scholarly journals Peptides identified on monocyte-derived dendritic cells: a marker for clinical immunogenicity to FVIII products

2019 ◽  
Vol 3 (9) ◽  
pp. 1429-1440 ◽  
Author(s):  
Wojciech Jankowski ◽  
Yara Park ◽  
Joseph McGill ◽  
Eugene Maraskovsky ◽  
Marco Hofmann ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Mhc Class Ii ◽  
Unmet Need ◽  
Specific Protein ◽  
Proteolytic Processing ◽  
T Cell Epitopes ◽  
Mhc Ii ◽  
Healthy Donors ◽  
Recombinant Fviii ◽  
Von Willebrand

Abstract The immunogenicity of protein therapeutics is an important safety and efficacy concern during drug development and regulation. Strategies to identify individuals and subpopulations at risk for an undesirable immune response represent an important unmet need. The major histocompatibility complex (MHC)–associated peptide proteomics (MAPPs) assay directly identifies the presence of peptides derived from a specific protein therapeutic on a donor’s MHC class II (MHC-II) proteins. We applied this technique to address several questions related to the use of factor VIII (FVIII) replacement therapy in the treatment of hemophilia A (HA). Although >12 FVIII therapeutics are marketed, most fall into 3 categories: (i) human plasma-derived FVIII (pdFVIII), (ii) full-length (FL)–recombinant FVIII (rFVIII; FL-rFVIII), and (iii) B-domain–deleted rFVIII. Here, we investigated whether there are differences between the FVIII peptides found on the MHC-II proteins of the same individual when incubated with these 3 classes. Based on several observational studies and a prospective, randomized, clinical trial showing that the originally approved rFVIII products may be more immunogenic than the pdFVIII products containing von Willebrand factor (VWF) in molar excess, it has been hypothesized that the pdFVIII molecules yield/present fewer peptides (ie, potential T-cell epitopes). We have experimentally tested this hypothesis and found that dendritic cells from HA patients and healthy donors present fewer FVIII peptides when administered pdFVIII vs FL-rFVIII, despite both containing the same molar VWF excess. Our results support the hypothesis that synthesis of pdFVIII under physiological conditions could result in reduced heterogeneity and/or subtle differences in structure/conformation which, in turn, may result in reduced FVIII proteolytic processing relative to FL-rFVIII.

scholarly journals B cell survival, surface BCR and BAFFR expression, CD74 metabolism, and CD8− dendritic cells require the intramembrane endopeptidase SPPL2A

2012 ◽  
Vol 210 (1) ◽  
pp. 31-40 ◽  
Author(s):  
Hannes Bergmann ◽  
Mehmet Yabas ◽  
Alanna Short ◽  
Lisa Miosge ◽  
Nadine Barthel ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
B Cell ◽  
B Lymphocyte ◽  
B Cell Lymphoma ◽  
Cell Types ◽  
Proteolytic Processing ◽  
Mhc Ii ◽  
Humoral Immunodeficiency ◽  
B Cell Subsets ◽  
New Treatments

Druggable proteins required for B lymphocyte survival and immune responses are an emerging source of new treatments for autoimmunity and lymphoid malignancy. In this study, we show that mice with an inactivating mutation in the intramembrane protease signal peptide peptidase–like 2A (SPPL2A) unexpectedly exhibit profound humoral immunodeficiency and lack mature B cell subsets, mirroring deficiency of the cytokine B cell–activating factor (BAFF). Accumulation of Sppl2a-deficient B cells was rescued by overexpression of the BAFF-induced survival protein B cell lymphoma 2 (BCL2) but not BAFF and was distinguished by low surface BAFF receptor and IgM and IgD B cell receptors. CD8-negative dendritic cells were also greatly decreased. SPPL2A deficiency blocked the proteolytic processing of CD74 MHC II invariant chain in both cell types, causing dramatic build-up of the p8 product of Cathepsin S and interfering with earlier steps in CD74 endosomal retention and processing. The findings illuminate an important role for the final step in the CD74–MHC II pathway and a new target for protease inhibitor treatment of B cell diseases.

scholarly journals Immunohistochemical and Molecular Characterization of the Human Periosteum

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Sönke Percy Frey ◽  
Hendrik Jansen ◽  
Stefanie Doht ◽  
Luis Filgueira ◽  
Rene Zellweger
Keyword(s):  
Stem Cells ◽  
Dendritic Cells ◽  
Mhc Class Ii ◽  
Immune Cells ◽  
Precursor Cells ◽  
Rt Pcr ◽  
Phenotypic Properties ◽  
Mhc Ii ◽  
Cambium Layer ◽  
Stromal Stem Cells

Purpose. The aim of the present study was to characterize the cell of the human periosteum using immunohistological and molecular methods.Methods. Phenotypic properties and the distribution of the cells within the different layers were investigated with immunohistochemical staining techniques and RT-PCR, focussing on markers for stromal stem cells, osteoblasts, osteoclasts and immune cells.Results. Immunohistochemical results revealed that all stained cells were located in the cambium layer and that most cells were positive for vimentin. The majority of cells consisted of stromal stem cells and osteoblastic precursor cells. The density increased towards the deeper layers of the cambium. In addition, cells positive for markers of the osteoblast, chondrocyte, and osteoclast lineages were found. Interestingly, there were MHC class II-expressing immune cells suggesting the presence of dendritic cells. Using lineage-specific primer pairs RT-PCR confirmed the immunofluorescence microscopy results, supporting that human periosteum serves as a reservoir of stromal stem cells, as well as cells of the osteoblastic, and the chondroblastic lineage, osteoclasts, and dendritic cells.Conclusion. Our work elucidates the role of periosteum as a source of cells with a high regenerative capacity. Undifferentiated stromal stem cells as well as osteoblastic precursor cells are dominating in the cambium layer. A new outlook is given towards an immune response coming from the periosteum as MHC II positive immune cells were detected.

scholarly journals Protection of mice against experimental cryptococcosis by synthesized peptides delivered in glucan particles

2021 ◽  
Author(s):  
Charles Specht ◽  
E. Jane Homan ◽  
Chrono K Lee ◽  
Zhongming Mou ◽  
Christina L Gomez ◽  
...  
Keyword(s):  
Mhc Class Ii ◽  
Peptide Sequence ◽  
T Cell Epitopes ◽  
Binding Region ◽  
Chitin Deacetylase ◽  
Lethal Challenge ◽  
Amino Acid Peptide ◽  
Mhc Ii ◽  
Strong Binding ◽  
Experimental Cryptococcosis

The high global burden of cryptococcosis has made development of a protective vaccine a public health priority. We previously demonstrated that a vaccine composed of recombinant Cryptococcus neoformans chitin deacetylase 2 (Cda2) delivered in glucan particles (GPs) protects BALB/c and C57BL/6 mice from an otherwise lethal challenge with a highly virulent C. neoformans strain. An immunoinformatic analysis of Cda2 revealed a peptide sequence predicted to have strong binding to the MHC Class II (MHC II) H2-IAd allele found in BALB/c mice. BALB/c mice vaccinated with GPs containing a 32 amino acid peptide (Cda2-Pep1) that included this strong binding region were protected from cryptococcosis. Protection was lost with GP-based vaccines containing versions of recombinant Cda2 protein and Cda2-Pep1 with mutations predicted to greatly diminish MHC II binding. Cda2 has homology to the three other C. neoformans chitin deacetylases, Cda1, Cda3 and Fpd1, in the high MHC II binding region. GPs loaded with homologous peptides of Cda1, Cda3 and Fpd1 protected BALB/c mice from experimental cryptococcosis, albeit not as robustly as the Cda2-Pep1 vaccine. Finally, seven other peptides were synthesized based on regions in Cda2 predicted to contain promising CD4+ T cell epitopes in BALB/c or C57BL/6 mice. While five peptide vaccines significantly protected BALB/c mice, only one protected C57BL/6 mice. Thus, GP-based vaccines containing a single peptide can protect mice against cryptococcosis. However, given the diversity of human MHC II alleles, a peptide-based Cryptococcus vaccine for use in humans would be challenging and likely need to contain multiple peptide sequences.

scholarly journals Trogocytosis of peptide–MHC class II complexes from dendritic cells confers antigen-presenting ability on basophils

2017 ◽  
Vol 114 (5) ◽  
pp. 1111-1116 ◽  
Author(s):  
Kensuke Miyake ◽  
Nozomu Shiozawa ◽  
Toshihisa Nagao ◽  
Soichiro Yoshikawa ◽  
Yoshinori Yamanishi ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Cell Differentiation ◽  
Cell Surface ◽  
Mhc Class Ii ◽  
Antigen Presenting Cells ◽  
Class Ii ◽  
Gm Csf ◽  
Mhc Ii ◽  
Th2 Cell ◽  
Antigen Presenting

Th2 immunity plays important roles in both protective and allergic responses. Nevertheless, the nature of antigen-presenting cells responsible for Th2 cell differentiation remains ill-defined compared with the nature of the cells responsible for Th1 and Th17 cell differentiation. Basophils have attracted attention as a producer of Th2-inducing cytokine IL-4, whereas their MHC class II (MHC-II) expression and function as antigen-presenting cells are matters of considerable controversy. Here we revisited the MHC-II expression on basophils and explored its functional relevance in Th2 cell differentiation. Basophils generated in vitro from bone marrow cells in culture with IL-3 plus GM-CSF displayed MHC-II on the cell surface, whereas those generated in culture with IL-3 alone did not. Of note, these MHC-II–expressing basophils showed little or no transcription of the corresponding MHC-II gene. The GM-CSF addition to culture expanded dendritic cells (DCs) other than basophils. Coculture of basophils and DCs revealed that basophils acquired peptide–MHC-II complexes from DCs via cell contact-dependent trogocytosis. The acquired complexes, together with CD86, enabled basophils to stimulate peptide-specific T cells, leading to their proliferation and IL-4 production, indicating that basophils can function as antigen-presenting cells for Th2 cell differentiation. Transfer of MHC-II from DCs to basophils was also detected in draining lymph nodes of mice with atopic dermatitis-like skin inflammation. Thus, the present study defined the mechanism by which basophils display MHC-II on the cell surface and appears to reconcile some discrepancies observed in previous studies.

scholarly journals CD83 increases MHC II and CD86 on dendritic cells by opposing IL-10–driven MARCH1-mediated ubiquitination and degradation

2011 ◽  
Vol 208 (1) ◽  
pp. 149-165 ◽  
Author(s):  
Lina E. Tze ◽  
Keisuke Horikawa ◽  
Heather Domaschenz ◽  
Debbie R. Howard ◽  
Carla M. Roots ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Mhc Class Ii ◽  
Molecular Mechanisms ◽  
Costimulatory Molecule ◽  
Class Ii ◽  
Interleukin 10 ◽  
Effective Vaccine ◽  
Vaccine Adjuvants ◽  
Mhc Ii ◽  
Tm Domain

Effective vaccine adjuvants must induce expression of major histocompatability (MHC) class II proteins and the costimulatory molecule CD86 on dendritic cells (DCs). However, some adjuvants elicit production of cytokines resulting in adverse inflammatory consequences. Development of agents that selectively increase MHC class II and CD86 expression without triggering unwanted cytokine production requires a better understanding of the molecular mechanisms influencing the production and degradation of MHC class II and CD86 in DCs. Here, we investigate how CD83, an immunoglobulin protein expressed on the surface of mature DCs, promotes MHC class II and CD86 expression. Using mice with an N-ethyl-N-nitrosourea–induced mutation eliminating the transmembrane (TM) region of CD83, we found that the TM domain of CD83 enhances MHC class II and CD86 expression by blocking MHC class II association with the ubiquitin ligase MARCH1. The TM region of CD83 blocks interleukin 10–driven, MARCH1-dependent ubiquitination and degradation of MHC class II and CD86 in DCs. Exploiting this posttranslational pathway for boosting MHC class II and CD86 expression on DCs may provide an opportunity to enhance the immunogenicity of vaccines.

scholarly journals CD11cloB220+ interferon-producing killer dendritic cells are activated natural killer cells

2007 ◽  
Vol 204 (11) ◽  
pp. 2569-2578 ◽  
Author(s):  
Christian A.J. Vosshenrich ◽  
Sarah Lesjean-Pottier ◽  
Milena Hasan ◽  
Odile Richard-Le Goff ◽  
Erwan Corcuff ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Nk Cells ◽  
Natural Killer ◽  
Mhc Class Ii ◽  
Nk Cell ◽  
Phenotypic Differences ◽  
Mhc Ii ◽  
Class Ii Mhc

Interferon-producing killer dendritic cells (IKDCs) are a recently described subset of CD11cloB220+ cells that share phenotypic and functional properties of DCs and natural killer (NK) cells (Chan, C.W., E. Crafton, H.N. Fan, J. Flook, K. Yoshimura, M. Skarica, D. Brockstedt, T.W. Dubensky, M.F. Stins, L.L. Lanier, et al. 2006. Nat. Med. 12:207–213; Taieb, J., N. Chaput, C. Menard, L. Apetoh, E. Ullrich, M. Bonmort, M. Pequignot, N. Casares, M. Terme, C. Flament, et al. 2006. Nat. Med. 12:214–219). IKDC development appears unusual in that cytokines using the interleukin (IL)-2 receptor β (IL-2Rβ) chain but not those using the common γ chain (γc) are necessary for their generation. By directly comparing Rag2−/−γc−/y, Rag2−/−IL-2Rβ−/−, Rag2−/−IL-15−/−, and Rag2−/−IL-2−/− mice, we demonstrate that IKDC development parallels NK cell development in its strict IL-15 dependence. Moreover, IKDCs uniformly express NK-specific Ncr-1 transcripts (encoding NKp46), whereas NKp46+ cells are absent in Ncr1gfp/+γc−/y mice. Distinguishing features of IKDCs (CD11cloB220+MHC-II+) were carefully examined on developing NK cells in the bone marrow and on peripheral NK cells. As B220 expression was heterogeneous, defining B220lo versus B220hi NK1.1+ NK cells could be considered as arbitrary, and few phenotypic differences were noted between NK1.1+ NK cells bearing different levels of B220. CD11c expression did not correlate with B220 or major histocompatibility complex (MHC) class II (MHC-II) expression, and most MHC-II+ NK1.1+ cells did not express B220 and were thus not IKDCs. Finally, CD11c, MHC-II, and B220 levels were up-regulated on NK1.1+ cells upon activation in vitro or in vivo in a proliferation-dependent fashion. Our data suggest that the majority of CD11cloB220+ “IKDC-like” cells represent activated NK cells.

scholarly journals Reorganization of multivesicular bodies regulates MHC class II antigen presentation by dendritic cells

2001 ◽  
Vol 155 (1) ◽  
pp. 53-64 ◽  
Author(s):  
Monique Kleijmeer ◽  
Georg Ramm ◽  
Danita Schuurhuis ◽  
Janice Griffith ◽  
Maria Rescigno ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Plasma Membrane ◽  
Mhc Class Ii ◽  
Class Ii ◽  
Multivesicular Bodies ◽  
Mhc Ii ◽  
Immature Dendritic Cells ◽  
Class Ii Mhc ◽  
Accessory Molecule ◽  
Class Ii Antigen

Immature dendritic cells (DCs) sample their environment for antigens and after stimulation present peptide associated with major histocompatibility complex class II (MHC II) to naive T cells. We have studied the intracellular trafficking of MHC II in cultured DCs. In immature cells, the majority of MHC II was stored intracellularly at the internal vesicles of multivesicular bodies (MVBs). In contrast, DM, an accessory molecule required for peptide loading, was located predominantly at the limiting membrane of MVBs. After stimulation, the internal vesicles carrying MHC II were transferred to the limiting membrane of the MVB, bringing MHC II and DM to the same membrane domain. Concomitantly, the MVBs transformed into long tubular organelles that extended into the periphery of the cells. Vesicles that were formed at the tips of these tubules nonselectively incorporated MHC II and DM and presumably mediated transport to the plasma membrane. We propose that in maturing DCs, the reorganization of MVBs is fundamental for the timing of MHC II antigen loading and transport to the plasma membrane.

Dendritic cells constitutively present self antigens in their immature state in vivo and regulate antigen presentation by controlling the rates of MHC class II synthesis and endocytosis

Blood ◽  
2004 ◽  
Vol 103 (6) ◽  
pp. 2187-2195 ◽  
Author(s):  
Nicholas S. Wilson ◽  
Dima El-Sukkari ◽  
José A. Villadangos
Keyword(s):  
Dendritic Cells ◽  
Antigen Presentation ◽  
Mhc Class Ii ◽  
Class Ii ◽  
Lymphoid Organ ◽  
Lymphoid Organs ◽  
Mhc Ii ◽  
Peptide Complexes ◽  
Self Antigens

Abstract Dendritic cells (DCs) change their antigen-presenting properties during maturation. Immature DCs efficiently capture antigens, but are reported to be impaired in their processing and presenting capacity. Upon an encounter with an inflammatory stimulus, DCs undergo a maturation process that leads to efficient presentation of antigens captured at the time of activation, but precludes processing of antigens encountered at later time points. The mechanisms that underlie these developmental changes are controversial. Thus, it is unclear whether immature DCs can present self antigens, and which are the checkpoints that regulate antigen presentation in immature and mature DCs. We have characterized these mechanisms using DCs derived directly from lymphoid organs. Immature lymphoid organ DCs constitutively presented self peptides bound to major histocompatibility complex class II (MHCII) molecules, but these MHCII-peptide complexes were degraded quickly after their transient expression on the cell surface. During maturation, MHC II endocytosis was down-regulated, so that newly generated MHC II–peptide complexes accumulated on the plasma membrane. Simultaneously, MHC II synthesis was down-regulated, thus preventing the turnover of the MHC II–peptide complexes that accumulated early during maturation. Our results demonstrate that immature DCs constitutively present self antigens in the lymphoid organs and characterize the molecular basis of the capacity of DCs to provide “antigenic memory” in vivo.

Blood dendritic cells in cattle infected with bovine leukemia virus (BLV): isolation and phenotyping

2012 ◽  
Vol 15 (4) ◽  
pp. 599-634
Author(s):  
M. Szczotka ◽  
J. Kuźmak ◽  
K. Kostro ◽  
D. Bednarek ◽  
M. Purzycka
Keyword(s):  
Flow Cytometry ◽  
Dendritic Cells ◽  
Immune Responses ◽  
Mhc Class Ii ◽  
Leukemia Virus ◽  
Mhc I ◽  
Mhc Ii ◽  
Antigen Presenting ◽  
Very High

AbstractDendritic cells (DCs) are most potent antigen presenting cells (APCs) with unique ability to prime effective immune responses. They express higher levels of MHC class II and accesory molecules on their surface, than other professional APCs. The investigations were performed on DCs generated from blood with the use of microbeads magnetically labeled with mouse anti human CD14. Flow cytometry was applied for determination of DCs immunophenotype in healthy and naturally infected with BLV cattle. For immunophenotyping mouse monoclonal antibodies anti bovine: CD11a, CD11b, CD11c, MHC-I and MHC-II were used. Our results demonstrated that dendritic cells infected with BLV expressed very high percentage of determinants: CD11a, CD11b, CD11c, MHC-I and MHC-II class. Leukaemic DCs exhibited DCs morphology and had a phenotype of mature DCs. The expression of gp51 glycoprotein of BLV on leukaemic DCs was detected in flow cytometry investigations.

Factor VIII Inhibitor Antibodies with C2 Domain Specificity Are Less inhibitory to Factor VIII Complexed with von Willebrand Factor

1996 ◽  
Vol 76 (05) ◽  
pp. 749-754 ◽  
Author(s):  
Suzuki Suzuki ◽  
Morio Arai ◽  
Kagehiro Amano ◽  
Kazuhiko Kagawa ◽  
Katsuyuki Fukutake
Keyword(s):  
Complex Formation ◽  
Factor Viii ◽  
Von Willebrand Factor ◽  
Domain Specificity ◽  
Factor Viii Inhibitor ◽  
C2 Domain ◽  
Recombinant Fviii ◽  
Von Willebrand ◽  
A2 Domain ◽  
Willebrand Factor

SummaryIn order to clarify the potential role of von Willebrand factor (vWf) in attenuating the inactivation of factor VIII (fVIII) by those antibodies with C2 domain specificity, we investigated a panel of 14 human antibodies to fVIII. Immunoblotting analysis localized light chain (C2 domain) epitopes for four cases, heavy chain (A2 domain) epitopes in five cases, while the remaining five cases were both light and heavy chains. The inhibitor titer was considerably higher for Kogenate, a recombinant fVIII concentrate, than for Haemate P, a fVIII/vWf complex concentrate, in all inhibitor plasmas that had C2 domain specificity. In five inhibitor plasmas with A2 domain specificity and in five with both A2 and C2 domain specificities, Kogenate gave titers similar to or lower than those with Haemate P. The inhibitory effect of IgG of each inhibitor plasma was then compared with recombinant fVIII and its complex with vWf. When compared to the other 10 inhibitor IgGs, IgG concentration, which inhibited 50% of fVIII activity (IC50), was remarkably higher for the fVIII/vWf complex than for fVIII in all the inhibitor IgGs that had C2 domain reactivity. Competition of inhibitor IgG and vWf for fVIII binding was observed in an ELISA system. In 10 inhibitors that had C2 domain reactivity, the dose dependent inhibition of fVIII-vWf complex formation was observed, while, in the group of inhibitors with A2 domain specificity, there was no inhibition of the complex formation except one case. We conclude that a subset of fVIII inhibitors, those that bind to C2 domain determinants, are less inhibitory to fVIII when it is complexed with vWf that binds to overlapping region in the C2 domain.

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