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Original Article

Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice

The Korean Journal of Parasitology 2007;45(4):255-266.
Published online: December 20, 2007

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Corresponding author (masouza@ufu.br)
• Received: June 20, 2007   • Accepted: November 12, 2007

Copyright © 2007 by The Korean Society for Parasitology

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Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice
Korean J Parasitol. 2007;45(4):255-266.   Published online December 20, 2007
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Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice
Korean J Parasitol. 2007;45(4):255-266.   Published online December 20, 2007
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Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice
Image Image Image Image Image
Fig. 1 ScLL effect in the lesion progression of L. amazonensis-infected BALB/c mice and parasite burden. BALB/c mice were immunized and infected with 106 stationary promastigotes of Leishmania and monitored for 10 weeks after infection. (A) Mice immunized with 25 µg/animal of SLA, SLA plus ScLL (10, 50 or 100 µg/animal), and control. (B) Mice immunized with 25 µg/animal of SLA, ScLL (10, 50 or 100 µg/animal), and control. Bars indicate the average and the standard deviation of differences between the average of the infected and uninfected footpads (n = 5). (C) Parasite burden in the footpad of immunized BALB/c mice with SLA and/or ScLL and infected by L. amazonensis. The results express the average and the standard deviation of the number of parasites by field into infected macrophages. ***P < 0.0001.
Fig. 2 DTH measurements. At the 10th week after infection with L. amazonensis, the immunized mice received 50 µg SLA antigen on the uninfected contralateral footpad. After 48 hr, the reading of the skin endurance was carried out as described in Materials and Methods. SLA (25 µg/animal) alone, SLA (25 µg/animal) plus ScLL (10, 50 or 100 µg/animal), PBS (control), and ScLL (10, 50 or 100 µg/animal) group mice. Bars indicate median for the measurement of the skin endurance in each group (n = 5). The line indicates 0.05 mm.
Fig. 3 Serum levels of specific anti-Leishmania immunoglobulins. At the 10th week after infection, the animals were bled and evaluated according to the level of specific antibodies by ELISA. Control (PBS) mice, and mice immunized with SLA (25 µg/animal) alone, SLA (25 µg/animal) plus ScLL (10, 50 or 100 µg/animal), and ScLL (10, 50 or 100 µg/animal) were analyzed. C (-) represents the sera of unimmunized and uninfected mice. (A) Levels of anti-Leishmania total IgG. (B) Levels of anti-Leishmania IgG1. (C) Levels of anti-Leishmania IgG2a. Bars express the average and the standard deviation of optical density (OD) at 492 nm in serum samples of each group (n = 5). *P < 0.05; **P < 0.0084; ***P < 0.0001.
Fig. 4 Histopathological features. (A) PBS group (skin) showing mononuclear exudate, neutrophils in the dermis and hypodermis, and parasitism of macrophages (arrowheads). (B) PBS group (popliteal lymph node) showing multinuclear giant cells with parasites, vacuoles with numerous amastigote forms in the periphery (arrowheads). (C) SLA group (skin) parasitism of the elongated cells of dermis compatible with fibroblasts (arrowheads). (D) SLA group (popliteal lymph node) parasitism of macrophages (arrowheads). (E) SLA + 100 µg/animal ScLL group (skin) parasitism of epidermal keratinocytes (arrowheads). (F) SLA + 100 µg/animal ScLL group (popliteal lymph node) eosinophilic exudate, and parasitism of macrophages (arrowheads). (G) 100 µg/animal ScLL group (skin) parasite-containing macrophages (arrowheads) and dermal fibroblasts. (H) 100 µg/animal ScLL group (popliteal lymph node) parasitism of macrophages (arrowheads). Data are representative of all immunized and infected mice with L. amazonensis. H-E stain. Original magnifications × 1,000.
Fig. 5 Expression and relative intensity of cytokines and iNOS by conventional RT-PCR assay in the lesion footpads of immunized mice and mice challenged with L. amazonensis. GAPDH expression was used as normal patterns of mRNA expression. Bars represent the relative intensity of the cytokines and iNOS expression. SLA corresponded to immunizing mice with SLA alone; SLA+10 µg, SLA+50 µg, SLA+100 µg corresponded to immunizing mice with SLA+ScLL 10 µg/animal, SLA+ScLL 50 µg/animal, SLA+ScLL 100 µg/animal, respectively; 10 µg, 50 µg, 100 µg, corresponded to immunizing mice with ScLL 10 µg/animal, ScLL 50 µg/animal, ScLL 100 µg/animal, respectively; and control corresponded to immunizing mice with PBS. C (-): negative reaction control.
Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice
Target Oligonucleotide sequence (5'-3')
Forward Reverse IL-4 TCATCGGCATTTTGAACGAG GCACCTTGGAAGCCCTACAG IL-12p35 AGACCAGAGACATGGAGTCATA TGCTTCACACTTCAGGAAAG IFN-γ GCAACAGCAAGGCGAAAAAG AAATTCAAATAGTGCTGGCAGAA TNF-α GATCTCAAGACAACCAACATGTG CTCCAGCTGGAAGACTCCTCCCAG iNOS GATGGTCAAGATCCAGAGAGGTCT AATTCGAGGCCACCCACC GAPDH CATGGCCTTCCGTGTTCCTA GGTCCTCAGTAGCCCAAGAT
Table 1. Primer sequences used in this study