Demonstration of Leishmania parasites in skin lesions of Indian post kala-azar dermal leishmaniasis (PKDL) cases.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to S K Kar.
Explore the source record for details and available documents.
The performance of direct agglutination test (DAT) was evaluated under field conditions in three endemic districts of Bihar. In three villages of these districts, 197 households were surveyed with a population of 1167 individuals. Out of 1167 individuals, 603 were clinically examined for VL and blood samples of each examined individual were collected for DAT. The proportion of DAT positivity in both males (32.33%) and females (32.94%) was statistically not significant (p > 0.05). The proportion of DAT positivity in all age groups was observed to be similar (p > 0.05). The GMRT was observed highest in active VL cases. Out of 20 DAT positive individuals having fever, 5 were examined parasitologically (bone marrow aspiration) and in three cases positivity was observed. This showed the relevance of DAT in early diagnosis of visceral leishmaniasis and usefulness of DAT in field conditions.
Knowing the exact breeding places inside the habitat is very important to plan the larval control strategy. Information regarding larval biology in relation to different seasons will be more useful to organize insecticide spray schedule at a particular month of maximum immature density to bring down the adult sandfly density. In the present study, maximum number of soil samples were found positive in the month of January and minimum in the month of September. Maximum positive soil samples were collected from cattle sheds, minimum in mixed dwellings and in case of human dwellings all soil samples were negative. Comparison of two methods for the isolation of immature stages showed that direct microscopic observation is superior to sugar flotation technique.
Explore the source record for details and available documents.
High titers of parasite antigen-specific IgG4 antibodies have been found to be circulating in the peripheral blood of chronic patients, asymptomatic microfilariae carriers, and endemic normals in bancroftian filariasis. But in contrast to this, the titers of antigen-specific IgG1, IgG2, and IgG3 isotype antibodies are much lower. Using soluble antigens of adult Setaria digitata, a cattle parasite which shows strong antigenic reactivity with filaria sera, we have identified, by immunoblot, 14- to 20-kDa antigens which are recognized only by the IgG4 isotype antibodies present in the sera of asymptomatic microfilariae carriers. These 14- to 20-kDa antigens, after fractionation by SDS-PAGE and transfer to nitrocellulose paper, when solubilized and tested in vitro, induced secretion of a higher quantity of IFN-gamma and a lower quantity of IL-4, IL-5, and IL-10 (differential Th1 and Th2 response) in the lymphocytes of endemic normals in comparison to what they induced in the lymphocytes of asymptomatic microfilariae carriers.
Cyclosporin A (CsA) at 120 mg/kg body weight when injected subcutaneously into BALB/c mice along with thyroglobulin emulsified in incomplete Freund's adjuvant (IFA) was found to suppress antigen-specific IgG titre by 86%. Isotyping revealed that both IgG1 and IgG2a titres were suppressed by 87% and 57%, respectively. But under identical conditions when complete Freund's adjuvant (CFA) was used, the suppression of antigen-specific IgG, IgG1 and IgG2a titres was 50%, 51% and 55%, respectively. Injection of anti-IL-1beta-neutralizing hamster monoclonal antibodies along with thyroglobulin and CsA emulsified in CFA increased the suppression of antigen-specific IgG titre. Under such conditions the IgG1 titre was suppressed more than the IgG2a titre. Recombinant human interleukin-1 receptor antagonist (rhuIL-1ra) also enhanced the suppression caused by CsA in the presence of CFA but control hamster immunoglobulin had no such effect. Recombinant human IL-1beta, when administered along with thyroglobulin and CsA emulsified in IFA, alleviated the suppression of antigen-specific IgG titre and the IgG1 titre was alleviated more than the IgG2a titre. Under identical conditions, rhuIL-1ra did not alleviate CsA-induced suppression. Lymphocytes from the lymph nodes of thyroglobulin-sensitized BALB/c mice when stimulated in vitro by thyroglobulin in the presence of CsA, secreted very little interferon-gamma (IFN-gamma) and IL-4, but on addition of an optimal dose of rhuIL-1beta, IFN-gamma and IL-4 secretion was partially restored.
Peripheral blood smears of 20 patients with Indian Kala-azar (KA) were collected throughout the day and night and examined for presence of Leishmania amastigotes. Amastigotes were detected in 46% of peripheral blood samples taken during the day (08:00-17:00 h) and 66% and during the night (18:00-07:00 h). This difference was statistically highly significant (P < 0.0001). The confirmation of amastigote diurnal periodicity is of interest, and the high percentage of parasitemic patients at night is likely to reflect the biting habit of the sandfly vector of Kala-azar Phlebotomus argentipes.
A pilot study for the control of Ph. argentipes, a known vector of kala-azar in India, was carried out using an ecological approach. Of the 15 houses selected for the study 10, including the cattle sheds and latrines, were plastered with a mixture of mud and lime, up to a height of 1.22 m taking care to seal all cracks and crevices. The remaining five houses were left unplastered and were considered as control areas. The pre-treatment and post-treatment resting densities of the sandfly were monitored both in treated and untreated houses. A sudden drop in the sandfly density was noticed in the treated houses, whereas there was no significant reduction in the check houses, suggesting an effective control.
Explore the source record for details and available documents.
We have analysed the suppressive effect of cyclophosphamide, when given in vivo, on the antibody response of BALB/c mice against Shigella dysenteriae antigens using ELISA and immunoblot. Out of various protocols tried, it was found that co-injection of cyclophosphamide at 150 mg/kg bodyweight, i.p., at the time of antigen administration and then after a lapse of 24 h during both primary and secondary immunizations, was the most effective in suppressing antibody response of mice. Analysis of sera by ELISA demonstrated the presence of some antibodies to S. dysenteriae antigens after secondary immunization, but immunoblot analysis using the same sera revealed complete suppression of antibody response. Animals whose antibody response was almost completely suppressed after two immunizations with co-injection of cyclophosphamide, when immunized again after the lapse of 14 days from the date of secondary immunization with Shigella antigens but without administration of cyclophosphamide, partially recovered their ability to respond to the same antigens. This protocol can now be used in mice to analyse the hierarchy of immunogenic epitopes present in a complex mixture of antigens.
A differential serological screen of a lambda gt11 cDNA expression library of Plasmodium falciparum was performed in an attempt to identify novel and putative host-protective antigens of the parasite. The screening was done with two categories of sera: (i) acute-phase sera obtained from smear-positive acutely infected P. falciparum patients from various regions in India and (ii) immune sera taken from healthy, permanent adult residents of P. falciparum-endemic rural districts of Orissa in eastern India. These adults had not suffered from any clinical malarial symptoms for at least the previous 3 years at the time of serum collection. Sixty-five clones obtained by screening the lambda gt11 library with two immune serum samples were analyzed extensively with a total of 70 acutely infected patient serum samples. Eight of these clones failed to react with any of the patient sera. Each of these eight clones, when tested individually with 92 serum samples from the immune group, reacted with a minimum of 43% of the samples from this category of sera. Thus, these eight epitopes may encode host-protective elements since they are not recognized by antibodies in the patient sera but react exclusively and extensively with the clinically immune set. Sequence analysis of two of these clones reveals that they are novel Plasmodium genes.
Explore the source record for details and available documents.
Side reactions following ivermectin treatment were evaluated in sixty males with high density bancroftian microfilaremia (GM 1388/ml). Following a single oral dose of ivermectin of different strengths (20, 50, 100 or 200 micrograms/kg), microfilariae clearance and side reactions were monitored in a double blind fashion. Microfilaria levels fell rapidly after ivermectin administration in all dosage groups and 98% of pretreatment microfilariae was cleared after 12 h of treatment. The rate of microfilaria (mf) clearance was slower with 20 micrograms/kg than with the highest dose (200 micrograms/kg) administered. Forty-six patients (77%) became amicrofilaraemic within 2 weeks of treatment. Side reactions were noted in 97% of cases. The most common reactions were fever, headache, weakness, myalgia and cough which appeared by 12 h and subsided by 72 h following treatment. The frequency and intensity of side reactions were related to pretreatment mf densities and were independent of the dose administered. Unusual side reactions were noted in a few patients with high density microfilaraemia. These included intense cough, shortness of breath, blood tinged mucoid expectoration associated with patchy pneumonitis of the lung. Itchy rashes, lymphatic nodules and raised alkaline phosphatase level were also observed in some patients. These side reactions were transient, self limiting and were not serious enough to warrant any treatment. These exaggerated unusual reactions were possibly due to allergic response of the susceptible host to rapid killing of large number of microfilariae.
Prevalence and clinical manifestation of lymphatic nodule in natural course of bancroftian filariasis was evaluated in a representative population of 2837 of a defined endemic community of Orissa. Detailed clinical and parasitological examination of study subjects revealed high disease (35.6%) and mf rate (15.8%) and mean microfilariae density (GM: 15.3 +/- 5.1 per 60 mm3 of blood) in the community. Lymphatic filarial nodule was manifested in 216 (7.6%) subjects in different spectrum of filarial infection including endemic normals and asymptomatic mf carriers. Significant proportion of cases with acute filarial disease in the community manifested nodule. Significant proportion of cases with nodule had microfilaraemia (33.3%) as compared to that of community. However highest frequency of nodule was observed amongst microfilaraemics with lowest density of microfilaraemia in the community (P < 0.05.). This possibly indicates microfilariae clearance from circulation as a consequence of death of adult parasite. The above observations suggest that lymphatic nodules may manifest as an early and/or the only clinical sign of bancroftian filariasis. This might have been mediated through either the attempt of the immuno-competent host to kill the adult parasite in a similar fashion that is effected through DEC or by death of the mature parasite in its natural process of attrition.
Using direct fluorescent antibody analysis it was shown that the sheath of live microfilariae of Wuchereria bancrofti has human albumin and the immunoglobulin G subclasses IgG1 and IgG4 on its surface.
Ivermectin treatment was evaluated for its efficacy and side reactions in sixty patients of Orissa with Bancroftian filarial infection and microfilaremia. Ivermectin was administered as a single oral dose at four dosage levels (20, 50, 100 and 200 micrograms/kg), and both microfilarial clearance and associated side reactions were monitored in a double blind fashion. Blood microfilariae were cleared in all patients at all dosages within 1 to 14 days. In most patients microfilariae reappeared by third month. The microfilaria appearance by third and sixth month averaged 12.2 to 44 percent of pretreatment values in the four study groups. Side reactions were encountered in almost all patients, the commonest being fever, headache, weakness, myalgia and cough which occurred most prominently 12 to 72 hours after treatment. Side reactions were more frequent and severe in patients with high microfilaria counts. Clinical reaction scores for each group were independent of the dose administered. The 200 micrograms dose group showed significantly more rapid microfilariae clearance and its delayed reappearance as compared with the other dosage groups and without inducing significantly greater clinical reaction scores.
To evaluate the possible role of ethnicity in susceptibility to filarial infection, a comparative study of the prevalence of filarial infection was initiated in an endemic village inhabited by two ethnic populations of mainlanders and tribals. An age and sex matched sampled population of 591 mainlanders and 106 tribals was studied by detailed clinical and parasitological (60 mm3 blood) examinations. Sera collected from both population groups (26 each) matched for clinical stage of infection were analysed for humoral immune responses such as antifilarial IgG, circulating filarial antigen and immune complex level. The overall prevalences of clinical disease and infection in both mainlanders (34.18 and 14.4%) and tribals (25.47 and 17.9%) were comparable. However, both annual average adenolymphangitic attack rate (1.77 year-1) and the prevalence of chronic filarial disease (22.6%) amongst tribals were significantly lower. No true elephantiasis was observed in tribals. No significant difference was observed in their humoral immune response, although the antifilarial antibody of IgG class in all stages of filarial infection was lower in tribals than in mainlanders. The results did not reveal any difference in susceptibility to filarial infection in the ethnic groups. The paucity of progressive lesions observed in tribals possibly reflects a difference in the anatomy of lymphatics or genetic or immunoregulatory mechanisms, that needs further study.
Two mouse monoclonal antibodies SKb1 and SKb6 were prepared by fusion of myeloma cells with spleen cells of female Balb/c mouse immunized with a mixture of bovine IgG1 and IgG2. In radioimmunoassay, SKb1 bound specifically to IgG2 but SKb6 reacted with both IgG1 and IgG2 molecules. In the competition experiments, heavy chain isolated from bovine IgG could inhibit the binding of 125I-IgG1 and 125I-IgG2 to SKb6, while it failed to inhibit the binding of 125I-IgG2 to SKb1. The epitope reacting with SKb1 was found to be present not only on bovine IgG2 but also on goat IgG and was not present on IgG molecules isolated from the serum of rabbit, rat, sheep, horse, human and monkey. Similarly, the epitope reacting to SKb6 was found to be present on bovine IgG1 and IgG2 and also on IgG molecules isolated from goat and sheep serum but was absent in the IgG molecules isolated from the serum of rabbit, rat, horse, human and monkey. The association constants of the interactions of SKb1 with 125I-IgG2 and of SKb6 with 125I-IgG1 and 125I-IgG2, determined by Scatchard analysis, Steward-Petty plot and Sips plot, were found to be in the order of 10(8)-10(10) L/M. The association constants were determined at varying temperatures to obtain the thermodynamic parameters. The enthalpy (delta H0) and entropy (delta S0) values for the above antigen-antibody interactions were in the range of 9.15-15.96 kcal/mole and 36.96-41.15 eu/mole respectively. The heterogeneity indices for similar interactions determined by Sips equation were consistent with the expected values for binding of monoclonal antibodies with homogeneous protein determinants.