scholarly journals AvrA effector protein of Salmonella enterica serovar Enteritidis is expressed and translocated in mesenteric lymph nodes at late stages of infection in mice

Microbiology ◽  
2014 ◽  
Vol 160 (6) ◽  
pp. 1191-1199 ◽  
Author(s):  
Mónica N. Giacomodonato ◽  
Mariángeles Noto Llana ◽  
María del Rosario Aya Castañeda ◽  
Fernanda R. Buzzola ◽  
Sebastián H. Sarnacki ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Culture Conditions ◽  
Effector Protein ◽  
Post Inoculation ◽  
Mesenteric Lymph Nodes ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Major Health Problem ◽  
Mesenteric Lymph

Salmonellosis is a major health problem worldwide. Salmonella enterica serovar Enteritidis (S. Enteritidis) has been a primary cause of Salmonella outbreaks in many countries. AvrA is an SPI-1 effector protein involved in the enteritis pathway, with critical roles in inhibiting inflammation and apoptosis. In this work, we constructed an AvrA-FLAG-tagged strain of S. Enteritidis to analyse the expression profile of AvrA in vitro, in cell culture and in vivo. AvrA expression and secretion were observed in vitro under culture conditions that mimicked intestinal and intracellular environments. In agreement, bacteria isolated from infected cell monolayers expressed and translocated AvrA for at least 24 h post-inoculation. For in vivo experiments, BALB/c mice were inoculated by the natural route of infection with the AvrA-FLAG strain. Infecting bacteria and infected cells were recovered from mesenteric lymph nodes (MLN). Our results showed that AvrA continues to be synthesized in vivo up to day 8 post-inoculation. Moreover, AvrA translocation was detected in the cytosol of cells isolated from MLN 8 days after infection. Interestingly, we observed that AvrA is secreted by both type three secretion system (T3SS)-1 and T3SS-2. In summary, these findings indicate that AvrA expression is not constrained to the initial host–bacteria encounter in the intestinal environment as defined previously. The AvrA effector may participate also in systemic S. Enteritidis infection.

scholarly journals Intraperitoneal Glucose Transport to Micrometastasis: A Multimodal In Vivo Imaging Investigation in a Mouse Lymphoma Model

2021 ◽  
Vol 22 (9) ◽  
pp. 4431
Author(s):  
Zsombor Ritter ◽  
Katalin Zámbó ◽  
Xinkai Jia ◽  
Dávid Szöllősi ◽  
Dániel Dezső ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Early Stage ◽  
B Cell Lymphoma ◽  
Post Inoculation ◽  
Advanced Stage ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Systemic Treatments ◽  
18F Fdg

Bc-DLFL.1 is a novel spontaneous, high-grade transplantable mouse B-cell lymphoma model for selective serosal propagation. These cells attach to the omentum and mesentery and show dissemination in mesenteric lymph nodes. We aimed to investigate its early stage spread at one day post-intraperitoneal inoculation of lymphoma cells (n = 18 mice), and its advanced stage at seven days post-inoculation with in vivo [18F]FDG-PET and [18F]PET/MRI, and ex vivo by autoradiography and Cherenkov luminescence imaging (CLI). Of the early stage group, nine animals received intraperitoneal injections, and nine received intravenous [18F]FDG injections. The advanced stage group (n = 3) received intravenous FDG injections. In the early stage, using autoradiography we observed a marked accumulation in the mesentery after intraperitoneal FDG injection. Using other imaging methods and autoradiography, following the intravenous injection of FDG no accumulations were detected. At the advanced stage, tracer accumulation was clearly detected in mesenteric lymph nodes and in the peritoneum after intravenous administration using PET. We confirmed the results with immunohistochemistry. Our results in this model highlight the importance of local FDG administration during diagnostic imaging to precisely assess early peritoneal manifestations of other malignancies (colon, stomach, ovary). These findings also support the importance of applying topical therapies, in addition to systemic treatments in peritoneal cancer spread.

scholarly journals Global vascular expression of murine CD34, a sialomucin-like endothelial ligand for L-selectin

Blood ◽  
1994 ◽  
Vol 84 (8) ◽  
pp. 2554-2565 ◽  
Author(s):  
S Baumhueter ◽  
N Dybdal ◽  
C Kyle ◽  
LA Lasky
Keyword(s):  
Lymph Nodes ◽  
Immune Surveillance ◽  
Nod Mice ◽  
Lymphoid Tissues ◽  
Mesenteric Lymph Nodes ◽  
High Endothelial Venules ◽  
Endothelial Cell Surface ◽  
Vascular Expression

Abstract Extravasation of leukocytes into organized lymphoid tissues and into sites of inflammation is critical to immune surveillance. Leukocyte migration to peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN) and Peyer's patches (PP) depends on L-selectin, which recognizes carbohydrate-bearing, sialomucin-like endothelial cell surface glycoproteins. Two of these ligands have been identified at the molecular level. One is the potentially soluble mucin, GlyCAM 1, which is almost exclusively produced by high endothelial venules (HEV) of PLN and MLN. The second HEV ligand for L-selectin is the membrane-bound sialomucin CD34. Historically, this molecule has been successfully used to purify human pluripotent bone marrow stem cells, and limited data suggest that human CD34 is present on the vascular endothelium of several organs. Here we describe a comprehensive analysis of the vascular expression of CD34 in murine tissues using a highly specific antimurine CD34 polyclonal antibody. CD34 was detected on vessels in all organs examined and was expressed during pancreatic and skin inflammatory episodes. A subset of HEV-like vessels in the inflamed pancreas of nonobese diabetic (NOD) mice are positive for both CD34 and GlyCAM 1, and bind to an L-selectin/immunoglobulin G (IgG) chimeric probe. Finally, we found that CD34 is present on vessels of deafferentiated PLN, despite the fact that these vessels are no longer able to interact with L-selectin or support lymphocyte binding in vitro or trafficking in vivo. Our data suggest that the regulation of posttranslational carbohydrate modifications of CD34 is critical in determining its capability to act as an L-selectin ligand. Based on its ubiquitous expression, we propose that an appropriately glycosylated form of vascular CD34 may act as a ligand for L-selectin-mediated leukocyte trafficking to both lymphoid and nonlymphoid sites.

scholarly journals Eosinophilia in transgenic mice expressing interleukin 5.

1990 ◽  
Vol 172 (5) ◽  
pp. 1425-1431 ◽  
Author(s):  
L A Dent ◽  
M Strath ◽  
A L Mellor ◽  
C J Sanderson
Keyword(s):  
Bone Marrow ◽  
Transgenic Mice ◽  
Lymph Nodes ◽  
Peyer's Patches ◽  
Peyer’S Patches ◽  
Mesenteric Lymph Nodes ◽  
Gene Encoding ◽  
Interleukin 5

Experiments in vitro suggest that although interleukin 5 (IL-5) stimulates the late stages of eosinophil differentiation, other cytokines are required for the generation of eosinophil progenitor cells. In this study transgenic mice constitutively expressing the IL-5 gene were established using a genomic fragment of the IL-5 gene coupled to the dominant control region from the gene encoding human CD2. Four independent eosinophilic transgenic lines have thus far been established, two of which with 8 and 49 transgene copies, are described in detail. These mice appeared macroscopically normal apart from splenomegaly. Eosinophils were at least 65- and 265-fold higher in blood from transgenics, relative to normal littermates, and approximately two- or sevenfold more numerous relative to blood from mice infected with the helminth Mesocestoides corti. Much more modest increases in blood neutrophil, lymphocyte, and monocyte numbers were noted in transgenics, relative to normal littermates (less than threefold). Thus IL-5 in vivo is relatively specific for the eosinophil lineage. Large numbers of eosinophils were present in spleen, bone marrow, and peritoneal exudate, and were highest in the line with the greatest transgene copy number. Eosinophilia was also noted in histological sections of transgenic lungs, Peyer's patches, mesenteric lymph nodes, and gut lamina propria but not in other tissues examined. IL-5 was detected in the sera of transgenics at levels comparable to those seen in sera from parasite-infected animals. IL-3 and granulocyte/macrophage colony-stimulating factor (GM-CSF) were not found. IL-5 mRNA was detected in transgenic thymus, Peyer's patches, and superficial lymph nodes, but not in heart, liver, brain, or skeletal muscle or in any tissues from nontransgenics. Bone marrow from transgenic mice was rich in IL-5-dependent eosinophil precursors. These data indicate that induction of the IL-5 gene is sufficient for production of eosinophilia, and that IL-5 can induce the full pathway of eosinophil differentiation. IL-5 may therefore not be restricted in action to the later stages of eosinophil differentiation, as suggested by earlier in vitro studies.

scholarly journals Stromal mesenteric lymph node cells are essential for the generation of gut-homing T cells in vivo

2008 ◽  
Vol 205 (11) ◽  
pp. 2483-2490 ◽  
Author(s):  
Swantje I. Hammerschmidt ◽  
Manuela Ahrendt ◽  
Ulrike Bode ◽  
Benjamin Wahl ◽  
Elisabeth Kremmer ◽  
...  
Keyword(s):  
T Cells ◽  
Stromal Cell ◽  
Tissue Tropism ◽  
Mesenteric Lymph Nodes ◽  
Activated T Cells ◽  
Mesenteric Lymph ◽  
In Vitro Activation ◽  
Lymph Node Cells

T cells primed in the gut-draining mesenteric lymph nodes (mLN) are imprinted to express α4β7-integrin and chemokine receptor CCR9, thereby enabling lymphocytes to migrate to the small intestine. In vitro activation by intestinal dendritic cells (DC) or addition of retinoic acid (RA) is sufficient to instruct expression of these gut-homing molecules. We report that in vivo stroma cells, but not DC, allow the mLN to induce the generation of gut tropism. Peripheral LN (pLN) transplanted into the gut mesenteries fail to support the generation of gut-homing T cells, even though gut-derived DC enter the transplants and prime T cells. DC that fail to induce α4β7-integrin and CCR9 in vitro readily induce these factors in vivo upon injection into mLN afferent lymphatics. Moreover, uniquely mesenteric but not pLN stroma cells express high levels of RA-producing enzymes and support induction of CCR9 on activated T cells in vitro. These results demonstrate a hitherto unrecognized contribution of stromal cell delivered signals, including RA, on the imprinting of tissue tropism in vivo.

STUDY OF LYMPHOCTYE INTERACTION IN HUMAN MESENTERIC LYMPH NODES IN VITRO

1984 ◽  
Vol 62 (5) ◽  
pp. 531-537
Author(s):  
AW Cripps ◽  
RL Clancy ◽  
H Chipchase ◽  
EJ Hennessy
Keyword(s):  
Lymph Nodes ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph

scholarly journals Analysis of Salmonella enterica Serotype-Host Specificity in Calves: Avirulence of S. enterica Serotype Gallinarum Correlates with Bacterial Dissemination from Mesenteric Lymph Nodes and Persistence In Vivo

2002 ◽  
Vol 70 (12) ◽  
pp. 6788-6797 ◽  
Author(s):  
Susan M. Paulin ◽  
Patricia R. Watson ◽  
Annette R. Benmore ◽  
Mark P. Stevens ◽  
Philip W. Jones ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Host Specificity ◽  
Intestinal Mucosa ◽  
Salmonella Enterica ◽  
Systemic Disease ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Oral Inoculation ◽  
Pass Through

ABSTRACT Host and bacterial factors that determine whether Salmonella serotypes remain restricted to the gastrointestinal tract or penetrate beyond the mucosa and cause systemic disease remain largely undefined. Here, factors influencing Salmonella host specificity in calves were assessed by characterizing the pathogenesis of different serotypes. Salmonella enterica serotype Dublin was highly virulent intravenously, whereas S. enterica serotype Choleraesuis was moderately virulent. Both serotypes were virulent in calves infected orally. In contrast, S. enterica serotypes Gallinarum and Abortusovis were avirulent by either route. Serotypes Dublin, Gallinarum, and Abortusovis colonized the intestinal tract 24 h after oral inoculation, yet only serotype Dublin was consistently recovered from systemic tissues. Serotypes Dublin and Gallinarum invaded bovine intestines in greater numbers and induced greater enteropathogenic responses than serotypes Choleraesuis and Abortusovis. However, only serotype Dublin was able to persist within the intestinal mucosa, and use of a novel cannulation model demonstrated that serotype Dublin was able to pass through the mesenteric lymph nodes in greater numbers than serotype Gallinarum. Together, these results suggest that initial interactions with the intestinal mucosa do not correlate with host specificity, although persistence within tissues and translocation via efferent lymphatics appear to be crucial for the induction of bovine salmonellosis.

scholarly journals Analysis of IL-2 Production in Hookworm Infected Hamsters Using Generated Polyclonal Antibodies

2012 ◽  
Vol 56 (1) ◽  
pp. 37-42
Author(s):  
Ewa Długosz ◽  
Jarosław Cendrowski ◽  
Piotr Bąska ◽  
Anna Siwińska ◽  
Halina Wędrychowicz ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lymph Nodes ◽  
Cell Cultures ◽  
Polyclonal Antibodies ◽  
Mesenteric Lymph Nodes ◽  
Ancylostoma Ceylanicum ◽  
Specific Antibodies ◽  
Mesenteric Lymph ◽  
Race Pcr

Abstract The aim of the study was cloning and analysis of the entire coding sequence of hamster IL-2 by the method of RACE-PCR, its expression in Escherichia coli cells, and production of IL-2 specific antibodies. These antibodies were used to determine in vitro IL-2 production by cells derived from the spleen and mesenteric lymph nodes of Ancylostoma ceylanicum infected hamsters. The highest concentration of IL-2 was noted in supernatants from cell cultures coming from the oldest, most resistant hamsters.

Systemically Transplanted Human Bone Marrow Mesenchymal Stem Cells Primarily Traffic to Mesenteric Lymph Nodes

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2580-2580
Author(s):  
Xin Li ◽  
Wen Ling ◽  
Sharmin Khan ◽  
Yuping Wang ◽  
Angela Pennisi ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Ex Vivo ◽  
Blue Dye ◽  
Mesenteric Lymph Nodes ◽  
Live Animal ◽  
Mesenteric Lymph ◽  
Animal Imaging ◽  
Intracardiac Injection ◽  
Live Animal Imaging

Abstract Abstract 2580 Intravenously administered mesenchymal stem cells (MSCs) are trapped in pulmonary vascular bed and only few MSCs home to bone or other tissues in physiological or pathological conditions. Following intracardiac injection MSCs pass the lung barrier but their homing to bone and tissue localization is uncertain. The aim of the study was to investigate trafficking and exact localization of human MSCs following intracardiac injection into unchallenged mice and a xenograft bone tumor model. MSCs were isolated from human fetal bones (ABR Inc, Alameda CA) and expanded in DMEM-LG medium supplemented with 10% FBS. Global gene expression profiling revealed that the cultured MSCs were devoid of hematopoietic cells and expressed typical mesenchymal markers such as CD166, CD146 and CD90. We have previously shown that these MSCs are capable of differentiation into osteoblasts and adipocytes and retain their differentiation potential after multiple passages (Haematologica 2006). The MSCs were transduced with a luciferase/GFP reporter in a lentiviral vector and were maximally passaged 8 times before used in vivo. Detection of MSCs in mice was determined by live-animal imaging and ex vivo bioluminescence activity using the IVIS system, by microscopic examination of GFP-expressing cells and by immunohistochemistry for GFP. MSCs (1×106 cells/mouse) were intracardiacly injected into unconditioned SCID mice (n=8) using Dovetail Slide Micromanipulator that ensures accurate injection. Following 2 or 7 days after MSC injection to SCID mice, live-animal imaging revealed bioluminescence activity mainly in the mice abdomen but not bone, while ex vivo examination detected MSCs in various abdominal organs, primarily in reproductive organs, intestine and pancreas. Careful microscopic examination revealed localization of MSCs in draining lymph nodes attached to these organs by connective tissue. Immunohistochemistry showed GFP-expressing MSCs in the adjacent mesenteric lymph nodes but not within the organs. To confirm our findings, MSCs were intracardially injected into C57BL6 mice (n=6) that harbor functional lymph nodes. Evans blue dye which is known to accumulate in and identify lymph nodes, was injected into the rear footpad or lateral tail base of the mice, 3 hours after MSC injection and 30 minutes prior to bioluminescence and florescence analyses. The Evans blue dye and GFP positivity were co-localized, indicating specific trafficking of MSCs to lymph nodes. Culturing of the dissected lymph nodes resulted in release of GFP-expressing MSCs which regained their in vitro morphology. For testing MSCs trafficking in a xenograft model, we used our SCID-rab system constructed by implanting a 4-weeks old rabbit bone into which human myeloma cells were directly injected (Leukemia 2004; Blood 2007). In this model myeloma cells grow restrictively in the implanted bone. MSCs injected intracardiacly into SCID-rab mice were mostly found in mesenteric lymph nodes but were also detected in the myelomatous bone 72 hours after MSCs injection, validating the ability of tumor cells to attract MSCs and that these MSCs are capable of transmigration. We conclude that MSCs primarily traffic to draining lymph nodes, partially explaining their in vivo immunomodulatory activity, and that understanding the mechanism by which MSCs traffic to lymph nodes may help develop approaches to shift their homing to desired organs. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Dissemination of Persistent Intestinal Bacteria via the Mesenteric Lymph Nodes Causes Typhoid Relapse

2011 ◽  
Vol 79 (4) ◽  
pp. 1479-1488 ◽  
Author(s):  
Amanda J. Griffin ◽  
Lin-Xi Li ◽  
Sabrina Voedisch ◽  
Oliver Pabst ◽  
Stephen J. McSorley
Keyword(s):  
Animal Model ◽  
Antibiotic Therapy ◽  
Lymph Nodes ◽  
Intestinal Bacteria ◽  
Magnetic Bead ◽  
Enteric Infection ◽  
Mesenteric Lymph Nodes ◽  
Mesenteric Lymph ◽  
Systemic Spread

ABSTRACTEnteric pathogens can cause relapsing infections in a proportion of treated patients, but greater understanding of this phenomenon is hindered by the lack of appropriate animal models. We report here a robust animal model of relapsing primary typhoid that initiates after apparently successful antibiotic treatment of susceptible mice. Four days of enrofloxacin treatment were sufficient to reduce bacterial loads below detectable levels in all major organs, and mice appeared otherwise healthy. However, any interruption of further antibiotic therapy allowed renewed fecal shedding and renewed bacterial growth in systemic tissues to occur, and mice eventually succumbed to relapsing infection.In vivoimaging of luminescentSalmonellaidentified the mesenteric lymph nodes (MLNs) as a major reservoir of relapsing infection. A magnetic-bead enrichment strategy isolated MLN-resident CD11b+Gr-1−monocytes associated with low numbers of persistentSalmonella. However, the removal of MLNs increased the severity of typhoid relapse, demonstrating that this organ serves as a protective filter to restrain the dissemination of bacteria during antibiotic therapy. Together, these data describe a robust animal model of typhoid relapse and identify an important intestinal phagocyte subset involved in protection against the systemic spread of enteric infection.

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