Oligomannose-coated Liposomes Research Articles

Importance of particle size of oligomannose-coated liposomes for induction of Th1 immunity

Oligomannose-coated liposomes (OMLs) comprised of dipalmitoylphosphatidylcholine,cholesterol and Man3-DPPE at a molar ratio of 1:1:0.1 and particle diameters of about 1000 nm can induce liposome-encased antigen-specific strong Th1 immunity. In this study, we evaluated the effect of particle sizes of OMLs on induction of Th1 immune responses in mice. Spleen cells obtained from mice immunized with antigen-encapsulating OMLs with 1000- and 800-nm diameters secreted remarkably high levels of IFN-γ upon in vitro stimulation. In addition, sera of mice that received these OMLs had significantly higher titers of antigen-specific IgG2a than those of IgG1, which are commonly associated with Th1 responses. In contrast, treatment with antigen-encapsulating OMLs with 400- and 200-nm diameters failed to induce IFN-γ secretion from spleen cells, although these OMLs did elicit elevation of antigen-specific IgGs. In addition, the titers of serum antigen-specific IgG2a were the same as those of IgG1 in mice that received 400-nm OMLs. Resident peritoneal mononuclear phagocytes (MNPs) treated with OMLs of diameter ≥ 600 nm secreted IL-12, which is essential for induction of Th1 immune responses, while those treated with OMLs of ≤ 400 nm failed to produce this cytokine. However, 400-nm OMLs did induce enhanced expression of MHC class II and costimulatory molecules on MNPs, similarly to OMLs of ≥ 600 nm. Taken together, these results strongly indicate that OMLs of diameter ≥ 600 nm are required to induce Th1 immune responses against OML-encased antigens, although OMLs of diameter ≤ 400 nm can activate MNPs.

Neospora GRA6 possesses immune-stimulating activity and confers efficient protection against Neospora caninum infection in mice

Vaccination has the potential to be the most cost-effective control measure for reducing the economic burden of neosporosis in cattle. In this study, the immune-stimulatory effect of recombinant Neospora caninum dense granule protein 6 (NcGRA6) was confirmed via its triggering of IL-12p40 production in murine macrophages. BALB/c mice were immunized with recombinant NcGRA6 fused with glutathione S-transferase (GST) protein with or without oligomannose-coated-liposomes (OMLs) as the potential adjuvant. Specific IgG1 antibody production was observed from 21 and 35 days after the first immunization in NcGRA6+GST- and NcGRA6+GST-OML-immunized mice, respectively. However, specific IgG2a was detected 1 week after the infection, and IgG2a levels of the NcGRA6+GST- group were higher than those of the NcGRA6+GST-OML-group. Moreover, spleen cell proliferation with concomitant interferon-gamma production was detected in mice immunized with NcGRA6+GST, indicating that a significant cellular immune response was induced. Mouse survival rates against N. caninum challenge infection were 91.7% for NcGRA6+GST and 83.3% for NcGRA6+GST-OML, which were significantly higher than those of control groups (GST-OML: 25%, phosphate-buffered saline: 16.7%). This indicates that naked NcGRA6+GST induced protective immunity. Thus, our findings highlight the immune-stimulating potential of NcGRA6 and the ability to induce protective immunity against N. caninum infection in mice.

Critical role of TLR2 in triggering protective immunity with cyclophilin entrapped in oligomannose-coated liposomes against Neospora caninum infection in mice

Neospora caninum is an intracellular protozoan parasite closely related to Toxoplasma gondii. N. caninum is thought to be a major cause of abortion in cattle worldwide. Given the current situation of drastic economic losses and a lack of efficient control strategies against such parasites, the challenge to develop potent vaccine candidates and technologies remains. We investigated the immune stimulating activity of N. caninum cyclophilin (NcCyp) with and without a formulation with oligomannose-coated-liposomes (OML) as the potential adjuvant. NcCyp-OML activated NF-κB in RAW 264.7 cells and triggered interleukin (IL)-12p40 production from murine peritoneal macrophages. In BALB/c mice, immunization with NcCyp-OML was associated with the production of specific antibodies (IgG1 and IgG2a). The specific antibody (IgG1) against naked NcCyp was also observed after the challenge infection, but it was significantly lower than those of NcCyp-OML. Moreover, significant cellular immune responses were induced, including spleen cell proliferation and interferon-gamma production. The immunization of mice with NcCyp-OML, and to lesser extent with naked NcCyp, induced significant protection against challenge with a lethal dose of N. caninum compared with the PBS control group. This protection was associated with a higher survival rate, slight changes in body weight, and lower clinical score of mice. In addition, the significant protective efficacy of NcCyp-OML was confirmed in another mouse strain, male C57BL/6 mice. The current study revealed the marked contribution of Toll-like receptor 2 (TLR2) to the protective immunity triggered by NcCyp-OML because higher numbers of TLR2−/− mice succumbed to a lethal dose of N. caninum compared with C57BL/6 mice. Moreover, prominent spleen cell proliferation and IFN-γ production was induced in NcCyp-OML-immunized mice by a TLR2-dependent mechanism.

In vitro uptake of oligomannose-coated liposomes leads to differentiation of inflammatory monocytes into mature antigen-presenting cells that can activate T cells

Oligomannose-coated liposomes (OMLs), containing entrapped antigens, serve as effective antigen delivery vehicles and as a novel adjuvant to induce antigen-specific cellular immune responses. However, in vitro activation of antigen-presenting cells (APCs) by OMLs has not yet been demonstrated. In this paper, we found that OMLs can deliver the antigens and the stimulatory signals into inflammatory monocytes in vitro, leading to differentiation of the cells to mature APCs. When OMLs were co-cultured with peripheral blood mononuclear cells from C57BL/6 mice in the presence of mouse serum, OMLs were preferentially incorporated into both Ly6Chigh monocytes and Ly6Clow monocytes, which are referred to as murine inflammatory and resident monocytes, respectively. The expression of CD11c, CD80, CD86, CCR7, and MHC class II on the Ly6Chigh monocytes was significantly enhanced during the 24 h after OML uptake, whereas upregulation of these molecules on the Ly6Clow monocytes was limited. In addition, the antigenic peptide of OVA encased in OMLs was presented on MHC class I of only Ly6Chigh monocytes. Furthermore, OVA-encasing OML-ingesting monocytes can activate CD8+ T cells from OT-1 mice, suggesting that antigens encapsulated in OMLs were cross-presented in inflammatory monocytes. Adoptive transfer of the monocytes that engulf OVA-encasing OMLs led to induction of an antigen-specific Th1 immune response in mice. Taken together, mature APCs can be generated from inflammatory monocytes in peripheral blood by ex vivo treatment of the cells with OMLs without any additional stimuli.

Requirement of TLR4 signaling for the induction of a Th1 immune response elicited by oligomannose-coated liposomes

We have previously demonstrated that administration of oligomannose-coated liposomes (OMLs), in which an antigen is encased, induce antigen-specific Th1 immune responses and CTLs. In the present study, we showed that TLR4 signaling is required for the induction of specific immune responses following OML administration. In C3H/HeJ mice, which express a dysfunctional TLR4, the antigen-specific Th1 immune response could not be elicited following intraperitoneal administration of OVA-encased OMLs (OML/OVA). However, OML uptake by peritoneal cells, the subsequent production of IL-12 and the upregulation of co-stimulatory molecules and MHC class II on the cells in response to OML uptake occurred in C3H/HeJ mice to the same extent as in wild type C3H/HeN mice. In addition, peritoneal phagocytic cells from TLR4−/− mice that ingest OML/OVA can activate CD4+ T cells from OT-II mice. On the other hand, the number of OML-ingesting peritoneal cells that migrated into mesenteric lymph nodes in C3H/HeJ mice was significantly less than that in C3H/HeN mice. Therefore, the chemotactic capability of OML-ingesting peritoneal phagocytes to the draining lymph nodes rather than the activation and maturation of the cells in response to OML uptake is impaired by lack of TLR4 signaling, and disorder of the Th1 immune response elicited by OMLs in mice, which lack TLR4 signaling, is due to the impairment of cell migration following OML uptake.

Novel immunotherapy approaches to food allergy.

Despite reaching high percentages of desensitization using allergen-specific immunotherapy (SIT) in patients with food allergy, recent studies suggest only a low number of patients to reach persistent clinical tolerance. This review describes current developments in strategies to improve safety and long-term efficacy of SIT. Modified allergens or tolerogenic peptides, ultimately optimized for human leukocyte antigen background of the patient, are explored for tolerance induction, whereas anti-IgE antibody (Omalizumab) may be used to facilitate SIT safety. Adjunct therapies to enhance efficacy may make use of TH1 polarizing agents, for example, CpG-oligodeoxynucleotides combined with modified allergen packaged in nanoparticles. Preclinical studies showed insulin-like growth factor-2, intravenous immunoglobulin, Tregitopes or allergen encased oligomannose-coated liposomes capable of inducing regulatory T-cells, recognized for their importance in clinical tolerance induction. Dietary intervention strategies utilizing herbal formula 2, VSL#3, nondigestible short-chain galacto-oligosaccharides and long-chain fructo-oligosaccharides (scGOS/lcFOS) plus Bifidobacterium breve M-16V or n-3 long-chain polyunsaturated fatty acids may facilitate safety and/or a favourable milieu for tolerance induction. Combining SIT using (adapted) allergens or tolerogenic peptides with adjunct therapy may be essential to improve safety and/or efficacy. Beyond using targeted approaches, specific dietary components may be explored to reduce side-effects and support clinical tolerance induction by SIT.

Plasmodium berghei circumsporozoite protein encapsulated in oligomannose-coated liposomes confers protection against sporozoite infection in mice.

BackgroundThe design and development of an effective malaria vaccine against the pre-erythrocytic and erythrocytic-stages of infection present a great challenge.MethodsIn the present study, protective efficacy of oligomannose-coated liposome (OML)-entrapped merozoite and sporozoite antigens against Plasmodium berghei challenge infection in BALB/c mice was evaluated.ResultsSubcutaneous immunization with truncated merozoite surface protein 1 entrapped with OML (OML-PbMSP1) prolonged survival, but failed to protect the mice from erythrocytic-stage infection, despite the antigen-specific antibody responses induced by the immunization regimen. In contrast, immunization with circumsporozoite protein entrapped with OML (OML-PbCSP) elicited antigen-specific humoral and cellular responses, which correlated with substantial protection against sporozoite challenge infections.ConclusionsThe current results represent the use of an oligomannose-coated liposome-based vaccine against pre-erythrocytic and erythrocytic stages malaria infection. This approach may offer a new vaccination strategy against malaria infection.

Pathophysiology and epidemiology of virus-induced asthma.

Many respiratory viruses are mainly responsible for common cold, bronchitis, bronchiolitis, and pneumonia. Furthermore, asthma and chronic obstructive pulmonary disease (COPD) are major cause of mortality. The prevalence of asthma in developed countries is approximately 10% in adults and even higher in children (Barnes, 2008). Thus, the medical costs for these diseases are a major burden in many countries. Respiratory virus infections also cause the most of acute exacerbation of asthma (virus-induced asthma) or COPD. Among them, human rhinoviruses (HRV) are detected in the two thirds of the cases with asthma exacerbations in children (Johnston et al., 1995). However, epidemiology and pathophysiology of asthma and COPD is not known. Furthermore, a few effective vaccines have been applied. Therefore, it may be important to better understand pathophysiology of virus-induced asthma or virus-induced COPD exacerbation. Both aspects of the virus agents and host defense systems including acute/chronic inflammation and airway tissue remodeling should be clarified. This e-book aims to review and discuss pathophysiology and epidemiology of virus-induced asthma and COPD focusing on new findings of the host immunity and virology. This Research Topic contains 7 review articles and 3 original articles regarding pathophysiology of virus-induced asthma. As the first article, Kudo et al. (2013) reviewed pathology of asthma. This article globally covers from molecular histopathology involved in cytokine networks of asthma. The readers may easily understand molecular immunopathology of virus-induced asthma. In the second issue, Okayama (2013) presents cellular and humoral immunity of asthma. Accumulating evidence implicates that the genetic and environmental factors may be associated with virus-induced asthma. This work focuses on the immunological mechanisms that may explain why asthma is associated with RSV-and HRV-infection. As the third review article, Kimura et al. (2013) present the molecular mechanisms between various cytokines and innate immunity of viral respiratory infections including virus-induced asthma. The authors also show the signaling pathways with regard to them. In the 4th review article, Tsukagoshi et al. (2013) discuss the genetic characteristics and molecular evolution of respiratory viruses, and epidemiology of asthma. They also show phylogenetic analysis of the detected viruses in the children with respiratory syncytial virus- (RSV) and/or HRV-associated wheezing and asthma. As the 5th review article, Inoue and Shimojo (2013) present epidemiology and pathophysiology of virus-induced asthma in children. They summarize the previous findings and discuss how clinicians can effectively intervene in these viral infections to prevent the development of asthma. Next, Kurai et al. (2013) and Saraya et al. (2014) present pathophysiology of virus-induced COPD and asthma in adults. They summarize current knowledge concerning exacerbation of both COPD and asthma by focusing on the clinical significance of associated respiratory virus infections. Furthermore, influenza A(H1N1)pdm09 virus have suddenly emerged in Mexico in the spring, 2009. The virus can cause influenza pandemy accompanying with pneumonia/wheezing. Obuchi et al. (2013) review essential reports with regard to asthma in patients infected with the virus, and they discuss the utility of influenza vaccines and antivirals. Although HPIV3 is an etiological agent for respiratory disorders such as pneumonia and asthma, there is no prophylactic human vaccine against the virus infection. In the 9th issue as original article, Senchi et al. (2013) present the development of an oligomannose-coated liposome (OML) nasal vaccine against HPIV3 in combination with an effective adjuvant Poly(I:C). They report that their newly-developed vaccine can successfully induce antigen-specific immunity with a small amount of antigen via the nasal route. These results highlight the utility of combining sophisticated systems in the development of a novel vaccine against HPIV3. In the final article, Matsunaga et al. (2014) present the development of monoclonal antibodies (MAbs) against hemagglutinin-neuraminidase (HN) of HPIV3. For synthesizing the antigen protein, they utilized the wheat germ cell-free system. This new cell-free system-based protocol for antigen production enabled to create the MAbs that can be applicable in various immune assays such as flowcytometry and immunoprecipitation analyses. The newly-developed MAbs could thus be a valuable tool for the study of HPIV3 infection as well as the several diagnostic tests of this virus. In conclusion, we believe that these works are well summarized and informative to glimpse the field of virus-associated asthma/COPD, and may help understanding the basic and clinical aspects of the disease. We would be happy if this collection of papers will offer new stimuli and perspectives for not only researchers but also clinicians working around the exciting and emerging the e-book.

SIGNR1-mediated phagocytosis, but not SIGNR1-mediated endocytosis or cell adhesion, suppresses LPS-induced secretion of IL-6 from murine macrophages

C-type lectin receptors (CLRs) serve as phagocytosis receptors for pathogens and also function as adhesion molecules and in the recognition and endocytosis of glycosylated self-antigens. In the present study, we demonstrated that phagocytosis mediated by a mouse mannose-binding CLR, SIGNR1 significantly suppressed the LPS-induced secretion of the specific pro-inflammatory cytokines from the resident peritoneal macrophages and the mouse macrophage-like cells that express SIGNR1 (RAW-SIGNR1). LPS-induced secretion of IL-6 from peritoneal macrophages suppressed in response to uptake of oligomannose-coated liposomes (OMLs), and the suppression was partly inhibited by treatment with an anti-SIGNR1 antibody. LPS-induced secretion of IL-6 from RAW-SIGNR1 cells was also clearly inhibited by treatment of the cells with OMLs >0.4μm in diameter, but treatment with OMLs <0.4μm in diameter did not affect the IL-6 secretion. In contrast, LPS-induced TNF-α secretion from the cells was not affected on treatment of the cells with OMLs. Suppression of the IL-6 secretion was not observed following treatment with oligomannose-containing soluble polymers or when cells were bound to an oligomannose-coated solid phase. Phagocytosis of oligomannose-coated liposomes did not interfere with the transcription of IL-6 mRNA, but did affect IL-6 mRNA stability, leading to suppression of IL-6 secretion. Interestingly, treatment of the cells with Ly290042, a PI3 kinase inhibitor, partly blocked the suppression of LPS-induced secretion of IL-6 by OML. Thus, we conclude that SIGNR1-mediated phagocytosis but not SIGNR1-mediated endocytosis and cell adhesion, suppresses the TLR4-mediated production of specific proinflammatory cytokines via PI3 kinase signaling.

Vaccination with profilin encapsulated in oligomannose-coated liposomes induces significant protective immunity against Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular parasite that can infect a variety of mammals and birds, causing toxoplasmosis. Several types of vaccines against T. gondii have been developed, but these have limitations in terms of their safety and inadequate efficacy. T. gondii profilin (TgPF) is a potential immunodominant antigen for a candidate vaccine. In this study, we encapsulated TgPF in oligomannose-coated liposomes (OMLs) to evaluate the immune response induced by this vaccine. C57BL/6 mice were immunized with TgPF-OML three times at 14-day intervals and challenged with T. gondii. TgPF-OML increased the survival of the mice and reduced the parasite burden in their brains after T. gondii infection. Immunization with TgPF-OML also induced TgPF-specific interferon-γ production and IgG antibodies in mice. Our results demonstrate that OML-encapsulated TgPF triggers strong humoral and cellular responses against T. gondii, and that TgPF-OML is a candidate vaccine that warrants further development.

C-type lectin receptor SIGNR1 expressed on peritoneal phagocytic cells with an immature dendritic cell-like phenotype is involved in uptake of oligomannose-coated liposomes and subsequent cell maturation

The mannose-binding C-type lectin receptor SIGNR1 appears to be a structural and functional murine homologue of human DC-SIGN, but expression of SIGNR1 and its function in induction of immune responses in dendritic cell (DC) lineages remains unclear. In this study, we demonstrated expression and function of SIGNR1 on mouse peritoneal phagocytic cells with an immature DC-like phenotype. Analysis of these cells with a series of cell lineage markers indicated that CD11b+F4/80− phagocytic cells expressed costimulatory molecules, the DC marker CD83, and MHC class II, suggesting an immature DC-like phenotype. These immature peritoneal DC-like cells expressed low levels of SIGNR1, in addition to another mannose-binding C-type lectin, CD206. The immature peritoneal DC-like cells ingested oligomannose- or Lewis antigen-coated liposomes in vitro through SIGNR1. Following in vitro uptake of oligomannose-coated liposomes, SIGNR1, but not CD206, disappeared rapidly from the surface of the cells. In response to in vitro uptake of OMLs, the peritoneal DC-like cells matured with increasing expression of CD11c, CD86, and MHC class II. Thus, low levels of SIGNR1 expressed on mouse peritoneal phagocytic cells with an immature DC-like phenotype are primarily involved in uptake of mannose- or fucose-decorated particles, and this uptake leads to cell maturation.

Effective stimulation of invariant natural killer T cells by oligomannose-coated liposomes

vThe in vitro and in vivo response of invariant natural killer T (iNKT) cells to alpha-galactosylceramide (αGC)-containing oligomannose-coated liposomes (αGC-OMLs) was examined to determine whether selective delivery of αGC to dendritic cells (DCs) and subsequent activation of iNKT cells could be achieved. Splenocytes stimulated with αGC-OMLs produced higher levels of IFN-γ compared to those stimulated with bare liposomes without an oligomannose coating (αGC-BLs). The ratio of IFN-γ/IL-4 produced from αGC-OML-treated splenocytes was higher than those produced from αGC-BL- and soluble αGC-treated cells. Depletion of CD3(+)-, DX5(+)- or CD11c(+)-cells from splenocytes almost completely abolished the αGC-OML-stimulated cytokine production, suggesting that both NKT cells and DCs were involved in the response to αGC-OML stimulation. In addition, αGC-OMLs were incorporated into both splenic and bone marrow-derived DCs more effectively than αGC-BLs. iNKT cells stimulated with DCs with ingested αGC-OMLs produced much higher levels of IFN-γ than those stimulated with DCs containing αGC-BLs or soluble αGC. Systemic administration of αGC-OMLs led to modification of the kinetics of IFN-γ production in vivo and also resulted in predominant production of IFN-γ from splenocytes over IL-4. In addition, iNKT cells proliferated and expanded upon in vivo activation of the cells with αGC-OMLs much more extensively than with αGC-BLs or soluble αGC. Collectively, our results suggest that αGC-OMLs can be used as a preferential delivery system for lipid antigens to DCs to activate iNKT cells in vivo and ex vivo.

Development of oligomannose-coated liposome-based nasal vaccine against human parainfluenza virus type 3

Human parainfluenza viruses (HPIVs) are the etiologic agents of lower respiratory infections and pneumonia in infants, young children and immunocompromised hosts. The overarching goal for the prevention of HPIV infection is the development of an effective vaccine against HPIVs. In the present study, we investigated the effectiveness of oligomannose-coated liposomes (OMLs) as an antigen-delivery system in combination with a synthetic double-stranded RNA analog for the induction of mucosal and systematic immunity against HPIV3. Full-length hemagglutinin-neuraminidase (HN) protein was synthesized using the wheat germ cell-free protein production system and then encapsulated into OML to serve as the antigen. Intranasal administration of the HN-filling OML (OML-HN) with the synthetic double-stranded RNA adjuvant, polyriboinosinic-polyribocytidylic acid [poly(I:C)] generated significant viral-specific systemic and mucosal immune responses as evidenced by the prominent induction of serum IgG and nasal wash IgA, respectively. On the other hand, no significant immune responses were observed in mice immunized with OML-HN without the adjuvant. Furthermore, serum from mice immunized with OML-HN plus poly(I:C) significantly suppressed viral infection in cell culture model. Our results provide the first evidence that intranasal co-administration of OML-encapsulated HN with the poly(I:C) adjuvant augments the viral-specific immunity against HPIV3.

Oligomannose-Coated Liposome as a Novel Adjuvant for the Induction of Cellular Immune Responses to Control Disease Status

Professional phagocytic cells, such as dendritic cells, are mainly responsible for phagocytosis, antigen presentation, and cytokine secretion, which induce subsequent activation of T cell-mediated immunity. Thus, strategies that deliver antigens and stimulatory signals to the cells have significant implications for vaccine design. In this paper, we summarize the potential for liposomes coated with the neoglycolipids containing oligomannose residues (OMLs) as a novel adjuvant for induction of Th1 immune responses and CTLs specific for the encased antigen. OMLs preferentially take up peripheral phagocytic cells. In response to OML uptake, the cells secrete IL-12 selectively, enhance the expression of costimulatory molecules, and migrate into lymphoid tissues from peripheral tissues. OMLs also have the ability to deliver encapsulated protein antigens to the MHC class I and class II pathways to generate antigen-specific CTLs and Th1 cells, respectively, and lipid antigen to CD1d to activate NKT cells. Since administration of OML-based vaccines can eliminate an established tumor, inhibit elevation of the serum IgE level, and prevent progression of protozoan infections in several murine, human, and bovine models, OML-based vaccines have revealed their potential for clinical use in vaccination for a variety of diseases in which CTLs and/or Th1 cells act as effector cells.

Novel Vaccine Adjuvants

Hindawi Publishing Corporation BioMed Research International Volume 2013, Article ID 835105, 2 pages http://dx.doi.org/10.1155/2013/835105 Editorial Novel Vaccine Adjuvants Anshu Agrawal, 1 Mohammad Owais, 2 and Udai P. Singh 3 Division of Basic and Clinical Immunology, Department of Medicine, University of California, Irvine, CA 92697, USA Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, Uttar Pradesh 202002, India Department of Pathology, Microbiology and Immunology, University of South Carolina, School of Medicine, Building 1, Room B 42, 6439 Garners Ferry Road, Columbia, SC 29208, USA Correspondence should be addressed to Anshu Agrawal; aagrawal@uci.edu Received 11 September 2013; Accepted 11 September 2013 Copyright © 2013 Anshu Agrawal et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Vaccines still remain the most successful method for pro- tection and eradication against diseases. However, to avoid harmful effects associated with whole organism vaccines, new vaccine candidates are composed of parts of an organism, and therefore these are weakly immunogenic. Adjuvants are essential components of vaccines that nonspecifically stim- ulate the immune system, particularly the innate immune system cells, to enhance the immunogenicity of vaccines. Initially the major function of adjuvants such as alum was to allow sustained presence of antigens by preventing their degradation in vivo. Recent advances have, however, demonstrated that success of alum as an adjuvant is also due to its ability to activate the innate immune system cells. Significant progress in the last decade has increased our understanding of the innate immune system which is highly complex and can be activated via a wide array of receptors to generate different immune responses. The nature of adaptive immune response, quality, and quantity are governed by how the innate immune responses are activated. Novel adjuvants are therefore targeted to receptors expressed on antigen-presenting cells (APCs) such as dendritic cells (DCs) to activate the innate immune system. DCs express an array of pathogen recognition receptors (PRRs) so that they can recognize any threat to the body. Examples of PRRs include the TLRs, CLRs, and NLRs. Apart from preventing infections, vaccinations are also being used as therapy against tumors. Targeting antigens to APCs along with adjuvants allows the induction of immune response against tumors. The manuscript by L. Arribillaga et al., in this tissue, demonstrates that fusion of an antigen to the extra domain A from fibronectin (EDA) targets antigens to TLR4-expressing cells such as DCs leading to their activation. The approach is also effective for cross-presentation as well as the induction of antiviral/tumor immunity. Furthermore, the approach is universal as conjugation of EDA to streptavidin allows any biotinylated antigens to be used as immunogen for vaccination purposes. In contrast to the approach above, N. Kojima et al. use oligomannose-coated liposomes to target and stimulate APCs. The oligomannose on the liposomes targets antigens to the C-type lectin receptor (CLR) and is effective in inducing CTLs and/or Th1 cells. This approach is also universal because both lipophilic and hydrophilic antigens can be incorporated in the liposomes. Contrary to these surface receptor targeting methodologies, C. A. Colaco et al. discuss the merits of using heat shock proteins (HSPs) as vaccine adjuvants because HSPs are not just stimulators of innate immunity but can also traffic antigens into APCs, facil- itating the induction of specific acquired immune responses. Since the new generations of adjuvants being developed are highly specific in the type of CD4T, CD8T, or B-cell response they can stimulate, therefore, recent approaches for vaccine development are focused on utilizing a combi- nation of adjuvants to activate the various components of immune system to generate an effective memory response. The manuscript by E. Mata et al. provides an excellent example of a combinatorial approach in which they discuss that a successful malaria vaccine may require a combination of adjuvants which can activate different parts of the immune system. The review also provides an excellent overview of various types and properties of adjuvants in both preclinical

Immunotherapy with oligomannose‐coated liposomes ameliorates allergic symptoms in a murine food allergy model

Allergen-specific immunotherapy has been anticipated to be a disease-modifying therapy for food allergies. We previously reported that CD8(+) regulatory T cells may prevent antigen-sensitized mice from developing allergic diarrhea. Because oligomannose-coated liposomes (OML) have been shown to induce MHC class I-restricted CD8(+) T cell responses, we analyzed the adjuvant activities of OML for inducing regulatory CD8(+) T cells and mucosal tolerogenic responses in allergen-sensitized mice. The BALB/c mice that were previously sensitized to ovalbumin (OVA) were intranasally immunized with OVA-encased in OML (OVA-OML) or OVA-encased in non-coated liposomes (OVA-NL). We assessed allergic diarrhea induced by oral OVA administration, OVA-specific immunoglobulin production, and cytokine production in the intestines and mesenteric lymph nodes (MLNs). Intranasal immunization with OVA-OML, but not OVA-NL, suppressed the development of allergic diarrhea. This was associated with in vitro Ag-induced IL-10 production and the in vivo expansion of CD8(+) CD28(-) and CD4(+) CD25(+) Foxp3(+) T cell populations among mesenteric lymph node mononuclear cells, and was significantly ablated by anti-SIGNR1 or anti-CR3 mAbs. Up-regulation of serum OVA-specific IgE was suppressed, whereas OVA-specific IgG1, IgG2a, and soluble IgA production were enhanced by intranasal administration of OVA-OML. Adoptive transfer of CD8(+) CD28(-) T cells but not CD28(+) CD8(+) T cells from the MLNs of OVA-OML-treated mice ameliorated the development of diarrhea. These results suggest that intranasal immunization with Ag-encased OML may be an effective immunotherapy for food allergies, as it induces a subset of regulatory CD8(+) T cells as well as CD4(+) CD25(+) Foxp3(+) T cell and modulates humoral immune responses in allergen-sensitized mice.

Liposomes and nanotechnology in drug development: focus on oncotargets

Nanotechnology is the development of an engineered device at the atomic, molecular, and macromolecular level in the nanometer range. Advances in nanotechnology have proven beneficial in therapeutic fields such as drug-delivery and gene/protein delivery. Antigen delivery systems are important for inducing and modifying immune responses. In cellular immunity, cytotoxic T lymphocytes (CTLs) are important in the host defense against tumors. Key to the development of CTL-inducible vaccines is the ability to deliver antigens to antigen-presenting cells efficiently and to induce the subsequent activation of T cell-mediated immunity without adjuvants, as they can induce excessive inflammation leading to systemic febrile disease. Since expression and cloning methods for tumor-associated antigens have been reported, cancer vaccines that induce effective cell immunity may be promising therapeutic candidates, but Th2 cells are undesirable for use in cancer immunotherapy. Peptide vaccines have immunological and economic advantages as cancer vaccines because CTL epitope peptides from tumor-associated antigens have high antigen-specificity. However, cancer vaccines have had limited effectiveness in clinical responses due to the ability of cancer cells to “escape” from cancer immunity and a low efficiency of antigen-specific CTL induction due to immunogenic-free synthetic peptides. In contrast, carbohydrate-decorated particles such as carbohydrate-coated liposomes with encapsulated antigens might be more suitable as antigen delivery vehicles to antigen-presenting cells. Oligomannose-coated liposomes (OML) can eliminate established tumors in mouse cancer models. In addition, OMLs with an encased antigen can induce antigen-specific CTLs from peripheral blood mononuclear cells obtained from patients. Feasibility studies of OML-based vaccines have revealed their potential for clinical use as vaccines for diseases where CTLs act as effector cells. Furthermore, use of the hepatitis B core particle, in which tumor-antigen epitopes are set, has consistently been shown to induce strong CTL responses without the use of an adjuvant. Thus, nanoparticles may provide a new prophylactic strategy for infectious disease and therapeutic approaches for cancer via the induction of T-cell immunity.

New Strategy of Adult T-cell Leukemia Treatment Targeted for Anti-tumor Immunity and a Longevity Gene-encoded Protein

Adult T-cell leukemia-lymphoma (ATL) is an aggressive peripheral T-cell neoplasm with a poor prognosis, developing after long-term infection with human T-cell leukemia virus-1 (HTLV-1). Multiple factors (e.g., virus, host cells, epigenetic aberrations, and immune factors) have been implicated in the development of ATL, although the underlying mechanisms of leukemogenesis have not been fully elucidated. Despite recent progress in both chemotherapy and supportive care for hematological malignancies, the prognosis of ATL is still poor; overall survival at 3 years is only 24%. New strategies for the therapy and prophylaxis of ATL (e.g., vaccines and novel molecular target agents) are still required. This article reviews new strategy of ATL treatment targeted for HTLV-1-specific cytotoxic T-lymphocytes (CTLs) and SIRT1, a longevity gene-encoded protein. HTLV-1-specific CTLs play a critical role in the host immune response against HTLV-1. We have described here the decreased frequency and function of HTLV-1-specific CD8+ T cells in ATL patients and the efficient induction of the HTLV-1-specific CTLs response in human leukocyte antigen-A* 0201-transgenic mice by the HTLV-1/hepatitis B core chimeric particle and oligomannose-coated liposomes encapsulating HTLV-1 epitope without adjuvant, suggesting that the efficient antigen delivery system and CTL induction can be exploited to develop a prophylactic vaccine model against tumors and infectious diseases. Furthermore, our studies suggest that SIRT1, a longevity gene-encoded protein, is a crucial anti-apoptotic molecule in ATL cells, and that SIRT1 inhibitors may be useful therapeutic agents for leukemia, especially in patients with ATL. These studies targeted for anti-tumor immunity such as vaccine and SIRT1 may support the new prophylactic and therapeutic approach for ATL.

Oligomannose‐coated liposomes efficiently induce human T‐cell leukemia virus‐1‐specific cytotoxic T lymphocytes without adjuvant

Human T-cell leukemia virus-1 (HTLV-1) causes adult T-cell leukemia/lymphoma, which is an aggressive peripheral T-cell neoplasm. Insufficient T-cell response to HTLV-1 is a potential risk factor in adult T-cell leukemia/lymphoma. Efficient induction of antigen-specific cytotoxic T lymphocytes is important for immunological suppression of virus-infected cell proliferation and oncogenesis, but efficient induction of antigen-specific cytotoxic T lymphocytes has evaded strategies utilizing poorly immunogenic free synthetic peptides. Here, we examined the efficient induction of an HTLV-1-specific CD8+ T-cell response by oligomannose-coated liposomes (OMLs) encapsulating the human leukocyte antigen (HLA)-A*0201-restricted HTLV-1 Tax-epitope (OML/Tax). Immunization of HLA-A*0201 transgenic mice with OML/Tax induced an HTLV-1-specific gamma-interferon reaction, whereas immunization with epitope peptide alone induced no reaction. Upon exposure of dendritic cells to OML/Tax, the levels of CD86, major histocompatibility complex class I, HLA-A02 and major histocompatibility complex class II expression were increased. In addition, our results showed that HTLV-1-specific CD8+ T cells can be efficiently induced by OML/Tax from HTLV-1 carriers compared with epitope peptide alone, and these HTLV-1-specific CD8+ T cells were able to lyse cells presenting the peptide. These results suggest that OML/Tax is capable of inducing antigen-specific cellular immune responses without adjuvants and may be useful as an effective vaccine carrier for prophylaxis in tumors and infectious diseases by substituting the epitope peptide.

Development of peritoneal macrophage along a dendritic cell lineage in response to uptake of oligomannose-coated liposomes

In this study, we investigate the potential of peritoneal macrophages to differentiate into dendritic cell (DCs) in response to preferential uptake of oligomannose-coated liposomes (OMLs). About 30% of peritoneal cells (PECs) preferentially took up OMLs that were administered into the peritoneal cavity. The OML-ingesting cells expressed CD11b and F4/80, but lacked CD11c expression, indicating that the OML-ingesting PECs with a CD11b highCD11c − phenotype are resident peritoneal macrophages. During in vitro cultivation, CD11c + cells arose among the PECs with ingested OMLs. CD11c + cells also developed among enriched peritoneal CD11b highCD11 − cells from OML-treated mice, and the resulting CD11c + cells expressed co-stimulatory molecules and MHC class II. In addition, OML-ingesting CD11b highCD11c + cells were found in spleen after the enriched peritoneal macrophages with ingested OMLs were transplanted in the peritoneal cavity of mice. These results show that a fraction of peritoneal macrophages can differentiate into mature DCs following uptake of OMLs.