[DESIGN AND EVALUATION OF A MULTIEPITOPIC PROTEIN AS A CANDIDATE FOR A CARRION DISEASE VACCINE]. / DISEÑO Y EVALUACIÓN DE UNA PROTEÍNA MULTIEPITÓPICA COMO CANDIDATA PARA VACUNA CONTRA LA ENFERMEDAD DE CARRIÓN.

OBJECTIVES.: To design and assess a multiepitopic protein as a candidate for a vaccine against Carrion disease. MATERIALS AND METHODS.: Using bioinformatics tools, epitopes of external membrane proteins were selected and a multiepitopic protein was designed. The multiepitopic protein gene was subcloned into the expression plasmid pET28b and transformed into E. coli BL21 pLys. The multiepitopic protein was expressed using isopropyl-ß-D-1-thiogalactopyranoside and purified using resin. This purified protein was used to immunize BALB/c mice obtaining polyclonal antibodies. In vitro invasion assays were conducted using a strain of Bartonella bacilliformis (B. bacilliformis) in human red blood cells. RESULTS.: The multiepitopic protein M1 presents preserved epitopes between isolates of B. bacilliformis with are non-toxic, and not homologous to human and surface proteins. Immunized mice presented IgG antibody levels capable of reducing in vitro the rate of invasion of B. bacilliformis into human red blood cells. CONCLUSIONS.: Multiepitopic protein M1 may serve as a candidate for a Carrion disease vaccine; however, more studies are needed to characterize the use of this antigen as a vaccine.

OBJETIVOS.: Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. MATERIALES Y MÉTODOS.: Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-ß-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. RESULTADOS.: La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. CONCLUSIONES.: La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.

Diseño y evaluación de una proteína multiepitópica como candidata para vacuna contra la enfermedad de carrión / Design and evaluation of a multiepitopic protein as a candidate for a carrion disease vaccine

RESUMEN Objetivos. Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. Materiales y métodos. Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-β-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. Resultados. La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. Conclusiones. La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.

ABSTRACT Objectives. To design and assess a multiepitopic protein as a candidate for a vaccine against Carrion disease. Materials and Methods. Using bioinformatics tools, epitopes of external membrane proteins were selected and a multiepitopic protein was designed. The multiepitopic protein gene was subcloned into the expression plasmid pET28b and transformed into E. coli BL21 pLys. The multiepitopic protein was expressed using isopropyl-β-D-1-thiogalactopyranoside and purified using resin. This purified protein was used to immunize BALB/c mice obtaining polyclonal antibodies. In vitro invasion assays were conducted using a strain of Bartonella bacilliformis (B. bacilliformis) in human red blood cells. Results. The multiepitopic protein M1 presents preserved epitopes between isolates of B. bacilliformis with are non-toxic, and not homologous to human and surface proteins. Immunized mice presented IgG antibody levels capable of reducing in vitro the rate of invasion of B. bacilliformis into human red blood cells. Conclusions. Multiepitopic protein M1 may serve as a candidate for a Carrion disease vaccine; however, more studies are needed to characterize the use of this antigen as a vaccine.

Epidemiological characteristics of chronic kidney disease of non-traditional causes in women of agricultural communities of El Salvador.

INTRODUCTION: In El Salvador end-stage renal disease (ESRD) was the first cause of hospital mortality overall, the first cause of hospital deaths in men, and the fifth cause of hospital mortality in women in 2013. In agricultural communities, chronic kidney disease (CKD) occurs predominantly in male agricultural workers, but it also affects women to a lesser degree, even those who are not involved in agricultural work. Internationally, most epidemiological CKD studies emphasize men and no epidemiological studies focused exclusively on women. OBJECTIVE: To describe the epidemiological characteristics of CKD in females in agricultural communities of El Salvador. METHODS: A cross-sectional epidemiological study was carried out in 2009 - 2011 based on active screening for CKD and risk factors in women aged ≥ 18 years in 3 disadvantaged populations of El Salvador: Bajo Lempa (Usulután Department), Guayapa Abajo (Ahuachapán Department), and Las Brisas (San Miguel Department). Epidemiological and clinical data were gathered through personal history, as well as urinalysis for renal damage markers, determinations of serum creatinine and glucose, and estimation of glomerular filtration rates. CKD cases were confirmed at 3 months. RESULTS: Prevalence of CKD was 13.9% in 1,412 women from 1,306 families studied. Chronic kidney disease of nontraditional causes (CKDu), not attributed to diabetes mellitus, hypertension, or proteinuric primary glomerulopathy (proteinuria > 1 g/L) was 6.6%. Prevalence of chronic renal failure was 6.8%. Prevalence of renal damage markers was 9.8% (microalbuminuria (30 - 300 mg/L) 5.7%; macroalbuminuria (> 300 mg/L) 2%; and hematuria, 2.1%. Prevalence of chronic kidney disease risk factors was: diabetes mellitus, 9.3%; hypertension, 23%; family history of CKD, 16%; family history of diabetes mellitus (DM), 18.7%; family history of hypertension (HT), 31.9%; obesity, 21%; central obesity, 30.7%; NSAID use, 84.3%; agricultural occupation, 15.2%; and contact with agrochemicals, 33.1%. CONCLUSIONS: CKD in women of Salvadoran agricultural communities is associated with disadvantaged populations, traditional (DM, HT, obesity) and non-traditional causes (environmental and occupational exposure to toxic agents and inadequate working conditions). Our results reinforce the hypotheses emerging from other studies, suggesting a multifactorial etiopathology including environmental and occupational nephrotoxic exposure.