LEISHMANIASIS
(Diagnosis, Diagnostics, Immunodiagnosis,
Immunodiagnostics, Pathogenesis, Vaccines
& Drugs)
1318.
Joshi AB. Singhasivanon P. Khusmith S.
Fungladda W. Nandy A. Evaluation of direct agglutination test
(DAT) as an immunodiagnostic tool for diagnosis of visceral leishmaniasis in
Nepal. Southeast Asian Journal of Tropical Medicine & Public Health.
30(3):583-5, 1999 Sep.
Abstract
Before field application of the direct agglutination test (DAT)
for leishmaniasis, it was assessed as a
diagnostic tool. Fifteen confirmed
visceral leishmaniasis cases (bone marrow aspiration positive), 120 tuberculosis, 58 leprosy, 15 malaria, 26
intestinal parasitic infection cases,
24 endemic healthy controls from adjacent to the study area, and 18 controls
from Kathmandu (who had never visited the VL endemic areas) were tested for
anti-leishmanial antibody agglutination titers. Two of the tuberculosis cases
were positive for anti-leishmanial agglutinating antibodies at 1:800. All the visceral leishmaniasis confirmed
cases were reactive to anti-leishmanial
antibody at > or = 1:3,200. Other specimens were negative for serology. The
sensitivity of the direct agglutination test was 100% and the specificity was
99.2%. The direct agglutination test had positive and negative predictive
values of 100% and 99.2% respectively.
The direct agglutination test has been found to be simple, rapid, reliable, economic, safe and
adaptable to micro-techniques using
microtiter plates. It is specific and sensitive. The direct
agglutination test is simple enough for it to be performed in a field
laboratory.
1737. Corral
RS. Petray PB. CpG DNA as a
Th1-promoting adjuvant in immunization against Trypanosoma cruzi. Vaccine. 19(2-3):234-42, 2000 Sep 15.
Abstract
Th1-type immune response
plays a critical role in resistance to Trypanosoma cruzi infection. We asked
whether a synthetic oligodeoxynucleotide that contains immunostimulatory CpG
motifs (CpG ODN), known to promote a Th1 response, could act as an adjuvant in
immunization with parasite antigens. Mice immunized with a whole homogenate
(WH) of T. cruzi antigens co-administered with CpG ODN presented high titers of
T. cruzi antibodies (IgG2a isotype), strong delayed type hypersensitivity and a
Th1-dominated (IFN-gamma and IL-12) cytokine profile. Furthermore, WH plus CpG
ODN protected mice from challenge with an otherwise lethal dose of bloodstream
trypomastigotes. As reported for leishmaniasis and malaria, CpG ODN holds
considerable promise as an adjuvant for future vaccines against T. cruzi.
1738. Handman
E. Noormohammadi AH. Curtis JM.
Baldwin T. Sjolander A. Therapy
of murine cutaneous leishmaniasis by DNA vaccination. Vaccine. 18(26):3011-7, 2000 Jul 1.
Abstract
Prophylactic DNA
vaccination protects mice against infection with Leishmania major by inducing
an exclusive Th1 immune response dominated by the production of IFN-gamma. Here
we show that DNA vaccines, initially designed to prevent infection, can also
have a significant therapeutic effect. In L. major infected mice, vaccination
with DNA encoding the Parasite Surface Antigen/gp46/M2 causes reduction in
lesion size and promotes healing in both genetically resistant C3H/He mice and
susceptible BALB/c mice. The therapeutic effect is underpinned by a shift in
the T cell-derived cytokine environment with an increase in the IFN-gamma
producing Th1 type cells. Application of such immunotherapy in conjunction with antiparasite drugs may
result in faster or more certain cure of the disease in humans.
1739. Hu
XS. Yang WT. Lu HG. Yan HP. Cheng JP.
Ma Y. Jin BQ. Zhang T.
Sequencing a specific kinetoplast DNA fragment of Leishmania donovani
for polymerase chain reaction
amplification in diagnosis of leishmaniasis in bone marrow and blood samples.
Journal of Parasitology. 86(4):822-6,
2000 Aug.
Abstract
A set of oligonucleotide
primers I and II was developed by analyzing the specificity of a cloned kinetoplast DNA (kDNA) fragment of
Leishmania donovani and sequencing the fragment. Polymerase chain reaction
(PCR) was conducted with the primers to amplify a minicircle kDNA fragment (297
bp) to detect L. donovani in the bone marrow (22 samples), whole blood (16
samples), and serum (17 samples) of 22 patients with visceral leishmaniasis.
All of 22 patients were diagnosed by microscopic identification. Control
samples of bone marrow, whole blood, and serum were obtained from patients with
leukemia and from healthy volunteers. In addition, 12 dogs were infected with
L. donovani promastigotes for the PCR test. The total number of patients
positive by PCR testing was 95.5%
(21/22), w ith 91.0% (20/22) from the bone marrow, 68.8% (11/16) from
the blood, and 29.4% (5/17) from the sera. Similar results were obtained in
infected dogs. No amplification products were seen in control samples from
humans or dogs. Our results suggest that PCR may be useful in detecting kDNA in
the bone marrow and blood of patients with visceral leishmaniasis.
1740. Kamau
SW. Hurtado M. Muller-Doblies UU. Grimm F. Nunez R. Flow
cytometric assessment of allopurinol susceptibility in Leishmania infantum
promastigote. Cytometry. 40(4):353-60, 2000 Aug 1.
Abstract
BACKGROUND: Leishmaniasis
is a major tropical and subtropical parasitic
disease. Sodium stibogluconate, N-methyl -D-glucamine antimoniate,
amphotericin B, pentamidine, and ketoconazole are drugs used to treat this
disease. Some of these drugs cause severe adverse side effects and treatment
failures are common. Allopurinol, a purine analog, has been used to treat
leishmaniasis, alone or combined with the previously mentioned drugs. Low cost,
ease of administration (oral), and lack of toxicity make allopurinol a
particularly appealing candidate. METHODS: The effect of allopurinol on
Leishmania infantum (MCAN/ES/89/IPZ229/1/89, zymodeme MON1) wild-type
promastigotes (wt-p229), and an altered form of these promastigotes (allo-p229)
resulting from long term in vitro exposure to allopurinol, was determined by
[(3)H]-thymidine incorporation assays and by diverse flow cytometric
approaches. RESULTS: Allopurinol arrested the proliferative capacity of wt-p229
promastigotes, reduced the proportion of
viable cells, and decreased their total protein content. In contrast,
allo-p229 promastigote proliferation was only slightly decelerated and the
proportion of viable cells and the protein content were not affected by the
allopurinol treatment. CONCLUSIONS: The flow cytometry approach allowed us to
demonstrate differences in allopurinol susceptibility of the two promastigote
forms, expanding the spectrum of flow cytometry applications in studies of
parasite resistance. Copyright 2000 Wiley-Liss, Inc.
1741. Kaul
P. Malla N. Kaur S. Mahajan RC. Ganguly NK. Evaluation of a 200-kDa
amastigote-specific antigen of L. donovani by enzyme-linked immunosorbent assay
(ELISA) for the diagnosis of visceral leishmaniasis. Transactions of the Royal
Society of Tropical Medicine & Hygiene. 94(2):173-5, 2000 Mar-Apr.
Abstract
A purified 200-kDa
antigenic fraction from Leishmania donovani axenic amastigotes was evaluated by
ELISA for the detection of antibody response
in visceral leishmaniasis (VL) patients, post kala-azar dermal
leishmaniasis (PKDL) patients and controls, for the diagnosis of visceral
leishmaniasis. A positive antibody response to the 200-kDa amastigote fraction
and to Leishmania amastigote soluble antigen (LASA) was found in 29 (96.6%) and 30 ((100%) confirmed VL
patients, respectively, by the use of ELISA. However, only 1 (10%) out of 10
PKDL patients had detectable antibody response to 200-kDa fraction while all
the 10 (100%) PKDL patients exhibited an immune response to LASA. Therefore,
use of the 200-kDa antigenic fraction for the detection of antibody response in
an ELISA follow-up (post treatment) of VL patients may have prognostic
significance, and it may also be useful for differentiating active VL and PKDL.
1742. Llorente
S. Gimeno L. Navarro MJ. Moreno
S. Rodriguez-Girones M. Therapy of visceral leishmaniasis in renal
transplant recipients intolerant to pentavalent antimonials.
Transplantation. 70(5):800-1, 2000 Sep
15.
Abstract
Visceral leishmaniasis
should be suspected in renal transplant recipients in whom a fever develops of
unknown origin. A 53-year-old renal transplant recipient developed pyrexia,
hepatosplenomegaly, and pancytopenia 4 years
after transplantation. Antileishmaniasis serology was negative, and the diagnosis was confirmed through bone marrow
examination. Treatment with glucantine
(N-methylglucamine antimoniate) led to acute pancreatitis, and treatment with ketoconazole plus
allopurinol for 21 days was effective to
eradicate Leishmania donovani.
1743.
Passos VM. Barreto SM.
Romanha AJ. Krettli AU. Volpini AC.
Lima e Costa MF.
American
cutaneous leishmaniasis: use of a skin test as a predictor of relapse after
treatment. Bulletin of the World Health Organization. 78(8):968-74, 2000.
Abstract
While relapses following
clinical cure of American cutaneous leishmaniasis are frequent, no test has been described until now to predict
such relapses. A cohort of 318 American cutaneous leishmaniasis patients was
followed up for two years after treatment with meglumine antimoniate, during
which time 32 relapses occurred, 30 in the first year and two in the second
(accumulated risk: 10.5%). No association was found between these relapses and
the parasite-specific antibody response before and after treatment, or between
the relapses and stratification by sociodemographic and clinical
characteristics. However when Leishmania was used as antigen, patients with a
negative skin test at the time of diagnosis presented a 3.4-fold higher risk
(hazard risk = 3.4; 95% confidence
interval, 1.7-7.0) of American cutaneous leishmaniasis relapse, compared with patients with a positive
response. This result shows that the skin test can be a predictor of American
cutaneous leishmaniasis relapse after treatment.
1744. Rais
S. Perianin A. Lenoir M.
Sadak A. Rivollet D. Paul M.
Deniau M. Sodium stibogluconate
(Pentostam) potentiates oxidant production in murine visceral leishmaniasis and in human blood. Antimicrobial Agents
& Chemotherapy. 44(9):2406-10, 2000
Sep.
Abstract
Sodium stibogluconate
(Sbb), a leishmanicidal drug, was studied for its in vivo effect on the formation of reactive oxygen species (ROS),
assessed by chemiluminescence (CL) in
the whole blood of mice infected with Leishmania infantum. Stimulation of ROS formation induced ex vivo by zymosan
particles or the protein kinase C activator phorbol myristate acetate (PMA) was
reduced by approximately 25% (P < 0.05) after infection of mice. Treatment
of infected mice with Sbb (50 to 400 mg/kg of body weight) enhanced the blood
CL induced by zymosan and PMA (47 to 96%, P < 0.01). The drug potentiation
effect also occurred in uninfected mice. In vitro treatment of normal human
blood with Sbb (1, 10, or 100 microg/ml) for 1 h primed the CL response to PMA
(29 to 54%). The priming effect of Sbb was also observed on the production of
superoxide by isolated polymorphonuclear leukocytes stimulated either by PMA
and zymosan or by the chemoattractants
N-formyl-Met-Leu-Phe and platelet-activating factor. These data provide the first evidence of priming of the
phagocyte respiratory burst by Sbb. This novel property of Sbb may contribute
to the drug's leishmanicidal effect.
1745. Vicandi
B. Jimenez-Heffernan JA. Lopez-Ferrer P. Ortega L. Viguer JM.
Cytologic diagnosis of leishmaniasis in HIV infection. A report of eight cases.
Acta Cytologica. 44(5):835-9, 2000
Sep-Oct.
Abstract
BACKGROUND: Leishmania
organisms are among the intracellular microorganisms with a tendency to develop
in patients with the acquired immunodeficiency syndrome (AIDS). With increasing
travel to endemic areas by patients with human immunodeficiency virus (HIV)
infection, it is becoming a more-frequent diagnosis in nonendemic areas. CASES:
Ten cytologic specimens from eight patients with leishmaniasis and AIDS were
reviewed. Eight samples were obtained from lymph nodes through fine needle
aspiration (FNA). Another sample was obtained after scraping a tongue ulcer.
The last one was an ascitic fluid specimen. Smears showed numerous parasitized
histiocytes with abundant intracellular Leishmania organisms (amastigotes).
Extracellular microorganisms were also abundant. Diff-Quik-stained smears allowed the clear recognition of the
characteristic morphologic appearance with a deep-staining area (nuclei) and
paranuclear zone (kinetoplast). Intracellular organisms were round, while
single, extracellular forms were a more elongated. CONCLUSION: The polymorphous
clinical manifestations usually seen in patients suffering from leishmaniasis
and AIDS constitute a diagnostic challenge that can be facilitated by
cytopathologic examination. Cytology permits easy and rapid identification of
Leishmania amastigotes, allowing a specific diagnosis and treatment.
2199. Attar ZJ. Chance ML. el-Safi S. Carney J. Azazy A. El-Hadi M. Dourado C. Hommel M. Latex agglutination test for the detection of urinary antigens in visceral leishmaniasis. Acta Tropica. 78(1):11-6, 2001 Jan 15.
Abstract
This paper describes a new latex agglutination test ('KATEX') for the detection of leishmanials antigen in the urine of patients with visceral leishmaniasis. In preliminary laboratory trials, using urine collected from well-defined cases and controls from Brazil, Yemen and Nepal, the test had 100% specificity and a sensitivity between 68 and 100%. When used in a time-course experiment in cotton rats infected with Leishmania donovani, the test became positive 1 week after inoculation and antigen levels in urine declined rapidly after chemotherapy (the test was negative before the end of the course of treatment). Finally, in an integrated study performed in Sudan, KATEX was compared to microscopy and four different serological tests in a group of 73 patients having presented with clinical manifestations suggestive of visceral leishmaniasis. Compared to microscopy, KATEX performed better than any single serological test in predicting positivity and a particularly good result was obtained by combining KATEX and the direct agglutination test (DAT).
2200. Babaloo Z. Kaye PM. Eslami MB. Interleukin-13 in Iranian patients with visceral leishmaniasis: relationship to other Th2 and Th1 cytokines. Transactions of the Royal Society of Tropical Medicine & Hygiene. 95(1):85-8, 2001 Jan-Feb.
Abstract
The role of interleukin (IL)-13, a Th2 cytokine sharing many of the features of IL-4, has not previously been examined in patients with visceral leishmaniasis (VL). We examined sera from Iranian patients with VL caused by Leishmania infantum. Serum IL-13 was detected in 50% (22/44) of patients with active primary disease. In comparison, IL-10 was detected in 79.5% (35/44), interferon gamma (IFN gamma) in 38.5% (17/44), and IL-4 in only 5% (2/44) of these patients. With few exceptions all 3 cytokines were undetectable after clinical recovery following antimony therapy. Five of 7 patients (71%) who failed antimony therapy and had relapsing disease had similar levels of IL-10 to patients with active primary disease. However, with only 1 exception, IL-13, IFN gamma and IL-4 were not detected in such patients. These data suggest that relapsing disease may result from defective cellular immunity, unrelated to immunosuppression mediated by IL-10.
2201. Baddini-Caramelli C. Matayoshi S. Moura EM. Araf D. Santo R. Voegels R. Kara-Jose N. Chronic dacryocystitis in American mucocutaneous leishmaniasis. Ophthalmic Plastic & Reconstructive Surgery. 17(1):48-52, 2001 Jan.
Abstract
PURPOSE: This study describes lacrimal tract involvement and surgical outcome in patients with mucocutaneous leishmaniasis. METHODS: Four patients, ages 20 to 75 years, had nasal lesions resulting from mucocutaneous leishmaniasis and sought treatment for chronic dacryocystitis. Each patient had had lacrimal symptoms since childhood or early adulthood, concomitantly with the development of upper airway lesions. Dacryocystography showed nasolacrimal duct stenosis in all cases on the affected side. Three patients underwent dacryocystorhinostomy (one bilaterally), and one patient had bilateral dacryocystectomy. RESULTS: Two patients had surgical fistula closure soon after surgery. A sequential endoscopic operation for remotion of a synechia between the fistula and the middle turbinate was successful in one of these patients. Histopathologic analysis of lacrimal sacs and nasal mucous membranes close to the anastomotic site revealed chronic nonspecific inflammatory process and negative immunohistochemistry for Leishmania. CONCLUSIONS: Dacryocystitis may result from nasal mucocutaneous leishmaniasis. The surgical outcome was unsatisfactory in one of the four patients.
2202. Bogdan C. Schonian G. Banuls AL. Hide M. Pratlong F. Lorenz E. Rollinghoff M. Mertens R. Visceral leishmaniasis in a German child who had never entered a known endemic area: case report and review of the literature. [Review] [35 refs] Clinical Infectious Diseases. 32(2):302-6, 2001 Jan 15.
Abstract
We describe a case of visceral leishmaniasis in a 15-month-old German child. Diagnosis was significantly delayed because the patient had no history of travel to known endemic areas. Congenital or blood transfusion-associated leishmaniasis was ruled out. Possible modes of transmission (including a potential new autochthonous focus of the disease in central Europe) are discussed. [References: 35]
2203. No abstract
2204. No abstract
2205. No abstract
2206. Fargnoli MC. Peris K. Cerroni L. Piccolo D. Chimenti S. A nodule on the forearm of a patient with Hodgkin disease. Archives of Dermatology. 137(1):85-90, 2001 Jan.
2207. Gicheru MM. Olobo JO. Anjili CO. Orago AS. Modabber F. Scott P. Vervet monkeys vaccinated with killed Leishmania major parasites and interleukin-12 develop a type 1 immune response but are not protected against challenge infection. Infection & Immunity. 69(1):245-51, 2001 Jan.
Abstract
Leishmania major is a protozoan parasite that causes chronic cutaneous lesions that often leave disfiguring scars. Infections in mice have demonstrated that leishmanial vaccines that include interleukin-12 (IL-12) as an adjuvant are able to induce protective immunity. In this study, we assessed the safety, immunopotency, and adjuvant potential of two doses of IL-12 when used with a killed L. major vaccine in vervet monkeys. The induction of cell-mediated immunity following vaccination was determined by measuring delayed-type hypersensitivity, in vitro lymphocyte proliferation, and gamma interferon (IFN-gamma) production. Protection was assessed by challenging the animals with L. major parasites and monitoring the course of infection. At low doses of IL-12 (10 microg), a small increase in the parameters of cell-mediated immunity was observed, relative to those in animals that received antigen without IL-12. However, none of these animals were protected against a challenge infection. At higher doses of IL-12 (30 microg), a substantial increase in Leishmania-specific immune responses was observed, and monkeys immunized with antigen and IL-12 exhibited an IFN-gamma response that was as great as that in animals that had resolved a primary infection and were immune. Nevertheless, despite the presence of correlates of protection, the disease course was only slightly altered, and protection was low compared to that in self-cured monkeys. These data suggest that protection against leishmaniasis may require more than the activation of Leishmania-specific IFN-gamma-producing T cells, which has important implications for designing a vaccine against leishmaniasis.
2208. Hussein MM. Mooij JM. Roujouleh HM. Hamour OA. Felemban H. Non-typhoid Salmonella septicemia and visceral leishmaniasis in a renal transplant patient. Transplantation. 71(3):479-81, 2001 Feb 15.
Abstract
BACKGROUND: We report on a renal transplant patient with recurrent attacks of fever, in which Salmonella septicemia as well as visceral leishmaniasis were diagnosed. PATIENT: The patient was a 62-year-old man with diabetic nephropathy and a living related kidney transplantation. RESULTS: Nearly 2 years after the transplantation, the patient developed recurrent attacks of fever, which were initially diagnosed as non-typhoid salmonellosis and improved after treatment. Three months later, he had relapses of fever. As the patient developed pancytopenia, a bone marrow aspiration was done, showing Leishmania parasites. The patient responded well to treatment with sodium stibogluconate. CONCLUSIONS: A high index of suspicion, together with better diagnostic assays to detect visceral leishmaniasis, is warranted in the diagnostic work-up of any fever of unknown origin in immunocompromised patients, especially in endemic areas.
2209. Kumar A. Mittal M. Prasad S. Treatment of leishmaniasis in pregnancy. International Journal of Gynaecology & Obstetrics. 72(2):189-90, 2001 Feb.
2210. Lachaud L. Chabbert E. Dubessay P. Reynes J. Lamothe J. Bastien P. Comparison of various sample preparation methods for PCR diagnosis of visceral leishmaniasis using peripheral blood. Journal of Clinical Microbiology. 39(2):613-7, 2001 Feb.
Abstract
We have compared various sample preparation methods for the PCR diagnosis of visceral leishmaniasis (VL) using peripheral blood samples and tested the influence of these protocols upon sensitivity. Four methods of lysis-DNA extraction were used with two types of blood samples: whole blood (WB) and buffy coat (BC). Comparisons were first carried out with seeded samples at various parasite concentrations. At high concentrations (> or = 1,000 parasites/ml), there were no significant differences in PCR sensitivity among the methods tested. At concentrations of < or = 100 parasites/ml, proteinase K (PK)-based methods proved clearly superior to guanidine-EDTA-based methods. Moreover, a 10-fold increase in sensitivity was observed for BC over that for WB. Thus, the best sensitivity was obtained with the BC prepared with PK-based methods. With this combination, the PCR reliably detected 10 parasites/ml but was inconsistent when the sample contained 1 parasite/ml of blood. The methods that yielded the highest sensitivities were evaluated with seven dogs and four human VL patients. Again, the utilization of the BC prepared with PK-based methods gave the best results. The optimization of each step of the assay (sample preparation, DNA extraction, and PCR conditions) yielded a highly sensitive tool for the diagnosis of VL using patient blood, thus avoiding more invasive diagnostic procedures and allowing the detection of low parasitemia during posttherapeutic follow-up.
2211. Marom D. Offer I. Tamary H. Jaffe CL. Garty BZ. Hemophagocytic lymphohistiocytosis associated with visceral leishmaniasis. Pediatric Hematology & Oncology. 18(1):65-70, 2001 Jan-Feb.
Abstract
A 2-year-old child presented with fever and hepatosplenomegaly. Laboratory findings showed pancytopenia, hypertriglyceridemia, hyperferritinemia, and high levels of soluble-IL2 receptors. Initial bone marrow aspiration and biopsy revealed mild hemophagocytosis. A diagnosis of hemophagocytic lymphohistiocytosis was made and appropriate treatment was begun. Repeated marrow aspiration performed because of lack of clinical response revealed Leishmania amastigotes in macrophages in addition to active hemophagocytosis. Treatment with liposomal amphotericin resulted with rapid recovery. Visceral leishmaniasis should be considered in the differential diagnosis of hemophagocytic syndrome.
2212. Pintado V. Martin-Rabadan P. Rivera ML. Moreno S. Bouza E. Visceral leishmaniasis in human immunodeficiency virus (HIV)-infected and non-HIV-infected patients. A comparative study. Medicine. 80(1):54-73, 2001 Jan.
Abstract
Visceral leishmaniasis is an endemic infection in Mediterranean countries, where it has become a frequent complication of acquired immunodeficiency syndrome (AIDS). The incidence of visceral leishmaniasis is increasing in Spain due to human immunodeficiency virus (HIV)-related cases, but some aspects of its epidemiology, clinical features, and management remain unknown. In addition, no comparative clinical studies about the disease in HIV-infected and non-HIV-infected patients have been reported. During a 24-year period, 120 cases of visceral leishmaniasis were diagnosed at our institution and 80 (66%) were associated with HIV infection. The mean age at diagnosis was higher in HIV-infected that in non-HIV-infected patients (33.2 versus 23.2 yr; p = 0.002), but the male/female ratio was similar in both groups. The main risk factor for HIV infection was intravenous drug abuse (78.7%). The clinical presentation of leishmaniasis was similar in both groups, but HIV-infected patients had a lower frequency of splenomegaly than HIV-negative individuals (80.8% versus 97.4%; p = 0.02). HIV-infected patients had a greater frequency and degree of leukopenia, lymphocytopenia, and thrombocytopenia. Most of them were profoundly immunosuppressed (mean CD4+ lymphocyte count, 90 cells/mm3) at the time of diagnosis of leishmaniasis, and 53.7% had AIDS. The sensitivity of serologic studies for Leishmania was significantly lower in HIV-infected than in non-HIV-infected patients (50% versus 80%; p < 0.001), but the diagnostic yield of bone marrow aspirate (67.1% versus 79.4%) and bone marrow culture (62.9% versus 66.6%) was similar in both groups. After initial treatment, the response rate was significantly lower in HIV-infected than in non-HIV-infected individuals (54.8% versus 89.7%; p = 0.001). The relapse rate was 46.2% and 7.5%, respectively (p < 0.001). Secondary prophylaxis with antimonial compounds or amphotericin B seems to be useful in preventing relapses in HIV-infected patients. The mortality rate was higher (53.7% versus 7.5%; p < 0.001) and the median survival time shorter (25 versus > 160 mo; p < 0.001) in AIDS patients than in HIV-negative individuals. Although leishmaniasis could contribute to death in a significant number of HIV-infected patients, it was the main cause of death in only a few of them. The CD4+ lymphocyte count and the use of highly active antiretroviral therapy and secondary prophylaxis for leishmaniasis were the most significant prognostic factors for survival in AIDS patients. Visceral leishmaniasis behaves as an opportunistic infection in HIV-infected individuals and should be considered as an AIDS-defining disease.
2213. Pizzuto M. Piazza M. Senese D. Scalamogna C. Calattini S. Corsico L. Persico T. Adriani B. Magni C. Guaraldi G. Gaiera G. Ludovisi A. Gramiccia M. Galli M. Moroni M. Corbellino M. Antinori S. Role of PCR in diagnosis and prognosis of visceral leishmaniasis in patients coinfected with human immunodeficiency virus type 1. Journal of Clinical Microbiology. 39(1):357-61, 2001 Jan.
Abstract
A group of 76 consecutive human immunodeficiency virus (HIV)-positive patients with fever of unknown origin (n = 52) or fever associated with pulmonary diseases was evaluated in order to assess the usefulness of PCR with peripheral blood in the diagnosis and follow-up of visceral leishmaniasis. We identified 10 cases of visceral leishmaniasis among the 52 patients with fever of unknown origin. At the time of diagnosis, all were parasitemic by PCR with peripheral blood. During follow-up, a progressive decline in parasitemia was observed under therapy, and all patients became PCR negative after a median of 5 weeks (range, 6 to 21 weeks). However, in eight of nine patients monitored for a median period of 88 weeks (range, 33 to 110 weeks), visceral leishmaniasis relapsed, with positive results by PCR with peripheral blood reappearing 1 to 2 weeks before the clinical onset of disease. Eight Leishmania infantum and two Leishmania donovani infections were identified by PCR-restriction fragment length polymorphism analysis. PCR with peripheral blood is a reliable method for diagnosis of visceral leishmaniasis in HIV-infected patients. During follow-up, it substantially reduces the need for traditional invasive tests to assess parasitological response, while a positive PCR result is predictive of clinical relapse.
2214. Probst P. Stromberg E. Ghalib HW. Mozel M. Badaro R. Reed SG. Webb JR. Identification and characterization of T cell-stimulating antigens from Leishmania by CD4 T cell expression cloning. Journal of Immunology. 166(1):498-505, 2001 Jan 1.
Abstract
Persistent immunity against Leishmania: infections in humans is mediated predominantly by CD4(+) T cells of the Th1 phenotype. Herein we report the expression cloning of eight Leishmania: Ags using parasite-specific T cell lines derived from an immune donor. The Ags identified by this technique include the flagellar proteins alpha- and beta-tubulin, histone H2b, ribosomal protein S4, malate dehydrogenase, and elongation factor 2, as well as two novel parasite proteins. None of these proteins have been previously reported as T cell-stimulating Ags from Leishmania: beta-tubulin-specific T cell clones generated against Leishmania: major amastigotes responded to Leishmania:-infected macrophages and dendritic cells. IFN-gamma enzyme-linked immunospot analysis demonstrated the presence of T cells specific for several of these Ags in PBMC from self-healing cutaneous leishmaniasis patients infected with either Leishmania: tropica or L. major. The responses elicited by Leishmania: histone H2b were particularly striking in terms of frequency of histone-specific T cells in PBMC (1 T cell of 6000 PBMC) as well as the percentage of responding donors (86%, 6 of 7). Ags identified by T cells from immune donors might constitute potential vaccine candidates for leishmaniasis.
2215. Reinecke P. Gabbart HE. Strunk W. Losche CC. Ocular scleromalacia caused by leishmaniasis: a rare cause of scleral perforation. British Journal of Ophthalmology. 85(2):240-1, 2001 Feb.
2216. No abstract
2217. Soares NM. Santiago MB. Pontes deCarvalho LC. An improved anti-C3/IgG ELISA for quantification of soluble immune complexes. Journal of Immunological Methods. 249(1-2):199-205, 2001 Mar 1.
Abstract
A semi-quantitative ELISA for complement-fixing, IgG-containing immune complexes (IC) is described. The assay is based on the insolubilization of IC by polyethyleneglycol, their capture by solid-phase anti-C3 antibodies, reaction with peroxidase-labeled anti-IgG antibodies and incubation with a chromogenic peroxidase substrate. It was markedly improved by the use of a single-step procedure which simultaneously washed and precipitated the insolubilized immune complexes. Intra-assay and inter-assay coefficients of variation were lower than 8.6 and 14.7%, respectively. As expected, higher levels of circulating immune complexes, in relation to healthy individuals, were found in patients with American visceral leishmaniasis (AVL), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), with prevalences comparable to those described in the literature. The ELISA can be quickly assembled from reagents and plasticware widely available commercially, detects immune complexes fulfilling three different criteria and is more sensitive than a previously published method based on the same principles (detection limit for complement-sensitized aggregated IgG of 2 microg ml(-1) as compared with a detection limit above 16 microg ml(-1)).
2218. Streit JA. Recker TJ. Filho FG. Beverley SM. Wilson ME. Protective immunity against the protozoan Leishmania chagasi is induced by subclinical cutaneous infection with virulent but not avirulent organisms. Journal of Immunology. 166(3):1921-9, 2001 Feb 1.
Abstract
Protective immunity against Leishmania major is provided by s.c. immunization with a low dose of L. major promastigotes or with dihydrofolate-thymidylate synthase gene locus (DHFR-TS) gene knockout L. major organisms. Whether these vaccine strategies will protect against infection with other Leishmania species that elicit distinct immune responses and clinical syndromes is not known. Therefore, we investigated protective immunity to Leishmania chagasi, a cause of visceral leishmaniasis. In contrast to L. major, a high dose s.c. inoculum of L. chagasi promastigotes was required to elicit protective immunity. Splenocytes from mice immunized with a high dose produced significantly greater amounts of IFN-gamma and lower TGF-beta than mice immunized with a low dose of promastigotes. The development of protective immunity did not require the presence of NK cells. Protection was not afforded by s.c. immunization with either attenuated L. chagasi or with L. major promastigotes, and s.c. L. chagasi did not protect against infection with L. major. Subcutaneous immunization with DHFR-TS gene knockouts derived from L. chagasi, L. donovani, or L. major did not protect against L. chagasi infection. We conclude that s.c. inoculation of high doses of live L. chagasi causes a subclinical infection that elicits protective immune responses in susceptible mice. However, L. chagasi that have been attenuated either by long-term passage or during the raising of recombinant gene knockout organisms do not elicit protective immunity, either because they fail to establish a subclinical infection or because they no longer express critical antigenic epitopes.
2219. Thakur CP. A single high dose treatment of kala-azar with Ambisome (amphotericin B lipid complex): a pilot study. International Journal of Antimicrobial Agents. 17(1):67-70, 2001 Jan.
Abstract
Thirty four patients with parasitologically confirmed visceral leishmaniasis were divided randomly into two groups of 17. Group A received Ambisome (amphotericin B lipid complex) at a dose of 15mg/kg body weight infused over 2h as a single dose; patients in group B received amphotericin B deoxycholate at a dose of 1mg/kg body infused for 2h for 20days. All 34 patients had a clinical, parasitological and ultimate cure. Ambisome was much better tolerated than amphotericin B, and adverse events were fewer in the Ambisome group. It was concluded that, if the cost of Ambisome were reduced, it would be a suitable first line drug. A longer study comparing three regimes of Ambisome: 15mg/kg body weight, 11mg/kg body weight and 7.5mg/kg body weight, should be undertaken.
2220. Thakur CP. Ahmed S. Observations on amphotericin B treatment of kala-azar given in a rural set up in Bihar, India. Indian Journal of Medical Research. 113:14-8, 2001 Jan.
Abstract
BACKGROUND
& OBJECTIVES: In a kala-azar endemic area in rural Bihar with a large
number of patients unresponsive to sodium antimony gluconate and pentamidine,
treatment with amphotericin B was tried in a rural set-up with the objective to
cure these patients, and to assess whether such a centre could be run
successfully in a rural set-up. METHODS: After thorough clinical examination
and biochemical investigations, parasitologically confirmed patients who had
haemoglobin above 5 g/dl, electrolyte imbalance if any, corrected, and ECG
changes suggestive of myocardial damage stabilised after 10 days of bed rest
were treated. Amphotericin B deoxycholate was infused at a dose of 1 mg/kg body
weight, daily for 20 days. The adverse events were closely monitored. RESULTS:
All 7 (100%) untreated patients of kala-azar, 258 (97%) of the 266 antimony and
pentamidine resistant patients, and 31 (86%) of the 36 patients who had
relapsed after a low dose regimen of amphotericin B, were cured with 20
infusions of amphotericin B. Eight (3%) patients of the antimony and
pentamidine resistant group and 5 (14%) patients who had relapsed after low
dose amphotericin B regimen required 25 infusions of amphotericin B to achieve
parasitological cure. INTERPRETATION & CONCLUSIONS: With some precautions
and proper management, all patients of kala-azar could be cured with
amphotericin B in a rural set-up. A significant (P < 0.05) percentage of
patients of group C relapsing after a low dose amphotericin B regime requiring
25 infusions for cure suggests that an adequate dose regime of amphotericin B should be given during the first course
of treatment to prevent emergence of drug resistance.
2814. Bottrel RL. Dutra WO. Martins FA. Gontijo B. Carvalho E. Barral-Netto M. Barral A. Almeida RP. Mayrink W. Locksley R. Gollob KJ. Flow cytometric determination of cellular sources and frequencies of key cytokine-producing lymphocytes directed against recombinant LACK and soluble Leishmania antigen in human cutaneous leishmaniasis. Infection & Immunity. 69(5):3232-9, 2001 May.
Abstract
Leishmaniasis, caused by infection with the
protozoan parasite Leishmania, affects millions of individuals worldwide,
causing serious morbidity and mortality. This study directly determined the
frequency of cells producing key immunoregulatory cytokines in response to the
recombinant antigen Leishmania homolog of receptors for activated kinase C
(LACK) and soluble leishmania antigen (SLA), and it determined relative
contributions of these antigens to the overall cytokine profile in individuals
infected for the first time with Leishmania braziliensis. All individuals
presented with the cutaneous clinical form of leishmaniasis and were analyzed
for proliferative responses to LACK antigen and SLA, frequency of lymphocyte subpopulations
(analyzed ex vivo), and antigen-induced (LACK and SLA) cytokine production at
the single-cell level (determined by flow cytometry). The following were
determined. (i) The Th1-type response previously seen in patients with
cutaneous leishmaniasis is due to gamma interferon (IFN-gamma) production by
several different sources, listed in order of contribution: CD4(+) T
lymphocytes, CD4(-), CD8(-) lymphocytes, and CD8(+) T lymphocytes. (ii) SLA
induced a higher frequency of lymphocytes producing IFN-gamma and tumor
necrosis factor alpha (TNF-alpha) than did LACK. (iii) LACK induced an
activation of monocyte populations as reflected by an increased percentage of
CD14-positive cells. (iv) Neither SLA nor LACK induced detectable frequencies
of cells producing interleukin-4 (IL-4) or IL-5. These data demonstrated a
multifaceted immune response to SLA in human leishmaniasis involving Th1 CD4(+)
T lymphocytes (IFN-gamma(+) and IL-10(-)/IL-4(-)), Tc1 CD8(+) T cells
(IFN-gamma(+), and IL-10(-)/IL-4(-)), and a high frequency of
TNF-alpha-producing lymphocytes. Moreover, it was determined that the
recombinant antigen LACK acts as a weak inducer of Th1-type lymphocyte
responses compared to SLA.
2815.
el-Hassan
AM. Zijlstra EE. Leishmaniasis in Sudan. Mucosal leishmaniasis.
Transactions of the Royal Society of Tropical Medicine & Hygiene. 95 Suppl 1:S19-26, 2001 Apr.
Abstract
Sudanese mucosal leishmaniasis is a chronic
infection of the upper respiratory tract and/or oral mucosa caused mainly by
Leishmania donovani. The disease occurs in areas of the country endemic for
visceral leishmaniasis, particularly among Masalit and other closely related
tribes in western Sudan. The condition may develop during or after an attack of
visceral leishmaniasis, but in most cases it is a primary mucosal disease.
Unlike South American mucocutaneous leishmaniasis, mucosal leishmaniasis in
Sudan is not preceded or accompanied by a cutaneous lesion. Pathologically, the
lesions show a mixture of macrophages, plasma cells and lymphocytes. An
epithelioid granuloma may also be found. Parasites are scanty. Diagnosis is
established by demonstration of parasites in smears or biopsies, by culture or
animal inoculation, or with the aid of the polymerase chain reaction. Most
patients give positive results in the direct agglutination test and leishmanin
skin test. Patients respond well to treatment with pentavalent antimony
compounds.
2816.
Nylen
S. Mortberg U. Kovalenko D. Satti I. Engstrom K. Bakhiet M.
Akuffo H. Differential induction of cellular responses by live and dead
Leishmania promastigotes in healthy donors.
Clinical & Experimental Immunology.
124(1):43-53, 2001 Apr.
Abstract
The most effective protection against human
leishmaniasis has been achieved following vaccination with live promastigotes.
Killed promastigotes + BCG can protect, albeit to a lower degree. To explore
what mechanisms may be involved in these differences, the ability of live and
dead promastigotes to induce immune responses were evaluated in vitro. The data
showed that live and dead promastigotes differ in their ability to induce
proliferation and cytokine production. Cytokine gene expression of Th1 related
cytokines (IL-12, IFNgamma and TNFalpha) in adult PBMC was more evident to live
than to heat killed promastigotes. This was coupled with significantly higher
number of IFNgamma secreting cells induced by live than killed promastigotes.
However, alpha-IL-12 antibodies did not block the IFNgamma response induced by
live promastigotes. Proliferative responses were variable. In contrast to adult
PBMC no IFNgamma secreting MNC could be detected in cord blood. However, in
these cells the live promastigotes consistently induced higher proliferative
response compared to dead. Implications of these findings are discussed.
2817.
Palmer
RA. Tran D. Hepburn NC. Ashton RE.
The management of cutaneous leishmaniasis from Belize. Clinical & Experimental
Dermatology. 26(1):16-20, 2001 Jan.
Abstract
We report 20 patients who contracted
cutaneous leishmaniasis in Central and South America, 18 of them in Belize. The
diagnosis was confirmed by the polymerase chain reaction (PCR) in 79% of those
tested; the corresponding figure for histology was 62%, touch smear 46%, and
culture 11%. Results of PCR can be falsely positive, so treatment should not be
based on PCR alone. Of the 20 cases 18 were healed 6 weeks after intravenous
sodium stibogluconate 20 mg/kg per day for 20 days. We present a management
protocol.
2818.
Zijlstra
EE. el-Hassan AM. Leishmaniasis in
Sudan. Visceral leishmaniasis. Transactions of the Royal Society of Tropical
Medicine & Hygiene. 95 Suppl
1:S27-58, 2001 Apr.
Abstract
From the early 1900s, visceral leishmaniasis
(VL; kala-azar) has been among the most important health problems in Sudan,
particularly in the main endemic area in the eastern and central regions.
Several major epidemics have occurred, the most recent--in Western Upper Nile
province in southern Sudan, detected in 1988--claiming over 100,000 lives. The
disease spread to other areas that were previously not known to be endemic for
VL. A major upsurge in the number of cases was noted in the endemic area. These
events triggered renewed interest in the disease. Epidemiological and
entomological studies confirmed Phlebotomus orientalis as the vector in several
parts of the country, typically associated with Acacia seyal and Balanites
aegyptiaca vegetation. Infection rates with Leishmania were high, but subject
to seasonal variation, as were the numbers of sand flies. Parasites isolated
from humans and sand flies belonged to three zymodemes (MON-18, MON-30 and
MON-82), which all belong to the L. donovani sensu lato cluster. Transmission
dynamics have not been elucidated fully; heavy transmission in relatively
scarcely populated areas such as Dinder national park suggested zoonotic
transmission whereas the large numbers of patients with post kala-azar dermal
leishmaniasis (PKDL) in heavily affected villages may indicate a human
reservoir and anthroponotic transmission. Clinical presentation in adults and
in children did not differ significantly, except that children were more
anaemic. Fever, weight loss, hepato-splenomegaly and lymphadenopathy were the
most common findings. PKDL was much more common than expected (56% of patients
with VL developed PKDL), but other post-VL manifestations were also found
affecting the eyes (uveitis, conjunctivitis, blepharitis), nasal and/or oral
mucosa. Evaluation of diagnostic methods showed that parasitological diagnosis
should still be the mainstay in diagnosis, with sensitivities for lymph node,
bone marrow and spleen aspirates of 58%, 70% and 96%, respectively. Simple,
cheap serological tests are needed. The direct agglutination test (DAT) had a
sensitivity of 72%, specificity of 94%, positive predictive value of 78% and
negative predictive value of 92%. As with other serological tests, the DAT
cannot distinguish between active disease, subclinical infection or past
infection. The introduction of freeze-dried antigen and control sera greatly
improved the practicality and accuracy of the DAT in the field. An
enzyme-linked immunosorbent assay using recombinant K39 antigen had higher
sensitivity than DAT (93%). The polymerase chain reaction using peripheral
blood gave a sensitivity of 70-93% and was more sensitive than microscopy of
lymph node or bone marrow aspirates in patients with suspected VL. The
leishmanin skin test (LST) was typically negative during active VL and
converted to positive in c. 80% of patients 6 months after treatment.
Immunological studies showed that both Th1 and Th2 cell responses could be
demonstrated in lymph nodes from VL patients as evidenced by the presence of
messenger ribonucleic acid for interleukin (IL)-10, interferon gamma and IL-2.
Treatment of peripheral blood mononuclear cells from VL patients with IL-12 was
found to drive the immune response toward a Th1 type response with the
production of interferon gamma, indicating a potential therapeutic role for
IL-12. VL responded well to treatment with sodium stibogluconate, which is
still the first line drug at a dose of 20 mg/kg intravenously or
intramuscularly per day for 15-30 d. Side effects and resistance were rare.
Liposomal amphotericin B was effective, with few side effects. Control measures
have not been implemented. Based on observations that VL does not occur in
individuals who have a positive LST, probably because of previous cutaneous
leishmaniasis, a vaccine containing heat-killed L. major promastigotes is
currently undergoing a phase III trial.