scholarly journals Local association of Trypanosoma cruzi chronic infection foci and enteric neuropathic lesions at the tissue micro-domain scale

2021 ◽  
Author(s):  
Archie A. Khan ◽  
Harry C. Langston ◽  
Fernanda C. Costa ◽  
Francisco Olmo ◽  
Martin C. Taylor ◽  
...  
Keyword(s):  
Chronic Infection ◽  
Digestive Disease ◽  
Clinical Manifestations ◽  
Imaging Systems ◽  
Immunofluorescence Analysis ◽  
Cell Level ◽  
Host Interactions ◽  
Chronic Chagas Disease

AbstractGastrointestinal (GI) disease affects a substantial subset of chronic Chagas disease (CD) patients, but the mechanism of pathogenesis is poorly understood. The lack of a robust, predictive animal model of chronic T. cruzi infection that exhibits functional digestive disease has held back research. To address this, we combined GI tracer assays and bioluminescence in vivo infection imaging systems for diverse parasite strains to discover models exhibiting chronic digestive transit dysfunction. We identified the colon as a specific site of both tissue parasite persistence and delayed transit. Digestive CD mice exhibited significant retention of faeces in both sated and fasted conditions. Histological and immunofluorescence analysis of the enteric nervous system (ENS) revealed a dramatic reduction in the number of neurons and a loss of immunoreactivity of the enteric neural network in the colon. This model therefore recapitulates key clinical manifestations of human digestive CD. We also exploited dual bioluminescent-fluorescent parasites to analyse rare chronic infection foci in the colon at the single cell level, revealing co-localisation with ENS lesions. This indicates that long-term T. cruzi-host interactions in the colon drive pathogenesis and thus chronic disease may be preventable using anti-parasitic chemotherapy.

scholarly journals Local association of Trypanosoma cruzi chronic infection foci and enteric neuropathic lesions at the tissue micro-domain scale

2021 ◽  
Vol 17 (8) ◽  
pp. e1009864
Author(s):  
Archie A. Khan ◽  
Harry C. Langston ◽  
Fernanda C. Costa ◽  
Francisco Olmo ◽  
Martin C. Taylor ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Chronic Infection ◽  
Ex Vivo ◽  
Clinical Investigation ◽  
Clinical Manifestations ◽  
Host Interactions ◽  
Dual Reporter ◽  
Enteric Neuropathy ◽  
Transgenic Parasites

Digestive Chagas disease (DCD) is an enteric neuropathy caused by Trypanosoma cruzi infection. The mechanism of pathogenesis is poorly understood and the lack of a robust, predictive animal model has held back research. We screened a series of mouse models using gastrointestinal tracer assays and in vivo infection imaging systems to discover a subset exhibiting chronic digestive transit dysfunction and significant retention of faeces in both sated and fasted conditions. The colon was a specific site of both tissue parasite persistence, delayed transit and dramatic loss of myenteric neurons as revealed by whole-mount immunofluorescence analysis. DCD mice therefore recapitulated key clinical manifestations of human disease. We also exploited dual reporter transgenic parasites to home in on locations of rare chronic infection foci in the colon by ex vivo bioluminescence imaging and then used fluorescence imaging in tissue microdomains to reveal co-localisation of infection and enteric nervous system lesions. This indicates that long-term T. cruzi-host interactions in the colon drive DCD pathogenesis, suggesting that the efficacy of anti-parasitic chemotherapy against chronic disease progression warrants further pre-clinical investigation.

scholarly journals Molecular characterization of NDP52, a novel protein of the nuclear domain 10, which is redistributed upon virus infection and interferon treatment.

1995 ◽  
Vol 130 (1) ◽  
pp. 1-13 ◽  
Author(s):  
F Korioth ◽  
C Gieffers ◽  
G G Maul ◽  
J Frey
Keyword(s):  
Protein Interactions ◽  
Leucine Zipper ◽  
Coiled Coil ◽  
Nuclear Staining ◽  
Interferon Treatment ◽  
Immunofluorescence Analysis ◽  
Host Interactions ◽  
Nuclear Domain ◽  
Novel Protein

The nuclear domain (ND)10 also described as POD or Kr bodies is involved in the development of acute promyelocytic leukemia and virus-host interactions. Immunofluorescence analysis using a variety of human autoimmune sera and monoclonal antibodies showed a typical dot like nuclear staining for ND10, suggesting that this structure consists of several proteins. Two of the ND10 proteins, Sp100 and PML are genetically characterized and show homology with several transcription factors. Here we describe NDP52, an additional novel protein of the ND10. We raised a new mAb C8A2, that specifically recognizes NDP52. Immunofluorescence analysis using this mAb showed a typical nuclear dot staining as it was described for ND10. Isolation and sequencing of the corresponding cDNA revealed that NDP52 has a predicted molecular mass of 52 kD. The deduced amino acid sequence exhibits an extended central coiled coil domain containing a leucine zipper motif. The COOH terminus of NDP52 shows homology with LIM domains, that have recently been described to mediate protein interactions, which let NDP52 appear as a suitable candidate for mediating interactions between ND10 proteins. In vivo, NDP52 is transcribed in all human tissues analyzed. Furthermore, we show that NDP52 colocalizes with the ND10 protein PML and can be redistributed upon viral infection and interferon treatment. These data suggest that ND10 proteins play an important role in the viral life cycle.

scholarly journals B Cell-Specific Expression of Ataxia-Telangiectasia Mutated Protein Kinase Promotes Chronic Gammaherpesvirus Infection

2017 ◽  
Vol 91 (19) ◽  
Author(s):  
Eric J. Darrah ◽  
Joseph M. Kulinski ◽  
Wadzanai P. Mboko ◽  
Gang Xin ◽  
Laurent P. Malherbe ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Ataxia Telangiectasia ◽  
Chronic Infection ◽  
Viral Latency ◽  
Viral Reactivation ◽  
Ataxia Telangiectasia Mutated

ABSTRACT Manipulation of host cellular pathways is a strategy employed by gammaherpesviruses, including mouse gammaherpesvirus 68 (MHV68), in order to negotiate a chronic infection. Ataxia-telangiectasia mutated (ATM) plays a unique yet incompletely understood role in gammaherpesvirus infection, as it has both proviral and antiviral effects. Chronic gammaherpesvirus infection is poorly controlled in a host with global ATM insufficiency, whether the host is a mouse or a human. In contrast, ATM facilitates replication, reactivation, and latency establishment of several gammaherpesviruses in vitro, suggesting that ATM is proviral in the context of infected cell cultures. The proviral role of ATM is also evident in vivo, as myeloid-specific ATM expression facilitates MHV68 reactivation during the establishment of viral latency. In order to better understand the complex relationship between host ATM and gammaherpesvirus infection, we depleted ATM specifically in B cells, a cell type critical for chronic gammaherpesvirus infection. B cell-specific ATM deficiency attenuated the establishment of viral latency due to compromised differentiation of ATM-deficient B cells. Further, we found that during long-term infection, peritoneal B-1b, but not related B-1a, B cells display the highest frequency of gammaherpesvirus infection. While ATM expression did not affect gammaherpesvirus tropism for B-1 B cells, B cell-specific ATM expression was necessary to support viral reactivation from peritoneal cells during long-term infection. Thus, our study reveals a role of ATM as a host factor that promotes chronic gammaherpesvirus infection of B cells. IMPORTANCE Gammaherpesviruses infect a majority of the human population and are associated with cancer, including B cell lymphomas. ATM is a unique host kinase that has both proviral and antiviral roles in the context of gammaherpesvirus infection. Further, there is insufficient understanding of the interplay of these roles in vivo during chronic infection. In this study, we show that ATM expression by splenic B cells is required for efficient establishment of gammaherpesvirus latency. We also show that ATM expression by peritoneal B cells is required to facilitate viral reactivation during long-term infection. Thus, our study defines a proviral role of B cell-specific ATM expression during chronic gammaherpesvirus infection.

scholarly journals Long-Term In Vivo Imaging of Multiple Organs at the Single Cell Level

PLoS ONE ◽  
2013 ◽  
Vol 8 (1) ◽  
pp. e52087 ◽  
Author(s):  
Benny J. Chen ◽  
Yiqun Jiao ◽  
Ping Zhang ◽  
Albert Y. Sun ◽  
Geoffrey S. Pitt ◽  
...  
Keyword(s):  
Single Cell ◽  
In Vivo Imaging ◽  
Single Cell Level ◽  
Cell Level ◽  
Multiple Organs

scholarly journals GvHD Kinetics after Haploidentical TK-Cells: In-Vivo HSV-TK Suicide Machinery Is Effective in GvHD Control and Provide a Long-Term Immune-Suppressive Treatment-Free Survival

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 548-548 ◽  
Author(s):  
Maria Teresa Lupo-Stanghellini ◽  
Chiara Bonini ◽  
Giacomo Oliveira ◽  
Attilio Bondanza ◽  
Luca Vago ◽  
...  
Keyword(s):  
Median Time ◽  
De Novo ◽  
Clinical Manifestations ◽  
High Rate ◽  
Phase Iii ◽  
High Dose ◽  
Concomitant Treatment ◽  
Free Survival

Abstract Introduction Extensive application of haploidentical SCT (haplo-SCT) is limited by high rate of late transplant mortality and relapse incidence associated with the delayed immune-reconstitution (IR) secondary to the procedures for severe graft-versus-host-disease (GvHD) prevention and treatment. In the past 20 years we deeply investigate the application of the paradigmatic herpes simplex virus thymidine kinase (TK) suicide gene strategy to allow the selective elimination of genetically modified donor T cells during GvHD, while sparing IR with effective graft-versus-leukemia and graft-versus-infectious effects. Aim of the study Here we report the incidence, characterization, stratification, treatment and outcome for both acute (a-) and chronic (c-) GvHD in haplo-SCT after TK-cells infusion. Methods We included for analysis 57 adult patients (pts, median age 53 years – r 17-66) who underwent an haplo-SCT according to TK-trial (Ciceri, Bonini et al, Lancet Oncol 2009; Phase III TK008, NCT0091462), between 2002 and 2014 at our Center. Data were collected from our Institutional database. A written consent was given by pts allowing the use of medical records for research in accordance with the Declaration of Helsinki. All consecutive pts receiving graft after selection of peripheral CD34+ cells - CliniMacs one-step procedure - were selected. No immune-suppression was introduced after SCT as GvHD prophylaxis. In vivo T-cell depletion with ATG (Fresenius) was administered in all pts. Donor lymphocytes genetically engineered to express the TK gene were infused in 34/57 pts (median 2 infusion/pts), 25/34 achieved IR (median time from SCT 84 days – r 18/182; median time from last TK-cells infusion 27 days – r 13/42). Results Twelve of 25 immune-reconstituted pts developed a-GvHD (grade I–IV; median time of onset 84 days post SCT – r 20/162; 19 days post last TK-infusion – r 8/54) and one developed c-GVHD. Direct association of TK-cells and GvHD was confirmed by vector-encoded protein immunostaining of lymphocytes infiltrating affected lesions. Eleven pts needed GvHD treatment: 4 pts received ganciclovir iv (GCV 5 mg/Kg/12h/14 days), 7 pts valganciclovir per os (VGCV 900 mg/12h/14 days). Both GCV and VGCV were effective in control clinical manifestations of GvHD in a median of 14 days (see figures 1. and 2.) and resulted in a significant reduction in numbers of circulating TK-cells, without reduction of CD3+ TK-negative lymphocytes maintaining long-term IR (see table). In 5 pts additional concomitant treatment with low-dose steroid (prednisone <0.5mg/kg per day for a median of 2 weeks) was given. A pt who presented severe gut and liver GvHD and one who received at SCT an high dose of unmanipulated lymphocytes (5.4x105/Kg) – were successfully treated with a combined therapy of prednisone and cyclosporine or rapamicine in association with GCV. Patient TK44 developed a severe classic de novo c-GVHD, with sclerodermatous lichenoid skin and mouth features plus moderate dry-eye symptoms that was successfully treated with VGCV and a transient course of mycophenolate mofetil (2 g per day) over a 2 months period. No cases of quiescent or progressive c-GvHD was observed after a median follow-up of 679 days (r 139/4035). Conclusion In our 12-years experience we can confirm that infusion of TK-cells is effective in accelerating IR while controlling GvHD, providing a long-term immunosuppressive therapy free survival in absence of GvHD related deaths or long-term complications. Abstract 548 Despite a consistent reduction in TK-cell numbers, the GCV/VGCV treatment of GVHD did not impair or prevent long-term IR. TK-cells infused / Kg x107 Time from SCT to GvHD (days) Time from last TK-cells to GvHD (days) Description (acute – chronic, grade) Before GCV/VGCV After GCV/VGCV GvHD outcome ,days after 1st dose of GCV/VGCV TK+ cells/mcl TK-cells/mcl % of TK+,tot CD3+cells TK+ cells/mcl TK-cells/mcl % of TK+, tot CD3+cells TK5 1 91 15 A II 36 248 12.7 7 378 1.8 CR 21 TK6 1 98 51 A I 213 270 44.1 / / / CR NA TK8 10 20 17 A IV 207 224 57.9 24 36 40.2 CR 20 TK16 2.2 20 8 A II 90 99 47.6 25 69 26.6 CR 4 TK20 2.4 30 28 A II 22 139 13.7 13 129 9.2 CR 29 TK25 0.4 162 14 A II 23 123 16 13 184 6.8 CR 7 TK38 1 110 54 A III 99 436 15.6 15 503 2.0 CR 10 TK44 1 159 146 C severe 12 399 2.9 0 303 0 CR 84 TK47 1 90 19 A II 351 422 45.4 34 222 13.3 CR 3 TK50 1 66 41 A II 355 238 59.9 26 88 22.8 CR 18 TK1007A 1.2 117 30 A I 20 146 12 / / / CR NA TK1011A 1.06 78 20 A II 11 97 11.3 7 131 5 CR 90 TK1014A 0.96 15 10 A II 11 41 26.8 2 85 2.3 CR 8 Figure 1 Figure 1. Figure 2 Figure 2. Disclosures Bonini: MolMed S.p.A.: Consultancy. Colombi:MolMed: Employment. Lambiase:MolMed S.p.A: Employment. Bordignon:MolMed: Employment.

scholarly journals Parallel Evolution in Pseudomonas aeruginosa over 39,000 Generations In Vivo

mBio ◽  
2010 ◽  
Vol 1 (4) ◽  
Author(s):  
Holly K. Huse ◽  
Taejoon Kwon ◽  
James E. A. Zlosnik ◽  
David P. Speert ◽  
Edward M. Marcotte ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Chronic Infection ◽  
Parallel Evolution ◽  
Chronic Infections ◽  
Content Type ◽  
Airway Infections ◽  
Chronic Colonization

ABSTRACTThe Gram-negative bacteriumPseudomonas aeruginosais a common cause of chronic airway infections in individuals with the heritable disease cystic fibrosis (CF). After prolonged colonization of the CF lung,P. aeruginosabecomes highly resistant to host clearance and antibiotic treatment; therefore, understanding how this bacterium evolves during chronic infection is important for identifying beneficial adaptations that could be targeted therapeutically. To identify potential adaptive traits ofP. aeruginosaduring chronic infection, we carried out global transcriptomic profiling of chronological clonal isolates obtained from 3 individuals with CF. Isolates were collected sequentially over periods ranging from 3 months to 8 years, representing up to 39,000in vivogenerations. We identified 24 genes that were commonly regulated by all 3P. aeruginosalineages, including several genes encoding traits previously shown to be important forin vivogrowth. Our results reveal that parallel evolution occurs in the CF lung and that at least a proportion of the traits identified are beneficial forP. aeruginosachronic colonization of the CF lung.IMPORTANCEDeadly diseases like AIDS, malaria, and tuberculosis are the result of long-term chronic infections. Pathogens that cause chronic infections adapt to the host environment, avoiding the immune response and resisting antimicrobial agents. Studies of pathogen adaptation are therefore important for understanding how the efficacy of current therapeutics may change upon prolonged infection. One notorious chronic pathogen isPseudomonas aeruginosa, a bacterium that causes long-term infections in individuals with the heritable disease cystic fibrosis (CF). We used gene expression profiles to identify 24 genes that commonly changed expression over time in 3P. aeruginosalineages, indicating that these changes occur in parallel in the lungs of individuals with CF. Several of these genes have previously been shown to encode traits critical forin vivo-relevant processes, suggesting that they are likely beneficial adaptations important for chronic colonization of the CF lung.

scholarly journals Intraspecific antagonism through viral toxin encoded by chronic Sulfolobus spindle-shaped virus

2021 ◽  
Vol 377 (1842) ◽  
Author(s):  
Samantha J. DeWerff ◽  
Changyi Zhang ◽  
John Schneider ◽  
Rachel J. Whitaker
Keyword(s):  
Competitive Advantage ◽  
Chronic Infection ◽  
Genetic Basis ◽  
Competitive Interactions ◽  
Theme Issue ◽  
Expression Plasmid ◽  
Host Interactions ◽  
Sulfolobus Islandicus ◽  
Nested Network

Virus–host interactions evolve along a symbiosis continuum from antagonism to mutualism. Long-term associations between virus and host, such as those in chronic infection, will select for traits that drive the interaction towards mutualism, especially when susceptible hosts are rare in the population. Virus–host mutualism has been demonstrated in thermophilic archaeal populations where Sulfolobus spindle-shaped viruses (SSVs) provide a competitive advantage to their host Sulfolobus islandicus by producing a toxin that kills uninfected strains. Here, we determine the genetic basis of this killing phenotype by identifying highly transcribed genes in cells that are chronically infected with a diversity of SSVs. We demonstrate that these genes alone confer growth inhibition by being expressed in uninfected cells via a Sulfolobus expression plasmid. Challenge of chronically infected strains with vector-expressed toxins revealed a nested network of cross-toxicity among divergent SSVs, with both broad and specific toxin efficacies. This suggests that competition between viruses and/or their hosts could maintain toxin diversity. We propose that competitive interactions among chronic viruses to promote their host fitness form the basis of virus–host mutualism. This article is part of the theme issue ‘The secret lives of microbial mobile genetic elements’.

Long term in vivo reactions to PTFE and polyester in the cardiovascular system - what will be the fate of septal defect occlusion devices?

2014 ◽  
Vol 62 (S 01) ◽  
Author(s):  
M. Sigler ◽  
S. Huell ◽  
R. Foth ◽  
W. Ruschewski ◽  
T. Tirilomis ◽  
...  
Keyword(s):  
Cardiovascular System ◽  
Septal Defect

The depressing (negative) effect of oestradiol benzoate on the in vitro secretion of LH and FSH by the pituitary gland of the LRH-pretreated long-term ovariectomized rat changes into the augmentative (positive) effect after discontinuation of the LRH-pretreatment

1985 ◽  
Vol 110 (3) ◽  
pp. 329-337 ◽  
Author(s):  
G. A. Schuiling ◽  
H. Moes ◽  
T. R. Koiter
Keyword(s):  
Depressing Effect ◽  
Ovx Rats ◽  
Gonadotrophin Secretion ◽  
Oestradiol Benzoate ◽  
Negative Effect ◽  
Positive Effect ◽  
Perifusion System

Abstract. The effect of pretreatment in vivo with oestradiol benzoate on in vitro secretion of LH and FSH was studied in long-term ovariectomized (OVX) rats both at the end of a 5-day continuous in vivo pretreatment with LRH and 4-days after cessation of such LRH pretreatment. Rats were on day 0 sc implanted with osmotic minipumps which released LRH at the rate of 250 ng/h. Control rats were implanted with a piece of silicone elastomer with the dimensions of a minipump. On days 2 and 4 the rats were injected with either 3 μg EB or with oil. On day 5 part of the rats were decapitated and the in vitro autonomous (i.e. non-LRH-stimulated) and 'supra-maximally' LRHstimulated release of LH and FSH was studied using a perifusion system. From other rats the minipumps were removed on day 5 and perifusion was performed on day 9. On the 5th day of the in vivo LRH pretreatment the pituitary LH/FSH stores were partially depleted; the pituitaries of the EB-treated rats more so than those of the oil-injected rats. EB alone had no significant effect on the content of the pituitary LH- and FSH stores. On day 9, i.e. 4 days after removal of the minipumps, the pituitary LH and FSH contents had increased in both the oil- and the EB injected rats, but had not yet recovered to control values. In rats not subjected to the 5-days pretreatment with LRH EB had a positive effect on the supra-maximally LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. EB had no effect on the non-stimulated secretion of FSH. After 5 days of in vivo pretreatment with LRH only, the in vitro non-stimulated and supra-maximally LRH-stimulated secretion of both LH and FSH were strongly impaired, the effect correlating well with the LRH-induced depletion of the pituitary LH/FSH stores. In such LRH-pretreated rats EB had on day 5 a negative effect on the (already depressed) LRH-stimulated secretion of LH (not on that of FSH). EB had no effect on the non-stimulated LH/FSH secretion. It could be demonstrated that the negative effect of the combined LRH/EB pretreatment was mainly due to the depressing effect of this treatment on the pituitary LH and FSH stores: the effect of oestradiol on the pituitary LRH-responsiveness (release as related to pituitary gonadotrophin content) remained positive. In LRH-pretreated rats, however, this positive effect of EB was smaller than in rats not pretreated with LRH. Four days after removal of the minipumps there was again a positive effect of EB on the LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. The positive effect of EB on the pituitary LRH-responsiveness was as strong as in rats which had not been exposed to exogenous LRH. The non-stimulated secretion of FSH was again not affected by EB. The results demonstrate that the effect of EB on the oestrogen-sensitive components of gonadotrophin secretion consists of two components: an effect on the pituitary LRH-responsiveness proper, and an effect on the pituitary LH/FSH stores. The magnitude of the effect of EB on the LRH-responsiveness is LRH dependent: it is very weak (almost zero) in LRH-pretreated rats, but strong in rats not exposed to LRH as well as in rats of which the LRH-pretreatment was stopped 4 days previously. Similarly, the effect of EB on the pituitary LH and FSH stores is LRH-dependent: in the absence of LRH, EB has no influence on the contents of these stores, but EB can potentiate the depleting effect of LRH on the LH/FSH-stores. Also this effect disappear after cessation of the LRH-pretreatment.

Enhanced Stability and Controlled Delivery of MOF Encapsulated Vaccines and Their Immunogenic Response In Vivo

2018 ◽  
Author(s):  
Michael Luzuriaga ◽  
Raymond P. Welch ◽  
Madushani Dharmawardana ◽  
Candace Benjamin ◽  
Shaobo Li ◽  
...  
Keyword(s):  
Viral Vector ◽  
Controlled Delivery ◽  
Conformational Structure ◽  
Spectroscopy Analysis ◽  
Zeolitic Imidazolate Framework ◽  
Structural Conformation ◽  
Depth Study ◽  
Viral Nanoparticle

<div><div><div><p>Vaccines have an innate tendency to lose their structural conformation upon environmental and chemical stressors. A loss in conformation reduces the therapeutic ability to prevent the spread of a pathogen. Herein, we report an in-depth study of zeolitic imidazolate framework-8 (ZIF-8) and its ability to provide protection for a model viral vector against dena- turing conditions. The immunoassay and spectroscopy analysis together demonstrate enhanced thermal and chemical stability to the conformational structure of the encapsulated viral nanoparticle. The long-term biological activity of this virus-ZIF composite was investigated in animal models to further elucidate the integrity of the encapsulated virus, the bio-safety, and immunogenicity of the overall composite. Additionally, histological analysis found no observable tissue damage in the skin or vital organs in mice, following multiple subcutaneous administrations. This study shows that ZIF-based protein composites are strong candidates for improved preservation of proteinaceous drugs, are biocompatible, and capable of controlling the release and adsorption of drugs in vivo.</p></div></div></div>

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