RESUMEN
Studies were conducted from 1986 through 1993 to further define the geographic distribution and relative importance of different species of Leishmania as a cause of leishmaniasis in Peru. Patients with a clinical diagnosis of cutaneous and/or mucosal or diffuse cutaneous leishmaniasis were enrolled at the Naval Medical Research Institute Detachment (NAMRID) Laboratory in Lima, the Tropical Disease Clinic at San Marcos University Daniel A. Carrión, the Central Military Hospital, and a Ministry of Health hospital in Cusco, Peru. Clinical features, lesion aspirates, and biopsy tissue were obtained from each patient. All specimens were collected and assayed separately, including multiple specimens from some of the same patients for Leishmania parasites by inoculating aliquots of either aspirates or biopsy tissue suspensions onto Senekji's blood agar medium. Stocks of Leishmania isolates were used to prepare promastigotes to produce extracts for identifying the Leishmania species by the cellulose acetate electrophoresis enzyme technique. A total of 351 isolates of Leishmania were obtained from 350 patients who were infected primarily in the low and high jungle of at least 15 different Departments of Peru. Of the 351 isolates, 79% were identified as L. (V.) braziliensis, 7% as L. (V.) guyanensis, 10% as L. (V.) peruviana, 2% as L. (V.) lainsoni, and 1.7% as L. (L.) amazonensis. The clinical form of disease varied depending on the species of Leishmania, with L. (V.) braziliensis being associated most frequently with cutaneous, mucosal ulcers and mixed cutaneous and mucosal disease, and L. (V) peruviana, L. (V.) guyanensis, L. (V.) lainsoni with cutaneous lesions. Leishmania (L.) amazonensis was isolated from six patients, three with cutaneous lesions, one with mucosal lesions, and two with diffuse cutaneous lesions. Among all of the leishmaniasis cases, males were affected more frequently, and cases occurred among patients less than 10 to more than 51 years of age. These data further defined the geographic distribution and the relative frequency of Leishmania species associated with different clinical forms of leishmaniasis in Peru.
Asunto(s)
Leishmania/clasificación , Leishmaniasis/epidemiología , Adolescente , Adulto , Distribución por Edad , Animales , Niño , Preescolar , Electroforesis en Acetato de Celulosa , Femenino , Geografía , Humanos , Lactante , Isoenzimas/análisis , Leishmania/enzimología , Leishmaniasis/parasitología , Masculino , Persona de Mediana Edad , Perú/epidemiología , Distribución por SexoRESUMEN
A randomized, open, controlled clinical trial was designed to evaluate the efficacy, tolerance, and safety of sodium stibogluconate plus allopurinol and sodium stibogluconate alone as treatment of patients with mucocutaneous leishmaniasis. In phase 1 of the study, all 22 patients with severe disease had improvement of their lesions, but only two had clinical cure (both of these patients received sodium stibogluconate alone). In phase 2, which included 59 patients with moderate disease, the cure rate among sodium stibogluconate recipients was 75% (21 of 28) compared with 63.6% (14 of 22) among the sodium stibogluconate plus allopurinol recipients. The rates of clinical adverse events were similar among both groups. Thrombocytopenia was more frequent in the sodium stibogluconate plus allopurinol recipients, but the difference was not statistically significant. Eight patients (two sodium stibogluconate recipients and six sodium stibogluconate plus allopurinol recipients) withdrew from the study because of severe thrombocytopenia. In this study, the addition of allopurinol to sodium stibogluconate provided no clinical benefit as treatment of mucocutaneous leishmaniasis.
Asunto(s)
Alopurinol/administración & dosificación , Gluconato de Sodio Antimonio/administración & dosificación , Antiprotozoarios/administración & dosificación , Leishmaniasis Mucocutánea/tratamiento farmacológico , Adulto , Alopurinol/efectos adversos , Gluconato de Sodio Antimonio/efectos adversos , Antiprotozoarios/efectos adversos , Quimioterapia Combinada , Tolerancia a Medicamentos , Humanos , Masculino , SeguridadRESUMEN
Surveys were conducted from 1986 through 1992 to define the etiology and geographic distribution of human leishmaniasis in Peru. Lesion aspirates and skin biopsies were obtained from clinically diagnosed cases of leishmaniasis and tested for promastigotes by standard culture techniques. The isozyme profile of the isolates was determined by the cellulose acetate electrophoresis technique. Data indicated that the isozyme profiles for Leishmania isolates from six patients were similar to that of reference strains of L. lainsoni. These results are the first reported evidence of L. lainsoni and the first association of this parasite with human cases of cutaneous leishmaniasis in Peru.
Asunto(s)
Leishmania/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Adolescente , Adulto , Animales , Biopsia , Electroforesis en Acetato de Celulosa , Humanos , Isoenzimas/análisis , Leishmania/clasificación , Leishmania/enzimología , Leishmaniasis Cutánea/epidemiología , Masculino , Perú/epidemiología , Piel/parasitologíaRESUMEN
The efficacy and toxicity of two regimens of antimony, 28 and 40 days of 20 mg of antimony/kg/day, were compared in the treatment of culture-positive mucosal leishmaniasis involving more than one anatomic site. Forty consecutive eligible Peruvians with infiltrative or ulcerative mucosal disease of the lips, nose, palate-uvula-pharynx, or larynx-epiglottis were randomized to receive either 28 days (P28) or 40 days (P40) of sodium stibogluconate (Pentostam). Treatment was prematurely terminated due to thrombocytopenia in three patients and two patients did not complete six months of follow-up. At one month post-treatment, 13% (2 of 16) of the P28 patients and 16% (3 of 19) of the P40 patients no longer had infiltrates or ulcers and were initially considered cured. During a further 11 months of follow-up, infiltrated lesions healed in eight more P28 patients and in 10 more P40 patients. The cure rate after 12 months of follow-up was therefore 63% for both groups (10 of 16 in the P28 group and 12 of 19 in the P40 group). The total of 13 patients who had infiltrates or ulcers at the 9-12-month follow-up were considered failures. All seven patients (three in the P28 group and four in the P40 group) whose lesions were culture-positive for Leishmania at some point in the 12 months after treatment, and who were thereby parasitologic failures, were also clinical failures.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Gluconato de Sodio Antimonio/uso terapéutico , Leishmaniasis Mucocutánea/tratamiento farmacológico , Enfermedades Profesionales/tratamiento farmacológico , Adulto , Gluconato de Sodio Antimonio/administración & dosificación , Gluconato de Sodio Antimonio/efectos adversos , Esquema de Medicación , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Perú , Resultado del TratamientoRESUMEN
Pentavalent antimony (Sbv), formulated as sodium stibogluconate or meglumine antimoniate, is the standard treatment for the leishmaniases. In 16 of 17 consecutive, prospectively observed patients in Washington D.C., serum levels of amylase and lipase rose to abnormal values after therapy with sodium stibogluconate was started; 12 of 17 had symptoms of pancreatitis. Sbv therapy was continued to completion in 7 of 17 patients and interrupted in 10 of 17. Pancreatitis improved in every patient after Sbv therapy was stopped. Sbv treatment was resumed after brief interruptions in 6 of 10 patients. All six of these patients had flares of pancreatitis, but each completed therapy. Subsequently, we measured amylase and lipase levels in stored sera from 32 patients treated in Peru with either sodium stibogluconate or meglumine antimoniate for mucosal leishmaniasis. In all 32 Peruvian patients, serum amylase and lipase rose to abnormal levels during Sbv therapy; 11 of 32 had symptoms of pancreatitis. Standard Sbv regimens induce pancreatitis in almost all patients, but continued therapy is often tolerated; pancreatitis subsides when therapy is stopped, and rechallenge may be tolerated after a brief halt in treatment.
Asunto(s)
Gluconato de Sodio Antimonio/efectos adversos , Pancreatitis/inducido químicamente , Adulto , Amilasas/sangre , Gluconato de Sodio Antimonio/administración & dosificación , Antiprotozoarios/administración & dosificación , Antiprotozoarios/efectos adversos , District of Columbia , Humanos , Leishmaniasis Mucocutánea/tratamiento farmacológico , Leishmaniasis Visceral/tratamiento farmacológico , Lipasa/sangre , Masculino , Meglumina/administración & dosificación , Meglumina/efectos adversos , Antimoniato de Meglumina , Persona de Mediana Edad , Compuestos Organometálicos/administración & dosificación , Compuestos Organometálicos/efectos adversos , Pancreatitis/enzimología , Perú , Estudios ProspectivosRESUMEN
Anopheles mosquitoes captured in Andoas, Peru, a Plasmodium vivax-endemic area in the Peruvian Amazon region, contained both VK210 and VK247 P. vivax circumsporozoite (CS) proteins. Approximately 0.9% of the 4,403 mosquitoes tested by enzyme-linked immunosorbent assay were positive; 28% and 72% of the positive mosquitoes contained VK210 and VK247 CS proteins, respectively. These findings correlate strongly with a recent report of the presence of antibodies in residents of this area that recognize the VK210 and VK247 repeats, clearly indicating that both P. vivax CS protein polymorphs are common in the region.
Asunto(s)
Anopheles/parasitología , Plasmodium vivax/química , Proteínas Protozoarias/análisis , Animales , Perú , Plasmodium vivax/aislamiento & purificaciónRESUMEN
Individuals living in a malaria-endemic area in northern Peru were found to have antibodies to the variant repeat sequence of the circumsporozoite (CS) protein of Plasmodium vivax. The presence of IgG antibody to the predominant repeat sequence GDRAA/DGPA represented by the recombinant protein NS1(81) V20 (V20), and the variant repeat sequence ANGAGNQPG contained in the synthetic peptide Pvk247, was determined by enzyme-linked immunosorbent assay. IgG antibodies to the repeats were present in 78 (26%) of 298 serum samples; 56% of the positive serum samples had antibodies to V20 and 60% had antibodies to Pvk247. These findings stress the importance of considering the variant epitope in designing a vaccine based on the repeat region of the vivax CS protein. In a malaria-endemic area such as the one in this study, in which exposure to the variant repeat epitope may be as frequent as exposure to the predominant repeat, a vaccine based solely on the predominant repeat epitope may be ineffective against the variant form.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Malaria Vivax/epidemiología , Plasmodium vivax/inmunología , Proteínas Protozoarias , Adolescente , Adulto , Factores de Edad , Secuencia de Aminoácidos , Animales , Antígenos de Protozoos/química , Antígenos de Protozoos/genética , Niño , Preescolar , Humanos , Inmunoglobulina G/sangre , Lactante , Malaria Vivax/inmunología , Datos de Secuencia Molecular , Perú/epidemiología , Plasmodium vivax/genética , PrevalenciaRESUMEN
The presence in the New World of a variant strain of Plasmodium vivax (VK247) containing a unique circumsporozoite (CS) repeat domain was determined by the detection of antibodies to the variant CS protein and by genetic analysis of the CS gene from field isolates. Whole blood specimens were collected on filter paper from patients infected with P. vivax in Mexico and Peru. Plasmodium vivax DNA was eluted from filter paper samples and the CS gene was amplified by the polymerase chain reaction (PCR) and analyzed for the presence of VK247 or VK210 DNA by oligoprobe hybridization. Sera eluted from a companion filter paper sample were screened for antibodies reactive with the predominant and variant repeat peptides by enzyme-linked immunosorbent assays (ELISA) and with sporozoites by the immunofluorescent antibody (IFA) test. All 24 patients were positive by PCR and oligoprobe hybridization for either VK210 (16 of 24), VK247 (3 of 24), or both (5 of 24). Mixed infections were common (5 of 7) in Peru, but were not observed in the Mexican isolates (0 of 17). All three VK247 infections from Mexico occurred in residents of the foothills above Tapachula (P = 0.02). Of patients with smear-positive P. vivax infection, 42% (10 of 24) had detectable antibodies eluted from dried blood dots that were reactive with the CS protein by IFA or ELISA. These findings establish the widespread distribution of the P. vivax variant CS protein in the New World and indicate that dried blood filter paper samples represent a valuable source of material for the serologic and molecular analysis of plasmodial infections.
Asunto(s)
Anticuerpos Antiprotozoarios/biosíntesis , Antígenos de Protozoos/genética , Variación Genética , Malaria Vivax/parasitología , Plasmodium vivax/inmunología , Proteínas Protozoarias , Animales , Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Secuencia de Bases , ADN Protozoario/análisis , ADN Protozoario/química , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Amplificación de Genes , Humanos , México , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos/química , Perú , Plasmodium vivax/genética , Reacción en Cadena de la PolimerasaRESUMEN
The circumsporozoite (CS) protein of Plasmodium vivax consists of a central repeat region flanked by highly conserved non-repeat regions. Serum samples from 33 individuals with naturally acquired infections of P. vivax were tested for antibodies to four antigens representing the vivax CS protein. Three recombinant proteins containing different overlapping sequences in the non-repeat regions and either the entire central repeat region (vivax-1 and vivax-2) or two of the repeat sequences (vivax-3) were used as antigens in an enzyme-linked immunosorbent assay (ELISA). Antibodies to two other proteins, one (NS1(81)V20) containing the entire predominant repeat region (GDRAA/DGQPA) and the other (Pvk247) containing the variant repeat sequence (ANGAGNQPG) that was recently reported from Thailand were also measured by ELISA. Immunoglobulin G antibodies to the antigen representing the predominant repeat were present in 15% of the patients on the first day of treatment (day 0) and in 24% of the patients two weeks later (post-treatment). Six and 12% of the patients had IgG antibodies to the antigen containing the variant repeat on day 0 and post-treatment, respectively. A larger proportion of the sera had antibodies to the three antigens containing the non-repeat sequences; on the first day of treatment and two weeks later, 79 and 97% of the patients, respectively, had antibodies to vivax-1, vivax-2, and vivax-3. In this sample of Peruvians naturally infected with P. vivax, the most prevalent antibody responses were targeted to epitopes in the non-repeat region of the CS protein rather than to epitopes in the repeat region.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Anticuerpos Antiprotozoarios/biosíntesis , Antígenos de Protozoos/inmunología , Malaria Vivax/inmunología , Plasmodium vivax/inmunología , Proteínas Protozoarias , Adulto , Animales , Anticuerpos Antiprotozoarios/sangre , Especificidad de Anticuerpos , Unión Competitiva , Ensayo de Inmunoadsorción Enzimática , Epítopos/inmunología , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Inmunoglobulina M/biosíntesis , Inmunoglobulina M/sangre , Proteínas Recombinantes/inmunología , Secuencias Repetitivas de Ácidos Nucleicos/inmunologíaRESUMEN
We studied the interaction of sera from residents of an area in northern Peru where vivax malaria is endemic with four recombinant DNA-derived circumsporozoite (CS) proteins of Plasmodium vivax. The antigens used in the enzyme-linked immunosorbent assay included one Escherichia coli-produced and three Saccharomyces cerevisiae-produced recombinant proteins. Three of the proteins (NS1(81)V20, Vivax-1, and Vivax-2) contain the entire central repeat region of the P. vivax CS protein, and one protein (Vivax-3) contains only two repeat sequences. Vivax-1, Vivax-2, and Vivax-3 contain different lengths of sequences flanking the repeats. A higher percentage of the sera had antibodies to Vivax-2 and Vivax-3, the two proteins containing the longest nonrepeat sequences, than to NS1(81)V20 or Vivax-1. Children less than 5 years of age did not have immunoglobulin G antibodies to NS1(81)V20; however, they had antibodies to Vivax-1, Vivax-2, and Vivax-3. The finding that individuals living in a malaria-endemic area produce antibodies to peptides containing nonrepeat regions of the CS protein emphasizes the need to characterize the immune response to these regions in naturally exposed and experimentally immunized humans.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Malaria/inmunología , Plasmodium vivax/inmunología , Proteínas Protozoarias , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Aminoácidos , Animales , Anticuerpos Antiprotozoarios/inmunología , Especificidad de Anticuerpos , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina G/inmunología , Lactante , Malaria/microbiología , Persona de Mediana Edad , Datos de Secuencia Molecular , Perú , Proteínas Recombinantes/inmunologíaRESUMEN
The global distribution of a newly described variant of the Plasmodium vivax circumsporozoite (CS) gene was determined by genetic analysis of wild isolates. Whole blood specimens were collected on filter paper from patients infected with P. vivax in South America. West Africa, and the Indian subcontinent. P. vivax DNA was released from the filter paper samples, and the CS gene was amplified by polymerase chain reaction and analyzed for genetic variation. Amplified DNA was probed with oligonucleotide probes that hybridize with the predominant CS repeat region (PV210) and the variant CS repeat region (PV247) of P. vivax. The PV247 variant was found in all three geographically diverse areas. In addition, five of six consecutive patients studied had simultaneous infection with both the predominant and variant forms of P. vivax. These findings suggest that a single-epitope vaccine based on the predominant CS domain is unlikely to be protective on even a regional basis.
Asunto(s)
Antígenos de Protozoos/genética , Variación Genética , Malaria/parasitología , Plasmodium vivax/genética , Proteínas Protozoarias/genética , África Occidental , Animales , Secuencia de Bases , Southern Blotting , ADN Protozoario/química , Humanos , India , Datos de Secuencia Molecular , Sondas de Oligonucleótidos/química , Reacción en Cadena de la Polimerasa , América del SurRESUMEN
OBJECTIVE: To determine the efficacy and toxicity of the World Health Organization's (WHO) recommended treatment for mucosal leishmaniasis: antimony, 20 mg/kg body weight per day for 28 days. DESIGN: Open trial with 12-month follow-up. SETTING: Inpatient unit of a regional referral hospital in a developing country. PATIENTS: Twenty-nine consecutive eligible patients with culture-confirmed infection of the mucosa with Leishmania species who were otherwise healthy. Eight patients (28%) had mild to moderate disease (confined to the nasal mucosa). Twenty-one patients (72%) had severe disease (including the oropharynx as well as the nasal mucosa). INTERVENTION: Antimony, 20 mg/kg body weight intravenously every day for 28 days. Patients received antimony in the form of sodium stibogluconate. MEASUREMENTS AND MAIN RESULTS: Initial results of therapy were as follows: 63 of 72 lesions (88%) healed or markedly improved; all lesions were culture-negative for parasites; and 18 of 29 patients (62%) showed complete clinical and parasitologic cure of all lesions. By the 12-month follow-up examinations, however, 37 lesions had recurred, 8 new lesions had appeared, and only 8 patients (30%) showed clinical cure of all lesions. Of the 8 patients with mild to moderate disease, 6 were cured compared with only 2 of the 21 patients with severe disease. Side effects of this treatment regimen included T-wave inversion on electrocardiogram (4 patients), abnormal liver function tests (10 patients), and musculoskeletal pain (24 patients). No side effects occurred during week 1 of therapy. CONCLUSIONS: The only recommended treatment for mucosal leishmaniasis is ineffective in patients with severe disease. The acceptable toxicity of the regimen suggests that longer courses of therapy with antimony, or that trials with other antileishmanial agents alone or combined with antimony be evaluated as initial therapy for this disease.
Asunto(s)
Gluconato de Sodio Antimonio/uso terapéutico , Leishmaniasis Mucocutánea/tratamiento farmacológico , Adulto , Gluconato de Sodio Antimonio/efectos adversos , Aspartato Aminotransferasas/efectos de los fármacos , Sangre/efectos de los fármacos , Electrocardiografía/efectos de los fármacos , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Perú , RecurrenciaRESUMEN
A case of diffuse cutaneous leishmaniasis (DCL) acquired in Peru is described. The causative agent was Leishmania mexicana amazonensis as determined by isoenzyme analysis and species-specific monoclonal antibody binding characteristics. Histological examination of biopsy material showed a large number of intracellular and extracellular amastigotes and few lymphocytes. Treatment with meglumine antimoniate (Glucantime) administered iv at a dosage of 20 mg antimony/kg body weight/day for 60 days resulted in visible improvement of the lesions, but not in clinical or parasitological cure.
Asunto(s)
Leishmaniasis/epidemiología , Adulto , Antimonio/uso terapéutico , Antiprotozoarios/uso terapéutico , Biopsia , Femenino , Humanos , Leishmaniasis/tratamiento farmacológico , Leishmaniasis/patología , Meglumina/uso terapéutico , Antimoniato de Meglumina , Compuestos Organometálicos/uso terapéutico , Perú/epidemiología , Piel/patologíaRESUMEN
When Leishmania species are grown in vitro, parasites from the stationary phase differ from those in log phase growth in being more infective and more resistant to complement and macrophage mediated killing. In the present study, log phase and stationary phase promastigotes of Leishmania braziliensis panamensis were compared at the molecular level. Differences in polypeptide and glycoprotein composition and antigenicity between log and stationary phase promastigotes of L. b. panamensis were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting; the former showed that two polypeptides were unique to log phase promastigotes and one was unique to stationary phase promastigotes. There were also differences in surface lectin binding characteristics of log and stationary phase promastigotes. Live stationary phase promastigotes bound more concanavalin and lentil lectin than log phase promastigotes, indicating a greater number of mannose residues on their surfaces.