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PubMed1977

Comparative study of skin reactions in leprosy patients to M. leprae-lepromin and to ICRC-in, an antigen from cultivable acid-fast bacilli from M. leprae isolated from lepromatous nodules.

Skin test antigens (Dharmendra type) were prepared from fresh M. leprae (lepromin) and from a culture of strain C-44 ICRC bacilli (ICRCin) grown 'in vitro' from M. leprae isolate from lepromatous nodules. Comparative study of skin reactivity to lepromin and ICRCin--both "early" and "late" reactions in 76 leprosy patients was conducted. In 29 lepromatous (LL) cases, 25 exhibited totally negative reaction at the end of third week. In tuberculoid (TT) 22 and 23 out of 31 were positive (greater than 4.5 mm) at 3 weeks to lepromin and ICRCin respectively. In the 16 BB group, the reactions were comparable in the same patient. The cellular reaction in tuberculoid cases consisted of lymphocytic infiltration, epitheloid giant cells and Langhan type cells and indistinguishable from each other. These data with characteristic total lack of reaction in 25/29 lepromatous leprosy cases and identical cellular reaction in TT patients, provide strong evidence that ICRC bacillus strain C-44 is antigenically identical with M. leprae.

Antigens, Bacterial↗

PubMed1979

Nature and familial character of lepromin sensitivity in 27 families and their siblings.

A group of 27 families consisting of 176 individuals had been investigated for the lepromin sensitivity (with Dharmendra antigen). The families were arranged under group A (9 families) in whom either of the parents or both were suffering from Lepromatous type of leprosy, group B numbering 4 families, of whom either of the parents or both were suffering from non-lepromatous type of leprosy and group C comprising 14 families where none of the parents was suffering from leprosy but some of the each family had the disease in their siblings. The present study points towards the possible genetic influence on lepromin sensitivity but at times may be influenced by the environmental factors. However the study does not permit to reach any valid conclusions; further elaborate investigations alone could prove the useful role of genetic influence in the propagation of lepromin sensitivity to the subsequent sibs.

Adolescent↗

PubMed1980

Conversion of reactions to leprolin and lepromin in patients with lepromatous leprosy by the transfer factor.

Conversion of leprolin and early lepromin reactions was achieved by two injections of transfer factor made of lymphocytes from lepromin-positive tuberculoid leprosy patients. However, the late reaction to lepromin remained unchanged. The importance of the degree of sensitivity of the cell donor was demonstrated, and a booster dose was also found to be useful. The feasibility of using transfer factor in treatment of lepromatous leprosy is briefly discussed.

Adult↗

PubMed1980

The granulomatous response of leprosy patients to lepromin and killed BCG.

The volunteers studied included 6 tuberculoid and 11 lepromatous leprosy patients (Ridley and Jopling). Each received intradermally 0.1 ml of lepromin (Wade method) and 0.1 ml of autoclaved BCG (grown on Dubos medium) at 2 separate sites on the left upper arm. Observations were made at weekly intervals, and biopsy specimens were taken at the fourth wk. Within 3 to 4 wk all 6 tuberculoid patients responded to lepromin with typical granulomas (5 of which measured 5 mm or more in diameter) composed of epithelioid cells, lymphocytes and giant cells. In contrast, 8 of the 11 lepromatous patients remained negative; the other 3 developed tiny papules less than 3 mm in diameter which disappeared after 2 wk. Biopsy specimens taken from injection sities at the fourth wk showed essentially normal histology. All 1 patients developed typical granulomas to heat-killed BCG (non were less than 5 mm in diameter and most were larger) consisting of epithelioid cells, many lymphocytes and a few giant cells. The reaction in tuberculoid leprosy was stronger than that in lepromatous leprosy and the reaction to BCG in tuberculoid leprosy was stronger than that to lepromin, but the differences were not very remarkable.

Adult↗

PubMed1980

A study of lepromin reaction in child contacts of leprosy patients and non contact children in Chengalpattu District Tamil Nadu.

In this study early lepromin reaction has been used to detect delayed hypersensitivity reaction to a prior infection with M. leprae, recent studies indicate that a certain number of contacts who develop sub-clinical infection remain anergic to the lepromin test. This is indicated by the Fluorescent leprosy antibody absorption (FLA-ABS) test of Abe (1979) which detects antibodies in the sera of infected individuals. Correlation of this serological test with lepromin response in contacts is in progress in the CJIL, Agra. These anergic contacts are likely to be susceptible to leprosy, a finding which would help in taking prophylactic measures.

Adolescent↗

PubMed1980

Assessment of diluted Dharmendra lepromin.

Twenty-one patients of lepromatous and tuberculoid type were tested with standard, 1:2 and 1:4 diluted Dharmendra lepromin. It was found that correlation exists between the degree of induration and the number of bacilli in the lepromin. Uniform and reproducible results were obtained with Dharmendra antigen containing 40 million bacilli/ml. This is of special significance in enhancing the present meagre supplies of lepromin.

Adult↗

PubMed1978

Leucocyte migration inhibition with Armadillo lepromin in human lymphocytes of healthy donors.

Armadillo lepromin activates human lymphocytes from normal donors to release leucocyte inhibiting factor. The above activity was expressed optimally when leucocytes were incubated with lepromin at 37 degrees C, and only partially when incubation was carried out at 30 degrees C or at 35 degrees C. The possible mechanism of the in vitro production of lymphokine from lymphocytes stimulated by armadillo lepromin is discussed.

Animals↗

PubMed1987

Lepromin-induced suppressor cells in lepromatous leprosy.

The presence or absence of suppressor cells in leprosy patients was investigated by measuring peripheral blood lepromin-induced suppression of the Con A response. Significant suppressor activity was measured in 15 of 15 untreated or recently treated patients with lepromatous leprosy and 3 of 5 patients with borderline lepromatous leprosy. In addition, in patients with lepromatous leprosy, suppressor cell activity was found in 10 of 14 patients that had been under treatment for more than 1 year but in only 2 of 27 patients who had active or thalidomide controlled erythema nodosum leprosum. Suppression was observed in only 5 of 29 tuberculoid leprosy patients, 1 of 6 patient contacts, and 0 of 11 normal controls. The differences between the lepromatous or borderline lepromatous group as compared with the tuberculoid group were statistically significant (P less than 0.001). Our findings confirm the presence of lepromin-triggered suppressor cells in the peripheral blood of patients with lepromatous leprosy. These suppressor cells may contribute to the selective unresponsiveness of lepromatous patients to the antigens of Mycobacterium leprae.

Concanavalin A↗

PubMed1979

Mitsuda-type lepromin reactions as a measure of host resistance in Mycobacterium lepraemurium infection.

The footpad reaction to autoclaved whole Mycobacterium lepraemurium organisms (MLM lepromin) in high-resistance (C57BL) and low-resistance (BALB/c) mice was studied. Infected C57BL mice gave a prolonged footpad response persisting for 4 weeks after skin testing with high and low doses of lepromin. This was accompanied by mononuclear cell infiltration. Uninfected C57BL mice gave no response. The majority of infected BALB/c mice gave no increase in footpad thickness. However, a high proportion of infected and control BALB/c mice tested with the high dose showed mononuclear cell infiltration which resembled that in C57BL mice. The low dose caused little infiltration in infected or control BALB/c mice. The course of infection in the two strains was different. Dissemination of organisms from the infected footpad was minimal in C57BL mice 5 months after infection. In BALB/c mice, dissemination to the draining lymph node and to some extent to the liver had occurred by 5 months. The draining lymph node of BALB/c mice showed histological evidence of local antibody formation, which uas not found in C57BL mice. On the basis of these findings, it was possible to fit murine leprosy in these two strains into a classification similar to that used for human leprosy.

Animals↗

PubMed1987

Immunohistologic comparison between armadillo-derived leprosin and standard lepromin skin tests in leprosy patients.

A comparison was made on the in situ immunological characteristics of dermal infiltrates of early (24-hour) and late (3-4 weeks) skin reactions in leprosy patients. The skin reactions were induced by armadillo-derived leprosin coupled to liposomes and standard Dharmendra lepromin. Most lymphocytes in the early reaction induced by both antigens were positive for Leu 4, Leu 3a, OKT8 and Ia like antigens indicating thereby the presence of activated T cells. The ratio of Leu 3a/OKT8+ cells were similar. In the late reaction elicited by both antigens, the lymphocytes in the granulomas were predominantly activated T lymphocytes expressing Leu 4, Leu 3a, OKT8 and Ia like antigens. Leu 3a+ cells were scattered diffusely amidst the epithelioid cells. In contrast, the OKT8+ cells were present mainly as 'a ring' in the periphery of the granuloma. A similar ratio of Leu 3a+/OKT8+ cells was observed in these granulomas. Macrophages in the granulomas expressed Ia like antigens. These observations indicate that the immunological characteristics of dermal infiltrates in the skin reaction induced by armadillo-derived leprosin coupled to liposomes and standard Dharmendra lepromin appear to be identical.

Animals↗

PubMed2004

Susceptibility to leprosy may be conditioned by an interaction between the NRAMP1 promoter polymorphisms and the lepromin response.

Controversial results have been achieved by attempting to associate the NRAMP1 gene with Mycobacterium leprae susceptibility as well as with the Mitsuda reaction, which represents a specific immune response to M. leprae. This study evaluated this association as well as the interaction of the polymorphism (GT)(n) in the promoter region of the NRAMP1 gene with a specific immune response to M. leprae measured by the intradermal Mitsuda test in leprosy patients and in non-consanguineous household contacts. The study aimed to evaluate the association of this gene polymorphism with resistance or susceptibility to the disease, and/or with clinical forms of the disease, in a population in an endemic area served by the State Reference Center in Sanitary Dermatology and Leprosy, Federal University of Uberlandia, MG, Brazil. Leprosy patients (90) were diagnosed according to Ridley and Jopling criteria and they grouped into multibacillary (MB) and paucibacillary (PB) patients. The control group consisted of 61 non-consanguineous contacts. NRAMP1 promoter genotypes were obtained through amplification by the polymerase chain reaction (PCR) followed by the detection through the low ionic-strength single strand conformational polymorphism (LIS-SSCP) electrophoretic technique. There were no significant differences in the allelic and genotypic frequencies for alleles 2, 3, and 4 in relation to the Mitsuda test among patients and household contacts, nor between those with MB and PB forms. However, individuals with a negative lepromin response associated with genotypes 22 and 23 presented a 7- and 8-fold greater chance of developing leprosy, respectively. Therefore, the NRAMP1 gene promoter polymorphism exhibited an interaction with the lepromin response, suggesting that allele 2 of the NRAMP1 promoter is an independent genetic factor that predisposes cells to enable pathogen survival, probably due to its low efficiency in iron transport. However, establishment of the infection and disease development may be conditioned by other immunological and genetic factors.

Alleles↗

PubMed1975

Specificity of the 48-hour reaction to Mitsuda antigen. Use of a soluble antigen from human and armadillo lepromin.

Two antigens were tested and compared in relation to the 48-h Fernandez reaction. They were obtained from standard human and from standard armadillo lepromin. All the tests were negative in patients with lepromatous leprosy and highly positive in those with tuberculoid leprosy and in lepromin-positive contacts. There was total agreement in all tests done with the two types of antigen. The antigenic component has the following basic properties: it precipitates with 80% saturated ammonium sulfate; it is not destroyed by autoclaving or by treatment with 0.4% phenol; it is non-dialysable; and it is destroyed by treatment with trypsin.

Animals↗

PubMed1990

ELISA & lepromin skin tests in household contacts of leprosy patients.

A total of 438 household contacts of leprosy patients and 228 non-contacts were studied for anti-PGL-I antibodies and cell mediated immune response to Mycobacterium leprae using ELISA and lepromin tests. The contacts showed relatively higher positivity to ELISA (29.4%) and lepromin (70.5%) as compared to non-contacts who showed 9.6 and 57 per cent positivity. In contacts as well as non-contacts the seropositivity was not found to be influenced by age and sex. However, seropositivity was higher (39.7%) in the contacts of patients with multibacillary leprosy as compared to contacts (27.7%) of patients with paucibacillary leprosy.

Adolescent↗

PubMed1983

Effect of dapsone (DDS) on T-cells and their response to PHA and lepromin in tuberculoid leprosy patients.

T and B cell counts and their response to PHA and lepromin were studied in 46 patients of polar tuberculoid leprosy (TT), divided into two age and sex matched groups. Twenty five normal healthy controls were similarly investigated. While one group consisted of 26 untreated TT patients, the other included 20 individuals who had taken dapsone, 50 mg daily, for 12 to 14 months. There was significant decrease in the number of T-lymphocytes and their response to PHA and lepromin was lower in the treated, as compared to the untreated patients and normal persons. No significant change was found in the number of B-lymphocytes in any of the groups.

B-Lymphocytes↗

PubMed1983

Variable lepromin response to Mycobacterium leprae in resistant armadillos.

Eight armadillos resistant to the infection of Mycobacterium leprae were lepromin tested. The tissue response was tuberculoid in 5, borderline in 2, and lepromatous in 1, thus showing a wide variation. It is seen that although cell-mediated immunity as evidenced by a tuberculoid granulomatous response to killed M. leprae is associated with resistance to the disease, there may be other yet unknown factors which protect armadillos from the infection. Lepromin responses were recognized histologically even at a low dose of 10(3) organisms, and the response increased with the dose up to 10(7) organisms. The tissue reaction to live organisms was the same as that to killed ones, and autoclaving of M. leprae produced no change in the tissue response to the antigens of M. leprae.

Animals↗

PubMed1982

[Comparative study of the lepromin reaction performed on medical students and on a control group].

The lepromin test was studied in 64 medical students and in a control group consisted of 40 patients from the orthopedic clinic. Both groups matched with respect to age and sex, but differed with regard to socio-economical conditions. The Fernandez reaction showed low frequency of positive results in both groups: 10% in the first group and 5,6% in the second one. This difference was not significant. The Mitsuda reaction showed high frequency of positive results: 98,4% in the medical students and 85,0% in the control group. This difference was significant. The authors discuss the factors that might have caused the high positivity of the lepromin test among the medical students.

Adolescent↗

PubMed1998

Why relapse occurs in PB leprosy patients after adequate MDT despite they are Mitsuda reactive: lessons form Convit's experiment on bacteria-clearing capacity of lepromin-induced granuloma.

It is amazing how after years of scientific research and therapeutic progress many simple and basic questions about protective immunity against Mycobacterium leprae remain unanswered. Although the World Health Organization (WHO) has recommended short-term multidrug therapy (WHO/MDT) for the treatment of paucibacillary (PB) leprosy patients, from time to time several workers from different parts of the globe have reported inadequate clinical responses in a few tuberculoid and indeterminate leprosy patients following adequate WHO/MDT despite the fact that they are Mitsuda responsive. A few borderline tuberculoid patients harbor acid-fast bacilli (AFB) in their nerves for many years even though they become clinically inactive following MDT, a fact which has been ignored by many leprosy field workers. Keeping these patients in mind, we have attempted to investigate the cause of the persistence of AFB in PB cases and have looked into the question of why Mitsuda positivity in tuberculoid and indeterminate leprosy patients, as well as in healthy contacts, is not invariably a guarantee for protectivity against the leprosy bacilli. We have: a) analyzed the histological features of lepromin-induced granulomas, b) studied the bacteria-clearing capacity of the macrophages within such granulomas, and c) studied the in vitro leukocyte migration inhibition factor released by the blood leukocytes of these subjects when M. leprae sonicates have been used as an elicitor. The results of these three tests in the three groups of subjects have been compared and led us to conclude that the bacteria-clearing capacity of the macrophages within lepromin-induced granuloma (positive CCB test) may be taken as an indicator of the capability of elimination of leprosy bacilli and protective immunity against the disease. This important macrophage function is not invariably present in all tuberculoid and indeterminate leprosy patients or in all contacts even though they are Mitsuda responsive and are able to show a positive leukocyte migration inhibition (LMI) test. It is likely but not certain that this deficit of the macrophage is genetically predetermined and persists after completion of short-term WHO/MDT. Thus, after discontinuation of treatment slow-growing, persisting M. leprae multiply within macrophages leading to relapse.

Adolescent↗

PubMed2002

Anti-leprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat-killed Mycobacterium leprae: lepromin skin test results.

Groups of rhesus monkeys (RM) were vaccinated and boosted with living Mycobacterium bovis Bacillus Calmette-Guerin (BCG) or BCG + low dose (LD) heat-killed Mycobacterium leprae (HKML) or high dose (HD) HKML or were unvaccinated. Animals vaccinated with BCG + LD and HD HKML were lepromin skin tested 2 weeks after boosting. All groups were lepromin tested 37 and 46 months after challenge with live M. leprae. Fernandez (72 h) and Mitsuda (28 day) responses were recorded. Ten of 10 rhesus monkeys in each of the two BCG + HKML-vaccinated groups significantly converted to strong positive Fernandez status within 2 weeks of boosting, compared to one of six positives in the unvaccinated unchallenged normal control group. Both BCG + HKML groups were significantly protected from clinical leprosy. Six of 10 in each of the two BCG + HKML groups significantly converted to Mitsuda positivity within 2 weeks of boosting compared to zero of six in the normal control group. The sizes of the Mitsuda responses were larger in the LD group than the HD HKML vaccinated/boosted group, suggesting suppression by vaccination with higher doses of HKML in combination with BCG. Fernandez responses were negative in normal RM as well as in the unvaccinated, ML-challenged group and the BCG-vaccinated, ML-challenged group at 37 or 46 months after ML inoculation, although the BCG-vaccinated group was significantly protected from leprosy and the unvaccinated group was not. In contrast, at 37 months the Fernandez reaction was positive in the BCG plus LD and the BCG plus HD HKML-vaccinated groups, both of which were significantly protected from clinical leprosy. By 46 months, the Fernandez responses were below significance in all groups. Thus, Fernandez reactivity is not a reliable correlate to protection from experimental leprosy in RM. Mitsuda responses became strongly positive in all four ML-challenged groups by 37 months and remained strongly positive at 46 months after ML inoculation, suggesting that strong Mitsuda reactivity reflects responses to living ML. BCG or BCG + LD or HD HKML vaccination/boosting of RM produced significant clinical protection from leprosy and there was a good correlation between protection from LL forms of leprosy and positive Mitsuda skin test responses after challenge with live ML. Positive Mitsuda responses were generated in essentially all individuals after challenge with live ML, and this response was primed by prior vaccination/boosting with BCG + HKML as shown by conversion to positivity 2 weeks after boosting. The data show that resistance to clinical leprosy is reflected by Mitsuda responses in ML-exposed RM, similar to results from human studies, and confirm the suitability of RM as a model for leprosy vaccine studies.

Animals↗