Background:
Paracoccidioidomycosis (PCM) is a severe granulomatous
disease
.
The hallmark of this mycosis is fibrin
degradation and granuloma formation as a result of a wound-healing process
in the context of excessive inflammation. Therefore, as the content of
collagen can be assessed by the methodology described in this manuscript, we
propose that the content of hydroxyproline (HYP) be employed as a new and
efficient measurement of granulomatous lesions developed. To estimate the
level of HYP the major byproduct of the degradation process, we hypothesized
that this simple and efficient technique could serve as a marker of disease
severity.
Methods:
Five B10.A female mice were infected with
P
.
brasiliensis
and, after 15 days, the omentum was
removed, subjected to histopathological analysis or processed (i.e.
deproteinized and derivatized), and further analyzed on a reverse phase HPLC
using a C-18 column. The omentum of five uninfected controls was also
collected and similarly analyzed.
Results:
Infected mice showed numerous, disseminated paracoccidioidomycotic lesions,
as well as marked collagen deposits, as observed in histopathologic
analysis, and high levels of HYP. Normal uninfected mice showed no
granulomas, little or no deposits of collagen fibers, and very low levels of
HYP, as evaluated by HPLC. Our results show that the disease intensity as
evaluated number and the morphology of the granulomatous lesions were
correlated to the HYP levels using small tissue samples from the omentum,
the main target organ of
P. brasiliensis
.
Conclusions:
Here we propose an alternative methodology to follow disease evolution and,
to some extent, fungal load in experimental
P. brasiliensis
infection and suggest its usefulness to other diseases with pronounced
fibrin degradation.