; Surveys; System; T-Lymphocyte; TLR1 gene; TLR2 gene; TLR4 gene; TNF gene; TNFRSF1A gene; TNFRSF5 gene; Therapeutic; Time; Tissue Microarray; Tosylphenylalanyl Chloromethyl Ketone; Tumor Antigens; Tumor Necrosis Factor Receptor; Tumor Tissue; Vaccines; Validation; Vertebral column; X Chromosome; Yang; antimicrobial peptide; base; beta-Defensins; cancer cell; chemokine; chemokine receptor; clinical application; clinically relevant; cytokine; cytotoxic; day; embryonic antigen; immunogenic; inhibitor/antagonist; interest; killings; macrophage; melanoma; novel; receptor; response; sperm cell; sperm protein; targeted delivery; tumor

An ideal vaccine carrier should deliver the target antigen to antigen- presenting cells (APCs), while, at the same time, promoting cellular activation. In this respect, we have hypothesized that utilization of the chemoattractant peptides as carriers for TAAs would fulfill these requirements, as they can both target and attract APCs, and in some cases (e.g., MBD2/mDF2b and HBD3) can also activate cells via TLRs. As we reported in several our papers (Biragyn et al. 1999; 2001; 2002, 2004; 2007; Schiavo et al., 2006), chemoattractants are indeed potent inducers of antitumor immunity against variety of tumors in mice by targeted delivery of TAAs to chemokine receptors on APCs. The vaccine did not require use of any adjuvants, but it was essential that tumor antigen was fused physically with a functionally active chemokine, since immunizations with unlinked free chemokine plus antigen did not induce immune responses. The breadth of the chemokine-based vaccines was in their ability to efficiently utilize the MHC processing pathways to elicit CD4+ T helper and CD8+ T cell responses. These results were published in Blood in two subsequent papers (Biragyn et al., Schiavo et al., 2006). 1) Based on these results, we have developed chemokine-based vaccine for human use. The vaccine targets an embryonic antigen OFA-iLRP, a highly conserved the 37-kDa oncofetal immature laminin receptor, that is specifically and highly expressed in a number of human malignancies. Our data indicate that their delivery to iDCs expressing CCR6 elicit potent therapeutic and protective antitumor immune responses against A20 lymphoma in mice. A20 tumor is the most difficult to cure model that closely mimics human B cell malignancy. Although OFA-ILRP shares the same protein backbone with 67-kDa laminin receptor protein, the vaccine did not induce any harmful autoimmunity in mice. Thus, we propose that the vaccine would also be effective for treatment of human malignancies which express OFA-iLRP. The study has been recently successfully completed and published (Biragyn et al., 2007). 2) SPANX-B (a sperm protein associated with the nucleus on the chromosome X -B) is novel antigen with unknown function. We have demonstrated that SPANX-B is cancer-testis antigen. The survey of a panel of human primary 145 tumors on a multi-tumor tissue microarray has revealed that SPANX-B is abundantly expressed in ovarian, colon, breast and lung cancers and melanoma. Moreover, at least in melanoma, its expression pattern was associated with the metastatic stage. We have also demonstrated that SPANX-B is an immunogenic antigen, as healthy humans contained circulating SPANX-B -specific T cell precursors that could be readily expanded to generate both helper CD4+ T cells and cytolytic CD8+ T cells. We have defined at least two immunodominant HLA-A2 restricted epitopes that can be used to elicit SPANX-B specific CD8+ CTLs. The CD8+ T cells, which were raised using these peptides, recognized and efficiently killed the HLA-matched human primary melanomas. On the other hand, we have also determined at least one HLA-DR-restricted and immunodominant epitope, which was used to generate the SPANX-B specific CD4+ T cells. Thus, SPANX-B is an immunogenic and clinically relevant antigen that has significant therapeutic potency as vaccine target. The results of this study has been recently submitted for publication (Almanzar et al., 2008). 3) Chemokine receptors are also used by a small family of antimicrobial peptides (AMPs), namely defensins. For example, human and murine b-defensins attract immature DCs utilizing chemokine receptor CCR6 (Yang et al., 1999; Biragyn et al., 2001). In fact, we have been first to discover that antimicrobial peptide mDF2b also functions as an endogenous immunomodulatory protein that activates APCs via TLR4 (Biragyn et al., 2002). A human counterpart for mouse mDF2b, which activates human DCs, is not found yet. However, others have recently confirmed our finding by demonstrating that human b-defensin 3 can also activate TLR1 and TLR2 (Funderburg et al., 2007). We report here that, besides DC maturation, mDF2b induces atypical death of APCs by exhibiting characteristic features of apoptosis and necrosis. Although mDF2b-treated APCs produced a number of potentially cytotoxic cytokines, the process was mediated by membrane-bound TNF. However, it did not utilize its apoptosis-inducing TNFR1 receptor, as mDF2b also efficiently killed APCs with deficient TNFRI. Instead, it was using TNFRII (a second TNF receptor that does not have death domains). To do this, mDF2b up regulated surface expression of both TNF and TNFRII presumably to facilitate their engagement and signaling. In support, mDF2b failed to induce cell death of APCs from mice with non-functional TNFRII or TNFa. Taken together, we have demonstrated that mDF2b induces two distinct processes separated by time: first, as we previously reported (Biragyn et al., 2002), it activates APCs inducing TLR-4/MyD88 signaling cascade that manifests in maturation of iDCs within one day. However, starting from the third day, the activated cells get self-destruction signaling from up regulated expression of membrane-bound TNF and TNFRII. This is p38, ERK1/2 and the NF-kB dependent process, as mDF2b failed to promote death in APCs from mice with non-functional p50 NF-kB, or in w.t. DCs treated in presence of inhibitors of NF-kB activity such as TPCK. The biological meaning of our observation remains unknown, though we speculate that it might be an important and general step for eliminating harmful long-term effects of activated or over-activated DCs and macrophages. These data have been recently published (Biragyn et al., 2008).

理想的疫苗载体应将靶抗原传递给抗原呈递细胞（APCs），同时促进细胞活化。在这方面，我们假设利用化学引诱肽作为taa的载体可以满足这些要求，因为它们既可以靶向也可以吸引apc，并且在某些情况下（例如MBD2/mDF2b和HBD3）也可以通过tlr激活细胞。正如我们在几篇论文中所报道的那样（Biragyn等人，1999;2001;2002,2004;2007；Schiavo等人，2006），通过靶向递送TAAs到apc上的趋化因子受体，趋化剂确实是抗肿瘤免疫的有效诱导剂。该疫苗不需要使用任何佐剂，但必须将肿瘤抗原与功能活跃的趋化因子物理融合，因为使用未连接的游离趋化因子加抗原进行免疫接种不会诱导免疫反应。基于趋化因子的疫苗的广度在于它们能够有效地利用MHC加工途径来引发CD4+ T辅助细胞和CD8+ T细胞反应。这些结果随后发表在《血液》杂志的两篇论文中（Biragyn et al., Schiavo et al., 2006）。

项目成果

Rationale for dendritic cell-based vaccines for acute myeloid leukemia.

基于树突状细胞的急性髓系白血病疫苗的基本原理。

• 发表时间: 2005
• 期刊: Haematologica
• 影响因子: 10.1
• 作者: Biragyn,Arya

Cloning of B cell lymphoma-associated antigens using modified phage-displayed expression cDNA library and immunized patient sera.

使用改良的噬菌体展示表达 cDNA 文库和免疫患者血清克隆 B 细胞淋巴瘤相关抗原。

• DOI: 10.1016/j.jim.2006.02.013
• 发表时间: 2006
• 期刊: Journal of immunological methods
• 影响因子: 2.2
• 作者: Cha,Soung-Chul;Kwak,LarryW;Ruffini,PierAdelchi;Qin,Hong;Neelapu,Sattva;Biragyn,Arya
• 通讯作者: Biragyn,Arya