M Ulrich

Central University of Venezuela, Caracas, Distrito Capital, Venezuela

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Publications (57)106.59 Total impact

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    ABSTRACT: Different experimental murine models have shown that protective immunity against Lesihmania depends upon T cells, cytokines, and antigen presenting cells. However, the role of cytokines in naturally-infected hosts like domestic dogs is controversial. Few studies have evaluated cytokines in dogs naturally-infected with Leishmania infantum/chagasi. Since the domestic dog is the main reservoir of the parasite, a study was conducted to determine cytokines in serum of 33 dogs with Canine Visceral Lesihmaniasis from endemic areas of the State of Nueva Esparta, Venezuela. Dogs were classified as symptomatic (SD) and asymptomatic (AD), according to the expression of three or more clinical signs and levels of antibodies for rK39 and rK26. Ten non-infected, rK39 negative controls were included from an endemic area (EA) and ten dogs from a non-endemic area were used as healthy controls (HC). The following cytokines (pg/mL) were measured in serum by flow cytometry (CBA Hu Th1/Th 2, BDTM kit): IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL-2. Results show a higher concentration (P<0.05) of IFN-γ (69.93±7.46), IL-4 (7.51±2.68), TNF-α (3.86±1.46), and IL-2 (39.85±3.84) in AD when compared with SD (60.8±10.6; 5.28±0.80; 2.76± 0.72; and 36.04±3.61, respectively); and HC (51±14; 4.65±0.2; 3.21±0.89, and 32.65±5.86, respectively). The AD also showed higher levels (P<0.01) of IL-6 (4.9±0.55) compared with HC (4.02±0.64). Results show that AD exhibit a higher proportion of cellular activation and proinflammatory cytokines. Results indicate that measuring of serum cytokines could reflect the immunological status in dogs in future clinical trials oriented to either vaccination or therapy.
    Revista de la Facultad de Ciencias Veterinarias. 06/2010; 51(1):43-50.
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    ABSTRACT: Different experimental murine models have shown that protective immunity against Lesihmania depends upon T cells, cytokines, and antigen presenting cells. However, the role of cytokines in naturally-infected hosts like domestic dogs is controversial. Few studies have evaluated cytokines in dogs naturally-infected with Leishmania infantum/chagasi. Since the domestic dog is the main reservoir of the parasite, a study was conducted to determine cytokines in serum of 33 dogs with Canine Visceral Lesihmaniasis from endemic areas of the State of Nueva Esparta, Venezuela. Dogs were classified as symptomatic (SD) and asymptomatic (AD), according to the expression of three or more clinical signs and levels of antibodies for rK39 and rK26. Ten non-infected, rK39 negative controls were included from an endemic area (EA) and ten dogs from a non-endemic area were used as healthy controls (HC). The following cytokines (pg/mL) were measured in serum by flow cytometry (CBA Hu Th1/Th 2, BD TM kit): IFN-g, TNF-a, IL-10, IL-6, IL-4 and IL-2. Results show a higher concentration (P<0.05) of IFN-g (69.93±7.46), IL-4 (7.51±2.68), TNF-a (3.86±1.46), and IL-2 (39.85±3.84) in AD
    Revista de la Facultad de Ciencias Veterinarias UCV Venezuela. 05/2010; 51(1):43-50.
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    ABSTRACT: Human visceral leishmaniasis is a serious public health problem in endemic countries because of its high potential lethality, particularly in children. Rapid diagnosis is essential to early treatment and control of visceral leishmaniasis. The aim was to compare three serodiagnostic tools for human visceral leishmaniasis. Three methods were compared: the rK39 dipstick (Kalazar detection test, Inbios International Inc.), ELISA rK26 and direct agglutination test (DAT) (KIT Biomedical Research). Fifty serum samples from patients positive for rK39 ELISA were compared from four endemic provinces in Venezuela: Nueva Esparta (Margarita island), Lara, Anzoátegui and Trujillo. Additional serum samples from 17 healthy volunteers and 25 patients with other diseases were included. The rK39 ELISA served as the baseline standard method. Sensitivity, specificity, positive predictive value, negative predictive value and likelihood ratio were calculated for each test. All methods had a positive correlation with rK39 ELISA (p<0.0001). They showed high sensitivity and specificity. The direct agglutination test and the rK39 dipstick showed high sensitivity values, 89.7% (95% CI: 81.34.0-98.2%) and 94.2% (95% CI: 87.7-100%), respectively, and high specificity, 81.0% (95% CI: 80.0-99.5%) and 100%. The rK26 ELISA showed good specificity, 99% (95% CI: 95.2-100%), but a very low sensitivity, 37% (95% CI: 23.4-50.2%). Overall results indicated that DAT and rK39 dipstick have the highest specificity and sensitivity. Both are simple, cost-effective and field applicable tests. Therefore, they are recommended for early and accurate diagnosis of visceral leishmaniasis.
    Biomédica: revista del Instituto Nacional de Salud 01/2010; 30(1):39-45. · 0.32 Impact Factor
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    ABSTRACT: Visceral leishmaniasis is the most severe clinical form of leishmaniasis and is often fatal without proper treatment. Therefore, early and accurate diagnosis is important, but often difficult in endemic areas. The aim was to evaluate a direct agglutination test as a potential visceral leishmaniasis diagnostic method in endemic areas of Venezuela. The performance of the direct agglutination test, based on freeze-dried Leishmania donovani antigen was evaluated under laboratory conditions using serum samples of humans and dogs from several Venezuelan visceral leishmaniasis endemic areas: Nueva Esparta (Margarita Island), Lara, Anzoátegui and Trujillo Status. The study included confirmed visceral leishmaniasis patients (n=30), visceral leishmaniasis suspected subjects (n=4), healthy controls (n=19) and patients with other confirmed diseases (n=20). In addition, 24 serum samples from dogs with confirmed visceral leishmaniasis and 18 healthy control dogs were tested. All serum samples of visceral leishmaniasis patients, either active or recovered, were positive. They showed anti-L. donovani titers above 1:1600. Three out of four suspected visceral leishmaniasis cases were also positive, while serum samples from endemic controls and patients with other diseases had titers lower than 1:800. A sensitivity of 100% was obtained for all threshold levels under consideration and 100% specificity at the threshold titer of 1:800 (95% confidence interval: 91-100%). A 93% sensitivity (95% confidence interval: 76-99%) was observed in dog samples, with 100% specificity (95% confidence interval: 79-100%) at the threshold titer of 1:200. The direct agglutination test seems suitable for use in epidemiological studies and for serological diagnosis of human visceral leishmaniasis and canine visceral leishmaniasis.
    Biomédica: revista del Instituto Nacional de Salud 10/2007; 27(3):447-53. · 0.32 Impact Factor
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    ABSTRACT: Diffuse cutaneous leishmaniasis (DCL), although rare, is profoundly incapacitating. At present there is no successful treatment for this progressive protozoan infection, which is associated with the absence of specific cell-mediated immunity (CMI) to Leishmania. This disease shares features with visceral leishmaniasis (VL), including specific CMI inactivity during active disease and a heavy parasitic burden, but VL responds well to treatment. Miltefosine is the first orally administered drug which has shown efficacy in the treatment of VL; it has not been adequately evaluated in the treatment of DCL. To evaluate the efficacy of miltefosine in the treatment of DCL, using clinical, parasitological, histopathological and immunological criteria. Sixteen patients with DCL were treated with miltefosine, 2.0-2.5 mg kg(-1) daily, for variable periods of time (75-218 days). Patients were hospitalized for the first month and evaluated every 2 weeks until the termination of treatment with routine laboratory chemistry, percentage clinical improvement, presence of parasites in skin smears, growth of parasites in culture medium and in hamsters, histopathological characteristics of the granulomas, adverse side-effects, and reactivity to leishmanin skin test antigen. Further cycles of treatment were given in some of these patients, particularly after suspension of treatment was followed by relapse. Patients showed dramatic clinical improvement and reduction in the parasite burden by day 15 after the initiation of treatment, which continued while treatment was maintained. By day 45, 15 patients showed 80-90% clinical improvement. Nevertheless, suspension of treatment was followed by the development of new lesions in all but one patient. Inoculation in hamsters was observed to be the most sensitive technique to detect persisting parasites. Adverse events were very mild. Miltefosine produced a dramatic clinical and parasitological response in patients with DCL and improvement continued during drug administration, but with a single exception all patients presented new lesions after suspension of treatment. There was no histological or skin test evidence to suggest the development of CMI during treatment, which may be an indispensable criterion for the evaluation of potentially effective drugs against DCL.
    British Journal of Dermatology 07/2007; 156(6):1328-35. · 3.76 Impact Factor
  • Jacinto Convit, Marian Ulrich
    Gaceta médica de Caracas 12/2006; 114(4):300-304.
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    ABSTRACT: We have evaluated biopsies from patients with atypical nodular and typical ulcerated lesions of cutaneous leishmaniasis, from leishmanin reactions and skin from normal individuals from Nicaragua, Honduras and Guatemala for the presence of inorganic particles using confocal microscopy with a polarised light source and conventional histopathological techniques. Analysis by semiquantitative confocal microscopy permitted the demonstration of significantly larger numbers of particles in atypical lesions. Silica and aluminium, important components of these particles, were less abundant in particles from normal skin. The histology of these atypical lesions, characterised by 'naked' sarcoidal granulomas with epithelioid differentiation but very few lymphocytes, was very similar to the histological reaction observed after 14 days in persisting inflammation at leishmanin skin test sites. The presence of these unusual lesions in areas of Central American countries characterised by the presence of large amounts of volcanic ash, as well the unexpectedly low prevalence of leprosy in Central America, suggest that environmental factors may contribute significantly to the frequency and clinical manifestations of these infections. Among possible environmental features, the presence of inorganic particles with immunomodulatory properties in the skin may be a significant factor.
    Transactions of the Royal Society of Tropical Medicine and Hygiene 09/2006; 100(8):734-9. · 1.82 Impact Factor
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    ABSTRACT: A 31-year-old man who has suffered since age 3 from diffuse cutaneous leishmaniasis (DCL), a disease with profound physical and psychosocial repercussions and no effective treatment at present, was treated with miltefosine. The patient was treated for 120 days, 100 mg/day for 1 week, then 150 mg/day subsequently. Lesions were free of parasites at 43 days, and no signs of infiltration were present at day 76. No adverse side effects were observed. The dramatic clinical effect of miltefosine in this patient appears to fully justify further evaluation of this experimental therapy in DCL.
    International Journal of Dermatology 07/2006; 45(6):751-3. · 1.34 Impact Factor
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    ABSTRACT: Biopsies of 71 cases of atypical cutaneous leishmaniasis from Costa Rican patients were evaluated by histopathological procedures and attempts were made to culture Leishmania from nine biopsies. Leishmanin skin tests were carried out in 31 patients and 112 healthy individuals. Additional biopsies from 19 patients from Nicaragua were evaluated by routine histopathology. Ten biopsies were studied by confocal and nine by scanning electron microscopy. Inorganic material was analysed using an electron probe for microanalysis. Leishmania parasites were isolated from only two biopsies, but 90.3% of the patients from Costa Rica were leishmanin-positive, as were 27.7% of healthy individuals. Routine histopathological studies revealed naked granulomas formed by differentiated macrophages. Abundant inorganic material was observed in sections examined by confocal microscopy. Electron probe analysis revealed that silica and aluminium were the predominant elements in large particles. We postulate that the presence of this inorganic material, possibly of volcanic origin, in the skin may modulate the immunological response to Leishmania and may inhibit visceralization in the cases caused by Leishmania chagasi.
    Transactions of the Royal Society of Tropical Medicine and Hygiene 02/2005; 99(1):13-7. · 1.82 Impact Factor
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    ABSTRACT: Severe mucocutaneous (MCL) and diffuse (DCL) forms of American cutaneous leishmaniasis (ACL) are infrequent in Venezuela. Chemotherapy produces only transitory remission in DCL, and occasional treatment failures are observed in MCL. We have evaluated therapy with an experimental vaccine in patients with severe leishmaniasis. Four patients with MCL and 3 with early DCL were treated with monthly intradermal injections of a vaccine containing promastigotes of Leishmania (Viannia) braziliensis killed by pasteurization and viable Bacillus Calmette- Guerin. Clinical and immunological responses were evaluated. Integrity of protein constituents in extracts of pasteurized promastigotes was evaluated by gel electrophoresis. Complete remission of lesions occurred after 5-9 injections in patients with MCL or 7-10 injections in patients with early DCL. DCL patients developed positive skin reactions, average size 18.7 mm. All have been free of active lesions for at least 10 months. Adverse effects of the vaccine were limited to local reactivity to BCG at the injection sites and fever in 2 patients. Extracts of pasteurized and fresh promastigotes did not reveal differences in the integrity of protein components detectable by gel electrophoresis. Immunotherapy with this modified vaccine offers an effective, safe option for the treatment of patients who do not respond to immunotherapy with vaccine containing autoclaved parasites or to chemotherapy.
    Memórias do Instituto Oswaldo Cruz 03/2004; 99(1):57-62. · 1.36 Impact Factor
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    ABSTRACT: To report recent data on the distribution of human and canine visceral leishmaniasis (VL) in Venezuela, and to highlight problems associated with effective control measures. We report the number of cases, incidence rate, age and sex distribution, and mortality rates for human VL (HVL) for the period of 1995 through 2000, based on National Registry of Leishmaniasis data. We carried out serological studies on a total of 3 025 domestic dogs from the 12 states in Venezuela reporting cases of human VL in this 1995-2000 period and also from the state of Yaracuy, where cases were reported earlier during the decade of the 1990s. From 1995 through 2000, 242 cases of HVL were reported from 12 states, in various sections of Venezuela. There was a relatively stable national incidence rate of 0.2 cases per 100 000 persons per year. Of the 242 cases, 26.0% were from Margarita Island, one of the three islands that make up the state of Nueva Esparta (Margarita Island was the only one of the Nueva Esparta islands that had HVL cases). Over the 1995-2000 period, the annual incidence rates for Nueva Esparta ranged from 1.7 to 3.8 cases per 100 000 population. Males in Venezuela were more frequently affected (59.5%) than were females (40.5%). In terms of age, 67.7% of the VL patients were </= 4 years of age, and 80.6% were younger than 15 years. The mortality rate among the persons with VL was 7.85% during the 1995-2000 period. Serological screening with rK39 antigen of 1 217 dogs from Margarita Island found a 28.5% positivity rate (testing of dogs was not done on the two other islands of Nueva Esparta). In contrast, the rate was 2.8% in the 1 808 samples from dogs from 12 states on the mainland. Human and canine VL are unevenly distributed in Venezuela. The distribution may reflect such factors as differences among the states in human population density, vector density, and the presence or absence of other trypanosomatidae. Particularly high infection rates in very young children as well as in domestic dogs occur in semiurban communities of Nueva Esparta, where other human-infecting trypanosomatidae have not been reported. Control measures related to limiting canine infection might contribute to disease control where VL infections are frequent. Reducing VL mortality requires increased awareness among medical professionals of the possibility of VL in the differential diagnosis of hepato-splenic syndromes, particularly in children.
    Revista Panamericana de Salud Pública 04/2003; 13(4):239-45. · 0.85 Impact Factor
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    ABSTRACT: Of a total of 11532 Venezuelan patients with American cutaneous leishmaniasis (ACL) receiving immunotherapy with a combined vaccine containing heat-killed Leishmania promastigotes and bacille Calmette-Guerin (BCG) during the period 1990-99, we evaluated 5341 from 4 widely separated geographical states. Clinical healing varied from 91.2 to 98.7%, with an average of 95.7%. Adverse reactions were mild and limited to those associated with BCG vaccination alone. Immunotherapy failures in 143 patients included 54.5% with typical localized ulcers and 45.5% with non-mucosal intermediate cutaneous leishmaniasis (ICL). Less than 2% of the patients in this study had lesions suggestive of ICL. The disproportionately large number of immunotherapy failures in the ICL group suggests that it should not be used as monotherapy in this group. Weaker reactivity to purified protein derivative in immunotherapy failures, while not statistically significant in the small group reported here, suggests the possibility that these patients develop a relatively torpid immune response. The high percentage of clinical cures achieved with immunotherapy, associated with few secondary effects and low cost, support the use of immunotherapy in the routine treatment of localized ACL.
    Transactions of the Royal Society of Tropical Medicine and Hygiene 01/2003; 97(4):469-72. · 1.82 Impact Factor
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    ABSTRACT: Sixty-five patients were diagnosed with visceral leishmaniasis (VL) on Margarita Island in the decade from 1990 to1999; 86.2% were <= 3 years old. All were leishmanin-negative at diagnosis. Evaluation of 23 cured patients in 1999 revealed that 22/23 had converted to leishmanin-positive; five had persisting antibodies to rK39 antigen, with no clinical evidence of disease. Leishmanin tests were positive in 20.2% of 1,643 healthy individuals from 417 households in endemic areas. Of the positive reactors, 39.8% were identified in 35 (8.4%) of the households, 15 of which had an antecedent case of VL, a serologically positive dog or both. Weak serological activity to rK39 antigen was detected in 3 of 488 human sera from the endemic areas. The presence of micro-foci of intense peri-urban transmission and the apparent absence of other Trypanosomatidae causing human disease offer a unique opportunity for the study of reservoirs, alternative vectors and evaluation of control measures on the Island.
    Memórias do Instituto Oswaldo Cruz 12/2002; 97(8):1079-83. · 1.36 Impact Factor
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    ABSTRACT: An increase in the incidence of human visceral leishmaniasis (HVL) has been detected in recent years on Margarita Island, located off the NE coast of Venezuela. Recent studies have revealed reactivity to rK39 antigen (Leishmania chagasi) in 20% of 541 sera from domestic dogs in endemic communities; PCR reactions were positive using primers for the L. donovani complex. Here we report that isolates from human and canine infection, identified by isoenzyme analysis, correspond to L. infantum, zymodeme MON-1. This appears to be the first isolation and identification of an isolate from HVL on Margarita Island and demonstrates the presence of this zymodeme in the canine population.
    Memórias do Instituto Oswaldo Cruz 11/2001; 96(7):901-2. · 1.36 Impact Factor
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    ABSTRACT: An increase in the incidence of human visceral leishmaniasis (HVL) has been detected in recent years on Margarita Island, located off the NE coast of Venezuela. Recent studies have revealed reactivity to rK39 antigen ( Leishmania chagasi ) in 20% of 541 sera from domestic dogs in endemic communities; PCR reactions were positive using primers for the L. donovani complex. Here we report that isolates from human and canine infection, identified by isoenzyme analysis, correspond to L. infantum, zymodeme MON-1. This appears to be the first isolation and identification of an isolate from HVL on Margarita Island and demonstrates the presence of this zymodeme in the canine population.
    Memórias do Instituto Oswaldo Cruz (ISSN: 1678-8060) Vol 96 Num 7. 01/2001;
  • E Rada, N Aranzazu, M Ulrich, J Convit
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    ABSTRACT: Humoral immune responses were studied in 24 leprosy patients treated with multidrug therapy (MDT) and 16 contacts. The patients were monitored for 2 to 3 years with repeated determination of IgG antibody levels directed to different mycobacterial proteins (Mycobacterium tuberculosis, Mt70; M. bovis, Mb65; M. leprae, Ml36, 28, 18, 10 kDa, and the complete protein M. leprae extract, MLSA). All recombinant antigens were used at 5 micrograms/ml concentration and the complete soluble M. leprae extract at 2 micrograms/ml. The results shown in this study reveal a clear decline in IgG antibodies directed toward mycobacterial proteins in the 12 multibacillary (MB) patients when they were submitted to MDT. Initially we found strong reactivity toward complete cytosolic protein and M. leprae membrane protein. The most reactive recombinant proteins in MB patients were Ml10, Ml36, Mt70 kDa and, finally, Ml18 kDa when compared to the paucibacillary (PB) group. After treatment was completed all lepromatous and borderline lepromatous patients showed low or undetectable levels as compared with their initial values before starting treatment.
    International Journal of Leprosy and Other Mycobacterial Diseases 01/2000; 67(4):414-21. · 0.22 Impact Factor
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    ABSTRACT: The frequency of American visceral leishmaniasis affecting humans on Margarita Island, Venezuela, has increased in recent years, and infected dogs appear to constitute the principal source of infection. ELISA tests with Leishmania donovani promastigotes and rK39 antigen from L. chagasi in serum from 541 dogs were positive in 33.1% and 21.6% of the samples, respectively. A second blood sample taken from 50 animals after 8-10 months revealed an increase from 24% to 40% of ELISA positivity to both antigens, suggesting high susceptibility and transmission in the canine population. Among 42 serologically positive dogs, 33% of which showed clinical signs of disease, 79% were positive in polymerase chain reactions using primers specific for the L. donovani complex. Control measures including epidemiological hypersurveillance, the humane sacrifice of infected dogs, and rapid diagnosis and treatment of human cases have been initiated.
    Transactions of the Royal Society of Tropical Medicine and Hygiene 01/2000; 94(5):484-7. · 1.82 Impact Factor
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    ABSTRACT: An endemic focus of American visceral leishmaniasis (AVL) in eastern Venezuela has been evaluated in terms of patients (n = 48), immunologic reactivity to Leishmania in household contacts (n = 187) and neighborhood controls (n = 170), detection of Leishmania (L. donovani complex) in dogs and wild animals by the polymerase chain reaction (PCR) and characteristics of the sandfly population. The male:female ratio of patients was 1.18:1; 89.6% were < or =12 years old. Serologic reactivity was significantly higher in household contacts than in controls (P = 0.0008), as was the size of leishmanin reactions in contacts < or =10 years of age (P = 0.0141). Leishmania donovani complex-specific PCRs were positive in dogs, an opossum (Didelphis marsupialis), and a black rat (Rattus rattus). Lutzomyia longipalpis and Lu. evansi, both implicated in the transmission of AVL, were identified among the 386 sand flies examined. These observations provide the bases for an active control program as well as further studies of reservoirs and vector-host relationships in this area.
    The American journal of tropical medicine and hygiene 12/1999; 61(6):945-50. · 2.53 Impact Factor
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    ABSTRACT: Multibacillary (MB) leprosy patients treated with multidrug therapy (MDT) or MDT + immunotherapy (IMT) with BCG + heat-killed Mycobacterium leprae were tested annually for their ability to proliferate in vitro to the mycobacterial antigens BCG, M. leprae soluble extract, and intact M. leprae. IgM antibody responses to phenolic glycolipid I (PGL-I) were measured, as well as serum nitrite levels in patients' sera, before, during and after treatment. Patients who received only MDT did not present cellular reactivity to intact M. leprae antigens, in contrast to the results obtained with BCG, which elicited reactivity at time zero, that increased after treatment. Regarding PGL-I antibody variations in relation to the initial value, we observed a statistically significant marked decrease at the end of 2 years which continued to fall in successive evaluations. MB patients showed high initial serum nitrite concentrations which dropped drastically with treatment. This decay was apparently associated with the bacillary load present in these patients. The group submitted to IMT + MDT showed high and long-lasting T-cell responses to mycobacterial antigens in a significant number of initially unresponsive MB patients. There was a marked increase to M. leprae soluble extract and BCG, as well as a more variable response to whole bacilli. The antibody levels in this group of patients are sustained for a somewhat longer period and decreased more slowly during the 5-year follow up.
    International Journal of Leprosy and Other Mycobacterial Diseases 10/1997; 65(3):320-7. · 0.22 Impact Factor
  • V Rodríguez, M Centeno, M Ulrich
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    ABSTRACT: American cutaneous leishmaniasis (ACL) presents a spectrum of clinical and immunological manifestations. Since the nature of the cellular response appears to play a fundamental role in determining the characteristics of the immunoglobulin isotype of specific antibody responses, we have compared the relative levels of specific antibodies of the four IgG isotypes against Leishmania in sera from patients with different clinical manifestations of ACL. Using a specific antibody capture assay, significant levels of antibodies of the IgG1, 2 and 3 isotypes were detected in localized cutaneous leishmaniasis (LCL); the average level of IgG4 antibodies was low and they were not detected in 10/20 sera. Sera from muco-cutaneous leishmaniasis (MCL) gave a comparatively strong IgG1 response. Sera from diffuse cutaneous leishmaniasis (DCL), the rare form characterized by antigen-specific anergy of cell-mediated immunity, showed highly significant levels of IgG4 antibodies compared to antibody levels of this isotype in the other groups; IgG1 and IgG2 levels were also elevated. Based on other studies of the relationship between the IgG isotype response and cell-mediated immunity, these results confirm a Th1-like CD4+ T cell response in LCL and MCL and a significant Th2-like response in DCL.
    Parasite Immunology 08/1996; 18(7):341-5. · 2.21 Impact Factor