scholarly journals Sublingual Immunization with Chimeric C1q/CD40 Ligand/HIV Virus-like Particles Induces Strong Mucosal Immune Responses against HIV

Vaccines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1236
Author(s):  
Dongliang Liu ◽  
Sheng Zhang ◽  
Ethan Poteet ◽  
Christian Marin-Muller ◽  
Changyi Chen ◽  
...  
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Sexual Transmission ◽  
Cd40 Ligand ◽  
Sublingual Administration ◽  
Epithelial Layer ◽  
Activation Markers ◽  
Effective Prevention ◽  
Mucosal Vaccines ◽  
Mucosal Immune Responses

Development of a vaccine that can elicit robust HIV specific antibody responses in the mucosal compartments is desired for effective prevention of HIV via sexual transmission. However, the current mucosal vaccines have either poor immunogenicity when administered orally or invite safety concerns when administered intranasally. Sublingual immunization has received more attention in recent years based on its efficiency in inducing systemic and mucosal immune responses in both mucosal and extra-mucosal tissues. To facilitate the transport of the immunogen across the sub-mucosal epithelial barrier, we found that CD91, the receptor of C1q, is prevalently expressed in the sublingual mucosal lining, and thus, a modified chimeric C1q surface conjugated CD40L/HIV VLP was generated. The ability of this chimeric C1q/CD40L/HIV VLP to bind, cross the epithelial layer, access and activate the sub-mucosal layer dendritic cells (DCs), and ultimately induce enhanced mucosal and systemic immune responses against HIV is evaluated in this study. We found that C1q/CD40L/HIV VLPs have enhanced binding, increased transport across the epithelial layer, and upregulate DC activation markers as compared to CD40L/HIV VLPs alone. Mice immunized with C1q/ CD40L/HIV VLPs by sublingual administration showed higher levels of IgA salivary antibodies against both HIV Gag and Env than mice immunized with CD40L/HIV VLPs. Moreover, sublingual immunization with C1q/CD40L/HIV VLPs induced more Env- and Gag-specific IFN-γ producing T cells than the CD40L/HIV VLPs group. Interestingly, C1q/CD40L/HIV VLP immunization can also induce more mucosal homing T cells than that in CD40L/HIV VLP group. Our data suggest that incorporation of C1q to CD40L/HIV VLPs is a promising novel strategy and that the sublingual immunization can be a favorite immunization route for HIV mucosal vaccines.

Age Influences Recovery of Systemic and Mucosal Immune Responses Following Acute Depletion of CD4 T Cells

1993 ◽  
Vol 69 (3) ◽  
pp. 285-291 ◽  
Author(s):  
Audrey L. Fleming ◽  
Elizabeth H. Field ◽  
Naser Tolaymat ◽  
John S. Cowdery
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Cd4 T Cells ◽  
Mucosal Immune Responses

scholarly journals Regulation of Mucosal Immune Responses – The Missing Link in IBD?

1996 ◽  
Vol 10 (2) ◽  
pp. 105-109
Author(s):  
Charles O Elson ◽  
Robert P Mccabe ◽  
Kenneth W Beagley ◽  
Almaz Sharmanov ◽  
Steven L Brandwein ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Immune Responses ◽  
Intestinal Inflammation ◽  
Bacterial Flora ◽  
Cell Subset ◽  
Experimental Models ◽  
Bacterial Antigens ◽  
Mucosal Immune Responses ◽  
Chronic Intestinal Inflammation

Although the etiology of inflammatory bowel disease (IBD) remains unknown, a major working hypothesis is that it represents a dysregulated immune response to common enteric bacterial antigens. Until recently there has been a relative dearth of experimental models to study this hypothesis. However, exciting developments in experimental models of colitis, including spontaneous, transgenic and knockout mice, now allow this and other hypotheses to be tested. The regulation of mucosal immune responses is not well understood in the normal animal, much less in those with chronic intestinal inflammation. Clearly the CD4 Th1 and Th2 pathways are important in the host response to microbial pathogens, and recent data indicate that the intestinal mucosa seems to be a site of preferential Th2 responses toward exogenous antigens. Deletion of certain cytokine genes involved in maintaining this Th1/Th2 balance (interleukin [IL]-2, IL-10) resulted in colitis, although deletion of others (IL-4, interferon-gamma) that are also involved did not. Whether these cytokine gene deletions cause a dysregulation of the mucosal immune response has yet to be shown. However, the importance of regulation can be demonstrated in a model in which a normal CD4+T cell subset (CD45Rbhigh) is transferred into syngeneic severe combined immunodeficiency syndrome recipients. This results in a striking colitis over the ensuing weeks with chronic diarrhea and wasting of the animals. If the reciprocal CD4+subset (CD45Rblow) is co-transferred or if whole CD4+T cells are transferred no colitis ensues. Therefore, T cells capable of causing colitis are present in normal animals but are prevented from doing so by immunoregulatory mechanisms. The antigens that drive the colitis in several of these models (IL-2 knockout mouse, human leukocyte antigen B27/β2M transgenic rat) appear to be those of the normal enteric bacterial flora because germ-free animals do not get the disease. Spontaneously colitic C3H/HeJBir mice also show prominent reactivity to enteric bacterial antigens. There are major differences among inbred mouse strains in susceptibility to colitis. The genes involved are not yet identified, but newly available technologies should allow that. In summary, these new models provide an experimental foundation to one of the major hypotheses on the cause of IBD, and will allow dissection of the genetic, environmental and immune components contributing to chronic colitis.

Development of CD40L-modified tumor small extracellular vesicles for effective induction of antitumor immune response

Nanomedicine ◽  
2020 ◽  
Vol 15 (17) ◽  
pp. 1641-1652
Author(s):  
Wen Liu ◽  
Yuki Takahashi ◽  
Masaki Morishita ◽  
Makiya Nishikawa ◽  
Yoshinobu Takakura
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Dendritic Cells ◽  
Immune Responses ◽  
Antigen Presentation ◽  
Extracellular Vesicles ◽  
Tumor Antigen ◽  
Tumor Antigens ◽  
Cd40 Ligand ◽  
Further Development

Aim: Tumor-derived small extracellular vesicles (TEVs) are considered for use in inducing tumor antigen-specific immune responses as they contain tumor antigens. The delivery of tumor antigens to the antigen presentation cells (especially dendritic cells [DCs]), and the activation of DCs are the main challenges of TEV therapy. Materials & methods: TEVs were modified with CD40 ligand (CD40L), which can target CD40 expressed on the surface of DCs and can activate them via CD40L-CD40 interactions. Results: It was found that CD40L-TEVs were efficiently taken up by DCs and also activated them. Moreover, tumor antigens were efficiently presented to the T cells by DCs treated with CD40L-TEVs. Conclusion: This study proved that CD40L-modification of TEVs will be helpful for further development of TEV-based tumor vaccination.

scholarly journals Profiling for Monocyte-Regulated T-Cell Immune Responses in Human PBMC Samples: Simultaneous Assessments of Cell Surface Markers and Cytokine Secretion

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5730-5730 ◽  
Author(s):  
Zhaoping Liu
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Cell Viability ◽  
Dose Response ◽  
Cell Activation ◽  
Conflicts Of Interest ◽  
Cytokine Secretion ◽  
Activation Markers ◽  
Specific Patient

Abstract Advances in immuno-oncology have accelerated the need for higher content cell analysis tools that are capable of reducing the time needed to actionable results. The complexity of immune responses necessitates that data should yield insight into both the context of mechanism (cell activation markers) and pathways (signaling molecules) for how patients would respond to a certain treatments, or which therapies are most effective for a specific patient. Traditionally, cellular endpoints and secreted cytokines have been measured using two separate assays on two different analysis platforms: cellular endpoints measured by flow cytometry or imaging, and secreted cytokines measured by plate-based or bead-based ELISA. IntelliCyt's iQue Screener PLUS with 3 lasers and 13 fluorescent channels, was built as an immunology profiling tool and can measure cellular activation markers and secreted cytokine levels in a single assay in under 20 minutes for a 384-well plate. In a highlighted example showing monocyte regulation of T cells, we profiled the immune responses of three different patients across seven different cell activation markers, cell viability, and secretion of IFNγ, TNF, and IL-10 in response to several immunogenic compounds. PBMCs from 3 individual healthy donors were treated with the monocyte activator lipopolysaccharide (LPS) and/or with T cell activator staphylococcal enterotoxin B (SEB) in duplicate dose response. Cell and compounds were incubated for 24 hours, then analyzed for viability, surface expression of CD3, CD4, CD8, CD14, CD25, CD45 and CD127, and cytokine secretion using the QBeads reagent kits (IntelliCyt Corp). ForeCyt software was utilized for data acquisition and analysis, where sequential gates were used to individually identify cells and beads from the samples based on size. The cells were further analyzed for viability and cell marker expression, and QBeads were further segregated into the individual detection beads for each cytokine. The media concentration of each cytokine was calculated via the use of a standard curve. Utilizing the built-in plate level analytics of ForeCyt, we generated 27 unique cytokine curves from a single 384 well plate, and were able to assess the differences in cytokine secretion for each of the 3 cytokines, across the 3 treatments (LPS alone, SEB alone, LPS+SEB), for all 3 donors. The cell-based measurements yielded 54 dose-response curves detailing the percentage of regulatory T-cells, T-helper cells, cytotoxic T-cells, monocytes, lymphocytes, and cell viability for each treatment/patient. The ability to simultaneously measure cell and bead based endpoints from a single sample reduces assay-to-assay variability and conserves the amount of patient cells required for testing. Taken together, these results highlight the power of the iQue Screener PLUS platform as an immunological profiling tool with not only the speed of sampling required for large-scale combinatorial studies, but the analysis power to quickly transform raw data into clinically relevant results for individual patients. Disclosures No relevant conflicts of interest to declare.

scholarly journals Incorporation of CD40 Ligand into the Envelope of Pseudotyped Single-Cycle Simian Immunodeficiency Viruses Enhances Immunogenicity

2008 ◽  
Vol 83 (3) ◽  
pp. 1216-1227 ◽  
Author(s):  
Fan-ching Lin ◽  
Yue Peng ◽  
Leslie A. Jones ◽  
Paulo H. Verardi ◽  
Tilahun D. Yilma
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Cd40 Ligand ◽  
Interleukin 12 ◽  
Vaccine Strategy ◽  
Aids Vaccine ◽  
Cytokines And Chemokines ◽  
Hla Dr ◽  
Aids Pandemic

ABSTRACT A vaccine for the prevention of human immunodeficiency virus (HIV) infection is desperately needed to control the AIDS pandemic. To address this problem, we developed vesicular stomatitis virus glycoprotein-pseudotyped replication-defective simian immunodeficiency viruses (dSIVs) as an AIDS vaccine strategy. The dSIVs retain characteristics of a live attenuated virus without the drawbacks of potential virulence caused by replicating virus. To improve vaccine immunogenicity, we incorporated CD40 ligand (CD40L) into the dSIV envelope. CD40L is one of the most potent stimuli for dendritic cell (DC) maturation and activation. Binding of CD40L to its receptor upregulates expression of major histocompatibility complex class I, class II, and costimulatory molecules on DCs and increases production of proinflammatory cytokines and chemokines, especially interleukin 12 (IL-12). This cytokine polarizes CD4+ T cells to Th1-type immune responses. DC activation and mixed lymphocyte reaction (MLR) studies were performed to evaluate the immunogenicity of CD40L-dSIV in vitro. Expression levels of CD80, CD86, HLA-DR, and CD54 on DCs transduced with the dSIV incorporating CD40L (CD40L-dSIV) were significantly higher than on those transduced with dSIV. Moreover, CD40L-dSIV-transduced DCs expressed up to 10-fold more IL-12 than dSIV-transduced DCs. CD40L-dSIV-transduced DCs enhanced proliferation and gamma interferon secretion by naive T cells in an MLR. In addition, CD40L-dSIV-immunized mice exhibited stronger humoral and cell-mediated immune responses than dSIV-vaccinated animals. The results show that incorporating CD40L into the dSIV envelope significantly enhances immunogenicity. As a result, CD40L-dSIVs can be strong candidates for development of a safe and highly immunogenic AIDS vaccine.

scholarly journals An Attenuated Duck Plague Virus (DPV) Vaccine Induces both Systemic and Mucosal Immune Responses To Protect Ducks against Virulent DPV Infection

2014 ◽  
Vol 21 (4) ◽  
pp. 457-462 ◽  
Author(s):  
Juan Huang ◽  
Renyong Jia ◽  
Mingshu Wang ◽  
Bing Shu ◽  
Xia Yu ◽  
...  
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Vaccine Strain ◽  
Severe Disease ◽  
Harderian Gland ◽  
Bursa Of Fabricius ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mucosal Immune Responses ◽  
Subcutaneous Immunization ◽  
Main Routine

ABSTRACTDuck plague (DP) is a severe disease caused by DP virus (DPV). Control of the disease is recognized as one of the biggest challenges in avian medicine. Vaccination is an efficient way to control DPV, and an attenuated vaccine is the main routine vaccine. The attenuated DPV vaccine strain CHa is a modified live vaccine, but the systemic and mucosal immune responses induced by this vaccine have been poorly understood. In this study, the immunogenicity and efficacy of the vaccine were evaluated after subcutaneous immunization of ducks. CD4+and CD8+T cells were counted by flow cytometry, and humoral and mucosal Ig antibodies were analyzed by enzyme-linked immunosorbent assay (ELISA). The results showed that high levels of T cells and Ig antibodies were present postimmunization and that there were more CD4+T cells than CD8+T cells. Titers of humoral IgG were higher than those of humoral IgA. Local IgA was found in each sample, whereas local IgG was found only in the spleen, thymus, bursa of Fabricius, harderian gland, liver, bile, and lung. In a protection assay, the attenuated DPV vaccine completely protected ducks against 1,000 50% lethal doses (LD50) of the lethal DPV strain CHv via oral infection. These data suggest that this subcutaneous vaccine elicits sufficient systemic and mucosal immune responses against lethal DPV challenge to be protective in ducks. This study provides broad insights into understanding the immune responses to the attenuated DPV vaccine strain CHa through subcutaneous immunization in ducks.

scholarly journals Flexible Antigen-Specific Redirection of Human Regulatory T Cells Via a Novel Universal Chimeric Antigen Receptor System

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3494-3494 ◽  
Author(s):  
Stefanie Koristka ◽  
Marc Cartellieri ◽  
Anja Feldmann ◽  
Claudia Arndt ◽  
Simon Loff ◽  
...  
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Target Cells ◽  
In Vitro Cultivation ◽  
Antigen Specificity ◽  
Activation Markers ◽  
Peptide Epitope ◽  
Vast Number ◽  
Treg Subset

Abstract Based on compelling evidence from a vast number of in vitro and in vivostudies, Tregs have become an attractive cell population to treat or even prevent auto- and alloimmunity including Graft-versus-Host disease (GvHD). However, several safety concerns still exist as for example the risk of global immunosuppression using polyclonal Tregs. In fact, experiments in mice showed that adoptive transfer or induction of antigen-specific Tregs is more potent regarding suppression of pathogenic immune responses when compared to polyclonal Treg populations. Unfortunately, the isolation and expansion of naturally occurring antigen-specific Tregs is technically difficult, labour-intensive, and time-consuming. An attractive way to overcome these limitations and to endow polyclonal Treg populations with a desired antigen-specificity is their engraftment with chimeric antigen receptors (CARs). In this context, CAR-modification represents a promising approach to redirect polyclonal Tregs in an antigen-specific manner to suppress ongoing self-destructive immune responses at the site of inflammation. Nevertheless, until now redirection of CAR-engineered T cells is limited to a single target antigen, restricting this approach to an unflexible monospecific therapy. Therefore, we developed a more flexible universal CAR (UCAR) platform that allows redirection of T cells to an in principal unrestricted number of surface antigens. T cells are engrafted with UCARs that bind to a small peptide epitope derived from a human nuclear protein. Cross-linkage to target cells is mediated by independent target modules that provide antigen-specificity and comprise the peptide epitope recognized by the UCAR. In order to target different tissue antigens, the target modules can easily be exchanged. Thereby, once established, the treatment strategy can easily be applied to various auto- and alloimmune diseases. At present, the CD45RA+ population is the Treg subset of choice for a clinical application as these cells have the highest capacity to maintain phenotypic and functional Treg properties upon prolonged ex vivo expansion. Here we show that highly pure, sorted CD4+CD25+CD127lowCD45RA+ Tregs can be genetically manipulated using lentiviral gene transfer, resulting in approximately 70 % of UCAR-expressing Treg cells. The transduction procedure itself did not affect the phenotype of UCAR-engineered Tregs as it was similar to non-transduced wildtype cells. Both Treg populations presevered FOXP3 expression even after prolonged in vitro cultivation (> 95 % FOXP3+). Upon incubation with antigen-positive target cells and a respective target module UCAR-engineered Tregs upregulate the activation markers CD69 and LAP demonstrating that the cells can be restimulated antigen-specifically. Most importantly, UCAR-engrafted Tregs were functionally activated upon antigen encounter, demonstrated by suppression of proliferation and expansion of cocultured autologous T effector cells. Taken together, our results pave the way towards an application of UCAR technology for a site-specific recruitment of CAR-modified Tregs into inflamed tissues aiming at re-establishing immune homeostasis. Due to its high flexibility UCAR-engrafted Tregs can easily and universally be used for treatment of various autoimmune diseases or GvHD just by exchanging the tissue-specific target modules. Disclosures Cartellieri: Cellex Patient Treatment GmbH: Employment. Ehninger:GEMoaB GmbH: Employment, Patents & Royalties. Ehninger:GEMoaB GmbH: Consultancy, Patents & Royalties. Bachmann:GEMoaB GmbH: Consultancy, Patents & Royalties.

scholarly journals A detailed analysis of innate and adaptive immune responsiveness upon infection with Salmonella enterica serotype Enteritidis in young broiler chickens

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Nathalie Meijerink ◽  
Robin H. G. A. van den Biggelaar ◽  
Daphne A. van Haarlem ◽  
J. Arjan Stegeman ◽  
Victor P. M. G. Rutten ◽  
...  
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Nk Cells ◽  
Salmonella Enterica ◽  
Broiler Chickens ◽  
Immune Modulation ◽  
Adaptive Immune Responses ◽  
Activation Markers ◽  
Adaptive Immune ◽  
Adaptive Immune Cells

AbstractSalmonella enterica serotype Enteritidis (SE) is a zoonotic pathogen which causes foodborne diseases in humans as well as severe disease symptoms in young chickens. More insight in innate and adaptive immune responses of chickens to SE infection is needed to understand elimination of SE. Seven-day-old broiler chickens were experimentally challenged with SE and numbers and responsiveness of innate and adaptive immune cells as well as antibody titers were assessed. SE was observed in the ileum and spleen of SE-infected chickens at 7 days post-infection (dpi). At 1 dpi numbers of intraepithelial cytotoxic CD8+ T cells were significantly increased alongside numerically increased intraepithelial IL-2Rα+ and 20E5+ natural killer (NK) cells at 1 and 3 dpi. At both time points, activation of intraepithelial and splenic NK cells was significantly enhanced. At 7 dpi in the spleen, presence of macrophages and expression of activation markers on dendritic cells were significantly increased. At 21 dpi, SE-induced proliferation of splenic CD4+ and CD8+ T cells was observed and SE-specific antibodies were detected in sera of all SE-infected chickens. In conclusion, SE results in enhanced numbers and activation of innate cells and we hypothesized that in concert with subsequent specific T cell and antibody responses, reduction of SE is achieved. A better understanding of innate and adaptive immune responses important in the elimination of SE will aid in developing immune-modulation strategies, which may increase resistance to SE in young broiler chickens.

scholarly journals Distinct Cytokine Regulation by Cholera Toxin and Type II Heat-Labile Toxins Involves Differential Regulation of CD40 Ligand on CD4+ T Cells

2001 ◽  
Vol 69 (7) ◽  
pp. 4486-4492 ◽  
Author(s):  
Michael Martin ◽  
Daniel J. Metzger ◽  
Suzanne M. Michalek ◽  
Terry D. Connell ◽  
Michael W. Russell
Keyword(s):  
T Cells ◽  
Cholera Toxin ◽  
Interleukin 2 ◽  
Cd40 Ligand ◽  
Type I ◽  
Type Ii ◽  
Activation Markers ◽  
Cytokine Profiles ◽  
Heat Labile ◽  
Tnf Α

ABSTRACT Cholera toxin (CT) and the type II heat-labile enterotoxins (HLT) LT-IIa and LT-IIb act as potent systemic and mucosal adjuvants and induce distinct T-helper (Th)-cell cytokine profiles. In the present study, CT and the type II HLT were found to differentially affect cytokine production by anti-CD3-stimulated human peripheral blood mononuclear cells (PBMC), and the cellular mechanisms responsible were investigated. CT suppressed interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-α), and IL-12 production by PBMC cultures more than either LT-IIa or LT-IIb. CT but not LT-IIa or LT-IIb reduced the expression of CD4+ T-cell surface activation markers (CD25 and CD69) and subsequent proliferative responses of anti-CD3-stimulated T cells. CT but not LT-IIa or LT-IIb significantly reduced the expression of CD40 ligand (CD40L) on CD4+ T cells. In a coculture system, CT-treated CD4+ T cells induced significantly less TNF-α and IL-12 p70 production by both autologous monocytes and monocyte-derived dendritic cells than either LT-IIa- or LT-IIb-treated CD4+ T cells. These findings demonstrate that CT, LT-IIa, and LT-IIb differentially affect CD40-CD40L interactions between antigen-presenting cells and T cells and help explain the distinct cytokine profiles observed with type I and type II HLT when used as mucosal adjuvants.

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