Production and analysis of immunomodulatory excretory-secretory products from the mouse gastrointestinal nematode Heligmosomoides polygyrus bakeri.
Heligmosomoides polygyrus bakeri (Hpb) infection in mice is a convenient model for studying the pathophysiology and immunology of gastrointestinal (GI) helminth infection. Hpb infection suppresses immune responses to bystander antigens and unrelated pathogens, and it slows the progression and modifies the outcome of immune-mediated diseases. Hpb-derived excretory-secretory (ES) products potently modulate CD4(+) helper T cell (TH) responses by inducing regulatory T cells, tolerogenic dendritic cells (DCs) and immunoregulatory cytokines. This observation has spiked interest in identifying the immunomodulatory molecules, especially proteins, in ES products from Hpb and other GI nematodes for development as novel therapies to treat individuals with immune-mediated diseases, such as inflammatory bowel diseases (IBDs). In this protocol, we describe how to (i) maintain Hpb in the laboratory for experimental infections, (ii) collect adult worms from infected mice to generate ES products and (iii) evaluate the modulatory effects of ES products on toll-like receptor (TLR) ligand-induced maturation of CD11c(+) DCs. The three major sections of the PROCEDURE can be used independently, and they require ∼6, 10 and 27 h, respectively. Although other methods use a modified Baermann apparatus to collect Hpb adult worms, we describe a method that involves dissection of adult worms from intestinal tissue. The protocol will be useful to investigators studying the host-parasite interface and identifying and analyzing helminth-derived molecules with therapeutic potential.
1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Department of Medicine, McGill University, Montreal, Quebec, Canada. [3] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Rajesh M
RM
Valanparambil
1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Mariela
M
Segura
1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Mifong
M
Tam
1] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada. [2] Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Armando
A
Jardim
1] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada. [2] Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Timothy G
TG
Geary
1] Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada. [2] Department of Medicine, McGill University, Montreal, Quebec, Canada. [3] Centre for Host-Parasite Interactions, Institute of Parasitology, McGill University, Ste.-Anne de Bellevue, Quebec, Canada.
Mary M
MM
Stevenson
D000818:Animals; D039521:CD11c Antigen / Q000378:metabolism; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D005243:Feces / Q000469:parasitology; D005260:Female; D006790:Host-Parasite Interactions / Q000276:immunology; D007155:Immunologic Factors / Q000302:isolation & purification / Q000494:pharmacology*; D007422:Intestines / Q000469:parasitology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009351:Nematospiroides dubius / Q000276:immunology* / Q000302:isolation & purification* / Q000502:physiology; D017206:Strongylida Infections / Q000276:immunology* / Q000469:parasitology
D039521:CD11c Antigen; D007155:Immunologic Factors