Expression of protein gene product 9.5 in lepromatous eyes showing ciliary body nerve damage and a "dying back" phenomenon in the posterior ciliary nerves.

BACKGROUND/AIM: Peripheral nerve destruction is the hallmark of leprosy. Ocular complications form a substantial part of the clinical manifestations but histopathology of nerve destruction within ocular structures has not been shown satisfactorily. The role of protein gene product (PGP) 9.5 in identifying nerve destruction in the ciliary body and posterior ciliary nerves of lepromatous eyes is shown. METHODS: Serial sections from two lepromatous eyes and two non-lepromatous eyes were stained with PGP 9.5. Histopathological comparison was done on the expression of the PGP 9.5 stain in nerves within the ciliary body, posterior ciliary nerves adjacent to the optic nerve, and nerves tracking through the sclera. RESULTS: In non-lepromatous eyes, PGP 9.5 was expressed in nerves within the ciliary body, the nerves within the sclera, and posterior ciliary nerves adjacent to the optic nerve. In lepromatous eyes no PGP 9.5 was expressed, signifying nerve destruction. CONCLUSIONS: Nerve destruction in lepromatous eyes has been confirmed histopathologically by the absence of or patchy staining with PGP 9.5. Nerve destruction in the ciliary body can extend to the posterior ciliary nerves by an ascending axonopathy. This "dying back" phenomenon is akin to the "glove and stocking" anaesthesia found in lepromatous leprosy.

Are viable Mycobacterium leprae present in lepromatous patients after completion of 12 months' and 24 months' multi-drug therapy?

A study was carried out to determine whether or not viable bacilli persist in MB patients treated with 12-month and 24-month multidrug therapy (MDT). In the first group, 60 untreated lepromatous patients who had an initial average bacterial index (BI) of 3+ or more were enrolled. At the completion of 12 months of MDT, skin biopsies were obtained and M. leprae concentrate was inoculated into the footpads of five thymectomized and irradiated (T900r) mice. Rees technique was used for the mouse footpad (MFP) experiment. Harvesting was done it the 6th, 9th and 12th months. Out of the 60 biopsies inoculated into mouse footpads to check the viability of bacilli, 2 skin biopsies (3.3%) showed significant growth and 10 (16%) showed equivocal growth. 27 patients also had nerve biopsies tested for growth in MFP studies. None of the inoculated nerve biopsies showed significant multiplication in the MFP experiments. However, 4 biopsies (14%) showed equivocal growth. In the second group, 20 patients had skin biopsies and 10 had nerve biopsies done at the end of 24 doses of MDT in order to test the viability of bacilli; none of the skin or nerve biopsies from these patients showed any growth. This study showed that M. leprae present in the tissues after 24 doses of MDT are not viable and the drug schedule of 24 doses is adequate to treat leprosy patients, irrespective of their BI. However, a small (3.3%) percentage of the patients with a high BI harbour viable bacteria in the skin after 12 doses of treatment. Since a large majority of the patients (38 patients) who had a high initial BI responded well to the treatment, it is important to find out the reason for the lack of response in two patients. One of the reasons may be the presence of drug-resistant strains. It is important to follow up on these patients for a longer duration to ascertain whether or not they would relapse.

Drug resistant-Mycobacterium leprae--results of mouse footpad studies from a laboratory in south India.

Out of 265 biopsies of leprosy patients received at the Experimental Pathology Laboratory of Schieffelin Leprosy Research and Training Centre from 1987 to 1997 for evaluating resistant strains of M. leprae, using the mouse footpad technique, 49 showed resistant strains of M leprae to varying concentrations of dapsone, rifampicin and clofazimine. 23 (47%) of these were from a control area. With 369 skin-smear positive multibacillary (MB) patients as the risk group (denominator), 23 (6.23%) were resistant to one or more drugs. 18 (4.88%) had dapsone resistance, 5 (1.36%) were resistant to rifampicin and 9 (2.44%) had resistance to low concentrations of clofazimine (0.0001%). Out of the 23 biopsies with drug resistance from the control area, primary dapsone resistance was seen in 7 (30%) biopsies and secondary dapsone resistance in 11 (48%). Primary rifampicin resistance was seen in 4 (17.4%) patients, secondary rifampicin resistance in 1 (4.35%) and primary clofazimine resistance in 7 (30%). 3 (13%) of the strains showed secondary clofazimine resistance. One biopsy had resistant strains to all the three drugs. In a control area where properly supervised effective multidrug therapy (MDT) was regularly administered over the years, the emergence of drug resistance is negligible. It may not be the case if the content, duration and regularity of the drug regimen were not satisfactory. Aware of the possible shortcomings in mass administration of MDT, it is emphasized that mouse footpad studies on drug resistance should be made available at least in endemic areas where the incidence of the disease has not changed despite good MDT coverage in order to monitor the emergence of drug resistance. Research into molecular biological identification of drug resistant-M.leprae should be intensified. These steps would help to institute timely measures to check the spread of any drug-resistant organisms in the community.

Drug resistant-Mycobacterium leprae--results of mouse footpad studies from a laboratory in south India

Out of 265 biopsies of leprosy patients received at the Experimental Pathology Laboratory of Schieffelin Leprosy Research and Training Centre from 1987 to 1997 for evaluating resistant strains of M. leprae, using the mouse footpad technique, 49 showed resistant strains of M leprae to varying concentrations of dapsone, rifampicin and clofazimine. 23 (47%) of these were from a control area. With 369 skin-smear positive multibacillary (MB) patients as the risk group (denominator), 23 (6.23%) were resistant to one or more drugs. 18 (4.88%) had dapsone resistance, 5 (1.36%) were resistant to rifampicin and 9 (2.44%) had resistance to low concentrations of clofazimine (0.0001%). Out of the 23 biopsies with drug resistance from the control area, primary dapsone resistance was seen in 7 (30%) biopsies and secondary dapsone resistance in 11 (48%). Primary rifampicin resistance was seen in 4 (17.4%) patients, secondary rifampicin resistance in 1 (4.35%) and primary clofazimine resistance in 7 (30%). 3 (13%) of the strains showed secondary clofazimine resistance. One biopsy had resistant strains to all the three drugs. In a control area where properly supervised effective multidrug therapy (MDT) was regularly administered over the years, the emergence of drug resistance is negligible. It may not be the case if the content, duration and regularity of the drug regimen were not satisfactory. Aware of the possible shortcomings in mass administration of MDT, it is emphasized that mouse footpad studies on drug resistance should be made available at least in endemic areas where the incidence of the disease has not changed despite good MDT coverage in order to monitor the emergence of drug resistance. Research into molecular biological identification of drug resistant-M.leprae should be intensified. These steps would help to institute timely measures to check the spread of any drug-resistant organisms in the community.

Epithelioid granuloma in the iris of a lepromatous leprosy patient; an unusual finding.

This case report depicts a case of histopathologically confirmed polar lepromatous (LL) leprosy with a bacterial index of 4+. He experienced recurrent episodes of erythema nodosum leprosum (ENL) in the first 5 years after diagnosis. Skin smears became negative after 6 years of dapsone monotherapy and have remained negative since that time. At 23 years after diagnosis, the patient had developed cataracts and underwent intracapsular cataract extractions with broad-based iridectomies. In one of the iris specimens, histopathologic examination revealed a focal granuloma composed of epithelioid cells. Subsequently a lepromin skin test showed a positive Mitsuda reaction with a borderline tuberculoid histopathology. This clearly illustrates the immunological upgrading of a polar lepromatous patient, perceived first in the iris tissue.

Pathology of a lepromatous eye.

Histopathological examination of an enucleated eye from a lepromatous leprosy patient showed the cornea, ciliary body, and part of the choroid to be infiltrated by macrophages filled with Mycobacterium leprae. The walls of blood vessels in the sclera, ciliary body and the anterior choroid demonstrated the presence of M. leprae, giving credence to the blood-borne entry of M. leprae into the eye. Unlike the eyes of experimental animals infected with M. leprae, histopathological study of this eye from a lepromatous leprosy patient demonstrated that M. leprae, although demonstrable in the anterior choroid, could not be found in the posterior parts of the eye, substantiating the claim that leprosy does not affect the posterior parts of the eye directly.

Lepromatous lymphadenopathy and concomitant tuberculous axillary lymphadenitis with sinus. A case report.

A 25-year-old male patient with florid lepromatous leprosy presented with right axillary lymphadenopathy and a discharging sinus. He also had scabies with chronic right otitis media. Histopathological examination of the lymph node revealed lepromatous lymphadenitis coexisting with tuberculosis. This unusual combination of two different clinical entities is recorded in this case report.

Nasal mucosa and skin of smear-positive leprosy patients after 24 months of fixed duration MDT: histopathological and microbiological study.

The skin and nasal mucosa of 10 lepromatous leprosy patients who had completed 24 doses of fixed duration multidrug therapy (MDT) but who continued to be skin-smear positive for acid-fast bacilli (AFB) were examined histopathologically. The nasal mucosa showed granuloma fractions that exceeded those seen in the skin specimens, signifying that activity in this region subsides much more gradually than the activity in the skin. Mouse foot pad studies done using T900r mice with an inoculum from the nasal mucosa biopsy specimens of these patients did not demonstrate any growth of Mycobacterium leprae, indicating that these bacilli were not viable. A skin specimen from one patient grew significant amounts of bacteria in the T900r mouse foot pad. These results show that 2 years of treatment with MDT would prevent dissemination of M. leprae from the nasal mucosa and, therefore, should preclude further transmission of the disease. It also indicates that viable bacteria might persist in the skin of patients, especially those with an initial bacterial index of > or = 4+ who have completed 24 doses of regular MDT. Therefore, a more cautious approach to administering only 12 doses of MDT to highly positive multibacillary patients is suggested.

Histopathological activity in paucibacillary leprosy patients after ROM therapy.

Histopathological activity was assessed in the skin tissue of 13 skin-smear negative, borderline tuberculoid leprosy patients after administration of a single dose of ROM (rifampin 600 mg, ofloxacin 400 mg and minocycline 100 mg) therapy. Biopsies taken just before therapy showed Mycobacterium leprae to be present in eight cases. After 6 months, only three showed granulomatous lesions and others showed only resolving or inactive lesions. Acid-fast bacilli (AFB) persisted in the nerves of three cases. At the end of 12 months, granulomas persisted in 2 out of 13 (15%) patients. No bacilli, however, were detected in any of them at the end of 12 months. This study demonstrated that 12 months after a single dose of ROM granuloma cleared in 85% of the patients and AFB were absent in all of them.

Clinical and histopathological activity in paucibacillary leprosy patients after fixed-duration multidrug therapy.

In 37 clinically-diagnosed borderline-tuberculoid (BT) leprosy patients skin biopsies were done prior to starting multidrug therapy (MDT) and at the end of 6 months therapy. Clinical and histopathological activity, graded as active, resolving and inactive, were studied at the end of 6 months of MDT. Of the 37 clinically-diagnosed BT patients 24 could be confirmed by histopathology as having BT leprosy, while the other 13 biopsies showed features of indeterminate (I) leprosy. After 6 months of MDT, out of the 24 histopathologically-confirmed BT patients, 4 (17%) showed clinical activity and 8 (33%) showed histopathological activity. Of the 13 histopathologically-diagnosed indeterminate cases all were clinically inactive but histological activity persisted in 3 cases (23%). Out of the 37 clinically-diagnosed BT patients 3 showed both clinical and histopathological activity at the end of MDT. This study emphasizes the importance of performing histopathological examinations on leprosy patients undergoing research studies for the confirmation of diagnosis and for proper classification of the disease. The histopathological activity that outlasts the MDT may be due to the bacillary fragments that persist but clinical activity coupled with histopathological activity seen in 3 patients at the end of 6 months may foreshadow a relapse and these patients and others like them need to be followed up for longer durations.

Pathology of iris in leprosy.

AIM: The histopathological features of the iris in leprosy were studied by light microscopy. METHOD: Formalin fixed and paraffin embedded iris tissue excised during cataract surgery from 20 leprosy patients were sectioned and studied with haematoxylin and eosin stain and modified Fite Faraco's stain for acid fast bacilli (AFB). RESULTS: Chronic inflammatory reactions were seen in the iris of 11 patients, seven of whom did not have any clinically demonstrable evidence of iridocyclitis. Smooth muscle disruption and destruction were seen in two specimens. AFB were found in the iris tissue of a polar lepromatous patient whose skin smears were negative for AFB and who had completed the WHO recommended antileprosy multidrug therapy (MDT). CONCLUSION: Histopathology discloses far more silent chronic iridocyclitis in leprosy patients than are diagnosed clinically. AFB can persist in the iris tissue even after completion of MDT. Smooth muscle disruption and destruction, a cause of the miotic pupil in leprosy has been conclusively demonstrated histopathologically.

Diffuse alopecia of the scalp in borderline-lepromatous leprosy in an Indian patient.

A case of borderline-lepromatous leprosy exhibiting alopecia of the scalp along with lepromatous lymphadenitis of suboccipital lymphnode is reported. To our knowledge generalized leprous alopecia of the scalp with lepromatous lymphadenitis of the suboccipital node is a rare occurrence in female Indian patients.