Biomedical subjects
Publications and source records attributed to B K Girdhar.
A case of suspected giant nerve abscess near the radial nerve in the upper arm of a patient with borderline leprosy is reported. On exploration, it turned out to be a deep seated lipoma. Consideration should be given to exploring nerve masses in leprosy.
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The characteristics of infiltrates in the dermal and neural granulomas from the same leprosy patients were compared by preparing a single cell suspension. Skin and nerve biopsies from 10 patients, 5 with tuberculoid and 5 with lepromatous leprosy were analysed. The granulomas contained lymphocytes and macrophages. Lymphocytes were the predominant infiltrating cells in the tuberculoid dermal and neural granulomas. A high proportion of lymphocytes in both the skin and nerve granulomas in these cases were activated T cells as they formed rosettes with sheep erythrocytes and expressed HLA-DR antigens. In contrast, lepromatous dermal and neural granulomas contained very few of these lymphocytes. Dermal and neural granulomas from both the types of leprosy contained mature macrophages as they were esterase positive, did not exhibit peroxidase activity and expressed HLA-DR antigens. These macrophages did not possess C3 surface receptors either. These findings suggest that the infiltrates in the skin and nerve granulomas of a given type of leprosy have similar characteristics.
Single cell suspension from the granulomas in nerves of leprosy patients were prepared for an in vitro study of the properties of infiltrating cells. Nerve biopsies from 17 patients with tuberculoid (n = 9) and lepromatous (n = 8) leprosy cases were analysed. The granulomas were found to contain lymphocytes and macrophages. Lymphocytes were the predominant infiltrating cells in the tuberculoid nerves. In contrast, lepromatous nerves contained very few of these cells. The majority of lymphocytes in tuberculoid granulomas were activated T cells as they formed rosettes with sheep erythrocytes, exhibited esterase dots in the cytoplasm and expressed HLA-DR antigens. A small proportion of the lymphocytes also formed rosettes with EAC. Most macrophages from both the granulomas were mature macrophages as they were esterase positive, did not exhibit peroxidase activity and expressed HLA-DR antigens. The macrophages did not possess C3 surface receptors.
Single cell suspension from dermal leprosy granulomas (10 tuberculoid and 10 lepromatous) was prepared and an assessment of the division and protein synthesis by the cells was made. The cells of tuberculoid granulomas showed a high incorporation of 3H-thymidine and 14C-leucine. On the contrary, the cells of the lepromatous granulomas exhibited poor division but their protein synthesis remained unimpaired. These observations suggest that the epithelioid cell granuloma of tuberculoid leprosy appears to be more active and secretory than the macrophage granuloma of lepromatous leprosy.
Two of the Mycobacterium leprae-specific assays--a serum antibody competition (for an epitope on 35-kDa protein) test (SACT) and an enzyme-linked immunosorbent assay (ELISA) for the disaccharide epitope of phenolic glycolipid-I (PGDS)--were comparatively evaluated as tools for monitoring chemotherapy in 125 lepromatous leprosy (LL/BL) patients. An adaptation of the SACT from a radioimmunoassay (RIA) to an ELISA procedure is also described. A moderate but statistically significant correlation was observed between the assays, although SACT appeared to be the more sensitive of the two. Levels of antibodies correlated better with the bacterial index (BI) than with the duration of treatment. However, wide individual variations in antibody levels (for a specific duration of treatment or BI) were seen in treated as well as untreated patients. Anti-PGDS antibody response of the IgG type was poorer than that of the IgM type and, apparently, it did not have a bearing on either treatment duration or the BI. Further studies will be needed to clarify whether the treated patients showing a negative (or low) BI and high antibody levels were harboring hidden foci of active infection, and whether treatment could safely be terminated in patients showing low values for both BI and antibody.
Five monoclonal antibodies (MAbs) directed against antigens of Mycobacterium leprae were tested for their ability to bind to components of tissue sections prepared from biopsies taken from patients with various forms of leprosy. Immunoperoxidase was the most successful marker system used, although immunofluorescence and alkaline phosphatase were also successful in certain cases. Positivity was high with all five antibodies successfully staining those sections containing a bacterial index of 3+ or more; sections with 0 bacterial counts also had areas staining positively with two of the MAbs. The positive staining in the tissues was confined to areas infiltrated by inflammatory cells; however it was not identifiable as being associated with individual bacteria. These findings suggest that immunostaining with specific monoclonal antibodies can help to identify leprosy in diagnostic samples in which acid-fast bacilli are not identifiable by standard histochemical means. Immunohistochemical techniques are likely to be valuable in studies of the distribution of M. leprae antigens and their association with individual tissue elements.
Palmar configurations of triradii and creases of 100 leprosy patients [50 lepromatous (BL/LL) and 50 tuberculoid (BT/LL)] were compared with those of 100 normal persons selected from families of these patients. The patterns of position of triradii were similar in controls and leprosy patients as such. But, the patterns in the two types of leprosy patients were different. As for palmar creases patterns, there was significant difference between those of controls and patients, double radial base crease occurring more often in patients. However, the differences between the two types of patients were not statistically significant.
Two infants, one 4 months old and the other 2 months old, having histologically confirmed indeterminate leprosy are reported. The route of infection, mode of transmission, and incubation period are discussed with reference to these two cases of infantile leprosy.
Clinico-bacteriological profile of 106 leprosy patients below 15 years of age was studied. Majority of the patients were males and fell in the 10-15 years age group (p less than 0.01). Nearly 89% had not received any prior treatment because of financial constraints. Seventy per cent gave a positive history of contact with adult patients who were mainly of the lepromatous variety (p less than 0.01). Skin lesions were present in 103 cases, mainly on the exposed areas and their number was found to increase significantly with advancing age (p less than 0.01). These lesions were hypopigmented patches in 71% of the children and erythematous in the rest. Cutaneous sensations were affected in most of the patients while nerve thickening was observed in 45. Positivity of the skin smears increased significantly as the number of skin lesions per patient increased (p less than 0.05). With advancing age, the disease moved from the tuberculoid end of the spectrum towards the lepromatous end (p less than 0.01).
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Regularity of DDS intake among 366 leprosy patients attending our out patient department voluntarily was assessed by urine spot test. It was found that only 54.6% of them had taken their last dose of drug within the previous three days. Those who kept their appointment showed better compliance than those who did not. Urinary DDS positivity was found to be unrelated to sex, occupation or the type of the disease. In the younger age group the compliance was low, as also among the patients coming from nearby places as compared to those who were residing in far off districts.
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Fifteen untreated leprosy patients were given rifampicin and dapsone for seven days, and then rifampicin, dapsone and clofazimine for seven days. Concentrations of rifampicin and dapsone were estimated in timed plasma specimens and in 24 h urine specimens on days 7 and 14. No significant differences in the pharmacokinetics of rifampicin and dapsone were observed between the two occasions of sampling.
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Single cell suspensions from granulomas of leprosy cases were prepared to enable an in vitro study on the characteristics of infiltrating cells. In all, biopsies from 21 untreated cases of tuberculoid leprosy and lepromatous leprosy were analyzed. The granulomas were found to contain lymphocytes and macrophages. The numbers of lymphocytes were higher in the suspensions of tuberculoid granulomas in comparison to lepromatous granulomas. A high percentage of lymphocytes from tuberculoid granulomas expressed OKT11 and Ia-like antigens, thereby indicating the presence of activated T cells. The proportion of Leu3a+ cells was greater in comparison to OKT8+ cells in these granulomas. In lepromatous granulomas, only a few positive lymphocytes expressing OKT11 or OKT8 antigens were observed. The ratio of Leu3a+:OKT8+ cells (2.79 +/- 0.61) was higher in the tuberculoid granulomas than in the lepromatous granulomas. Most macrophages from both types of granulomas expressed Ia-like antigens.
A study was made on the Langerhans' cells at the sites of contact sensitivity skin reactions in 45 untreated leprosy patients. The skin reaction was induced by 2,4-dinitrochlorobenzene (DNCB). Langerhans' cells were quantitated using OKT6 monoclonal antibody and indirect immunofluorescence. Clinically, the skin reaction in the tuberculoid patients was positive at 4, 24, and 48 hr, while the lepromatous patients failed to respond at any of the time intervals. Sequential histological analysis of the skin reaction showed predominantly mononuclear cell infiltrates around the blood vessels and neurovascular bundles in both the tuberculoid and lepromatous patients. Time kinetic assessment showed no difference in the numbers and distribution of OKT6+ epidermal Langerhans' cells at the site of the DNCB skin reactions among the tuberculoid and lepromatous patients. This, therefore, suggests that either there is a functional defect in Langerhans' cells or some other mechanism(s) such as a T-cell abnormality is responsible for the lack of clinical reaction in lepromatous patients.