Exogenous Stimulation of Type I Interferon Protects Mice with Chronic Granulomatous Disease from Aspergillosis through Early Recruitment of Host-Protective Neutrophils into the Lung
Authors: Seyedmousavi Seyedmojtaba , Davis Michael J. , Sugui Janyce A. , Pinkhasov Tzvia , Moyer Shannon , Salazar Andres M. , Chang Yun C. , Kwon-Chung Kyung J. 
Published: March 2018

Invasive aspergillosis (IA) remains the primary cause of morbidity and mortality in chronic granulomatous disease (CGD) patients, often due to infection by
Aspergillus
species refractory to antifungals. This motivates the search for alternative treatments, including immunotherapy. We investigated the effect of exogenous type I interferon (IFN) activation on the outcome of IA caused by three
Aspergillus
species,
A. fumigatus
,
A. nidulans
, and
A. tanneri
, in CGD mice. The animals were treated with poly(I):poly(C) carboxymethyl cellulose poly-
l
-lysine (PICLC), a mimetic of double-stranded RNA, 24 h preinfection and postinfection. The survival rates and lung fungal burdens were markedly improved by PICLC immunotherapy in animals infected with any one of the three
Aspergillus
species. While protection from IA was remarkable, PICLC induction of type I IFN in the lungs surged 24 h posttreatment and returned to baseline levels by 48 h, suggesting that PICLC altered early events in protection against IA. Immunophenotyping of recruited leukocytes and histopathological examination of tissue sections showed that PICLC induced similar cellular infiltrates as those in untreated-infected mice, in both cases dominated by monocytic cells and neutrophils. However, the PICLC immunotherapy resulted in a marked earlier recruitment of the leukocytes. Unlike with conidia, infection with
A. nidulans
germlings reduced the protective effect of PICLC immunotherapy. Additionally, antibody depletion of neutrophils totally reversed the protection, suggesting that neutrophils are crucial for PICLC-mediated protection. Together, these data show that prophylactic PICLC immunotherapy prerecruits these cells, enabling them to attack the conidia and thus resulting in a profound protection from IA.

IMPORTANCE
Patients with chronic granulomatous disease (CGD) are highly susceptible to invasive aspergillosis (IA). While
Aspergillus fumigatus
is the most-studied
Aspergillus
species, CGD patients often suffer IA caused by
A. nidulans
,
A. tanneri
, and other rare species. These non-
fumigatus Aspergillus
species are more resistant to antifungal drugs and cause higher fatality rates than
A. fumigatus
. Therefore, alternative therapies are needed to protect CGD patients. We report an effective immunotherapy of mice infected with three
Aspergillus
species via PICLC dosing. While protection from IA was long lasting, PICLC induction of type I IFN surged but quickly returned to baseline levels, suggesting that PICLC was altering early events in IA. Interestingly, we found responding immune cells to be similar between PICLC-treated and untreated-infected mice. However, PICLC immunotherapy resulted in an earlier recruitment of the leukocytes and suppressed fungal growth. This study highlights the value of type I IFN induction in CGD patients.