Regression line representing the serum levels variation of anti-<i>Paracoccidioides brasiliensis</i> antibodies as a function of antifungal treatment time, by using 122 serum samples from 8 patients who did not relapse with double agar gel immunodiffusion test (DID) positive to admission, before treatment.

<p>A – Line standing for double agar gel immunodiffusion test (DID) (p<0.0001) evolution. B – Line standing for enzyme-linked immunosorbent assay (ELISA) (p<0.0001) evolution. Compared under F test, such regressions did not show any difference (p = 0.96).</p

Level of agreement between methods, evaluated 2×2, for serological diagnosis of relapse in paracoccidioidomycosis-patients.

<p>Evaluation of the double agar gel immunodiffusion test (DID), the enzyme-linked immunosorbent assay (ELISA) and <i>immunoblotting</i> (IB) for identification of the glycoproteins of 43 kDa and 70 kDa. Binomial test and kappa coefficient.</p

Qualitative results of serological tests performed in 20 paracoccidioidomycosis-patients with relapse.

<p>Comparison among the double agar gel immunodiffusion test (DID), the enzyme-linked immunosorbent assay (ELISA), and the <i>immunoblotting</i> (IB) for identification of the glycoproteins of 43 kDa (IBgp43) and 70 kDa (IBgp70).</p><p>— Patients without serum to conduct these tests; NR - nonreactive +: reactive.</p

Results of the specific antibodies serum levels determined by the enzyme-linked immunosorbent assay (ELISA) and by the double agar gel immunodiffusion test (DID) in 21 patients with relapse of paracoccidioidomycosis.

<p>Comparison of the serum levels observed before treatment and at the time of relapse.</p><p>— serum not available; NR: nonreactive; cut off/ELISA = 0.710; cut off/DID = NR.</p><p>Evaluation of pretreatment/sensitivity: DID = 76.1% ELISA = 76.1% p = 0.25.</p><p>Comparison of sensitivity between the pretreatment and time to relapse:</p><p>DID: pretreatment = 80% relapse = 45% p = 0.017.</p><p>ELISA: pretreatment = 80% relapse = 65% p = 0.125.</p

Peritoneal and alveolar macrophages from naïve Swiss and BALB/c mice react differently to heat-killed <i>R</i>. <i>oryzae</i>.

(A) Nitric oxide (NO), (B) hydrogen peroxide (H2O2), (C) tumor necrosis factor-alpha (TNF-α), and (D) interleukin 10 (IL-10) levels in cell-free supernatants of peritoneal macrophages (PMΦ). (E) NO (F) H2O2, and (G) TNF-α levels in cell-free supernatants of alveolar macrophages (AMΦ) of non-infected Swiss, and BALB/c mice co-cultured or not co-cultured with heat-killed spores of R. oryzae. Student’s t-test; n = 5–7; *p < 0.05, **p< 0.01, ***p< 0.001.</p

Increased peripheral blood TCD4+ counts and serum SP-D levels in patients with chronic paracoccidioidomycosis, during and after antifungal therapy

<div><p> BACKGROUND The main clinical forms of paracoccidioidomycosis (PCM) are the acute/subacute form (AF) and the chronic form (CF), and they both display considerable clinical variability. The immune responses of PCM patients, during and after treatment, remain neglected, mainly in the case of CF patients, due to the high prevalence of pulmonary sequelae. OBJECTIVE To evaluate the distribution of whole blood T cell subsets, serum cytokines, and biomarkers of pulmonary fibrosis in PCM patients, according to the clinical form and at different time points, during the antifungal therapy. METHODS Eighty-seven PCM patients, from an endemic area in Brazil, were categorised into groups, according to the clinical form (AF or CF) and the moment of treatment. The peripheral blood T lymphocyte subsets of these patients were analysed using fluorescence-activated cell sorting. The serum levels of cytokines, basic fibroblast growth factor and surfactant protein-D (SP-D) were also analysed. FINDINGS In the CF patients, an expansion of the peripheral blood TCD4+ cells was observed during the treatment, and this persisted even after two years of antifungal treatment. In addition, these patients showed high serum levels of SP-D. CONCLUSION Our findings highlight the immunological changes CF patients undergo, during and after treatment, possibly due to the hypoxia triggered by pulmonary fibrosis and emphysema.</p></div

Increased PMΦ production of H₂O₂ by peritoneal macrophages (PMΦ) at the end of disseminated infection is related to efficient fungal clearance in BALB/c mice.

(A) Total fungal load (B) Nitric oxide (NO), (C) hydrogen peroxide (H2O2), (D) tumor necrosis factor-alpha (TNF-α), and (E) interleukin-10 (IL-10) levels in cell-free supernatants of PMΦ from Swiss or BALB/c mice co-cultured with heat-killed spores of R. oryzae. Linear regression analysis between H2O2 levels and total fungal load in BALB/c (F) and Swiss (G) mice. The infected group was composed of mice inoculated intravenously with 3 × 104 spores of R. oryzae and evaluated after 7 and 30 days. Any significant differences relative to infected samples compared to different times post-infection (letters) and different strains (*) are indicated. Student’s t-test; n = 5–7; *p < 0.05, **p< 0.01, ***p< 0.001.</p

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AMΦ from BALB/c mice display higher production of nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), and interleukin-10 (IL-10) than AMΦ from Swiss mice after 7 days of <i>R</i>. <i>oryzae</i> pulmonary infection.

(A) Total fungal load. (B) NO, (C) H2O2, (D) TNF-α, and (E) IL-10 levels in cell-free supernatants of AMΦ from Swiss or BALB/c mice co-cultured with heat-killed spores of R. oryzae. The infected group comprised mice inoculated intratracheally with 2 ×106 spores of R. oryzae and evaluated after 7 and 30 days. Significant differences relative to infected samples compared to different times post-infection (letters) and to different strains (*) are indicated. Student´s t-test; n = 5–7; *p < 0.05, **p< 0.01, ***p< 0.001.</p

Immunomodulatory effect of crude leaf extract of <i>Piper gaudichaudianum</i> on LPS-stimulated human monocytes.

<p>The results of the controls (medium and ketoprofin, unstimulated and LPS stimulated) were used in the analyses for all extract evaluated. Data are expressed as median ± SEM (pg/mL). ANOVA with Bonferroni post-test; p < 0.05; *: significantly different from placebo treatment, #: significantly different from ketoprofen treatment.</p