Article
Characterization of lung inflammation and
its impact on macrophage function in aging
Cynthia H. Canan,1 Nandan S. Gokhale,1,2 Bridget Carruthers,3 William P. Lafuse,
Larry S. Schlesinger, Jordi B. Torrelles, and Joanne Turner4
Department of Microbial Infection and Immunity, College of Medicine, and Center for Microbial Interface Biology, The Ohio
State University, Columbus Ohio, USA
RECEIVED FEBRUARY 13, 2014; REVISED MAY 28, 2014; ACCEPTED JUNE 1, 2014. DOI: 10.1189/jlb.4A0214-093RR
ABSTRACT
Systemic inflammation that occurs with increasing age
(inflammaging) is thought to contribute to the increased
susceptibility of the elderly to several disease states.
The elderly are at significant risk for developing pulmo-
nary disorders and infectious diseases, but the contri-
bution of inflammation in the pulmonary environment
has received little attention. In this study, we demon-
strate that the lungs of old mice have elevated levels of
proinflammatory cytokines and a resident population of
highly activated pulmonary macrophages that are re-
fractory to further activation by IFN-␥. The impact of this
inflammatory state on macrophage function was deter-
mined in vitro in response to infection with M.tb. Macro-
phages from the lungs of old mice secreted more pro-
inflammatory cytokines in response to M.tb infection
than similar cells from young mice and also demon-
strated enhanced M.tb uptake and P-L fusion. Sup-
plementation of mouse chow with the NSAID ibupro-
fen led to a reversal of lung and macrophage inflam-
matory signatures. These data indicate that the
pulmonary environment becomes inflammatory with
increasing age and that this inflammatory environ-
ment can be reversed with ibuprofen. J. Leukoc. Biol.
96: 473– 480; 2014.
Introduction
Increasing age is associated with an abundance of circulating
inflammatory cytokines that are considered to be a significant
contributing factor to the susceptibility of the elderly to nu-
merous disease states, including diabetes, heart disease, and
Abbreviations: 15d-PDJ2:⫽15-deoxy-delta 12,14-PG D2,
APC⫽allophycocyanin, BSL3⫽biosafety level 3, CIITA⫽class II, MHC, trans-
activator, CMIF⫽Campus Microscopy and Imaging Facility, COPD⫽chronic
obstructive pulmonary disease, COX⫽cyclooxygenase, CT⫽cycle thresh-
old, DPBS⫽Dulbecco’s PBS, GFP-M.tb⫽GFP expressing M.tb strain Erd-
man, IRF-1⫽IFN regulatory factor 1, IRGM-1⫽immunity-related GTPase
family M member 1, LAMP-1⫽lysosomal-associated membrane protein 1,
M.tb⫽Mycobacterium tuberculosis, MOI⫽multiplicity of infection,
MR⫽mannose receptor, NSAID⫽nonsteroidal anti-inflammatory drug,
OADC⫽oleic albumin dextrose catalase, P-L⫽phago-lysosomal, PGH2⫽PG
H2, qPCR⫽quantitative PCR, RU⫽relative unit(s)
0741-5400/14/0096-473 © Society for Leukocyte Biology
chronic pulmonary disorders, such as emphysema, bronchitis,
and COPD [1– 4]. This systemic increase in basal inflammatory
cytokines in old age has been termed inflammaging and is
broadly attributed to altered immune cell function, particu-
larly within the myeloid lineage [5]. However, studies of
monocyte or macrophage function in vitro have been contra-
dictory with some studies showing that the capacity of myeloid
cells to produce inflammatory cytokines can be impaired in
old age [6, 7], whereas others have shown that inflammatory
cytokine secretion can be enhanced [8, 9]. These disparities
have often been attributed to the source of monocytes or de-
rived/resident macrophages, as well as the specific stimulus
that has been used [5].
Our own studies have focused on resident tissue-derived pul-
monary macrophages and have shown previously that macro-
phages from the lungs of old mice are capable of secreting
abundant TNF-␣ and IL-12 in response to in vitro or in vivo
stimulus with the virulent pulmonary pathogen M.tb [10 –12].
M.tb is a significant global threat, accounting for ⬎9 million
cases and 1.4 million deaths from tuberculosis each year [13,
14]. M.tb is also responsible for significant morbidity and mor-
tality in the elderly [15], and the aged mouse model has been
used extensively to understand innate and adaptive immune
function in the context of this infectious disease [16 –18]. The
accumulated knowledge of immune cell function in vitro and
in vivo in response to M.tb in humans and animal models [19]
makes this an ideal and globally relevant model system to in-
vestigate macrophage function in old age.
Here, we extend our studies on pulmonary macrophage
function in old age to characterize further the functional state
of macrophages ex vivo and in response to in vitro infection
with M.tb. Our data demonstrate that resident pulmonary mac-
rophages from old mice are phenotypically and functionally
1. These authors contributed equally.
2. Current address: Dept. of Molecular Genetics and Microbiology, Duke
University, Durham, NC 27710, USA.
3. Current address: Office of Environmental Health and Safety, The Ohio
State University, Columbus, OH 43210, USA.
4. Correspondence: Dept. of Microbial Infection and Immunity, College of
Medicine, and Center for Microbial Interface Biology, The Ohio State
University, 786 Biomedical Research Tower, 460 West 12th Ave., Colum-
bus, OH 43210, USA. E-mail: joanne.turner@osumc.edu
Volume 96, September 2014 Journal of Leukocyte Biology 473