Distinct Roles for IL-4 and IL-10 in Regulating T2 Immunity during Allergic Bronchopulmonary Mycosis This information is current as Yadira Hernandez, Shikha Arora, John R. Erb-Downward, of October 1, 2021. Roderick A. McDonald, Galen B. Toews and Gary B. Huffnagle J Immunol 2005; 174:1027-1036; ; doi: 10.4049/jimmunol.174.2.1027 http://www.jimmunol.org/content/174/2/1027 Downloaded from References This article cites 58 articles, 32 of which you can access for free at: http://www.jimmunol.org/content/174/2/1027.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 1, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Distinct Roles for IL-4 and IL-10 in Regulating T2 Immunity during Allergic Bronchopulmonary Mycosis1 Yadira Hernandez,*† Shikha Arora,* John R. Erb-Downward,*† Roderick A. McDonald,* Galen B. Toews,* and Gary B. Huffnagle2*† Pulmonary Cryptococcus neoformans infection of C57BL/6 mice is an established model of an allergic bronchopulmonary mycosis that has also been used to test a number of immunomodulatory agents. Our objective was to determine the role of IL-4 and IL-10 in the development/manifestation of the T2 response to C. neoformans in the lungs and lung-associated lymph nodes. In contrast to wild-type (WT) mice, which develop a chronic infection, pulmonary clearance was significantly greater in IL-4 knockout (KO) and IL-10 KO mice but was not due to an up-regulation of a non-T cell effector mechanism. Pulmonary eosinophilia was absent in both IL-4 KO and IL-10 KO mice compared with WT mice. The production of IL-4, IL-5, and IL-13 by lung leukocytes from IL-4 KO and IL-10 KO mice was lower but IFN-␥ levels remained the same. TNF-␣ and IL-12 production by lung leukocytes was Downloaded from up-regulated in IL-10 KO but not IL-4 KO mice. Overall, IL-4 KO mice did not develop the systemic (lung-associated lymph nodes and serum) or local (lungs) T2 responses characteristic of the allergic bronchopulmonary C. neoformans infection. In contrast, the systemic T2 elements of the response remained unaltered in IL-10 KO mice whereas the T2 response in the lungs failed to develop indicating that the action of IL-10 in T cell regulation was distinct from that of IL-4. Thus, although IL-10 has been reported to down-regulate pulmonary T2 responses to isolated fungal Ags, IL-10 can augment pulmonary T2 responses if they occur in the context of fungal infection. The Journal of Immunology, 2005, 174: 1027–1036. http://www.jimmunol.org/ he fungal pathogen Cryptococcus neoformans enters the involves the up-regulation of T2 immunity. C. neoformans chronic body through inhalation and establishes a primary pulmo- infection stays primarily localized in the lungs of C57BL/6 mice, T nary infection. Inbred mouse models of pulmonary cryp- with minimal dissemination to extrapulmonary sites such as the tococcosis indicate a genetic component to the response. CBA/J spleen and the CNS. When compared with CBA/J or C.B-17 mice, and C.B-17 mice clear the infection faster than BALB/c mice. susceptibility of C57BL/6 mice does not correlate with the level of C57BL/6 mice can develop a chronic pulmonary infection (1–4). inflammation at the site of infection, but does correlate with high Clearance of C. neoformans in resistant hosts involves the devel- ␥ levels of IL-5 secretion, low levels of IFN- , and low levels of by guest on October 1, 2021 opment of T1 cell-mediated immunity (5–9). T cells recruit and IL-2 production (2–4). Elevated levels of IL-5 in the lungs of activate phagocytic effector cells, such as macrophages, against C. C57BL/6 mice promote the development of pulmonary eosino- neoformans inducing a granulomatous reaction and the develop- philia, which results in eosinophil-mediated tissue damage in the ment of delayed-type hypersensitivity responses (10). Many cyto- lungs, including deposition of eosinophilic YM1 crystals (4, 24). kines, mainly T1 cytokines, have been shown to play important The level of susceptibility in C. neoformans infection correlates roles in clearance of C. neoformans. Among these cytokines are with the number of eosinophils infiltrating the lungs (4). Suscep- ␥ ␣ IFN- , IL-2, IL-12, IL-18, TNF- , and IL-15, and the CC chemo- tible C57BL/6 mice have a large number of eosinophils in their kine ligands type 2 and type 3 (3, 9, 11–22). lungs, moderately resistant BALB/c mice have transient influx of Chronic fungal infections can develop when the T1/T2 balance eosinophils, and highly resistant CBA/J mice have only a few eo- of cellular immunity is shifted away from T1 toward T2 immune sinophils in their lungs (4). Thus, low dose infection of 6- to 8-wk- responses (23). C57BL/6 mice, 6- to 8-wk-old at the time of in- old C57BL/6 mice with C. neoformans strain 24067 produces a fection, develop a nonresolving pulmonary fungal infection, which chronic allergic bronchopulmonary mycosis (ABPM).3 This model has been used to address the role of immunomodulatory agents such as OX40, Mycobacterium bacillus Calmette-Gue´rin, ␣-galac- *Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine and †Department of Microbiology and Immunology, University of Michigan Medical tosylceramide (a CD1 ligand), IL-5 antagonists and anti-capsular School, Ann Arbor, MI 48109 Abs in addition to antifungal drugs in modulating immunity and Received for publication January 23, 2004. Accepted for publication November promoting protective host responses (25–32). Because both IL-4 2, 2004. and IL-10 can play significant regulatory roles in T2 responses to The costs of publication of this article were defrayed in part by the payment of page purified allergens (33, 34), our objective was to investigate the role charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. of IL-4 and IL-10 in the development and manifestation of the T2 1 This work was supported by Grants from the National Institutes of Health R01- response to C. neoformans in the lungs and lung-associated lymph HL065912 and R01-AI059201 (to G.B.H.), R01-HL051082 (to G.B.T.). G.B.H. was nodes (LALN) in this model of ABPM. also supported in part by a New Investigator Award in Molecular Pathogenic My- cology from the Burroughs-Wellcome Fund. Y.H. was supported in part by a Rack- ham Graduate Fellowship from the University of Michigan, Ann Arbor, MI. 2 Address correspondence and reprint requests to Dr. Gary B. Huffnagle, Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, 6301 Med- 3 Abbreviations used in this paper: ABPM, allergic bronchopulmonary mycosis; ical Sciences Research Building III, Box 0642, University of Michigan Medical LALN, lung-associated lymph nodes; KO, knockout; WT, wild type; HKC, heat- School, Ann Arbor, MI 48109-0642. E-mail address: [email protected] killed C. neoformans. Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 1028 IL-4 AND IL-10 IN T2 IMMUNITY DURING ABPM Materials and Methods plete medium. Total lung leukocyte numbers were assessed in the presence Mice of trypan blue using a hemocytometer. Female IL-4 knockout (KO), IL-10 KO, and wild-type (WT) mice on a Lung leukocyte subsets Ϯ C57BL/6 background (16 2 g) were obtained from The Jackson Labo- Macrophages, neutrophils, and eosinophils were visually counted in ratory. Mice were 6- to 8-wk-old at the time of infection. Mice were housed Wright-Giemsa-stained samples of lung cell suspensions cytospun onto in sterilized cages covered with a filter top. Food and water were given ad glass slides (Thermo Shandon Cytospin). For Wright-Giemsa staining, the libitum. The mice were maintained by the Unit for Laboratory Animal slides were fixed for 2 min with a one-step methanol-based Wright-Giemsa Medicine, University of Michigan, in accordance with regulations ap- stain (Harleco; EM Diagnostics Systems) followed by steps two and three proved by the University of Michigan Committee on the Use and Care of of the Diff-Quik whole blood stain kit (Diff-Quik; Baxter Scientific Prod- Animals. ucts). A total of 200–300 cells were counted from randomly chosen high C. neoformans culture power microscope fields for each sample. The percentage of a leukocyte subset was multiplied by the total number of leukocytes to give the abso- C. neoformans strain 24067 (52D) was obtained from the American Type lute number of that type of leukocyte in the sample. Culture Collection. For injection, yeast were grown to stationary phase Numbers of B, CD4, and CD8 T cells were determined by flow cytom- 5 (48–72 h) at 37°C in Sabouraud dextrose broth (1% neopeptone and 2% etry. Lung leukocytes (5 ϫ 10 ) were incubated for 30 min on ice in a total ␮ dextrose; Difco) on a shaker.