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Natural killer (NK) cells are important immune effector cells that directly recognize and kill<jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>via a perforin-dependent cytotoxic mechanism. We previously showed that NK cells from HIV-infected patients have aberrant anticryptococcal killing and that interleukin-12 (IL-12) restores the activity at least partially through restoration of NKp30. However, the mechanisms causing this defect or how IL-12 restores the function was unknown. By examining the sequential steps in NK cell killing of<jats:italic>Cryptococcus<\/jats:italic>, we found that NK cells from HIV-infected patients had defective binding of NK cells to<jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>. Moreover, those NK cells that bound to<jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>failed to polarize perforin-containing granules to the microbial synapse compared to healthy controls, suggesting that binding was insufficient to restore a defect in perforin polarization. We also identified lower expression of intracellular perforin and defective perforin release from NK cells of HIV-infected patients in response to<jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>. Importantly, treatment of NK cells from HIV-infected patients with IL-12 reversed the multiple defects in binding, granule polarization, perforin content, and perforin release and restored anticryptococcal activity. Thus, there are multiple defects in the cytolytic machinery of NK cells from HIV-infected patients, which cumulatively result in defective NK cell anticryptococcal activity, and each of these defects can be reversed with IL-12.<\/jats:p><jats:p><jats:bold>IMPORTANCE<\/jats:bold>The mechanisms by which NK cells bind directly to pathogens and deploy their deadly cytolytic machinery during microbial host defense are only beginning to be elucidated. With the goal of understanding this process, we used NK cells from HIV-infected patients, which were known to have a defect in killing of<jats:named-content content-type=\"genus-species\">Cryptococcus neoformans<\/jats:named-content>. Taking advantage of previous studies that had shown that IL-12 restored killing, we used the cytokine as a gain-of-function approach to define the relevance of multiple steps in the recognition and cytolytic pathway. We demonstrated that NK cells from HIV-infected patients failed to kill<jats:italic>Cryptococcus<\/jats:italic>due to defects in perforin expression, granule polarization, and release of perforin. Additionally, IL-12 restored recognition of<jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>through binding of the NK-activating receptor NKp30. These observations identify important mechanisms used by NK cells to kill microbes and determine that defects in NK cells from HIV-infected patients are reversible.<\/jats:p>","DOI":"10.1128\/mbio.00878-16","type":"journal-article","created":{"date-parts":[[2016,8,24]],"date-time":"2016-08-24T00:58:21Z","timestamp":1472000301000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["Mechanisms by Which Interleukin-12 Corrects Defective NK Cell Anticryptococcal Activity in HIV-Infected Patients"],"prefix":"10.1128","volume":"7","author":[{"given":"Stephen K.","family":"Kyei","sequence":"first","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Ogbomo","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"ShuShun","family":"Li","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Timm-McCann","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard F.","family":"Xiang","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaunna M.","family":"Huston","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anutosh","family":"Ganguly","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pina","family":"Colarusso","sequence":"additional","affiliation":[{"name":"The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada"},{"name":"Department of Physiology and Pharmacology, University of Calgary, Calgary, Alberta, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. 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