{"id":26,"date":"2017-01-05T19:57:18","date_gmt":"2017-01-05T19:57:18","guid":{"rendered":"http:\/\/wordpress.uchospitals.edu\/basu-lab\/?page_id=26"},"modified":"2017-03-23T00:25:41","modified_gmt":"2017-03-23T00:25:41","slug":"host-pathogen","status":"publish","type":"page","link":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/host-pathogen\/","title":{"rendered":"Host-Pathogen Interaction at Controlled MOI"},"content":{"rendered":"
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There is considerable heterogeneity in outcome in host-pathogen interaction. Deciphering the underlying cause of this heterogeneity from population-level studies is hard because of uncontrolled multiplicity of Infection (MOI). Recent studies using FACS and microfluidics, along with <\/span>single-cell RNA-seq studies also indicate significant heterogeneity in<\/span> transcription of critical immune genes within seemingly homogeneous cell populations. <\/span><\/p>\n

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We study controlled infections of murine macrophages with Candida albicans,<\/em> a medically important opportunistic fungal pathogen using reverse emulsion microfluidics. Preliminary work with this system suggests infection results in two distinct outcomes: in one, phagosome maturation successfully checks growth and results in cell death of C. albicans<\/em> (GROUP A); in the other, C. albicans<\/em> successfully filaments and grows within the macrophage phagosome (GROUP B), eventually killing the macrophage.<\/p>\n

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Using emulsion microfluidics to create unique, isolated micro-environment (see left), we track C. albicans<\/em> and macrophage interaction at a pre-defined MOI. We then simultaneously incubate and image the host-pathogen cells in the droplets as the infection progresses, using a live-cell imaging system.<\/p>\n

We note some interesting results: there is an intrinsic heterogeneity in the C. albicans<\/em> population (yellow), as seen in the benign pseudo-hyphal cell (a) as well as the more virulent hyphal phenotype (b).<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/article>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n

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(a) (b)<\/address>\n
\"\"(c) (d) (e) (f)<\/address>\n

There are also important differences seen in the infection outcome, viz., successful phagocytosis of C. albicans <\/em>(yellow) by macrophage (red) (c), different amounts of pseudo-hyphal growth in C. albicans<\/em> (d & e); hyphal filaments from the phagosome growing out the macrophage, resulting in macrophage death (f).<\/p>\n

Based on fluorescence and morphologic signal, we will sort subpopulations based on outcome using microfluidic droplet sorter. Our final aim is to perform population-level and single-cell RNA-Seq on sorted macrophages and engulfed C. albicans <\/em>to transcriptionally profile cell sub-populations based on outcome- of more efficient macrophage killers (GROUP A) as well as C. albicans<\/em> of greater virulence (GROUP B).<\/p>\n","protected":false},"excerpt":{"rendered":"

There is considerable heterogeneity in outcome in host-pathogen interaction. Deciphering the underlying cause of this heterogeneity from population-level studies is hard because of uncontrolled multiplicity of Infection (MOI). Recent studies using FACS and microfluidics, along with single-cell RNA-seq studies also indicate significant heterogeneity in transcription of critical immune genes within seemingly homogeneous cell populations. We … <\/p>\n

Continue reading “Host-Pathogen Interaction at Controlled MOI”<\/span><\/a><\/p>\n","protected":false},"author":233,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-26","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/pages\/26","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/users\/233"}],"replies":[{"embeddable":true,"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/comments?post=26"}],"version-history":[{"count":4,"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/pages\/26\/revisions"}],"predecessor-version":[{"id":304,"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/pages\/26\/revisions\/304"}],"wp:attachment":[{"href":"https:\/\/medpress.bsd.uchicago.edu\/basu-lab\/wp-json\/wp\/v2\/media?parent=26"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}