Trichinella spiralis-induced mastocytosis and erythropoiesis are simultaneously supported by a bipotent mast cell/erythrocyte precursor cell

Five highly soluble, chymotrypsin-like, neutral serine proteases, with molecular masses in the range 30-33 kDa, were isolated from Trichinella spiralis-infected mouse small intestine. These enzymes were closely related antigenically on Western blotting and by Ouchterlony double diffusion using a polyclonal, cross-absorbed, sheep antibody raised against mouse mast cell protease-1 (MMCP-1) and on the basis of N-terminal amino acid sequence analysis, were identified as variant forms of MMCP-1. Substrate and inhibitor analysis confirmed that the five variants (MMCP-1 A-E) had similar characteristics, although highly significant (P = 0.025 to P < 0.0001) variations in Km and kcat, were detected. Against human alpha 1-proteinase inhibitor the Ki for MMCP-1C (45 pM) was significantly (P < 0.0001) greater than those for the other proteases (0.76-2.2 pM). The differences in electrophoretic mobility are probably a result of variable glycosylation, since removal of N-linked carbohydrate produced a polypeptide of approx. 28 kDa in each case which was, like the native enzyme, immunoreactive on Western blotting. A much less soluble 28 kDa enzyme was isolated from serosal mast cells and identified as MMCP-4 by N-terminal amino acid sequencing. Like MMCP-1 it has chymotrypsin-like substrate specificities with activity at neutral pH. However, it was antigenically distinct from MMCP-1 and, using sheep anti-MMCP-1, was not detected on Western blotting or by Ouchterlony double diffusion, e.l.i.s.a. or immunohistochemistry. This last technique established that the MMCP-1 variants were uniquely present in enteric mast cells, thereby providing a highly selective means of distinguishing the mucosal and connective tissue mast cell subsets in the mouse.

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Absence of intestinal mast cell response in congenitally athymic mice during Trichinella spiralis infection

Nature ◽

10.1038/264258a0 ◽

1976 ◽

Vol 264 (5583) ◽

pp. 258-260 ◽

Cited By ~ 181

Author(s):

E. J. RUITENBERG ◽

ANNEKE ELGERSMA

Keyword(s):

Mast Cell ◽

Cell Response ◽

Trichinella Spiralis ◽

Athymic Mice

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Expression Profiling Reveals Novel Innate and Inflammatory Responses in the Jejunal Epithelial Compartment during Infection with Trichinella spiralis

Infection and Immunity ◽

10.1128/iai.72.10.6076-6086.2004 ◽

2004 ◽

Vol 72 (10) ◽

pp. 6076-6086 ◽

Cited By ~ 39

Author(s):

Pamela A. Knight ◽

Alan D. Pemberton ◽

Kevin A. Robertson ◽

Douglas J. Roy ◽

Steven H. Wright ◽

...

Keyword(s):

Mast Cell ◽

Goblet Cell ◽

Expression Profiling ◽

Time Course ◽

Inflammatory Responses ◽

Trichinella Spiralis ◽

Mucosal Mast Cell ◽

Screening Process ◽

Mucin Gene ◽

Enhanced Production

ABSTRACT Infection with intestinal nematodes induces profound pathological changes to the gut that are associated with eventual parasite expulsion. We have applied expression profiling as an initial screening process with oligonucleotide microarrays (Affymetrix MG-U74AV2 gene chips) and time course kinetics to investigate gene transcription triggered by the intraepithelial nematode Trichinella spiralis in jejunal epithelium from BALB/c mice. Of the 4,114 genes detected, 2,617 were present in all uninfected and T. spiralis-infected replicates, 8% of which were notably upregulated, whereas 12% were downregulated at the time of worm expulsion (day 14 postinfection). Upregulation of goblet cell mucin gene transcripts intestinal mucin gene 3 (MUC3), calcium chloride channel 5 (CLCA5), and goblet cell gene 4 (GOB4) is consistent with enhanced production and alteration of mucus, whereas a 60- to 70-fold upregulation of transcripts for mast cell proteases 1 and 2 (MCPT-1 and -2) is consistent with intraepithelial mucosal mast cell recruitment. Importantly, there was novel expression of sialyltransferase 4C (SIAT4C), small proline-rich protein 2A (SPRR2A), and resistin-like molecule β (RELMβ) on day 14 postinfection. In contrast, DNase I and regenerating protein 3 (REG3) transcripts were substantially downregulated. Time course analyses revealed early (within 48 h of infection) induction of Siat4c, Sprr2A, and Relmβ and later (within 120 h) induction of Mcpt-1 and -2. The findings demonstrate early innate responses and later inflammatory changes within the epithelium. The early epithelial responses may be associated both with repair (Sprr2A) and with the development of innate immunity (Siat4c and Relmβ).

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Effect of mast cell stabilization on angiogenesis in primary and secondary experimental Trichinella spiralis infection

Parasites & Vectors ◽

10.1186/s13071-021-05075-9 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Marwa A. EL-Dardiry ◽

Amany A. Abdel-Aal ◽

Magda S. A. Abdeltawab ◽

Mona El-Sherbini ◽

Marwa A. Hassan ◽

...

Keyword(s):

Mast Cell ◽

Inflammatory Response ◽

Structural Damage ◽

Trichinella Spiralis ◽

Histopathological Examination ◽

Cell Formation ◽

Parasite Load ◽

Drug Regimen ◽

Parasite Count ◽

Angiogenic Potential

Abstract Background Mast cells are known to affect the primary and secondary immune responses against parasites, and this effect is partially mediated through the release of pro-angiogenic mediators. The aim of this study was to explore the effect of the mast cell stabilizer (MCS), ketotifen, with and without albendazole, an anti-parasitic prescription medicine, on the inflammatory response against Trichinella spiralis, with the overall aim to investigate its effect on angiogenesis accompanying nurse cell formation. Methods The effect of ketotifen and albendazole was explored in eight groups of female BALB/c mice. Four groups were sensitized with a small dose of T. spiralis larvae. The drug regimen was then applied to both sensitized (challenged) and non-sensitized mice. The parasite load was assessed by histopathological examination of the small intestine and muscle tissue, and angiogenesis was assessed by immunohistochemistry to determine the expression of vascular endothelial growth factor (VEGF). Results Sensitized mice showed a significantly lower parasite load and a more pronounced inflammatory response than mice receiving a single infective dose of T. spiralis larvae. All treated groups showed a significant reduction in parasite count compared to the control groups (groups IAa and IBa), reaching approximately an 98.8% reduction in adult parasite count in the sensitized group treated with albendazole (groups IIAb and IIBb). MCS significantly decreased the parasite count during both the intestinal or muscular phases, reduced tissue inflammation, and decreased local VEGF expression, both in the non-sensitized and sensitized groups. Conclusion Sensitization with a low dose of T. spiralis larvae was found to confer a partial protective immunity against re-infection and to positively affect the study outcomes, thus underlining the importance of vaccination, but after extensive studies. The anti-angiogenic effect of MCS protects against larval encystation during the muscle phase. The anti-angiogenic potential of albendazole suggests that the action of this anti-helminthic during trichinellosis is not confined to structural damage to the parasite cuticle but includes an effect on host immunopathological response. Graphical Abstract

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Mucosal mast cells and nematode infection: strain-specific differences in mast cell precursor frequency revisited

Journal of Helminthology ◽

10.1079/joh2002160 ◽

2003 ◽

Vol 77 (2) ◽

pp. 155-161 ◽

Cited By ~ 9

Author(s):

J.K. Brown ◽

S.H. Wright ◽

H.R.P. Miller

Keyword(s):

Bone Marrow ◽

Mast Cell ◽

Mast Cells ◽

Transforming Growth Factor ◽

Bone Marrow Cells ◽

Trichinella Spiralis ◽

Cell Precursor ◽

Mucosal Mast Cells ◽

Bone Marrow Cultures ◽

Marrow Cultures

AbstractMucosal mast cells (MMC) play an important role in the immune response against selected species of intestinal nematode. The kinetics with which different strains of inbred mice resolve infection withTrichinella spiraliscorrelates with their ability to mount MMC responses in the intestinal mucosa. Homologues of MMC that express and constitutively secrete abundant amounts of the granule chymase, mouse mast cell protease-1 (mMCP-1), can be generatedin vitrofrom bone marrow cultures supplemented with interleukins-3 and -9, stem cell factor and transforming growth factor-β1. Using the enhanced growth characteristics of these MMC homologues, a novel limiting dilution assay for mast cell precursor (MCp) frequency has been developed. The assay is highly specific, in that cultures containing mast cells are identified with mMCP-1 specific antibody, and almost three-fold more sensitive than previously published systems. MCp frequencies were compared in BALB/c and C57/BL10 strains of mice that, respectively, respond rapidly and slowly to infection withT. spiralis. MCp frequency (1/378 bone marrow cells) was significantly greater (P<0.05) in BALB/c than C57/BL10 mice (frequency: 1/751). Similarly the rate of growth of MMC homologues and the production of mMCP-1 was significantly (P<0.05) greater in BALB/c than in C57/BL10 bone marrow cultures.

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Delayed Expulsion of the Nematode Trichinella spiralisIn Mice Lacking the Mucosal Mast Cell–Specific Granule Chymase, Mouse Mast Cell Protease-1

Journal of Experimental Medicine ◽

10.1084/jem.192.12.1849 ◽

2000 ◽

Vol 192 (12) ◽

pp. 1849-1856 ◽

Cited By ~ 211

Author(s):

Pamela A. Knight ◽

Steven H. Wright ◽

Catherine E. Lawrence ◽

Yvonne Y.W. Paterson ◽

Hugh R.P. Miller

Keyword(s):

Mast Cell ◽

Serine Proteases ◽

Trichinella Spiralis ◽

Gastrointestinal Nematodes ◽

Intestinal Lumen ◽

Mucosal Mast Cell ◽

Mast Cell Protease ◽

Muscle Larvae ◽

Mouse Mast Cell

Expulsion of gastrointestinal nematodes is associated with pronounced mucosal mast cell (MMC) hyperplasia, differentiation, and activation, accompanied by the systemic release of MMC granule chymases (chymotrypsin-like serine proteases). The β-chymase mouse mast cell protease-1 (mMCP-1) is expressed predominantly by intraepithelial MMCs, and levels in the bloodstream and intestinal lumen are maximal at the time of worm expulsion in parasitized mice. To address the in vivo functions of MMC-specific β-chymases, we have generated transgenic mice that lack the mMCP-1 gene. They were backcrossed onto a congenic BALB/c background to investigate the response to nematode infection. The deletion of the mMCP-1 gene is associated with significantly delayed expulsion of Trichinella spiralis and increased deposition of muscle larvae in BALB/c mice despite the presence of normal and sometimes increased numbers of MMCs. Neither worm fecundity nor worm burdens were altered in Nippostrongylus-infected mMCP-1−/− BALB/c mice. These data demonstrate, for the first time, that the ablation of an MMC-derived effector molecule compromises the expulsion process.

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Influence of variation in host strain and parasite isolate on inflammatory and antibody responses toTrichinella spiralisin mice

Parasitology ◽

10.1017/s0031182000067111 ◽

1993 ◽

Vol 106 (4) ◽

pp. 371-378 ◽

Cited By ~ 28

Author(s):

P. K. Goyal ◽

D. Wakelin

Keyword(s):

Mast Cell ◽

Trichinella Spiralis ◽

Serum Levels ◽

Antibody Responses ◽

High Responder ◽

Worm Recovery ◽

Response Phenotype ◽

Igg Isotypes ◽

Low Responder ◽

Antibody Levels

SUMMARYVariation in the immunogenicity of 3 isolates ofTrichinella spiraliswas assessed by the parameters of adult worm recovery, mast cell, eosinophil and antibody responses in mice of defined response phenotype. The levels of the protective, inflammatory and immune responses induced by infection differed between the isolates. Isolates showed considerable variation in the capacity to elicit mast cell and eosinophil responses. All induced increases in parasite-specific antibody, levels of total (IgGAM) antibody and of IgM and IgG isotypes rose steadily after infection, but there were significant differences in levels of response. The IgGAM response was correlated with the number of worms present, i.e. the greatest response was seen in low responder (C57BL/10) mice infected with the longest-surviving isolates. All isolates elicited specific IgG1 and IgG2a antibodies after infection, although, again, there were isolate-specific differences in the levels and kinetics of response. Levels of these isotypes were always higher, although not significantly so, in high-responder NIH mice. Low-responder mice showed higher IgE serum levels than high-responder mice after infection, one isolate giving much higher IgE values than the other two.

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Dendritic cell expression of the transcription factor T-bet regulates mast cell progenitor homing to mucosal tissue

Journal of Experimental Medicine ◽

10.1084/jem.20060626 ◽

2007 ◽

Vol 204 (2) ◽

pp. 431-439 ◽

Cited By ~ 34

Author(s):

Pilar Alcaide ◽

Tatiana G. Jones ◽

Graham M. Lord ◽

Laurie H. Glimcher ◽

Jenny Hallgren ◽

...

Keyword(s):

Transcription Factor ◽

Mast Cell ◽

Adhesion Molecule ◽

Trichinella Spiralis ◽

Cellular Adhesion ◽

Cell Trafficking ◽

Peripheral Tissues ◽

Cellular Adhesion Molecule

The transcription factor T-bet was identified in CD4+ T cells, and it controls interferon γ production and T helper type 1 cell differentiation. T-bet is expressed in certain other leukocytes, and we recently showed (Lord, G.M., R.M. Rao, H. Choe, B.M. Sullivan, A.H. Lichtman, F.W. Luscinskas, and L.H. Glimcher. 2005. Blood. 106:3432–3439) that it regulates T cell trafficking. We examined whether T-bet influences homing of mast cell progenitors (MCp) to peripheral tissues. Surprisingly, we found that MCp homing to the lung or small intestine in T-bet−/− mice is reduced. This is reproduced in adhesion studies using bone marrow–derived MCs (BMMCs) from T-bet−/− mice, which showed diminished adhesion to mucosal addresin cellular adhesion molecule–1 (MAdCAM-1) and vascular cell adhesion molecule–1 (VCAM-1), endothelial ligands required for MCp intestinal homing. MCp, their precursors, and BMMCs do not express T-bet, suggesting that T-bet plays an indirect role in homing. However, adoptive transfer experiments revealed that T-bet expression by BM cells is required for MCp homing to the intestine. Furthermore, transfer of WT BM-derived dendritic cells (DCs) to T-bet−/− mice restores normal MCp intestinal homing in vivo and MCp adhesion to MAdCAM-1 and VCAM-1 in vitro. Nonetheless, T-bet−/− mice respond vigorously to intestinal infection with Trichinella spiralis, eliminating a role for T-bet in MC recruitment to sites of infection and their activation and function. Therefore, remarkably, T-bet expression by DCs indirectly controls MCp homing to mucosal tissues.

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Prolonged expulsion of adultTrichinella spiralisand eosinophil infiltration in mast cell-deficient W/Wvmice

Journal of Helminthology ◽

10.1017/s0022149x00008002 ◽

1985 ◽

Vol 59 (3) ◽

pp. 233-239 ◽

Cited By ~ 20

Author(s):

M. Kamiya ◽

Y. Oku ◽

H. Itayama ◽

M. Ohbayashi

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Trichinella Spiralis ◽

Specific Ige ◽

Passive Cutaneous Anaphylaxis ◽

Eosinophil Infiltration ◽

Serum Antibodies ◽

Indirect Haemagglutination ◽

Specific Serum

ABSTRACTEosinophil infiltrations were observed in the intestine and the muscle of bothTrichinella spiralis-infected (WBxC57BL/6)F1-W/Wvmice and their littermates, WBB6F1−+/+, +/W, +/Wv, almost to the same extent. W/Wvmice did not show infiltration of subepithelial mast cells and globule leucocytes in response toT. spiralisinfection. Increased numbers of these cells were observed in their littermates. Worms in W/Wvmice were retained for longer periods than those in littermates. Also, no difference was noted in the production of specific serum antibodies between W/Wvmice and their littermates, as determined by passive cutaneous anaphylaxis (PCA) for specific IgE and by indirect haemagglutination (IHA). These results suggest a possible participation of SMC, GL and eosinophils in the expulsion of adultT. spiralis.

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In vivo exit of c-kit+/CD49dhi/β7+ mucosal mast cell precursors from the bone marrow following infection with the intestinal nematode Trichinella spiralis

Blood ◽

10.1182/blood-2003-09-3146 ◽

2004 ◽

Vol 103 (7) ◽

pp. 2655-2660 ◽

Cited By ~ 35

Author(s):

Joanne L. Pennock ◽

Richard K. Grencis

Keyword(s):

Bone Marrow ◽

Mast Cell ◽

Mast Cells ◽

Trichinella Spiralis ◽

Mucosal Mast Cell ◽

Hematopoietic Tissue ◽

In Vivo Experiments ◽

Intestinal Nematode

Abstract We have used the parasite helminth Trichinella spiralis to study the generation and differentiation of mast cell progenitors in the bone marrow of mice, as this infection triggers an intestinal mastocytosis which correlates with parasite expulsion. C-kit+ mast cell progenitors have previously been defined by methylcellulose colony-forming units and by limiting dilution assays in vitro. In vivo experiments have demonstrated the essential requirement by mast cells for specific integrin expression. We have defined 2 c-kit+ populations in the bone marrow, one of which coexpresses CD49d/β7 integrin, a marker essential for small intestine immigration. We have confirmed the phenotype of these cells by using antagonistic anti-c-kit antibody in vivo. Our data show that the loss of c-kit+/β7+ cells from the bone marrow correlates with their appearance in the blood and precedes detection of mature mast cells in the gut by 3 days. This exit correlates with an increase in soluble stem cell factor (SCF) in the serum, suggesting that the c-kit/SCF interaction may be chemotactic or haptotactic in nature. This study shows that during infection the bone marrow environment generates mast cells destined for the intestinal mucosa before their exit into the periphery, indicating a clear interplay between infection site and hematopoietic tissue. (Blood. 2004;103:2655-2660)

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