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PubMed1988

Antigenic profile of ICRC bacilli with special reference to isolation of immunogenic subunit.

A vaccine containing ICRC bacilli induces persistent immune conversion in lepromatous leprosy (LL) patients and lepromin-negative healthy subjects, in association with upgrading of the tissue responses in the former. With an idea to isolate the immunogenic 'subunit(s)', antigenicity of ICRC sonicate and its fractions were tested, with reference to both B- and T-cell responses. A very high molecular weight glycolipoprotein, named PP-I with an apparent molecular weight of 1,000,000, has been isolated using gel permeation high performance liquid chromatography (HPLC). PP-I, which focussed as a single band at pH 5 in an LKB isoelectric focussing column, quantitatively interacted with 80% of the circulating antibodies in pooled LL sera, and also induced a late (3 weeks) Mitsuda-type skin response which shows excellent correlation with host immunity against Mycobacterium leprae. These observations suggest that PP-I is a complex bifunctional antigen containing epitopes for both B and T cells. The PP-I fraction of ICRC and a similar high molecular weight HPLC fraction of M. leprae produced a line of identity against rabbit anti-ICRC serum in Ouchterlony gel diffusion and gave comparable skin responses in healthy volunteers in leprosy endemic areas. The data indicate that the PP-I fractions from the two organisms are antigenically closely related. Preliminary studies in human volunteers showed that administration of PP-I of ICRC resulted in immune conversion in lepromin-negative healthy subjects. PP-I thus appears to be the appropriate immunogen that could be used in preparation of a 'subunit' antileprosy vaccine.

Antigens, Bacterial↗

PubMed1974

Leukocyte antimicrobial function in patients with leprosy.

Patients with lepromatous leprosy are unresponsive to lepromin skin-test material and possess defective lymphocyte function in vitro, including impaired mitogenesis in response to antigens of Mycobacterium leprae. It has been claimed that their macrophages cannot digest M. leprae in vitro; such a defect could explain both lepromin nonreactivity and impaired lymphocyte function on the basis of failure of the afferent limb of the immune response (i.e., defective macrophage "processing" of M. leprae). The present studies indicate that macrophages from patients with lepromatous and tuberculoid leprosy and from normal donors do not differ in their ability to digest heat-killed M. leprae in vitro, or in their ability to sustain the viability of M. leprae in tissue culture; that monocytes, macrophages, and polymorphonuclear leukocytes of leprosy patients and controls possess equivalent microbicidal activity against Listeria monocytogenes, Escherichia coli, Proteus vulgaris, Staphylococcus aureus, and Candida albicans; and that polymorphonuclear leukocytes from patients with lepromatous leprosy iodinate ingested bacteria normally. Whether the basic immune defect leading to the development of lepromatous leprosy resides in the lymphocyte or in the macrophage remains to be determined. However, the present study shows that phagocytic cells from patients with either principal form of leprosy function normally in a variety of sophisticated tests of antimicrobial function.

Candida albicans↗

PubMed1978

Leprosy in a chimpanzee. Morphology of the skin lesions and characterization of the organism.

Microscopic features of lepromatoid lesions in a chimpanzee inoculated with bovine leukemia virus consisted of diffuse dermal infiltrations of foamy histiocytes, except for a subepidermal "clear zone" devoid of infiltrating cells. Acid-fast organisms were in histiocytes, dermal nerves and a branch of the left radial nerve. The organisms were more intensely acid-fast with the Fite-Faraco than with the Ziehl-Neelsen stain. The organisms did not oxidize 3,4-dihydroxyphenylalanine (dopa) to indole but their acid-fast staining quality was removed by pretreatment with pyridine. Human leprosy patients, inoculated simultaneously with chimpanzee and human lepromin, reacted similarly to both lepromins. Organisms, when inoculated into mouse footpads, multipllied in 6 months to a quantity that was compatible with their identification as Mycobacterium leprae.

Animals↗

PubMedDate not supplied

[Assessment of the Mitsuda reaction in lepromatous patients inactive before and after immunotherapy].

In this study the immunopotentiator levamisole as well as a mixture of BCG/Mycobacterium leprae were investigated in inactive lepromatous leprosy patients by using the Mitsuda reaction as a parameter. Twenty lepromatous patients ten years ago classified as histologically negative for Mitsuda's test were divided into three groups: five patients that were only retested with Mitsuda antigen; eight patients that received oral levamisol and seven patients that received a mixture of alive BCG plus autoclaved M. leprae. The results indicated that: 1) the levamisole did not alter the reactivity to lepromin in any of the patients studied; 2) neither the changes in the reactivity to lepromin by using the mixture (3 cases) nor those that occurred spontaneously (3 cases) were clear. They properly reflected the natural variation of patients with some degree of resistance to Mycobacterium leprae.

Adjuvants, Immunologic↗

PubMed1999

An immunotherapeutic vaccine for multibacillary leprosy.

On January 30, 1998, a vaccine for leprosy based on Mycobacterium w (the code word under which this species hitherto unspecified was investigated) was launched for public use for therapeutic purposes. The vaccine has completed phase III immunotherapeutic trials as an adjunct to chemotherapy in urban and rural leprosy control centres and has received the authorization from the Drugs Controller of India for industrial manufacture. It will be made available by M/s Cadila Pharmaceuticals, Ahmedabad. As an adjunct to chemotherapy, the vaccine expediates bacterial clearance and accelerates clinical regression of lesions. It shortens significantly the period for release from treatment (RFT) of patients. It is effective in inducing a fall of bacterial index (BI) in multibacillary patients who are either nonresponders or slow responders to the standard multidrug therapy and who have persistent BI over long periods. An additional benefit of immunization with this vaccine is the conversion of >60% of LL, 71% of BL and 100% of BB patients from lepromin negativity to lepromin positivity status. A significant number of vaccinated patients showed histopathological upgrading and eventually attainment of a state of nonspecific infiltration without dermal granulomas. The vaccine was well tolerated and the incidence of Type 2 reactions and their severity was less in combined immuno cum chemotherapy group than in the group receiving only chemotherapy. This review describes the nature of the vaccine and the way it was developed.

Animals↗

PubMed1991

Localized borderline lepromatous leprosy.

A 48-year-old soldier presented with 3 small leprosy lesions localized over the flexor area of the forearm. There was no nerve thickening and clinically the lesions looked like borderline-tuberculoid leprosy. However, these lesions demonstrated a bacteriological index (BI) of 4+ while no acid-fast bacilli (AFB) could be demonstrated from any other site of the body. A lepromin test was negative. Histologically evidence of borderline lepromatous leprosy was conspicuous. The case was diagnosed as localized borderline lepromatous-leprosy and treated with multidrug therapy. After 1 year of treatment, the lesions regressed, a lepromin test was positive (5 mm) and the BI from the lesions fell to 1+.

Forearm↗

PubMed1993

Reversal reaction in multibacillary leprosy patients following MDT with and without immunotherapy with a candidate for an antileprosy vaccine, Mycobacterium w.

Immunotherapy with a candidate for an antileprosy vaccine, Mycobacterium w, was given in addition to standard multidrug therapy (MDT) to 53 multibacillary lepromin negative patients belonging to BB, BL and LL types of leprosy (vaccine group). An equal control group received MDT and injections of micronized starch as placebo. Both the vaccine and placebo were administered intradermally every 3 months. The patients were evaluated at determined intervals by clinical, bacteriological and histopathological parameters and lepromin testing. Reactional episodes were analysed with reference to incidence, onset, frequency and severity during and after release from treatment (RFT). Incidence of reversal reaction (RR) was marginally higher in the vaccine group (22.6% vaccine group vs 15% control group). All cases with a history of downgrading type 1 reaction developed RR during therapy. Most episodes occurred within the 1st year of the commencement of therapy--50% developing within 3 months. Late reversal reaction (after RFT) were observed in 3.8% of cases in both groups, and 50% of the reactors in the control group and 33% in the vaccine group had repeated reactional episodes. Incidence of neuritis associated with RR as well as isolated neuritis was similar in both groups.

Bacterial Vaccines↗

PubMed1998

Experimental leprosy in rhesus monkeys: transmission, susceptibility, clinical and immunological findings.

A total of 46 Rhesus monkeys (RM) was inoculated with Mycobacterium leprae (ML) and followed clinically and immunologically for extended periods. Twenty-one (45.7%) of the RM developed leprosy spanning the known leprosy spectrum, with six of 21 (28.6%) having disease in the borderline lepromatous to lepromatous area of the spectrum. RM with paucibacillary forms of leprosy produced predominantly IgG anti-phenolic glycolipid (PGL-I) antibodies and positive lepromin skin test and/or in vitro blastogenesis responses; IgM anti-PGL-I predominated in animals with BB-LL leprosy and correlated with negative immune responses to lepromin. IgG anti-PGL-I antibodies persisted in a number of RM for several years without histopathological evidence of leprosy, suggesting possible persisting subclinical infection. The data show that RM are a valuable model for the study of leprosy. Eleven of the 46 RM were inoculated with ML from sources infected with simian immunodeficiency virus (SIV), the monkey counterpart to the human immunodeficiency virus (HIV). The possible effect of SIV on the clinical outcome of ML infection could not be determined due to insufficient numbers of animals to yield statistically significant results.

Animals↗

PubMed1999

Disabilities in multibacillary leprosy following multidrug therapy with and without immunotherapy with Mycobacterium w antileprosy vaccine.

A vaccine based on autoclaved Mycobacterium w was administered, in addition to standard multidrug therapy (MDT), to 157 bacteriologically positive, lepromin-negative, multibacillary leprosy patients supported by a well-matched control group of 147 patients with similar type of disease who received a placebo injection in addition to MDT. The MDT was given for a minimum period of 2 years and continued until skin-smear negativity, while the vaccine/placebo was given at 3-month intervals up to a maximum of 8 doses in the initial 2 years. The overall incidence of type 1 and type 2 reactions and neuritis during treatment and follow up was nearly equal in the patients in the vaccine and placebo groups; the differences were not statistically significant. The occurrence of disabilities, such as anesthesia, trophic ulcers, claw hand and grade 3 deformities, were not different statistically in the vaccine and placebo groups, an observation valid both for deformities present at induction and for those which developed during the course of therapy and surveillance. A statistically significant difference was observed in the recovery of newly developed trophic ulcers; recovery was quicker in the vaccine group. The recovery rate for motor deformities was marginally higher in the vaccine group, although not significant (p = 0.068) statistically. There was a statistically significant reduction in the incidence of grade 3 deformities following MDT with and without immunotherapy. To conclude, the addition of vaccine to MDT did not precipitate neuritis or deformities over and above that encountered with MDT alone, although it did accelerate bacteriological clearance, histopathological upgrading, conversion to lepromin positivity, and clinical improvement.

Anti-Inflammatory Agents, Non-Steroidal↗

PubMed2000

Antileprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat-killed Mycobacterium leprae: immunologic observations.

Groups of rhesus monkeys were vaccinated and boosted with Mycobacterium bovis bacillus Calmette Guerin (BCG) or BCG plus low-dose (LD) or high-dose (HD) heat-killed M. leprae (HKML), or were unvaccinated. Prior to and following vaccination-boosting and subsequent M. leprae (ML) challenge, these and unvaccinated, unchallenged control monkeys were observed longitudinally for approximately 3 years. Vaccination with BCG plus HKML initially stimulated significant in vitro blood mononuclear cell blastogenic responses to lepromin, which returned to baseline post-boosting and post-live-ML-challenge, minimally reappearing significantly 2 years post-ML-challenge. Vaccination with BCG failed to stimulated positive blastogenic responses to lepromin before ML-challenge but small, marginally positive, intermittent responses were seen post-ML-challenge. Compared to the unvaccinated ML-challenged group, significant increases in the numbers of blood CD4+ and CD8+ T-cell subsets and an increased CD4+:CD8+ ratio were observed in both BCG plus HKML-vaccinated, ML-challenged groups, but not in the BCG-only-vaccinated, ML-challenged group. CD4+CD29+ and CD4+CD45RA+ subset numbers increased significantly over time in only the BCG plus LD HKML-vaccinated, ML-challenged group. Compared to unvaccinated, ML-challenged groups, vaccination with BCG or BCG plus HKML followed by ML-challenge produced lower IgM:IgG antiphenolic glycolipid-I (PGL-I) serum antibody ratios and protected rhesus monkeys from clinical leprosy, consistent with prior observations that low IgM:IgG anti-PGL-I responses correlated with resistance to and protection from leprosy.

Adjuvants, Immunologic↗

PubMed2000

Efficacy of single-dose ROM therapy plus low-dose convit vaccine as an adjuvant for treatment of paucibacillary leprosy patients with a single skin lesion.

The recent World Health Organization multicentric field study on the treatment of paucibacillary (PB) leprosy patients with single skin lesion (SSL) and a single dose of rifampin-ofloxacin-minocycline (ROM) brought new hope to those who are engaged in the eradication of leprosy from India. Being encouraged by the WHO report, we undertook the present hospital-based study and found that PB leprosy patients with SSL were morphologically and histopathologically heterogeneous. The histological spectrum of SSL ranged from indeterminate through tuberculoid (TT) to borderline tuberculoid (BT) leprosy, and most patients had active BT leprosy. Ninety new, untreated PB leprosy patients with SSL were included in the present study for comparative assessment of the efficacies of ROM and ROM plus Convit vaccine therapies. Children, pregnant women, lactating mothers and patients with any thickening of nerves were excluded. All patients were bacteriologically negative (skin-smear test) but lepromin reactive. The patients were divided into two groups after proper matching for morphological and histological status of SSL: a) The test group included 60 patients and the control group included 30 patients. The test group was given a single dose of ROM initially and two injections of low-dose Convit vaccine, one initially and the other at the end of 3 months. b) The control group was given only a single dose of ROM initially. Both groups were followed clinically every 2 weeks for 6 months and retested for histological, bacteriological and lepromin status at the end of 6 months. Thereafter, they were followed clinically every month for another 6 months. In the test group, the SSL resolved in 33.3%, regressed in 48.3%, and remained active in 18.3% of the patients, while the granuloma disappeared in 70% of the cases. Only one patient developed neuritis, and in another patient the disease relapsed on the eighth month. On the other hand, the SSL in the control patients resolved, regressed and remained active in 13.3%, 63.3% and 23.3% of the cases, respectively, while the granuloma disappeared in 53.3% of the cases. In the seven patients who remained active, the disease course was progressive, and two of them developed neuritis. The clinical outcome of the patients treated with ROM plus low-dose Convit vaccine was statistically superior to those treated with single-dose ROM therapy alone.

Adjuvants, Immunologic↗

PubMedDate not supplied

An attempt to produce experimental tuberculoid leprosy in the nine-banded armadillo.

In an attempt to produce experimental tuberculoid leprosy, three nine-banded armadillos, two with borderline tuberculoid lepromin reaction, and one with tuberculoid lepromin reaction, were chosen. They were injected subcutaneously in a four square centimetre area in the abdominal skin with saline suspension of 6.5 x 10(7) M. leprae. Induration of skin at the injected site appeared in 24 hours and persisted for 6 months in one and for 18 months in the other two animals. Histopathological examination of the infected site at 6 weeks, 18 and 20 months showed progressively decreasing granulomatous inflammation; but the cutaneous nerves were uninvolved. Autopsy examination of the three animals failed to show disseminated disease. Since there was no evidence of nerve involvement, experimental transmission of tuberculoid leprosy to armadillos could not be established in this study.

Animals↗

PubMed1989

Indeterminate leprosy in a population survey and in the subsequent follow-ups of children in Burma.

The paper discusses various aspects of indeterminate (I) leprosy in the initial survey undertaken in the Burma BCG trial (69,242 inhabitants), and in the annual examinations of 28,220 children in the trial followed up over periods of five to eight years. Age-specific rates in the initial mass survey are presented. In total 1914 cases were detected (6.2% I, 76% T, 16% L and 1.8% B). Among the children in the BCG trial 768 cases were detected: 255 of them had the I form and their proportion (33%) was much higher than in the population survey. Of these 255 I cases only 4.3% had a negative or doubtful lepromin reaction. Two-thirds of these 255 cases evolved to the tuberculoid pole in less than one year. No L cases appeared in the trial population until ten and eleven years after the start of the trial. It is concluded that whereas a high proportion of indeterminate cases regress spontaneously or evolve towards the T pole, the indeterminate lepromin negative cases are important in the dynamics of the disease, because a proportion of them, if untreated, tend to envolve towards the L form. This stresses the importance of detection and treatment of I cases at an early stage in an effective strategy for controlling leprosy.

Adolescent↗

PubMed1984

Relapse rates among nonlepromatous patients released from control.

A total of 1701 nonlepromatous patients treated with dapsone monotherapy for at least 4 1/2 years and released from control were followed up and examined for evidence of relapse. They contributed a total of 5254 person years of risk, and there were 51 relapses (3%), giving an overall relapse rate of 9.7/1000 person years of risk. This paper examines the effect of various factors on the risk of relapses, such as age, sex, and classification of the disease; duration and regularity of treatment; percentage of attendance; deformity grade; number of patches and lepromin status. Some of the factors studied, such as age, sex, classification, percentage of attendance, and the number of patches in association with the lepromin status, were found to significantly influence the risk of relapse in these patients. Therapy in nonlepromatous leprosy is discussed in light of these findings.

Adolescent↗

PubMedDate not supplied

Induction of delayed-type hypersensitivity by ICRC anti-leprosy vaccine and the adoptive transfer of cell-mediated immunity in mice.

The immunogenic potency of anti-leprosy ICRC vaccine preparations was determined by foot pad enlargement in mice. Female Balb/c mice were sensitized by I.D. route with irradiated, heat-killed or live antigens of 2 strains (C-44 and C-75) of ICRC bacilli and tested against lepromin and ICRC-antigens. ICRC antigens sensitized mice against lepromin; similarly, BCG and M. leprae sensitized mice, which showed good cross reactivity with ICRC antigens. A mixture of irradiated ICRC bacilli and live BCG was not more immunogenic to mice than its single constituents. The immunity induced by ICRC antigens could be adoptively transferred to syngeneic recipients by sensitized spleen cells.

Animals↗

PubMed1984

Oral zinc as an adjunct to dapsone in lepromatous leprosy.

Oral zinc was tried in 15 cases of multibacillary leprosy as an immunostimulant in addition to conventional antileprosy drugs. Results were compared with those in ten similar cases treated with dapsone alone. Cases treated with zinc showed faster clinical improvement, regrowth of eyebrows, rapid fall in the bacterial index (BI) and in the bacterial index in the granuloma (BIG), early and greater influx in lymphocytes in the granuloma, and neovascularization. Upgrading occurred in 6 out of 15 patients taking zinc, but in only 1 out of 10 patients in the control group. Five out of six patients who showed upgrading in the treated group became lepromin positive. Only one patient (BL) showed lepromin conversion in the control group.

Adult↗

PubMed1977

Recent advances in microbiology in leprosy.

The recent advances in microbiology of leprosy are reviewed. Till now the leprosy bacillus had not been cultivated in laboratory media; the recent claims of success have not been confirmed. There has been a breakthrough in the experimental transmission of leprosy to experimental animals--the white mice, the immune depressed white mice, and the nine-banded armadillo. Apart from providing definite proof for the causative relationship of the bacillus discovered by Hansen and the disease leprosy, the experimental transmission to animals have considerably advances our knowledge about the disease. The mouse has provided a suitable model for screening of antileprosy drugs, detecting development of drug resistance, ascertaining viability of the bacilli and determining the genuineness of a supposed culture of the leprosy bacillus. The armadillo has provided a model for making basic studies of the disease as it occurs in man. Further, the generalised infection in armadillo has provided large amounts of leprosy bacilli for preparing standardised lepromin, for preparing a specific skin-testing antigen containing the active protein fraction of the bacillus, and a step towards the production of a vaccine against leprosy. It is interesting to note that the fact that the protein fraction of the bacillus was responsible for the positive lepromin reaction was discovered by Dharmendra far back in 1941.

Animals↗

PubMed1992

Association of HLA antigens with differential responsiveness to Mycobacterium w vaccine in multibacillary leprosy patients.

Leprosy patients undergoing phase II trials in two hospitals of New Delhi, India, were HLA typed to see the association of HLA with differential responsiveness to Mycobacterium w vaccine. The vaccine comprises an atypical, nonpathogenic mycobacterium, Mycobacterium w, which has cross-reactive antigens with M. leprae. Multibacillary patients who are lepromin negative are vaccinated at an interval of 3 months. Considerable improvement is evident in the patients in terms of a decline in bacterial indices and histopathological and immunological upgrading. But all the patients do not respond to the vaccine in the same manner; some are slow responders, while others are good responders. HLA-A28 and DQw3 (DQw8 + 9) were found to be associated with slow responsiveness, while DQw1 and DQw7 were found to be associated with a more rapid responsiveness to the M. w vaccine. However, these associations were not significant after P correction for the number of antigens tested for each locus except for HLA-DQw3 (DQw8 and DQw9) and DQw7. DQw7, a new defined split of HLA-DQw3, seems to be associated with the responsiveness to M. w vaccine.

Cross Reactions↗