Anti-IL-4 mAb treatment reduces lesion thickness in LTCP393(R)-inoculated BALB/c mice.

<p>The animals were inoculated with 1×10⁶<i>L. braziliensis</i> stationary phase promastigotes, isolates (A) LTCP393(R) or (B) LTCP15171(S), in the right ear dermis, treated with anti-IL-4 monoclonal antibody (black signs) or rat IgG (white signs), and the course of lesion development was monitored for 7 weeks. Lesion thickness was determined as the difference between the infected ear and the contralateral one, noninfected. Results are expressed as mean ± standard error of mean, and are representative of 2 independent experiments. *p<0,05.</p

Anti-IL-4 mAb treatment reduces parasite loads in LTCP393(R)-inoculated BALB/c mice lesions.

<p>The animals were inoculated at right ear dermis with 1×10⁶ stationary phase promastigotes of <i>L. braziliensis</i>, isolates LTCP393(R) (black bars) or LTCP15171(S) (white bars), treated with anti-IL-4 monoclonal antibody (hatched bars) or rat IgG (full bars), and the parasite load estimated at 7 weeks post infection in the ears (A) and draining lymph nodes (B). Results are expressed as mean ± standard error of mean, and are representative of 2 independent experiments. Statistical analysis: ANOVA and Tukey multiple comparision test. *p<0,05.</p

LTCP393(R)-inoculated BALB/c mice present higher parasite loads in the lesion sites.

<p>Parasite load estimate in ears (A) and lymph nodes (B) of BALB/c mice inoculated at right ear dermis with 1×10⁶ stationary phase promastigotes of <i>L. braziliensis</i>, isolates LTCP393(R) (black bars) or LTCP15171(S) (white bars), at 3, 5, 7 and 12 weeks post infection. Results are expressed as mean ± standard error of mean, and are representative of 3 independent experiments. n.d. = non-detected. *p<0,05.</p

Flow cytometry of infected BALB/c mice ears inflammatory infiltrate.

<p>The animals were inoculated with 1×10⁶ stationary phase <i>L. braziliensis</i> promastigotes, isolates LTCP393(R) (black bars) or LTCP15171(S) (white bars). Analyses were conducted at 3, 5, 7 and 12 weeks post infection, FoxP3, CD103, CTLA-4 and GITR expression on CD4⁺CD25⁺ cells were evaluated at 7 and 12 weeks post infection. Results are expressed as total number of cells in the lesion (A, B, C and D), and as percentage of cells inside CD4⁺CD25⁺ gate (E and F). Results are expressed as mean ± standard error of mean, and are representative of 3 independent experiments. *p<0,05.</p

Cytokines and enzymes mRNA expression in the ears from LTCP393(R) and LTCP15171(S)-inoculated mice.

<p>The animals were inoculated with 1×10⁶ stationary phase <i>L. braziliensis</i> promastigotes, isolates LTCP393(R) (black bars) or LTCP15171(S) (white bars). mRNA was extracted from the ears at 3, 5, 7 and 12 weeks post infection, and the specific mRNAs for IFN-γ (A), IL-4 (B), IL-10 (C), TGF-β (D), TNF-α (E), iNOS (F) and Arg I(G) were detected by real time RT PCR. The specific molecules mRNA expression was normalized based in the endogenous expression of the mRNAm for β-actin. Results are listed as the mean of expression in the ears of infected animals in relation to ears of uninfected ones ± standard error of mean, and are representative of 3 independent experiments. *p<0,05.</p

LTCP393(R)-inoculated BALB/c mice present larger lesions.

<p>(A) The animals were inoculated with 1×10⁶<i>Leishmania braziliensis</i> stationary phase promastigotes, isolates LTCP393(R) (black squares) or LTCP15171(S) (white squares) in the right ear dermis and the course of lesion development was monitored for 15 weeks. Lesion thickness was determined as the difference between the infected ear and the contralateral one, noninfected. Results are expressed as mean ± standard error of mean, and are representative of 2–3 independent experiments. (B) Photographs of mouse ears at 3, 5, 7 and 12 weeks post infection (w.p.i.). The arrow indicates ulceration in the ear of an animal infected with LTCP393(R) isolate at 7 weeks post infection, and the arrowhead indicates remaining lesion at 12 weeks post infection in animals infected with LTCP393(R) isolate. *p<0,05.</p