Cultivation of &lt;em&gt;Heligmosomoides Polygyrus:&lt;/em&gt; An Immunomodulatory Nematode Parasite and its Secreted Products

Johnston, Chris J. C.; Robertson, E. Graeme; Harcus, Yvonne; Grainger, John; Coakley, Gillian; Smyth, Danielle J.; McSorley, Henry J.; Maizels, Rick M.

doi:10.3791/52412

Cited by 79 publications

(72 citation statements)

References 47 publications

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“…A bacterial monoculture and nematode growth media are sufficient to generate culturesterile L3 Hpb larvae In a laboratory setting, infective (L3) stage Hpb are normally grown using a copra-culture method, with very little dissection of growth parameters. In short, the feces of infected mice (containing Hpb eggs) are cultured in a moist, dark environment for several days 9 . Therein, the eggs hatch and develop through two free-living stages of larvae (L1 and L2), up to the infective L3 stage 7 .…”

Section: Resultsmentioning

confidence: 99%

The gut microbiota limits Th2 immunity toHeligmosomoides polygyrus bakeriinfection

Russell

Faubert

Verdú

et al. 2020

Preprint

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Helminth-induced alterations to the gut microbiota have been shown to affect immune responses at local and peripheral sites. Studies examining helminth-microbiota interactions, however, have been limited due to the practical constraints of performing germ-free experiments with parasites that thrive in microbial-rich conditions to complete their development. The infectious (L3) larvae of the murine helminth Heligmosomoides polygyrus bakeri (Hpb), for example, are normally reared using a fecal-culture method and therefore are inherently unsuitable for germ-free studies in vivo. Herein, we detail an adapted methodology for rearing effectively germ-free Hpb larvae that are able to maintain the axenic status of a germ-free host during infection. We validate that these larvae do not display any fitness defects relative to fecal-grown larvae and evoke a comparable immune response in vivo. Characterization of axenic Hpb infection reveals that the commensal microbiota play a multifaceted role during infection -curbing the anti-Hpb Th2 response and directing the resolution of tissue granulomas, while simultaneously promoting parasite fitness. Overall these data demonstrate a mutualistic relationship between commensal microbes, enteric helminths and the infected host.

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Section: Resultsmentioning

confidence: 99%

The gut microbiota limits Th2 immunity toHeligmosomoides polygyrus bakeriinfection

Russell

Faubert

Verdú

et al. 2020

Preprint

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“…Heligmosomoides polygyrus bakeri life cycle was maintained in house and infective third-stage larvae (L3) were obtained as described elsewhere [51]. Mice were infected with 200 H .…”

Section: Methodsmentioning

confidence: 99%

Macrophage origin limits functional plasticity in helminth-bacterial co-infection

et al. 2017

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Rapid reprogramming of the macrophage activation phenotype is considered important in the defense against consecutive infection with diverse infectious agents. However, in the setting of persistent, chronic infection the functional importance of macrophage-intrinsic adaptation to changing environments vs. recruitment of new macrophages remains unclear. Here we show that resident peritoneal macrophages expanded by infection with the nematode Heligmosomoides polygyrus bakeri altered their activation phenotype in response to infection with Salmonella enterica ser. Typhimurium in vitro and in vivo. The nematode-expanded resident F4/80high macrophages efficiently upregulated bacterial induced effector molecules (e.g. MHC-II, NOS2) similarly to newly recruited monocyte-derived macrophages. Nonetheless, recruitment of blood monocyte-derived macrophages to Salmonella infection occurred with equal magnitude in co-infected animals and caused displacement of the nematode-expanded, tissue resident-derived macrophages from the peritoneal cavity. Global gene expression analysis revealed that although nematode-expanded resident F4/80high macrophages made an anti-bacterial response, this was muted as compared to newly recruited F4/80low macrophages. However, the F4/80high macrophages adopted unique functional characteristics that included enhanced neutrophil-stimulating chemokine production. Thus, our data provide important evidence that plastic adaptation of MΦ activation does occur in vivo, but that cellular plasticity is outweighed by functional capabilities specific to the tissue origin of the cell.

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“…CBA x C57BL/6 F1 (CBF1) mice were infected with 400 L3 infective-stage H. bakeri larvae by gavage and adult nematodes were collected from the small intestine 14 days post infection. The nematodes were washed and maintained in serum-free media in vitro as described previously (17). For genomic analyses, DNA was collected from adult worms immediately following harvest from the gut, with extensive washing to remove host material followed by purification using Zymo Research Genomic DNA Clean & Concentrator kit following manufacturer's instructions (further details in Supplementary Methods).…”

Section: Nematode Lifecycle and Sample Collectionmentioning

confidence: 99%

Secretion of an Argonaute protein by a parasitic nematode and the evolution of its siRNA guides

Chow

Koutsovoulos

Ovando-Vázquez³

et al. 2018

Preprint

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Cultivation of <em>Heligmosomoides Polygyrus:</em> An Immunomodulatory Nematode Parasite and its Secreted Products

Cited by 79 publications

References 47 publications

The gut microbiota limits Th2 immunity toHeligmosomoides polygyrus bakeriinfection

The gut microbiota limits Th2 immunity toHeligmosomoides polygyrus bakeriinfection

Macrophage origin limits functional plasticity in helminth-bacterial co-infection

Secretion of an Argonaute protein by a parasitic nematode and the evolution of its siRNA guides

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