Cloning and expression of Leishmania major superoxide dismutase B1: a potential target antigen for serodiagnosis of Leishmaniasis

Leishmaniasis- a neglected public health problem- is a group of diseases affecting an estimated 12 million people worldwide. In the present study, recombinant Leishmania major superoxide dismutase B1 [rLmSODB1] has been utilized as a potential antigen for the serodiagnosis of human cutaneous [CL] and visceral leishmaniasis [VL] in the endemic regions of southern part of Iran. Additionally, the sensitivity and specificity of ELISA-based serodiagnosis using rLmSODB1 and the soluble Leishmania antigen [SLA] were compared. For the first time, rLmSODB1 has been cloned successfully and used for ELISA-based serodiagnosis. Sera from 30 CL and 24 VL cases were included in this study. Additional studies were also done for the evaluation of cross-reactivity using sera from 41 endemic controls including normal endemic donors [n= 20], systemic lupus erythematosus patients [n=5], rheumatoid arthritis patients [n= 5], and patients with tuberculosis [n=11]. Analysis indicated that rLmSODB1 was recognized by62.5% and 13.3% of sera from patients with VL and CL, showing a sensitivity of 72.7% and 53.6%, respectively. However 95.8% of VL and 30% of CL sera reacted with SLA, revealing sensitivities of 96% and 58.8%, respectively. Additionally, from 41 sera collected either from healthy subjects or patients affected with other diseases, 97.5% were negative with SLA or rLmSODB1 [specificity 97.6%]. These results show that rLmSODB1 almost does not react with sera from patients with tuberculosis and autoimmune diseases and may be considered as a candidate antigen for the specific immunodiagnosis of visceral leishmaniasis

Impaired superoxide dismutase level in patients with acquired immunodeficiency syndrome

Infection with the human immunodeficieney virus [HIV] is assumed to induce oxidative stress with consequent depletion of antioxidant molecules. The aim of this study was to determine the superoxide dismutase [SOD] level in HIV patients, as a way of determining its possible role in the pathogenesis of HIV disease. We prospectively studied 233 HIV-antibody negative apparently healthy controls and 175 consecutive acquired immunodeficiency syndrome [AIDS] patients for levels of SOD by standard methods. [Randox, UK]. The AIDS patients who were mainly distributed in the age grade of 20-29 years and 30-39 years, had mean SOD concentration of 96.83 +/- 20.44 U L. The control had mean SOD concentration of 187.33 +/- 21.12 U/L. The difference in these results was statistically significant [P<0.05]. SOD correlated negatively but not significantly for age in controls [r = -0.0468, p = 0.5878], and positively but not significantly in AIDS patients [r = 0.0109, p = 0.8965]. SOD is significantly depleted in the AIDS patients compared to controls. This result suggests that SOD which is a component of the total antioxidant system is greatly consumed in advanced HIV disease and is predicative of abnormalities in its replenishing mechanism. Regular measurement of SOD levels in HIV infection could serve as an adjunct in monitoring disease progression

The pattern of immune cell infiltration in chromoblastomycosis: involvement of macrophage inflammatory protein-1 alpha/CCL3 and fungi persistence

Chromoblastomycosis (CR) is a subcutaneous chronic mycosis characterized by a granulomatous inflammatory response. However, little is known regarding the pattern of leukocyte subsets in CR and the pathways involved in their recruitment. The objective of this study was to assess the cellular subsets, chemokine, chemokine receptors and enzymes in CR. The inflammatory infiltrate was characterized by immunohistochemistry using antibodies against macrophages (CD68), Langerhans'cells (S100), lymphocytes (CD3, CD4, CD8, CD45RO, CD20 and CD56) and neutrophils (CD15). The expression of MIP-1alpha (Macrophage inflammatory protein-1alpha), chemokine receptors (CXCR3 and CCR1) and enzymes (superoxide dismutase-SOD and nitric oxide synthase-iNOS) was also evaluated by the same method. We observed an increase in all populations evaluated when compared with the controls. Numbers of CD15+ and CD56+ were significantly lower than CD3+, CD4+, CD20+ and CD68+ cells. Statistical analysis revealed an association of fungi numbers with CD3, CD45RO and iNOS-positive cells. Furthermore, MIP-1alpha expression was associated with CD45RO, CD68, iNOS and CXCR3. Our results suggest a possible role of MIP-1alpha and fungi persistence in the cell infiltration in CR sites.

A cromomicose é micose subcutânea crônica sistêmica caracterizada por resposta inflamatória crônica granulomatosa. No entanto, existem poucos dados a respeito do padrão de subtipos de leucócitos na cromomicose e sobre as vias envolvidas no recrutamento destas células. O objetivo deste trabalho foi avaliar os tipos celulares, bem como a expressão de quimiocinas, receptores de quimiocinas e enzimas em lesões de cromomicose. O infiltrado inflamatório foi caracterizado por meio de técnica imuno-histoquímica utilizando os seguintes marcadores CD68 (macrófagos), S100 (células de Langerhans), CD3, CD4, CD8, CD45RO, CD20 e CD56 (linfócitos) e CD15 (neutrófilos). A expressão de MIP-1alfa (Proteína Inflamatória do Macrófago-1alfa), receptores de quimiocinas (CXCR3 e CCR1) e enzimas (superóxido dismutase-SOD e óxido nítrico sintase induzida-iNOS) foi avaliada pelo mesmo método. Observou-se um aumento de todas as populações celulares avaliadas em relação às amostras controle. As populações de células CD15+ e CD56+ foram significativamente menores que células CD3+, CD4+, CD20+ e CD68+. A análise estatística revelou uma correlação positiva entre o número de fungos com as células CD3, CD45RO e iNOS-positivas. A expressão de MIP-1alfa foi também associada às populações de células CD45RO, CD68, iNOS e CXCR3 positivas. Nossos resultados apontam para um possível papel de MIP-1alfa e da persistência fúngica na infiltração de células inflamatórias nos sítios de cromomicose.