In vivo DCs Depletion with Diphtheria Toxin and MARCO+/MOMA1+ Cells Depletion with Clodronate Liposomes in B6.CD11c-DTR Mice

BIO-PROTOCOL ◽  
2016 ◽  
Vol 6 (15) ◽  
Author(s):  
Henrique Silva ◽  
Carlos Tadokoro ◽  
Maria D’Império-Lima
Keyword(s):  
Diphtheria Toxin ◽  
Clodronate Liposomes

scholarly journals Inducible ablation of mouse Langerhans cells diminishes but fails to abrogate contact hypersensitivity

2005 ◽  
Vol 169 (4) ◽  
pp. 569-576 ◽  
Author(s):  
Clare L. Bennett ◽  
Erwin van Rijn ◽  
Steffen Jung ◽  
Kayo Inaba ◽  
Ralph M. Steinman ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Steady State ◽  
Diphtheria Toxin ◽  
Langerhans Cells ◽  
Contact Hypersensitivity ◽  
Relative Importance ◽  
Skin Immunity

Langerhans cells (LC) form a unique subset of dendritic cells (DC) in the epidermis but so far their in vivo functions in skin immunity and tolerance could not be determined, in particular in relation to dermal DC (dDC). Here, we exploit a novel diphtheria toxin (DT) receptor (DTR)/DT-based system to achieve inducible ablation of LC without affecting the skin environment. Within 24 h after intra-peritoneal injection of DT into Langerin-DTR mice LC are completely depleted from the epidermis and only begin to return 4 wk later. LC deletion occurs by apoptosis in the absence of inflammation and, in particular, the dDC compartment is not affected. In LC-depleted mice contact hypersensitivity (CHS) responses are significantly decreased, although ear swelling still occurs indicating that dDC can mediate CHS when necessary. Our results establish Langerin-DTR mice as a unique tool to study LC function in the steady state and to explore their relative importance compared with dDC in orchestrating skin immunity and tolerance.

scholarly journals In vivo tracking of transplanted macrophages with near infrared fluorescent dye reveals temporal distribution and specific homing in the liver that can be perturbed by clodronate liposomes

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0242488
Author(s):  
Satoshi Nishiwaki ◽  
Shigeki Saito ◽  
Kyosuke Takeshita ◽  
Hidefumi Kato ◽  
Ryuzo Ueda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Normal State ◽  
Near Infrared ◽  
Temporal Distribution ◽  
Organ Damage ◽  
The Other ◽  
Acquired Immunity ◽  
Activated Macrophages ◽  
Clodronate Liposomes

Macrophages play an indispensable role in both innate and acquired immunity, while the persistence of activated macrophages can sometimes be harmful to the host, resulting in multi-organ damage. Macrophages develop from monocytes in the circulation. However, little is known about the organ affinity of macrophages in the normal state. Using in vivo imaging with XenoLight DiR®, we observed that macrophages showed strong affinity for the liver, spleen and lung, and weak affinity for the gut and bone marrow, but little or no affinity for the kidney and skin. We also found that administered macrophages were still alive 168 hours after injection. On the other hand, treatment with clodronate liposomes, which are readily taken up by macrophages via phagocytosis, strongly reduced the number of macrophages in the liver, spleen and lung.

scholarly journals Altered ratio of dendritic cell subsets in skin-draining lymph nodes promotes Th2-driven contact hypersensitivity

2021 ◽  
Vol 118 (3) ◽  
pp. e2021364118
Author(s):  
Hannah L. Miller ◽  
Prabhakar Sairam Andhey ◽  
Melissa K. Swiecki ◽  
Bruce A. Rosa ◽  
Konstantin Zaitsev ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Lymph Nodes ◽  
Diphtheria Toxin ◽  
Fluorescein Isothiocyanate ◽  
Skin Inflammation ◽  
Contact Hypersensitivity ◽  
Type I ◽  
Th2 Response ◽  
Draining Lymph Nodes

Plasmacytoid dendritic cells (pDCs) specialize in the production of type I IFN (IFN-I). pDCs can be depleted in vivo by injecting diphtheria toxin (DT) in a mouse in which pDCs express a diphtheria toxin receptor (DTR) transgene driven by the human CLEC4C promoter. This promoter is enriched for binding sites for TCF4, a transcription factor that promotes pDC differentiation and expression of pDC markers, including CLEC4C. Here, we found that injection of DT in CLEC4C-DTR+ mice markedly augmented Th2-dependent skin inflammation in a model of contact hypersensitivity (CHS) induced by the hapten fluorescein isothiocyanate. Unexpectedly, this biased Th2 response was independent of reduced IFN-I accompanying pDC depletion. In fact, DT treatment altered the representation of conventional dendritic cells (cDCs) in the skin-draining lymph nodes during the sensitization phase of CHS; there were fewer Th1-priming CD326+ CD103+ cDC1 and more Th2-priming CD11b+ cDC2. Single-cell RNA-sequencing of CLEC4C-DTR+ cDCs revealed that CD326+ DCs, like pDCs, expressed DTR and were depleted together with pDCs by DT treatment. Since CD326+ DCs did not express Tcf4, DTR expression might be driven by yet-undefined transcription factors activating the CLEC4C promoter. These results demonstrate that altered DC representation in the skin-draining lymph nodes during sensitization to allergens can cause Th2-driven CHS.

scholarly journals Interleukin 2-diphtheria toxin fusion protein can abolish cell-mediated immunity in vivo.

1988 ◽  
Vol 85 (11) ◽  
pp. 3980-3984 ◽  
Author(s):  
V. E. Kelley ◽  
P. Bacha ◽  
O. Pankewycz ◽  
J. C. Nichols ◽  
J. R. Murphy ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Diphtheria Toxin ◽  
Interleukin 2 ◽  
Cell Mediated Immunity

scholarly journals Endosomal proteolysis of diphtheria toxin without toxin translocation into the cytosol of rat liver in vivo

FEBS Journal ◽  
2008 ◽  
Vol 275 (8) ◽  
pp. 1708-1722 ◽  
Author(s):  
Tatiana El Hage ◽  
Paulette Decottignies ◽  
François Authier
Keyword(s):  
Rat Liver ◽  
Diphtheria Toxin

scholarly journals INACTIVATION OF BACTERIAL EXOTOXINS AND ENDOTOXIN BY IRON

1960 ◽  
Vol 112 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Aaron Janoff ◽  
Benjamin W. Zweifach
Keyword(s):  
Guinea Pigs ◽  
Diphtheria Toxin ◽  
Reticuloendothelial System ◽  
Partial Recovery ◽  
Alpha Toxin ◽  
Lethal Effects ◽  
Partial Restoration ◽  
Manganese Zinc

After incubation with various agents in vitro, the lethal effects of aqueous solutions of Cl. perfringens alpha toxin, C. diphtheriae toxin, and E. coli endotoxin were tested in mice, guinea pigs, and rabbits, respectively. Iron, copper, cysteine, ascorbic acid, and versene counteracted the lethal effects of alpha toxin in mice, while magnesium, manganese, zinc, and citrate did not. Iron also counteracted the lethal effects of diphtheria toxin in guinea pigs. After incubation of endotoxin with iron, its lethal effects and tissue-necrotizing actions in rabbits were counteracted. However, the pyrogenic properties of the toxin were not affected. The solubilities of perfringens alpha toxin and diphtheria toxin were markedly reduced after incubation with detoxifying metals, and resolubilization of these toxins with chelators resulted in partial restoration of toxicity. Addition of versene to detoxified endotoxin also resulted in partial recovery of lethal effectiveness. The inactivation of bacterial toxins by iron under in vitro conditions is not specific to this metal, is a reversible process, and may be due to desolubilization, reduction, or to competition by the metal for sites on the toxin normally bound by other cations in vivo. Although no evidence is presented in this paper to support the view that there is a relationship between the inactivation of endotoxin and the storage iron in the reticuloendothelial system of shocked animals, the observation of an in vitro inactivation of endotoxin by inorganic iron warrants consideration of such a mechanism.

scholarly journals Diphtheria-toxin based anti-human CCR4 immunotoxin for targeting human CCR4+ cells in vivo

2015 ◽  
Vol 9 (7) ◽  
pp. 1458-1470 ◽  
Author(s):  
Zhaohui Wang ◽  
Min Wei ◽  
Huiping Zhang ◽  
Hongyuan Chen ◽  
Sharon Germana ◽  
...  
Keyword(s):  
Diphtheria Toxin

scholarly journals Failure of Diphtheria Toxin Model to Induce Parkinson-like Behavior in Mice

2021 ◽  
Vol 22 (17) ◽  
pp. 9496
Author(s):  
Lucie Valek ◽  
Irmgard Tegeder
Keyword(s):  
Substantia Nigra ◽  
Diphtheria Toxin ◽  
Behavioral Correlates ◽  
Open Field Behavior ◽  
Neuronal Marker ◽  
Transgenic Expression ◽  
Protein Levels ◽  
Behavioral Abnormalities ◽  
Toxin Receptor

Rodent models of Parkinson’s disease are based on transgenic expression of mutant synuclein, deletion of PD genes, injections of MPTP or rotenone, or seeding of synuclein fibrils. The models show histopathologic features of PD such as Lewi bodies but mostly only subtle in vivo manifestations or systemic toxicity. The models only partly mimic a predominant loss of dopaminergic neurons in the substantia nigra. We therefore generated mice that express the transgenic diphtheria toxin receptor (DTR) specifically in DA neurons by crossing DAT-Cre mice with Rosa26 loxP-STOP-loxP DTR mice. After defining a well-tolerated DTx dose, DAT-DTR and DTR-flfl controls were subjected to non-toxic DTx treatment (5 × 100 pg/g) and subsequent histology and behavioral tests. DAT protein levels were reduced in the midbrain, and tyrosine hydroxylase-positive neurons were reduced in the substantia nigra, whereas the pan-neuronal marker NeuN was not affected. Despite the promising histologic results, there was no difference in motor function tests or open field behavior. These are tests in which double mutant Pink1−/−SNCAA53T Parkinson mice show behavioral abnormalities. Higher doses of DTx were toxic in both groups. The data suggest that DTx treatment in mice with Cre/loxP-driven DAT-DTR expression leads to partial ablation of DA-neurons but without PD-reminiscent behavioral correlates.

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