Peptide Aggregation Induced Immunogenic Rupture (PAIIR)

Under the influence of stress and membrane damage, cells undergo immunogenic cell death (ICD), which involves the release of damage associated molecular patterns (DAMPs), natural adjuvants for enhancing an immune response. In the presence of an antigen, released DAMPs can determine the type and magnitude of the immune response, and therefore the longevity and efficacy of an antigen-specific immunity. In the last decade, the immune response effect of ICD has been shown, yet there is no tool that can induce controlled ICD with predictable results, regardless of the cell type. We designed a peptide-based tool, called [II], for controlled damage to cell membrane to induce ICD and DAMPs release. Herein we describe a series of experiments that determine that the mechanism of action of [II] includes a caspase-dependent ICD and subsequent release of immune stimulating DAMPs, on various cell types. Moreover, we tested the hypothesis that controlled DAMP release via [II] in vivo was associated with enhancement of antigen-specific adaptive immunity with influenza hemagglutinin (HA) subunit vaccine. HA and [II] showed significantly higher HA specific IgG1 and IgG2a antibodies, compared to HA-only immunized mice, while the peptide itself did not elicit antibodies. In this paper, we demonstrate the first peptide-aggregation induced immunogenic rupture (PAIIR) approach as vaccine adjuvants for increasing both humoral and cellular immunity. In consideration of its ability to enhance IgG2a responses that are associated with heterosubtypic influenza virus protection, PAIIR is a promising adjuvant to promote universal protection upon influenza HA vaccination.

Download Full-text

The Role of Syk/CARD9-Coupled C-Type Lectin Receptors in Immunity toMycobacterium tuberculosisInfections

Clinical and Developmental Immunology ◽

10.1155/2010/567571 ◽

2010 ◽

Vol 2010 ◽

pp. 1-9 ◽

Cited By ~ 27

Author(s):

Mohlopheni Jackson Marakalala ◽

Lisa M. Graham ◽

Gordon D. Brown

Keyword(s):

Immune Response ◽

Pattern Recognition ◽

Mycobacterium Tuberculosis ◽

Immune Cells ◽

Innate Immune ◽

Innate Immune Cells ◽

Lectin Receptors ◽

Molecular Patterns

There is increasing interest in understanding the mechanisms underlying the interactions that occur betweenMycobacterium tuberculosisand host innate immune cells. These cells express pattern recognition receptors (PRRs) which recognise mycobacterial pathogen-associated molecular patterns (PAMPs) and which can influence the host immune response to the infection. Although many of the PRRs appear to be redundant in the control ofM. tuberculosisinfectionin vivo, recent discoveries have revealed a key, nonredundant, role of the Syk/CARD9 signalling pathway in antimycobacterial immunity. Here we review these discoveries, as well as recent data investigating the role of the Syk/CARD9-coupled PRRs that have been implicated in mycobacterial recognition, including Dectin-1 and Mincle.

Download Full-text

Agonists of Receptors of the Innate Immunity and Defective Viral Particles as New Generation of Adjuvants

Epidemiology and Vaccinal Prevention ◽

10.31631/2073-3046-2018-17-1-76-86 ◽

2018 ◽

Vol 17 (1) ◽

pp. 76-86

Author(s):

O. A. Svitich ◽

V. F. Lavrov ◽

P. I. Kukina ◽

A. A. Iskandaryan ◽

L. V. Gankovskaya ◽

...

Keyword(s):

Immune Response ◽

Innate Immunity ◽

Poly I:C ◽

Vaccine Adjuvants ◽

Toll Like Receptor ◽

Traditional Methods ◽

Viral Particles ◽

Pathogen Recognition Receptors ◽

Molecular Patterns ◽

New Generation

Vaccines for many years act as one of the most effective and successfully used medicines. Vaccines obtained by traditional methods contain in their composition live, weakened or killed microorganisms (bacteria, viruses, etc.). Now more often, modern, split, subunit, recombinant, polyvalent and some other types of vaccines are being used. The addition of adjuvants to vaccines generally increases the immune response to their administration. It was established that the formation of postvaccinal immunity begins immediately after the introduction of the vaccine, by activating the factors of innate immunity in the interaction of pathogen-associated molecular patterns (PAMPs), in vaccines, with the pathogen-recognition receptors (PRRs) of the immunocompetent cells of the recipient. It is also shown that PRRs activators, including TOLL-like receptor agonists (TLRs) and poly (I:C) polynucleotide oligomers of inosine and cytidylic acids, have the ability to substantially increase the immunogenicity of vaccines, and attempts are being made to use them creation of new types of adjuvants. Defective interfering viral particles (D-particles, DIPs) are also classed as effective stimulants of innate immunity and can also be considered promising vaccine adjuvants.

Download Full-text

Targeted Nanotherapy for Cognitive Impairment: Blocking Amyloid-β-Induced Membrane Damage in Brain Tissue

10.20944/preprints201808.0204.v1 ◽

2018 ◽

Author(s):

Joseph D'Arrigo

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Human Subjects ◽

Membrane Damage ◽

Cell Types ◽

Cubic Phase ◽

Type I ◽

Co Morbidity ◽

Cns Penetration

A frequent co-morbidity of cerebrovascular pathology and Alzheimer's disease pathology has been observed over past decades. Accordingly, much evidence has been reported which indicates that microvascular endothelial dysfunction, due to cerebrovascular risk factors (e.g., atherosclerosis, obesity, diabetes, smoking, hypertension, aging), precedes cognitive decline in Alzheimer's disease and contributes to its pathogenesis. By incorporating appropriate drug(s) into biomimetic (lipid cubic phase) nanocarriers, one obtains a multitasking combination therapeutic which targets certain cell-surface scavenger receptors, mainly class B type I (i.e., SR-BI), and crosses the blood-brain barrier (BBB). Such targeting allows for various Alzheimer's-related cell types to be simultaneously searched out, in vivo, for localized drug treatment. This in vivo targeting advantage may be particularly important for repurposing an FDA-approved drug, especially one which has shown the added ability to restore some cognitive functions in certain animal models of Alzheimer's disease (e.g., the anticancer drug bexarotene); this (candidate repurposing) drug up to now, by itself (i.e, without nanocarrier), displayed poor CNS penetration in human subjects.

Download Full-text

Selective Lympholysis: A Unique Method Using the Double-Dye Technique.

Blood ◽

10.1182/blood.v108.11.3884.3884 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3884-3884

Author(s):

William J. Owens ◽

Thomas C. Cesario ◽

Edward Shanbrom

Keyword(s):

Methylene Blue ◽

Whole Blood ◽

Mononuclear Cells ◽

Cell Types ◽

The Novel ◽

Treated Sample ◽

Unique Method ◽

Series Of Experiments ◽

Lymphocytic Cell Line

Abstract Many methods are utilized to destroy mononuclear cells (primarily lymphocytes) either for neoplasm or immunosuppression. Neither radiation nor chemotherapy are truly selective or completely successful. The concept of “Double-Dye” treatment of blood for transfusion has been developed in order to inactivate parasites, bacteria and viruses (J Thromb Haemost2003; 1 Supplement 1 July: P1114). In recent studies, it has been observed that this same “Double-Dye” concept presents the possibility of very selectively eliminating lymphocytes (mononuclear cells) without affecting neutrophils in whole blood. To demonstrate the selectivity of dyes for lymphocytic/mononuclear cell types, two sets of experiments were performed. In the first, 0.3% (w/v) of the “Double Dye’ solution was added to several normal citrated whole blood samples to assess the effect on normal cells, compared to an untreated control. At 24hours post treatment, the lymphocyte count in the treated sample had dropped more than 80%, while little effect on neutrophils was noted. The control counts showed little change for either lymphocytes or neutrophils. Table 1. Lymphocyte reduction by 0.3%(w/v) Double-dye solution (units: cells/mm3). 0hr Control 0.3% Dye WBC 6850 ± 560 6920 ± 630 Neut 5030 ± 375 5100 ± 420 Lymph 1485 ± 220 1490 ± 265 24hr Control 0.3% Dye WBC 6700 ± 480 5300 ± 505 Neut 5025 ± 360 5025 ± 420 Lymph 1474 ± 240 275 ± 75 In the second series of experiments, 0.3%(w/v) “Double-Dye” solution or 0.15%(w/v) Crystal Violet or 0.15%(w/v) Methylene Blue were added to two T-cell leukemia lines (Jurkat, L1210), with a non-malignant, non-lymphocytic cell line (WISH) for the control. The combination of dyes showed the most potent activity against the lymphocytic lines, while the control was virtually unaffected. Table 2: Viability of cell lines after 24 hour exposure to dye solutions. Jurkat L1210 WISH Control 100% ± 2% 100% ± 4% 99% ± 2% 0.3% Double Dye 37% ± 5% 12% ± 4% 88% ± 9% 0.15% Meth. Blue 58% ± 12% 52% ± 9% 100% ± 3% 0.15% Cr. Violet 90% ± 12% 92% ± 10% 99% ± 4% The novel use of these dyes reported here coincided with the recent interest in utilizing methylene blue to increase transfusion safety, but recognizing that the concurrent need to photoactivate was too toxic to certain proteins and didn’t inactivate all pathogens (Transfusion2003; 43(9): 1238–47). Studies investigating the in-vivo efficacy of these novel immunosuppressive and chemotherapeutic methods are currently underway.

Download Full-text

Squalene-Based Influenza Vaccine Adjuvants and Their Impact on the Hemagglutinin-Specific B Cell Response

Pathogens ◽

10.3390/pathogens10030355 ◽

2021 ◽

Vol 10 (3) ◽

pp. 355

Author(s):

Phuong Nguyen-Contant ◽

Mark Y. Sangster ◽

David J. Topham

Keyword(s):

Immune Response ◽

Influenza Vaccine ◽

B Cell ◽

Rational Design ◽

Influenza Vaccines ◽

Vaccine Adjuvants ◽

Design And Optimization ◽

Cell Responses ◽

Influenza Hemagglutinin ◽

B Cell Responses

Influenza infections continue to cause significant annual morbidity and mortality despite ongoing influenza vaccine research. Adjuvants are administered in conjunction with influenza vaccines to enhance the immune response and strengthen protection against disease. Squalene-based emulsion adjuvants including MF59, AS03, and AF03, are registered for administration with influenza vaccines and are widely used in many countries. Squalene-based emulsion adjuvants induce a strong innate immune response, enhancing antigen presentation both quantitively and qualitatively to generate strong B cell responses and antibody production. They also diversify the reactivity profiles and strengthen the affinities of antibodies against the influenza hemagglutinin, increasing protection across virus clades. In this review, we consider the mechanisms of the enhancement of innate and adaptive immune responses by squalene-based emulsionSE adjuvants and the resulting increase in magnitude and breadth of hemagglutinin-specific B cell responses. We relate observed effects of SE adjuvants and current mechanistic understandings to events in responding lymph nodes. These insights will guide the rational design and optimization of influenza vaccines to provide broad and effective protection.

Download Full-text

T- and B-cell immunosuppressive activity of novel α-santonin analogs with humoral and cellular immune response in Balb/c mice

MedChemComm ◽

10.1039/c6md00527f ◽

2017 ◽

Vol 8 (1) ◽

pp. 211-219 ◽

Cited By ~ 6

Author(s):

Nisar A. Dangroo ◽

Jasvinder Singh ◽

Nidhi Gupta ◽

Shashank Singh ◽

Anapurna Kaul ◽

...

Keyword(s):

Immune Response ◽

B Cell ◽

Cellular Immunity ◽

Cellular Immune Response ◽

Immunosuppressive Activity ◽

Humoral And Cellular Immunity ◽

Cellular Immune

Herein we report the synthesis of α-santonin analogs, and identification of potent immunosuppressant molecules. In vivo investigation on BALB/c mice revealed that compound 4e suppresses both humoral and cellular immunity.

Download Full-text

Adenovirus fibre exchange alters cell tropism in vitro but not transgene-specific T CD8+ immune responses in vivo

Journal of General Virology ◽

10.1099/vir.0.79846-0 ◽

2004 ◽

Vol 85 (5) ◽

pp. 1227-1236 ◽

Cited By ~ 5

Author(s):

S. Mercier ◽

S. Verhaagh ◽

J. Goudsmit ◽

A. Lemckert ◽

M. Monteil ◽

...

Keyword(s):

Immune Response ◽

Endothelial Cells ◽

Dendritic Cells ◽

Cytotoxic T Lymphocyte ◽

Cell Types ◽

Relative Role ◽

Intramuscular Inoculation ◽

Lymphocyte Responses

Gene transfer with recombinant adenoviruses (rAds) is a powerful means of inducing an immune response against a transgene product. However, little is known about the mechanisms that underlie the induction of the immune response after intramuscular inoculation of adenovirus and, in particular, the relative role of the different cell types transduced. Several studies have suggested that CD8+ cytotoxic T lymphocyte responses elicited after inoculation of adenoviruses (Ads) are induced both by direct transduction of antigen presenting cells (APCs) and by cross-priming. In the present study, a library of fibre-chimeric rAds was screened in order to identify rAds with distinct capacities to express transgene product in murine cell types naturally found in muscle, i.e. myoblasts, endothelial cells (both representing non-APCs) and dendritic cells (representing APCs). Four selected pseudotypes, differing in their ability to infect muscular cells were used to immunize C57BL/6 mice. The relationship between the capacity to transduce non-APC or APC in vitro and the ability to induce humoral and cellular responses against the β-galactosidase antigen after intramuscular inoculation were studied. Results indicate that CD8+ T cell responses against the β-galactosidase antigen were similar after inoculation of the four viruses, thus revealing no direct relationship with their ability to transduce myoblasts, endothelial cells or dendritic cells in vitro.

Download Full-text

In vivo reprogramming of murine host immune response genes following Leishmania major infection.

10.1101/2021.10.05.463063 ◽

2021 ◽

Author(s):

Gopinath Venugopal ◽

Jordan T. Bird ◽

Charity L. Washam ◽

Hayden Roys ◽

Anne Bowlin ◽

...

Keyword(s):

Gene Expression ◽

Immune Response ◽

Antigen Presentation ◽

Rna Sequencing ◽

Cell Types ◽

Host Immune Response ◽

Rna Seq ◽

Major Infection ◽

Presentation Pathway

Leishmania parasites cause cutaneous leishmaniasis (CL), a pathologic disease characterized by disfiguring, ulcerative skin lesions. Both parasite and host gene expression following infection with various Leishmania species has been investigated in vitro, but global transcriptional analysis following L. major infection in vivo is lacking. Thus, we conducted a comprehensive transcriptomic profiling study combining bulk RNA sequencing (RNA-Seq) and single-cell RNA sequencing (scRNA-Seq) to identify global changes in gene expression in vivo following L. major infection. Bulk RNA-Seq analysis revealed that host immune response pathways like the antigen processing and presentation pathway were significantly enriched amongst differentially expressed genes (DEGs) upon infection, while ribosomal pathways were significantly downregulated in infected mice compared to naive controls. scRNA-Seq analyses revealed cellular heterogeneity including distinct resident and recruited cell types in the skin following murine L. major infection. Within the individual immune cell types, several DEGs indicative of many interferon induced GTPases and antigen presentation molecules were significantly enhanced in the infected ears including macrophages (Gbp2, H2-K1, H2-Aa, H2-Ab1), resident macrophages (H2-K1, H2-D1, Gbp4, Gbp8, Gbp2), and inflammatory monocytes (Gbp2, Gbp5, Gbp7, Gbp3). Ingenuity Pathway Analysis of scRNA-Seq data indicated the antigen presentation pathway was increased with infection, while EIF2 signaling is the top downregulated pathway followed by eIF4/p70S6k and mTOR signaling in multiple cell types including macrophages, BECs, and LECs. Altogether, this transcriptomic profile highlights known recruitment of myeloid cells to lesions and recognizes a previously undefined role for EIF2 signaling in murine L. major infection in vivo.

Download Full-text

Ultrastructural Study of a differentiating human colon carcinoma cell line

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100158583 ◽

1990 ◽

Vol 48 (3) ◽

pp. 208-209

Author(s):

Sylvie Polak-Charcon ◽

Mehrdad Hekmati ◽

Yehuda Ben Shaul

Keyword(s):

Cell Line ◽

Morphological Changes ◽

Secretory Granules ◽

Human Colon ◽

Cell Types ◽

Culture Conditions ◽

Morphological Evidence ◽

Human Colon Carcinoma Cell ◽

Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

Download Full-text

Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162132 ◽

1990 ◽

Vol 48 (3) ◽

pp. 914-915

Author(s):

D.J.P. Ferguson ◽

A.R. Berendt ◽

J. Tansey ◽

K. Marsh ◽

C.I. Newbold

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Umbilical Vein ◽

Cell Types ◽

Amelanotic Melanoma ◽

Cytoplasmic Process ◽

Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

Download Full-text