Botting, J. The History of Thalidomide. Drug News Perspect. 15(9) (2002) 604–611.

The first paper describing the pharmacological actions of thalidomide was published in 1956. The drug, then designated as K17, was thought to have sedative effects superior to those of comparator drugs and was thought to be virtually nontoxic. Only 2 years after thalidomide’s launch as Contergan in Germany, its alleged lack of toxicity came into question, with reports of the drug causing numerous side effects. Shortly thereafter, thalidomide was connected with an epidemic of horrific deformities in children whose mothers had taken the drug during pregnancy. This disaster brought on by thalidomide’s teratogenic effects was responsible for the institution of some regulatory bodies, such as the United Kingdom’s Committee on the Safety of Drugs, and for the strengthening of others, such as the U.S. Food and Drug Administration. An objective examination of published papers and contemporary accounts confirms that the preclinical tests on thalidomide were superficial, and there is no doubt that it was never administered to pregnant animals prior to its use in patients. Within a short time after its withdrawal from the market due to its suspected association with fetal abnormalities, the drug was shown to produce fetal toxicity in laboratory animals. Had there been more extensive testing on laboratory animals before the drug was launched, the disaster could have been avoided.—Author’s Abstract

Dolev, E. [The thalidomide story: a new chapter] Harefuah 142(3) (2003) 186– 187, 239, 238. [Article in Hebrew]

Thalidomide, a medication the use of which was banned about forty years ago due to its teratogenic effect, has returned into use due to its anti-angiogenic and immunomodulatory virtues. It is used now as a novel treatment in several malignant diseases as well as in the treatment of various inflammatory conditions. Recently it has been found that thalidomide may antagonize corneal blood vessels growth which is a late results of exposure to mustard gas. The story of thalidomide, which is very unusual, may serve as the snake symbol in medicine, to remind us about the possible double face of every medication, of which every physician should be aware.—Author’s Abstract

Fusegawa, H., Wang, B. H., Sakurai, K., Nagasawa, K., Okauchi, M., and Nagakura, K. Outbreak of tuberculosis in a 2000-year-old Chinese population. Kansenshogaku Zasshi. 77(3) (2003) 146–149.

The molecular identification of Mycobacterium tuberculosis DNA in ancient human remains has been achieved mainly in mummies with macroscopic changes but not in the skeletons without bone tuberculosis. Using polymerase chain reaction studies, we identified mycobacterial DNA in 2000-year-old human skeletons without pathological changes. Our findings suggest that these people suffered from an outbreak of tuberculosis. Molecular examinations for mycobacterial DNA in the bone marrow of skeletons may contribute to the clarification of ancient diseases in old human populations.—Authors’ Abstract

Gomez i Prat, J. and de Souza, S. M. Prehistoric tuberculosis in America: adding comments to a literature review. Mem. Inst. Oswaldo Cruz. 98 (Suppl 1) (2003) 151–159.

Tuberculosis is a prehistoric American human disease. This paper reviews the literature and discusses hypotheses for origins and epidemiological patterns of prehistoric tuberculosis. From the last decades, 24 papers about prehistoric tuberculosis were published and 133 cases were reviewed. In South America most are isolated case studies, contrary to North America where more skeletal series were analyzed. Disease was usually located at the deserts of Chile and Peru, Central Plains in USA, and Lake Ontario in Canada. Skeletal remains represent most of the cases, but 16 mummies have also been described. Thirty individuals had lung disease, 19 of them diagnosed by the ribs. More then 100 individuals had osseous tuberculosis and 26 also had it in other organs. As today, transmission of the infection and establishment of the disease were favored by cultural and life-style changes such as sedentarization, crowding, undernutrition, use of dark and insulated houses, and by the frequency of interpersonal contacts. The papers confirm that despite previous perceptions, tuberculosis seems to have occurred in America for millennia. It only had epidemiological expression when special conditions favored its expansion. Occurring as epidemic bursts or low endemic disease, it had differential impact on groups or social segments in America for at least two millennia.—Authors’ Abstract

Mori, S., Kato, S., Yokoyama, H., Tanaka, U., and Kaneda, S. [The history of Yunosawa village and the policy of leprosy of Japan. II] Nihon Hansenbyo Gakkai Zasshi. 72(1) (2003) 27–44. [Article in Japanese]

There was a village which was called Yunosawa, lots of leprosy patients lived, existed from 1887 to 1941, Kusatu town, Gunma Prefecture, Japan. It was the only place continued securing self-government to the last as area was free from the isolation policy of State in prewar days there. The aim of this study will make clear the dynamism of “The protection from the tension of the society of leprosy patient currently persecuted” to “The defense of the society from the leprosy patient who is a source of infection.” In this study, explained the history of the Yunosawa village and the shift of the policy of leprosy by State had relation to the village. In addition, the effort of residents and Christianity persons’ activity are drawn in this paper. Moreover also drew what is desired how it is going to live under adverse circumstances, and showed worth of free medical-treatment area here.—Authors’ Abstract

Nwosu, M. C. and Nwosu, S. N. Leprosy control in the post leprosaria abolition years in Nigeria: reasons for default and irregular attendance at treatment centres. West Afr. J. Med. 21(3) (2002) 188–191.

A questionnaire was administered to all patients with leprosy seen at the four leprosy clinics in Anambra State in a face to face interview. The questions covered, among other items, the clinic attendance behaviour and the single most important reason, monthly, for absenteeism in the preceding year. The total and individual frequencies of the reasons for absenteeism were determined for the various behavioural subgroups. The differences in frequencies and associations were analysed. Values of p <0.05 were considered as significant. The results showed that 27 females and 26 males were interviewed. 39.6% of the patients were irregular attenders 735% were defaulters. Attendance at meetings (p <.001); work at home (p <0.01) fear/shame/indignation (p <0.05); no confidence in treatment (p <0.025) were significant reasons for absenteeism among irregular attenders inter-current illnesses as reasons for absenteeism did not differ significantly between regular and irregular attendees. The association between clinic attendance behaviour and lesion location (revealed Vs concealed) was not statistically significant (X(2)0.3). The findings in this study indicate that in the post leprosaria abolition years, default and irregular clinic attendance by patients with leprosy are numerically large and may compound the problems of control programmes, and thus negate the realization of the global goal of intercepting leprosy transmission.—Authors’ Abstract

Sato, H. and Narita, M. Politics of leprosy segregation in Japan: the emergence, transformation and abolition of the patient segregation policy. Soc. Sci. Med. 56(12) (2003) 2529–2539.

The segregation of leprosy patients, a practice introduced early in the 20th century, was maintained in Japan after World War II. It locked in the viability of subsequent policy choices, and patients’ isolation was sustained long after it was proven to be scientifically unnecessary. For leprologists and leprosarium directors, there was little opportunity to conceptualize and test the epidemiological validity and effectiveness of outpatient services as alternatives to the existing policy, since most of the patients were already hospitalized. Since leprosy was no longer a threat to the general public, bureaucratic officials, as well as legislators, lacked strong incentives to reformulate the overall policy. Within the Ministry of Health and Welfare, daily tasks were largely transferred to the section for leprosarium management, and the search for other options lost importance. For patients, long institutionalization elevated their dependency on life in leprosaria. These conditions must be emphasized as policy legacies, the results of past policies, since they posed obstacles to effective policy innovation in accordance with changing scientific knowledge. To make policies reflective of scientific knowledge, it is essential to understand and foresee the effect of policy legacy, when introducing and appraising public health policies.—Authors’ Abstract

Wolf, R. and Wolf, D. A tattooed butterfly as a vector of atypical Mycobacteria. J. Am. Acad. Dermatol. 48(5 Suppl) (2003) S73–S74.

We report the first case of cutaneous inoculation of atypical Mycobacteria secondary to tattooing. The diagnosis of atypical Mycobacteria infection of the skin was confirmed on the basis of the clinical and histologic appearance, the detection of acid-fast bacilli on Ziehl-Neelsen stain, and positive polymerase chain reaction. The medical complications of tattooing, which are manifold, are briefly summarized. This case emphasizes the need for federal regulation of tattooing, which is an invasive procedures associated with infectious and noninfectious complications.—Authors’ Abstract

Aberg, J. A., Williams, P. L., Liu, T., Lederman, H. M., Hafner, R., Torriani, F. J., Lennox, J. L., Dube, M. P., MacGregor, R. R., Currier, J. S., and AIDS Clinical Trial Group 393 Study Team. A study of discontinuing maintenance therapy in human immunodeficiency virus-infected subjects with disseminated Mycobacterium avium complex: AIDS. J. Infect. Dis. 187(8)(2003) 1346.

The present nonrandomized prospective study evaluated whether antimycobacterial therapy for disseminated Mycobacterium avium complex (MAC) could be withdrawn from human immunodeficiency virus-infected subjects who experienced immunologic recovery while receiving highly active antiretroviral therapy (HAART). Eligible subjects had received macrolide-based therapy for least 12 months, were asymptomatic for MAC, had received HAART for at least 16 weeks, and had CD4+ T cell counts >100 cells/microL. Forty-eight subjects were enrolled, with a median CD4+ T cell count of 240 cells/microL at the time of discontinuation of MAC therapy. Forty-seven subjects remained MAC free, whereas 1 subject developed localized MAC osteomyelitis. The median duration of follow-up while not receiving therapy was 77 weeks, and the incidence of MAC infection was 1.44/100 person-years (95% confidence interval, 0.04–8.01). Withdrawal of anti-MAC therapy appears to be safe in patients who have been treated with a macrolide-based regimen for at least 1 year and have an immunologic response on HAART.—Authors’ Abstract

Brown-Elliott, B. A., Crist, C. J., Mann, L. B., Wilson, R. W., and Wallace, R. J. Jr. In Vitro Activity of Linezolid against Slowly Growing Nontuberculous Mycobacteria. Antimicrob. Agents Chemother. 47(5) (2003) 1736–1738.

MICs of linezolid in broth microdilutions were tested against 341 slowly growing nontuberculous mycobacteria (NTM) belonging to 15 species. The proposed linezolid susceptibility MICs for all Mycobacterium marinum, Mycobacterium szulgai, Mycobacterium kansasii, Mycobacterium malmoense, and Mycobacterium xenopi isolates and for 90% of Mycobacterium gordonae and Mycobacterium triplex isolates were 8 micro g/ml. Linezolid has excellent therapeutic potential against most species of NTM.—Authors’ Abstract

Das, A. R., Dattagupta, N., Sridhar, C. N., and Wu, W. K. A novel thiocationic liposomal formulation of antisense oligonucleotides with activity against Mycobacterium tuberculosis. Scand. J. Infect. Dis. 35(3) (2003) 168–174.

This study describes the development of a novel thiocationic (OBEHYTOP) lipid-based formulation of phosphorothioate antisense oligonucleotides (PAOs) showing inhibitory activity against Mycobacterium tuberculosis (mTB) as measured by an in vitro BACTEC 460TB assay. PAOs were designed based on sequences complementary to essential regions of the mycobacterial genome from published nucleic acid databases in GenBank. These included the superoxide dismutase sod A gene (TBS3), catalase-peroxidase katG gene (TBK1, TBK10), RNA polymerase beta-subunit rpo B gene (TBR5) and diaminopimelate decarboxylase lys A gene (TBL5). The effect of PAOs (TBS3, K1, K10, R5 and L5) alone on mTB was not significant compared with the no-drug control over a period of exposure of 150 hr (ranges of –11.8 to +23.58% at 72 hr; 15.26 to +25.82% at 96 hr and –5.51 to +24.00% at 150 hr). Liposomal formulations (10:5:2 OBEHYTOP : oleic acid:vitamin D3) of PAOs resulted in statistically significant (p <0.05 in all cases) inhibition (ranges of –51.45 to –63.00% at 72 hr; –56.75 to –67.96% at 96 hr; –51.45 to –60.26% at 150 hr) compared with PAOs alone, thiocationic liposomal control and liposomal components. Positive controls of streptomycin and isoniazid used at their minimum inhibitory concentrations of 2.00 and 0.10 microM, respectively, resulted in average % inhibition values of –94% and –97.36%, respectively, indicating that these thiocationic lipid-formulated PAOs showed inhibitory activity directed against mTB in vitro.—Authors’ Abstract

Dhople, A. M. and Namba, K. Activities of sitafloxacin (DU-6859a), either singly or in combination with rifampin, against Mycobacterium ulcerans infection in mice. J. Chemother. 15(1) (2003) 47–52.

Efficacy of a new fluoroquinolone, sitafloxacin (DU-6859a), against Mycobacterium ulcerans was evaluated in vivo using the mouse footpad system. The growth of M. ulcerans in mouse footpads was completely inhibited when mice were fed with sitafloxacin at a dose of 25 mg/kg body weight per day; on the other hand similar effects were observed with ofloxacin at a dose of 100 mg/kg body weight per day. In the presence of rifampin, the above dose of sitafloxacin could be reduced by 75% to achieve total inhibition, while, under similar circumstances, the dose of ofloxacin could be reduced by only 50%. Either used singly or in combination with rifampin, the effects of sitafloxacin were bactericidal. The results suggest that sitafloxacin should be evaluated as a chemotherapeutic agent against M. ulcerans infection.—Authors’ Abstract

Dhople, A. M. and Namba, K. In-vitro activity of sitafloxacin (DU-6859a), either singly or in combination with rifampin analogs, against Mycobacterium leprae. J. Infect. Chemother. 9(1) (2003) 12–15.

The in-vitro antibacterial activity of sitafloxacin (DU-6859a) against Mycobacterium leprae was evaluated and compared with those of ofloxacin, levofloxacin, and ciprofloxacin. Two biochemical indicators (intracellular ATP and uptake of [(3)H]-thymidine) were used to measure the in-vitro growth of M. leprae in Dhople-Hanks (DH) medium. Sitafloxacin was found to be more potent than the other three commonly used fluoroquinolones, with the minimum inhibitory concentration (MIC) against M. leprae being 0.1875 microg/ml and the action being bactericidal. The MIC’s of ofloxacin, levofloxacin, and ciprofloxacin were 1.5, 0.75, and 3.0 microg/ml, respectively. Similar to ofloxacin and levofloxacin, sitafloxacin also exhibited synergistic activity when combined with either rifabutin or KRM-1648, but not with rifampin. Thus, further studies on the incorporation of sitafloxacin in multidrug therapy regimens in treating leprosy patients are suggested.—Authors’ Abstract

Gordon, J. N. and Goggin, P. M. Thalidomide and its derivatives: emerging from the wilderness. Postgrad. Med. J. 79(929) (2003) 127–132.

Forty years on from its worldwide withdrawal, thalidomide is currently undergoing a remarkable renaissance as a novel and powerful immunomodulatory agent. Over the last decade it has been found to be active in a wide variety of inflammatory and malignant disorders where conventional therapies have failed. Recently, considerable progress has been made in elucidating its complex mechanisms of action, which include both anticytokine and antiangiogenic properties. However, in addition to its well known teratogenic potential, it has a significant side effect profile that leads to cessation of treatment in up to 30% of subjects. In response to this, two new classes of potentially safer and non-teratogenic derivatives have recently been developed. This review summarises the biological effects, therapeutic applications, safety profile, and future potential of thalidomide and its derivatives.—Authors’ Abstract

Hall, V. C., El-Azhary, R. A., Bouwhuis, S., and Rajkumar, S. V. Dermatologic side effects of thalidomide in patients with multiple myeloma. J. Am. Acad. Dermatol. 48(4) (2003) 548–552.

BACKGROUND: Thalidomide, an antiangiogenic agent, was approved by the Food and Drug Administration in 1998 for the treatment of erythema nodosum leprosum. Although its teratogenic and neurologic side effects are well known, its dermatologic side effects continue to be defined. OBJECTIVE: We report the dermatologic side effects in 87 patients with multiple myeloma enrolled in a comparative, open-label, clinical trial treated with thalidomide alone (50 patients) or thalidomide and dexamethasone (37 patients). METHOD: We reviewed the records of all patients enrolled in the clinical trial. The frequency, type, severity, and time of onset of all skin eruptions that were temporally related to thalidomide treatment were recorded. RESULTS: Minor to moderate skin eruptions were noted in 46% of patients taking thalidomide alone and in 43% of those taking thalidomide and dexamethasone. These included morbilliform, seborrheic, maculopapular, or nonspecific dermatitis. Severe skin reactions (exfoliative erythroderma, erythema multiforme, and toxic epidermal necrolysis) that required hospitalization and withdrawal of thalidomide developed in 3 patients receiving thalidomide and dexamethasone. CONCLUSION: The prevalence of dermatologic side effects of thalidomide appear to be higher than previously reported. Although in most patients they were minor, in a few patients they were quite severe, particularly when given in conjunction with dexamethasone for newly diagnosed myeloma. Further studies are needed to verify the extent of the interaction between thalidomide and dexamethasone in this group of patients.—Authors’ Abstract

Haslett, P. A., Hanekom, W. A., Muller, G., and Kaplan, G. Thalidomide and a thalidomide analogue drug costimulate virus-specific CD8+ T cells in vitro. J. Infect. Dis. 187(6) (2003) 946–955.

CD8(+) T cell immunity is critical for protection from viral disease, such as that caused by the human immunodeficiency virus (HIV) or cytomegalovirus (CMV). It is therefore important to identify therapies that can boost antiviral immunity. The recent finding that thalidomide acts as a T cell costimulator suggested that this drug may boost antiviral CD8(+) T cell responses. In this in vitro study, in a human autologous CD8(+) T cell/dendritic cell (DC) coculture system, thalidomide and a potent thalidomide analogue were shown to enhance virus-specific CD8(+) T cell cytokine production and cytotoxic activity. The drug-enhanced antiviral activity was noted in cells from both healthy donors and persons chronically coinfected with HIV and CMV. This stimulatory effect was directed at CD8(+) T cells, and not DCs. These results suggest an application for thalidomide and the thalidomide analogue as a novel immune-adjuvant therapy in chronic viral infections.—Authors’ Abstract

Hegarty, A., Hodgson, T., and Porter, S. Thalidomide for the treatment of recalcitrant oral Crohn’s disease and orofacial granulomatosis. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 95(5) (2003) 576–585.

It has been suggested that thalidomide may be effective in the management of Crohn’s disease, including the associated oral lesions. We detail the clinical response to low-dose thalidomide of 5 patients with clinical features of orofacial granulomatosis or oral Crohn’s disease recalcitrant to recognized immunosuppressant therapy. All patients had clinical resolution of their symptoms and signs. Transient somnolence was the only reported adverse effect. Remission was maintained by extending the period between thalidomide doses. Thalidomide should be considered an effective therapy for the short-term treatment of severe orofacial granulomatosis in appropriately counseled patients.—Authors’ Abstract

Matthews, S. J. and McCoy, C. Thalidomide: a review of approved and investigational uses. Clin. Ther. 25(2) (2003) 342–395.

BACKGROUND: Thalidomide is best known as a major teratogen that caused birth defects in up to 12,000 children in the 1960s. More recently, this agent has been approved by the US Food and Drug Administration for the treatment of erythema nodosum leprosum (ENL) through a restricted-use program. Its immunomodulatory, anti-inflammatory, and antiangiogenic properties are currently under study in a number of clinical conditions. OBJECTIVE: This article reviews the pharmacology of thalidomide; its approved and off-label uses in dermatologic, oncologic, and gastrointestinal conditions; and adverse events associated with its use. METHODS: Relevant articles were identified through searches of MEDLINE (1966–June 2002), International Pharmaceutical Abstracts (1970–June 2002), and EMBASE (1990– June 2002). Search terms included but were not limited to thalidomide, pharmacokinetics, pharmacology, therapeutic use, and teratogenicity, as well as terms for specific disease states and adverse events. Further publications were identified from the reference lists of the reviewed articles. Abstracts of recent symposia were obtained from the American Society of Clinical Oncology Web site. RESULTS: Thalidomide is thought to exert its therapeutic effect through the modulation of cytokines, particularly tumor necrosis factor-alpha. In addition to its approved indication for ENL, thalidomide has been studied in various other conditions, including graft-versus-host disease, discoid lupus erythematosus, sarcoidosis, relapsed/refractory multiple myeloma, Waldenstrom’s macroglobulinemia, myelodysplastic syndromes, acute myeloid leukemia, myelofibrosis with myeloid metaplasia, renal cell carcinoma, malignant gliomas, prostate cancer, Kaposi’s sarcoma, colorectal carcinoma, oral aphthous ulcers, Behcet’s disease, Crohn’s disease, and HIV/AIDS-associated wasting. Adverse events most frequently associated with its use include somnolence, constipation, rash, peripheral neuropathy, and thromboembolism. CONCLUSIONS: Use of thalidomide is limited by toxicity, limited efficacy data, and restricted access. Evidence of its efficacy in conditions other than ENL awaits the results of controlled clinical trials.—Authors’ Abstract

Mlambo, G. and Sigola, L. B. Rifampicin and dexamethasone have similar effects on macrophage phagocytosis of zymosan, but differ in their effects on nitrite and TNF-alpha production. Int. Immunopharmacol. 3(4) (2003) 513–522.

The antibiotic rifampicin is used extensively in the treatment of mycobacterial and other infections. It has previously been suggested that rifampicin binds to and activates the glucocorticoid receptor potentially leading to pharmacological glucocorticoid-like effects such as host immunosuppression (Calleja, et al.). This study compares the effects of rifampicin with the synthetic glucocorticoid dexamethasone, on macrophage phagocytosis of zymosan particles and production of nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha) by splenocytes or macrophages. Rifampicin and dexamethasone, partially suppressed zymosan phagocytosis by macrophages, respectively, and both effects were ameliorated by the glucocorticoid receptor antagonist RU486. In other experiments, rifampicin had no effects on NO responses; however, dexamethasone inhibited NO in an RU486-sensitive manner. At high doses, rifampicin moderately suppressed TNF-alpha while dexamethasone inhibited it in a dose-dependent manner, which was ameliorated by the presence of RU486. These findings suggest that rifampicin has differential immunomodulatory effects on these innate immune mechanisms.—Authors’ Abstract

Okafor, M. C. Thalidomide for erythema nodosum leprosum and other applications. Pharmacotherapy. 23(4) (2003) 481–493.

Thalidomide, administered as a sedative and antiemetic decades ago, was considered responsible for numerous devastating cases of birth defects and consequently was banned from markets worldwide. However, the drug remarkably has resurfaced with promise of immunomodulatory benefit in a wide array of immunologic disorders for which available treatments were limited. It is approved by the Food and Drug Administration for erythema nodosum leprosum (ENL). Although the relative paucity of leprosy and ENL worldwide may perceivably limit interest in and knowledge about thalidomide, increasing numbers of new and potential uses expand its applicability widely beyond ENL. Thalidomide, an inhibitor of tumor necrosis factor a, is the best known agent for short-term treatment of ENL skin manifestations, as well as postremission maintenance therapy to prevent recurrence. For this indication, it is effective as monotherapy and as part of combination therapy with corticosteroids. Studies of thalidomide in chronic graft-versus-host disease showed benefit in children and adults as treatment, but not as prophylaxis. The agent has been administered successfully for treatment of cachexia related to cancer, tuberculosis, and human immunodeficiency virus infection, although evidence of efficacy is inconclusive. Thalidomide monotherapy effectively induced objective response in trials in patients with both newly diagnosed and advanced or refractory multiple myeloma. Combination therapy with thalidomide and corticosteroids was also effective in these patients, as well as in treatment of aphthous and genital ulcers. Limited evidence supports the drug’s benefit in treatment of Kaposi’s sarcoma. Other thalidomide applications include Crohn’s disease, rheumatoid arthritis, and multiple sclerosis. Somnolence, constipation, and rash were the most frequently cited adverse effects in studies, but thalidomide-induced neuropathy and idiopathic thromboembolism were critical causes for drug discontinuation. Thalidomide is still contraindicated in pregnant women, women of childbearing age, and sexually active men not using contraception. Clinicians should be conversant with thalidomide in ENL (its primary application) in the natural course of leprosy, as well as in the agent’s other applications.—Author’s Abstract

Pannikar, V. The Return of Thalidomide: New Uses and Renewed Concerns. WHO Pharmaceuticals Newsletter, No. 2, 2003 (available at www.who-umc.org)

Today, a large number of thalidomide babies continue to be born each year possibly reflecting regulatory insufficiency and widespread use under inadequate supervision. In Brazil, which has more than 1000 registered thalidomide victims, the last officially known case was born in 1995. There is evidence that second generation babies with similar deformities are being born to thalidomide victims. In the US, Celgene Corporation has had FDA approval to market the drug since 1998 for the cutaneous manifestations of moderate to severe erythema nodosum leprosum. In Europe, the US company Pharmion Corp and French rival Laphal have both secured orphan drug status for thalidomide and have applied to market the drug as a therapy for multiple myeloma and for ENL in the EU. The EU is currently holding discussions on the re-launch of thalidomide. Whatever the outcome of the EU discussions, it cannot be over emphasized that any potential benefit with thalidomide must be balanced with the known toxicity and the accompanying ethical and legal constraints on its use. Experience has shown that it is virtually impossible to develop and implement a fool-proof surveillance mechanism to combat misuse of thalidomide.—Author’s Abstract

Pukrittayakamee, S., Prakongpan, S., Wanwimolruk, S., Clemens, R., Looareesuwan, S., and White, N. J. Adverse effect of rifampin on quinine efficacy in uncomplicated falciparum malaria. Antimicrob. Agents Chemother. 47(5) (2003) 1509–1513.

The effects of adding rifampin to quinine were assessed in adults with uncomplicated falciparum malaria. Patients were randomized to receive oral quinine either alone (N = 30) or in combination with rifampin (N = 29). Although parasite clearance times were shorter in the quinine-rifampin-treated patients (mean ± standard deviation, 70 ± 21 versus 82 ± 18 hr; p = 0.023), recrudescence rates were five times higher (N = 15 of 23; 65%) than those obtained with quinine alone (N = 3 of 25; 12%), p <0.001. Patients receiving rifampin had significantly greater conversion of quinine to 3-hydroxyquinine and consequently considerably lower concentrations of quinine in their plasma after the second day of treatment (median area under the plasma drug concentration-time curve from day zero to day 7 = 11.7 versus 47.5 micro g/ml. day, p <0.001). Rifampin significantly increases the metabolic clearance of quinine and thereby reduces cure rates. Rifampin should not be combined with quinine for the treatment of malaria, and the doses of quinine should probably be increased in patients who are already receiving rifampin treatment.—Authors’ Abstract

Schafer, P. H., Gandhi, A. K., Loveland, M. A., Chen, R. S., Man, H. W., Schnetkamp, P. P., Wolbring, G., Govinda, S., Corral, L. G., Payvandi, F., Muller, G. W., and Stirling, D. I. Enhancement of Cytokine Production and AP-1 Transcriptional Activity in T Cells by Thalidomide-Related Immunomodulatory Drugs. J. Pharmacol. Exp. Ther. 305(3) (2003) 1222–1232.

CC-4047 (ACTIMID(TM)) and CC-5013 (REVIMID(TM)) belong to a class of thalidomide analogs, collectively known as the immunomodulatory drugs (IMiDs(TM)), that are currently being assessed in the treatment of patients with multiple myeloma (MM) and other cancers. IMiDs potently enhance T cell and natural killer (NK) cell responses and inhibit TNF-alpha, IL-1beta, and IL-12 production from LPS-stimulated PBMC. However, the molecular mechanism of action for these compounds is unknown. Herein we report on the ability of the IMiDs to upregulate production of IL-2 from activated human CD4(+) and CD8(+) peripheral blood T cells, production of IL-2 and IFN-gamma from Th1-type cells, and production of IL-5 and IL-10 from Th2-type cells. Elevation of IL-2 production from Jurkat T cells was observed as early as 6 hours post-stimulation and correlated with an increase in IL-2 promoter activity that was dependent upon the proximal but not the distal AP-1 binding site. The IMiDs enhanced AP-1-driven transcriptional activity 2 to 4-fold after 6 hours of T cell stimulation, and their relative potencies for AP-1 activation correlated with their potencies for increased IL-2 production in Jurkat T cells and in CD4(+) or CD8(+) human peripheral blood T cells. The most potent of these IMiDs, CC-4047, had no effect on NFAT transcriptional activity, calcium signaling, or phosphorylation of ERK1/2, JNK1/2, p38 MAPK, or c-Jun/Jun D in Jurkat T cells. These data suggest that IMiDs increase T cell cytokine production by potentiating AP-1 transcriptional activity.—Authors’ Abstract

Valerio, G., Bracciale, P., Manisco, V., Quitadamo, M., Legari, G., and Bellanova, S. Long-term tolerance and effectiveness of moxifloxacin therapy for tuberculosis: preliminary results. J. Chemother. 15(1) (2003) 66–70.

The resurgence of tuberculosis is a major problem. Increasing multiple resistance to current drugs used for therapy, non-compliance to therapy or co-morbidity are challenging problems that do not allow use of standard therapy in all patients. Quinolones are claimed to be active drugs in TB infection. Moxifloxacin shows the highest intracellular concentration in vitro and in experimental animals, but long-term tolerability is unknown. Our aim was to observe in compliant patients, not eligible for standard therapy, the effect of 6 months of therapy with moxifloxacin, isoniazid and rifampin. Nineteen patients, a control group, were observed for the same period under therapy with streptomycin, pirazinamide, rifampin, isoniazid. The patients were affected by indolent miliary pattern and concomitant lymphoma or leukemia in 3 cases; rare nodular involvement with genitourinary diseases in 3 others; segmental to lobar involvement in 4 others with concomitant multidrug resistance, bone localization, hepatitis. The control group was more uniform and showed segmental to lobar nodular involvement with pleuritis in 3 patients, together with hepatitis in 3. Monthly checks of blood gas analysis, chest X-ray, functional testing, serum titers of antibodies against antigen 60, sputum slides and complete chemical analysis were performed. A follow-up visit was performed 1 month after therapy. Patients under moxifloxacin therapy experienced no toxicity, almost complete sterilization and remission of the disease. Sterilization was obtained in 15 days. Patients under standard therapy also had a good clinical outcome, although therapy was delayed in 3 cases because of increased transaminases within the first 15 days of therapy. Moxifloxacin seems to be well tolerated and combination therapy including moxifloxacin for TB seems to be as active as the standard therapy in patients with complex illness.—Authors’ Abstract

Bari, M. M. Surgical reconstruction of leprotic foot drop. Mymensingh Med. J. 12(1) (2003) 11–12.

We have operated 152 patients for correction of foot-drop due to leprosy from March 1992 to July 1999. The method used was circumtibial transfer of the tibial is posterior to the tendons of extensor hellucis longus and the extensor digitorum longus in the foot together with lengthening of the Achilles tendon. The results were satisfactory in 135 of these cases as judged by adequate restoration of heel—toe gait and of active dorsiflexion. The follow up period ranged from 6 months to 8 years. Inadequate post-operative physiotherapy was the reason for unsatisfactory results in seventeen cases.—Author’s Abstract

Davis, S. V., Shenoi, S. D., Balachandran, C., and Pai, S. B. A fatal case of erythema necroticans. Indian J. Lepr. 74(2) (2002) 145–149.

Erythema nodosum leprosum (ENL) classically presents as tender, erythematous nodules over the face, arms and legs. Severe ENL can become vesicular or bullous and break-down and is termed erythema necroticans (Jopling and McDougall, 1996) and is treated with corticosteroids. The causes of death in a majority of leprosy patients are the same as in the general population, with the exception of renal damage in lepromatous leprosy. There is possible increased mortality from side-effects of antileprosy drugs, steroids, or other drugs used in reactions, from toxaemia in severe reactions, and from asphyxia due to glottic oedema (Jopling and McDougall, 1996). We report here a case of erythema necroticans, the cause of death being septicaemia, secondary to skin ulcers and urinary tract infection, precipitated by corticosteroids.—Authors’ Abstract

Girdhar, A., Chakma, J. K., and Girdhar, B. K. Pulsed corticosteroid therapy in patients with chronic recurrent ENL: a pilot study. Indian J. Lepr. 74(3) (2003) 233–236.

A pilot study has been undertaken to compare the efficacy of small dose pulsed betamethasone therapy with need based oral steroids in chronic recurrent erythema nodosum leprosum (ENL) patients. Though this mode of therapy was well tolerated, no advantage with intermittent steroid administration was observed. This could have been on account of small dose of steroid given monthly. Treatment of chronic recurrent erythema nodosum leprosum (ENL) patients continues to be unsatisfactory, particularly, because of nonavailability of thalidomide. Though corticosteroids are effective in suppressing all the manifestations and even restoring partially or fully the functional impairment, their side effects and dependence are equally troublesome. Based on (a) the reported efficacy and safety of intermittent use of corticosteroids in several immune complex mediated disorders (Cathcart, et al. 1976, Kimberly, et al. 1979), Liebling, et al 1981 and Pasricha and Gupta 1984) and (b) ENL (type II) reactions having similar pathology, a pilot study has been undertaken to see the efficacy and the tolerance of pulsed steroids in chronic ENL patients.—Authors’ Abstract

Keita, S., Faye, O., Konare, H. D., Sow, S. O. Ndiaye, H. T., and Traore, I. [Evaluation of the clinical classification of new cases of leprosy. Study conducted at the Marchoux Institute in Bamako, Mali] Ann. Dermatol. Venereol. 130(2) (2003) 184–186. [Article in French]

INTRODUCTION: The difficulties related to the bacilloscopic diagnosis of leprosy, providing a more reliable classification of cases, in 1995 led the WHO to recommend the use of a new classification, in endemic countries, based on clinical criteria alone, in order to simplify the poly-chemotherapeutic regimens. According to our experience in the Marchoux Institute, this classification may lead to errors in diagnosis through overzealous or mis-interpretation of the two forms of leprosy. The aim of our study was to evaluate the concordance between this clinical classification and that based on a bacilloscopic examination. PATIENTS AND METHODS: We conducted a descriptive study of new cases of leprosy seen at the Marchoux Institute, without distinction in gender or age, from January to December 2000. All the patients included underwent clinical examination and a bacilloscopic exploration to provide a double classification. The concordance between the two classifications was assessed using the Kappa test. RESULTS: Two hundred new cases of leprosy were included. Out of 126 clinically multi-bacillary cases, 61 were confirmed bacteriologically, and 65 were false positives. Out of 74 clinical cases with few bacilli, 2 were bacteriologically multi-bacilli. The concordance between the two classifications was average (Kappa = 0.40). There was a significant difference between the percentages of multi-bacilli observed in both classifications (p <10(–8)). DISCUSSION: The clinical classification may well overestimate the multi-bacillary form. In the absence of a reliable bacilloscopic apparatus, a more detailed clinical classification of leprosy forms must be developed.—Authors’ Abstract

Khan, T., Awan, A. A., Kazmi, H. S., Shah, A. A., Muhammad, S., and Muhammad, S. Frequency of ocular complications of leprosy in institutionalized patients in NWFP Pakistan. J. Ayub. Med. Coll. Abbottabad 14(4) (2002) 29–33.

BACKGROUND: There is no systemic disease, which so frequently gives rise to disorders of the eye as leprosy does. The study was conducted to determine the prevalence and gravity of ocular complications in institutionalized leprosy patients in NWFP. It is important to provide necessary information to leprosy health workers and general physicians in order to sensitize them to early detection and treatment or referral to appropriate centre. METHODS: A prospective study of ocular complications of leprosy patients was conducted at the leprosy centre of Lady Reading Hospital Peshawar and the Leprosy Hospital Balakot, district Mansehra. The study included a record of the name, age, sex, type, duration of disease and completion of multi-drug therapy (MDT). Classification of the patients was done according to Ridley and Jopling 5-group system. Visual acuity was tested by Snellen chart and those patients having a vision of less than 3/60 were labelled as blind. Ocular adnexa were examined by naked eye and lacrimal sac regurgitation test was done. Slit lamp biomicroscopy was done for anterior segment examination and direct ophthalmoscope was used for fundoscopy. RESULTS: The authors studied 143 patients in the above mentioned leprosy centres. Out of these, 59 had lepromatous leprosy, 39 borderline tuberculoid leprosy, 9 tuberculoid leprosy, 33 borderline lepromatous leprosy, and 33 borderline leprosy. The majority of patients came from the northern districts of NWFP, including Malakand division and district Mansehra. The male to female ratio was 4:1. The age of the patients ranged from 14 to 80 years and the duration of the disease ranged from 1 year to 48 years. Ocular complications were found in 73% of the patients. These complications included loss of eyebrows in 57 patients, loss of eyelashes in 37, corneal changes (including opacity, ulceration, and/or anaesthesia) in 44, iridocyclitis in 31, lagophthalmos in 36, ectropion in 13, and chronic dacryocystitis in 3. Of the total of 15 (11%) patients who went blind from ocular complications, 16 eyes did so due to corneal opacities, 6 eyes due to cataract, 5 eyes due to chronic anterior uveitis and one eye due to corneal ulcer, panophthalmitis and phthisis bulbi each. CONCLUSIONS: A significant number of leprosy patients (73%) have ocular complications. The frequency of ocular complications increases with the increasing age and duration of disease of the patients.—Authors’ Abstract

Malaviya, G. N. Median nerve palsy following claw finger correction in leprosy: effect of M. leprae or a consequence of surgery. Indian J. Lepr. 74(3) (2002) 217–220.

Median nerve palsy, though not a frequent occurrence after claw finger correction, does exist as a post-operative complication after claw finger correction. A retrospective study was carried out to examine the occurrence of post-operative median palsy, in cases of isolated ulnar palsy, where the transferred motor tendon was routed through the carpal tunnel. We noted that six patients developed median nerve palsy following claw finger correction. Median palsy developed at different times after surgery—the “early onset” type developing within three weeks post-operatively, “reactional” type developed when patient was undergoing physiotherapy exercises and learning to use the transfer and “delayed insidious” type presenting six months or more after operation. We could not succeed to get the true prevalence of such occurrences because all the operated hands could not be re-examined.—Author’s Abstract

Malaviya, G. N. Some more about unfavourable results after corrective surgery as seen in leprosy. Indian J. Lepr. 74(3) (2002) 243–257.

The paper describes unfavourable outcomes of some of the commonly performed surgical procedures in leprosy affected persons and the underlying causes. An awareness about unfavourable outcomes of surgery is helpful to the beginners because they can anticipate the problems and take appropriate measures to prevent that and failing which prepare themselves to face and sort that out. Careful pre-operative evaluation of the patient is an important first step.—Author’s Abstract

Misra, U. K., Kalita, J., Mahadevan, A., and Shankar, S. K. Pseudoathetosis in a patient with leprosy. Mov. Disord. 18(5) (2003) 598–601.

A 35-year-old man with borderline tuberculoid leprosy developed Type I lepra reaction 12 days after anti-leprosy treatment. There was acute worsening of neuropathic symptoms and skin lesions. He developed severe sensory ataxia and pseudoathetosis resulting in marked disability. His symptoms significantly improved on corticosteroid therapy.—Authors’ Abstract

Ozkan, T., Ozer, K., Yukse, A., and Gulgonen, A. Surgical reconstruction of irreversible ulnar nerve paralysis in leprosy. Lepr. Rev. 74(1) (2003) 53–62.

Twenty-five patients with irreversible leprotic ulnar nerve palsy having undergone lumbrical replacement with two different tendon transfer techniques were assessed 6–120 months after surgery. Nineteen patients were reconstructed with the flexor digitorum four-tail procedure (FDS-4T), and six with Zancolli’s lasso procedure (ZLP). Mean paralysis times were 103 months for FDS-4T, and 68 months for ZLP. Mean age of the patients was 36 years (21–57). Grip strength measurements, improvement in active range of motion at the PIP joints, patients’ ability to open and close their hands fully, as well as sequence of phalangeal flexion, were noted. Mean grip strength measurements during follow-up were 76% of the contralateral extremity in the FDS-4T group and 82% in the ZLP group. Comparison of the follow-up grip strength with the preoperative value revealed 1% improvement in the FDS-4T group and 20% in the ZLP group. Claw hand deformity was completely corrected in 12 patients in FDS-4T group, and in five patients in the ZLP group. Residual flexion contracture remained in five patients after surgery. Swan-neck deformity subsequently developed in seven fingers. Age, sex, mean follow-up and surgical technique did not relate statistically to the functional outcome. However, preoperative extensor lag of the PIP joint and mean paralysis time significantly affected the functional outcome. ZLP was found to be a more effective procedure in restoring grip strength, whereas FDS-4T was more effective in correcting claw hand deformity.—Authors’ Abstract

Ramu, G. and Desikan, K. V. Reactions in borderline leprosy. Indian J. Lepr. 74(2) (2002) 115–128.

This is a retrospective study of 276 patients consisting of 157 active and 119 reactive patients of borderline leprosy. They were followed up for 10 years after sulphone monotherapy. The presenting symptoms were carefully examined from the records and systematically presented. Frequency of reactions was least in BT cases and most in BL cases. Risk factors of reaction appear to be the type of leprosy, multiplicity of lesions, high BI and, possibly, psychological stress. Biopsy of skin lesions was performed in all cases initially, and at the subsidence of the disease. Histological findings closely correlated with clinical classification. While all the cases showed clinical subsidence, histological subsidence was found in 200 (73%) cases, and the condition was static in 36 cases (13%). Immunological upgrading was seen in 110%, while 4% showed downgrading. Bacteriological status and lepromin reaction of active and reactive cases were compared. All these factors need to be taken into consideration for instituting prompt and proper treatment.—Authors’ Abstract

Richard, B. M. Location of the extracranial extent of leprous facial nerve pathology may allow leprous facial palsy to be reanimated by free muscle transfer. Br. J. Plast. Surg. 56(1) (2003) 14–19.

Leprosy is a mycobacterial nerve and skin infection, which can be eradicated by antibiotics. Some patients affected by leprosy, once cured, have residual nerve impairment with paralysis and sensory neuropathy. A series of patients with facial nerve paralysis, investigated using clinical, histological and electrophysiological techniques, demonstrated that the nerve pathology was distal to the section of main trunk prior to its bifurcation. Facial reanimation was achieved with a free gracilis-muscle transfer, coapting its motor nerve to the ipsilateral facial nerve trunk proximal to the site of the leprosy pathology, with a moderate clinical result.—Author’s Abstract

Thappa, D. M., Dave, S., Karthikeyan, K., Laxmisha, C., and Jayanthi, S. Localized lepromatous leprosy presenting as a painful nodule in a muscle. Indian J. Lepr. 74(3) (2002) 237–242.

Lepromatous leprosy is a generalized disease usually presenting with numerous macules, papules, nodules or plaques involving wide areas of the skin. It is generally believed that in India lepromatous leprosy often originates from the borderline spectrum (Jha, et al., 1991). Localized lepromatous or borderline lepromatous disease is a rare variant of multibacillary leprosy (Yoder, et al., 1985; Jha, et al., 1991; Pfaltzgraff and Ramu, 1994; Vijaikumar, et al., 2001). This variant usually presents as a single nodule or a localized area of nodules and papules, while most of the body surface appears normal (Pfaltzgraff and Ramu, 1994; Vijaikumar, et al., 2001). Its occurrence in our case as a single painful nodule in the bicep muscle of left forearm was indeed intriguing, such presentation being rarely reported in the literature.—Authors’ Abstract

Yowan, P., Danneman, K., Koshy, S., Richard, J., and Daniel, E. Knowledge and practice of eye-care among leprosy patients. Indian J. Lepr. 74(2) (2002) 129–135.

In one hundred and thirty leprosy patients attending the Schieffelin Leprosy Research and Training Center, Karigiri, Tamil Nadu, India, the knowledge, attitude and practice of eye-care were ascertained using a questionnaire developed by Mathews and Mangalam. 74.6% the patients surveyed were aware of the disease, 60% knew about the early signs of leprosy, 74.6% considered leprosy curable and 36.9% knew the duration of treatment with MDT. Less than half of the patients (40.8%) knew that blindness occurred in leprosy and was preventable. More males had this knowledge (46.5%) than females (22.6%) (p = 0.001). Knowledge on how to take care of the eyes (26.9%), that eyes become anaesthetic due to leprosy (27.7%), and that precautions should be taken if sensation is lost (27. 7%) was very poor. Knowledge on prevention of damage in eyes (57.7%) and the fact that rubbing eyes could cause damage (55.4%) was found in more than half the patients. More males (64.6%) had knowledge on the prevention of damage in eyes than females (35.5%) (p = 0.008). Only 25.4% of the patients tried some measures to prevent eye injury, 21.5% used home remedies and all had the help of family members in their eye-care. More males (26.3%) used home remedies than females (6.5%). The older age group had better knowledge on taking care of the eyes than those aged 40 and below (p = 0.026). Although more patients with existing complications knew to take care of their eyes than those who did not have complications, the knowledge and practice of eye-care in both these groups were poor. Knowledge of leprosy in illiterate patients was not different from those who had some formal schooling, but the practice of eye-care differed significantly (p = 0.02). Health education must be undertaken to increase the knowledge of eye-care among leprosy patients, especially among illiterate persons, women and younger patients.—Authors’ Abstract

Actor, J. K., Indrigo, J., Beachdel, C. M., Olsen, M., Wells, A., Hunter, Jr. R. L., and Dasgupta, A. Mycobacterial Glycolipid Cord Factor Trehalose 6,6-Dimycolate Causes a Decrease in Serum Cortisol during the Granulomatous Response. Neuroimmunomodulation 10(5) (2003) 270–282.

Serum cortisol levels were evaluated in mice following intravenous administration of purified mycobacterial glycolipid trehalose 6,6-dimycolate (TDM). C57BL/6 mice develop lung granulomas in response to TDM, while A/J mice are deficient in this process. Administration of TDM to C57BL/6 mice led to a rapid reduction in serum cortisol, concurrent with initiation of the granulomatous response and cytokine and chemokine mRNA induction. Cortisol levels were lowest on day 5 after TDM administration, but there was significant production of IL-6, TNF-alpha and IL-1beta messages. Granuloma formation and full immune responsiveness to TDM were only apparent upon a sufficient decrease in levels of systemic cortisol. Treatment of the C57BL/6 mice with hydrocortisone abolished inflammatory responses. Histologically nonresponding A/J mice exhibited higher constitutive serum cortisol and demonstrated different kinetics of cortisol reduction upon administration of TDM. A/J mice demonstrated hyperplastic morphology in the suprarenal gland with a high degree of vacuolization in the medullary region and activation of cells in the zona fasciculata and zona reticularis. The A/J mice were dysregulated with respect to cytokine responses thought to be necessary during granuloma formation. The high constitutive serum cortisol in the A/J mice may therefore contribute to pulmonary immunoresponsiveness and the establishment of an environment counterproductive to the initiation of granulomatous responses. The identification of a mycobacterial glycolipid able to influence serum cortisol levels is unique and is discussed in relation to immunopathology during tuberculosis disease.—Authors’ Abstract

Bhattacharyya, A., Pathak, S., Basak, C., Law, S., Kundu, M., and Basu, J. Execution of macrophage apoptosis by Mycobacterium avium through apoptosis signal-regulating kinase 1/p38 mitogen-activated protein kinase signaling and caspase 8 activation. J. Biol. Chem. 278(29) (2003) 26517–26525.

Macrophage apoptosis is an important component of the innate immune defense machinery (against pathogenic mycobacteria) responsible for limiting bacillary viability. However, little is known about the mechanism of how apoptosis is executed in mycobacteria-infected macrophages. Apoptosis signal-regulating kinase 1 (ASK1) was activated in M. avium-treated macrophages and in turn activated p38 mitogen-activated protein (MAP) kinase. M. avium-induced macrophage cell death could be blocked in cells transfected with a catalytically inactive mutant of ASK1 or with dominant negative p38 MAP kinase arguing in favor of a central role of ASK1/p38 MAP kinase signaling in apoptosis of macrophages challenged with M. avium. ASK1/p38 MAP kinase signaling was linked to the activation of caspase 8. At the same time, M. avium triggered caspase 8 activation and cell death occurred in a Fas-associated death domain (FADD)-dependent manner, suggesting the involvement of death receptor signaling. FADD did not exert its effect through ASK1 activation. The death signal induced upon caspase 8 activation linked to mitochondrial death signaling through the formation of truncated Bid (t-Bid), its translocation to the mitochondria and release of cytochrome c (cyt c). Caspase 8 inhibitor (z-IETD-FMK) could block the release of cytochrome c as well as the activation of caspases 9 and 3. The final steps of apoptosis probably involved caspases 9 and 3, since inhibitors of both caspases could block cell death. Of foremost interest in the present study was the finding that ASK1/p38 signaling was essential for caspase 8 activation linked to M. avium-induced death signaling. It provides the first elucidation of a signaling pathway in which ASK1 plays a central role in innate immunity.—Authors’ Abtstract

Chiu, B. C., Freeman, C. M., Stolberg, V. R., Komuniecki, E., Lincoln, P. M., Kunkel, S. L., and Chensue, S. W. Cytokine-Chemokine Networks in Experimental Mycobacterial and Schistosomal Lung Granuloma Formation. Am. J. Respir. Cell. Mol. Biol. 29(1) (2003) 106–116.

Type-1 and type-2 lung granulomas respectively elicited by bead immobilized Mycobacteria bovis (PPD) and Schistosoma mansoni egg (SEA) Ags display different patterns chemokine expression. This study tested the hypothesis that chemokine expression patterns were related to upstream cytokine signaling. Using quantitative transcript analysis we defined expression profiles for 16 chemokines and then examined the in vivo effects of neutralizing antibodies against interferon-gamma (IFNgamma), interleukin-4 (IL-4), IL-10, IL-12, and IL-13. Transcripts for CXCL2, 5, 9, 10 and 11 and the CCL chemokines, CCL3 and lymphotactin (XCL1) were largely enhanced by Th1-related cytokines, IFN gamma or IL-12. Transcripts for CCL11, CCL22, CCL17 and CCL1 were enhanced largely by Th2-related cytokines, IL-4, IL-10 or IL-13. Transcripts for CCL4, CCL2, CCL8, CCL7, and CCL12 were potentially induced by either Th1- or Th2-related cytokines although some of these showed biased expression. IFNgamma and IL-4 enhanced the greatest complement of transcripts and their neutralization had the greatest anti-inflammatory effect on type-1 and type-2 granulomas, respectively. Th1/Th2 cross-regulation was evident since endogenous Th2 cytokines inhibited type-1, whereas Th1 cytokines inhibited type-2 biased chemokines. These findings reveal a complex cytokine-chemokine regulatory network that dictates profiles of local chemokine expression during T cell-mediated granuloma formation. —Authors’ Abstract

Mendez-Samperio, P., Ayala, H., and Vazquez, A. NF-kappaB Is Involved in Regulation of CD40 Ligand Expression on Mycobacterium bovis Bacillus Calmette-Guerin-Activated Human T Cells. Clin. Diagn. Lab. Immunol. 10(3) (2003) 376–382.

Interaction between CD40L (CD154) on activated T cells and its receptor CD40 on antigen-presenting cells has been reported to be important in the resolution of infection by mycobacteria. However, the mechanism(s) by which Mycobacterium bovis bacillus Calmette-Guerin (BCG) up-regulates membrane expression of CD40L molecules is poorly understood. This study was done to investigate the role of the nuclear factor kappaB (NF-kappaB) signaling pathway in the regulation of CD40L expression in human CD4(+) T cells stimulated with BCG. Specific pharmacologic inhibition of the NF-kappaB pathway revealed that this signaling cascade was required in the regulation of CD40L expression on the surface of BCG-activated CD4(+) T cells. These results were further supported by the fact that treatment of BCG-activated CD4(+) T cells with these pharmacological inhibitors significantly down-regulated CD40L mRNA. In this study, inhibitor kappaBalpha (IkappaBalpha) and IkappaBbeta protein production was not affected by the chemical protease inhibitors and, more importantly, BCG led to the rapid but transient induction of NF-kappaB activity. Our results also indicated that CD40L expression on BCG-activated CD4(+) T cells resulted from transcriptional up-regulation of the CD40L gene by a mechanism which is independent of de novo protein synthesis. Interestingly, BCG-induced activation of NF-kappaB and the increased CD40L cell surface expression were blocked by the protein kinase C (PKC) inhibitors 1-[5-isoquinolinesulfonyl]-2-methylpiperazine and salicylate, both of which block phosphorylation of IkappaB. Moreover, rottlerin a Ca(2+)-independent PKC isoform inhibitor, significantly down-regulated CD40L mRNA in BCG-activated CD4(+) T cells. These data strongly suggest that CD40L expression by BCG-activated CD4(+) T cells is regulated via the PKC pathway and by NF-kappaB DNA binding activity.—Authors’ Abstract

Venkatesh, J., Kumar, P., MuraliKrishna, P. S., Manjunath, R., and Varshney, U. Importance of uracil DNA glycosylase in Pseudomonas aeruginosa and Mycobacterium smegmatis, G+C-rich bacteria, in mutation prevention, tolerance to acidified nitrite and endurance in mouse macrophages. J. Biol. Chem. 278(27) (2003) 24350–24358.

Uracil DNA glycosylase (Ung or UDG), initiates the excision repair of an unusual base, uracil in DNA. Ung is a highly conserved protein found in all organisms. Paradoxically, loss of this evolutionarily conserved enzyme has not been seen to result in severe growth phenotypes in the cellular life forms. In this study, we chose G+C rich genome containing bacteria (Pseudomonas aeruginosa and Mycobacterium smegmatis), as model organisms to investigate the biological significance of ung. Ung deficiency was created either by expression of a highly specific inhibitor protein, Ugi and/or by targeted disruption of the ung gene. We show that abrogation of Ung activity in P. aeruginosa and M. smegmatis confers upon them an increased mutator phenotype and sensitivity to reactive nitrogen intermediates generated by acidified nitrite. Also, in a mouse macrophage infection model, P. aeruginosa (Ung–) shows a significant decrease in its survival. Infections of the macrophages, with M. smegmatis, show an initial increase in the bacterial counts, which remain at this level for up to 48 hr before a decline. Interestingly, abrogation of Ung activity in M. smegmatis results in nearly a total abolition of their multiplication and much-decreased residency in macrophages stimulated with IFN. These observations suggest Ung as a useful target to control growth of G+C rich bacteria.—Authors’ Abstract

Venketaraman, V., Dayaram, Y. K., Amin, A. G., Ngo, R., Green, R. M., Talaue, M. T., Mann, J., and Connell, N. D. Role of glutathione in macrophage control of mycobacteria. Infect. Immun. 71(4) (2003) 1864–1871.

Reactive oxygen and nitrogen intermediates are important antimicrobial defense mechanisms of macrophages and other phagocytic cells. While reactive nitrogen intermediates have been shown to play an important role in tuberculosis control in the murine system, their role in human disease is not clearly established. Glutathione, a tripeptide and antioxidant, is synthesized at high levels by cells during reactive oxygen intermediate and nitrogen intermediate production. Glutathione has been recently shown to play an important role in apoptosis and to regulate antigen-presenting-cell functions. Glutathione also serves as a carrier molecule for nitric oxide, in the form of S-nitrosoglutathione. Previous work from this laboratory has shown that glutathione and S-nitrosoglutathione are directly toxic to mycobacteria. A mutant strain of Mycobacterium bovis BCG, defective in the transport of small peptides such as glutathione, is resistant to the toxic effect of glutathione and S-nitrosoglutathione. Using the peptide transport mutant as a tool, we investigated the role of glutathione and S-nitrosoglutathione in animal and human macrophages in controlling intracellular mycobacterial growth.—Authors’ Abstract

Worku, S. and Hoft, D. F. Differential effects of control and antigen-specific T cells on intracellular mycobacterial growth. Infect. Immun. 71(4) (2003) 1763–1773.

We investigated the effects of peripheral blood mononuclear cells expanded with irrelevant control and mycobacterial antigens on the intracellular growth of Mycobacterium bovis bacillus Calmette-Guerin (BCG) in human macrophages. More than 90% of the cells present after 1 week of in vitro expansion were CD3(+). T cells were expanded from purified protein derivative-negative controls, persons with latent tuberculosis, and BCG-vaccinated individuals. T cells expanded with nonmycobacterial antigens enhanced the intracellular growth of BCG in suboptimal cultures of macrophages. T cells expanded with live BCG or lysates of Mycobacterium tuberculosis directly inhibited intracellular BCG. Recent intradermal BCG vaccination significantly enhanced the inhibitory activity of T cells expanded with mycobacterial antigens (p <0.02), consistent with the induction of memory-immune inhibitory T-cell responses. Selected mycobacterial antigens (Mtb41 > lipoarabinomannan > 38kd > Ag85B > Mtb39) expanded inhibitory T cells, demonstrating the involvement of antigen-specific T cells in intracellular BCG inhibition. We studied the T-cell subsets and molecular mechanisms involved in the memory-immune inhibition of intracellular BCG. Mycobacteria-specific gammadelta T cells were the most potent inhibitors of intracellular BCG growth. Direct contact between T cells and macrophages was necessary for the BCG growth-enhancing and inhibitory activities mediated by control and mycobacteria-specific T cells, respectively. Increases in tumor necrosis factor alpha, interleukin-6, transforming growth factor beta, and vascular endothelial growth factor mRNA expression were associated with the enhancement of intracellular BCG growth. Increases in gamma interferon, FAS, FAS ligand, perforin, granzyme, and granulysin mRNA expression were associated with intracellular BCG inhibition. These culture systems provide in vitro models for studying the opposing T-cell mechanisms involved in mycobacterial survival and protective host immunity.—Authors’ Abstract

Buhrer-Sekula, S., Smits, H. L., Gussenhoven, G. C., Van Leeuwen, J., Amador, S., Fujiwara, T., Klatser, P. R., and Oskam, L. Simple and fast lateral flow test for classification of leprosy patients and identification of contacts with high risk of developing leprosy. J. Clin. Microbiol. 41(5) (2003) 1991–1995.

The interruption of leprosy transmission is one of the main challenges for leprosy control programs since no consistent evidence exists that transmission has been reduced after the introduction of multidrug therapy. Sources of infection are primarily people with high loads of bacteria with or without clinical signs of leprosy. The availability of a simple test system for the detection of antibodies to phenolic glycolipid-I (PGL-I) of Mycobacterium leprae to identify these individuals may be important in the prevention of transmission. We have developed a lateral flow assay, the ML Flow test, for the detection of antibodies to PGL-I which takes only 10 min to perform. An agreement of 91% was observed between enzyme-linked immunosorbent assay and our test; the agreement beyond chance (kappa value) was 0.77. We evaluated the use of whole blood by comparing 539 blood and serum samples from an area of high endemicity. The observed agreement was 85.9% (kappa = 0.70). Storage of the lateral flow test and the running buffer at 28 degrees C for up to 1 year did not influence the results of the assay. The sensitivity of the ML Flow test in correctly classifying MB patients was 97.4%. The specificity of the ML Flow test, based on the results of the control group, was 90.2%. The ML Flow test is a fast and easy-to-perform method for the detection of immunoglobulin M antibodies to PGL-I of M. leprae. It does not require any special equipment, and the highly stable reagents make the test robust and suitable for use in tropical countries.—Authors’ Abstract

Hernandez, M. O., Neves, I., Sales, J. S., Carvalho, D. S., Sarno, E. N., and Sampaio, E. P. Induction of apoptosis in monocytes by Mycobacterium leprae in vitro: a possible role for tumour necrosis factor-alpha. Immunology. 109(1) (2003) 156–164.

A diverse range of infectious organisms, including mycobacteria, have been reported to induce cell death in vivo and in vitro. Although morphological features of apoptosis have been identified in leprosy lesions, it has not yet been determined whether Mycobacterium leprae modulates programmed cell death. For that purpose, peripheral blood mononuclear cells obtained from leprosy patients were stimulated with different concentrations of this pathogen. Following analysis by flow cytometry on 7AAD/CD14+ cells, it was observed that M. leprae induced apoptosis of monocyte-derived macrophages in a dose-dependent manner in both leprosy patients and healthy individuals, but still with lower efficiency as compared to M. tuberculosis. Expression of tumour necrosis factor-alpha (TNF-alpha), Bax-alpha, Bak mRNA and TNF-alpha protein was also detected in these cultures; in addition, an enhancement in the rate of apoptotic cells (and of TNF-alpha release) was noted when interferon-gamma was added to the wells. On the other hand, incubation of the cells with pentoxifylline impaired mycobacterium-induced cell death, the secretion of TNF-alpha, and gene expression in vitro. In addition, diminished bacterial entry decreased both TNF-alpha levels and the death of CD14+ cells, albeit to a different extent. When investigating leprosy reactions, an enhanced rate of spontaneous apoptosis was detected as compared to the unreactive lepromatous patients. The results demonstrated that M. leprae can lead to apoptosis of macrophages through a mechanism that could be at least partially related to the expression of pro-apoptotic members of the Bcl-2 protein family and of TNF-alpha. Moreover, while phagocytosis may be necessary, it seems not to be crucial to the induction of cell death by the mycobacteria.—Authors’ Abstract

Kiszewski, A. E., Becerril, E., Baquera, J., Ruiz-Maldonado, R., and HernAndez Pando, R. Expression of cyclooxygenase type 2 in lepromatous and tuberculoid leprosy lesions. Br. J. Dermatol. 148(4) (2003) 795–798.

BACKGROUND: Leprosy is an infectious disease with two polar forms, tuberculoid leprosy (TL) and lepromatous leprosy (LL), which are dominated by T-helper (Th) 1 and Th2 cells, respectively. High concentrations of prostaglandin E2 produced by the inducible enzyme cyclooxygenase type 2 (COX-2) in LL could inhibit Th1 cytokine production, contributing to T-cell anergy. Objectives To compare the COX-2 expression in LL and TL. Methods Skin biopsies from 40 leprosy patients (LL, N = 20; TL, N = 20) were used to determine by immunohistochemistry and automated morphometry the percentage of COX-2 immunostained cells. Results Most COX-2-positive cells were macrophages; their percentages in the inflammatory infiltrate located in the papillary dermis, reticular dermis and periadnexally were significantly higher in LL than TL (p <0.001 by Student’s t-test). CONCLUSIONS: The high expression of COX-2 in LL may be related to high prostaglandin production contributing to T-cell anergy.—Authors’ Abstract

Krutzik, S. R., Ochoa, M. T., Sieling, P. A., Uematsu, S., Ng, Y. W., Legaspi, A., Liu, P. T., Cole, S. T., Godowski, P. J., Maeda, Y., Sarno, E. N., Norgard, M. V., Brennan, P. J., Akira, S., Rea, T. H., and Modlin, R. L. Activation and regulation of Toll-like receptors 2 and 1 in human leprosy. Nat. Med. 9(5) (2003) 525–532.

The expression and activation of Toll-like receptors (TLRs) was investigated in leprosy, a spectral disease in which clinical manifestations correlate with the type of immune response mounted toward Mycobacterium leprae. TLR2-TLR1 heterodimers mediated cell activation by killed M. leprae, indicating the presence of triacylated lipoproteins. A genome-wide scan of M. leprae detected 31 putative lipoproteins. Synthetic lipopeptides representing the 19-kD and 33-kD lipoproteins activated both monocytes and dendritic cells. Activation was enhanced by type-1 cytokines and inhibited by type-2 cytokines. In addition, interferon (IFN)-gamma and granulocyte-macrophage colony-stimulating factor (GM-CSF) enhanced TLR1 expression in monocytes and dendritic cells, respectively, whereas IL-4 downregulated TLR2 expression. TLR2 and TLR1 were more strongly expressed in lesions from the localized tuberculoid form (T-lep) as compared with the disseminated lepromatous form (L-lep) of the disease. These data provide evidence that regulated expression and activation of TLRs at the site of disease contribute to the host defense against microbial pathogens.—Authors’ Abstract

Lee, S. B., Kim, B. C., Jin, S. H., Park, Y. G., Kim, S. K., Kang, T. J., and Chae, G. Missense mutations of the interleukin-12 receptor beta 1(IL12RB1) and interferon-gamma receptor 1 (IFNGR1) genes are not associated with susceptibility to lepromatous leprosy in Korea. Immunogenetics 55(3) (2003) 177–181.

Interleukin-12 receptor beta 1 (IL12RB1), interleukin-12 receptor beta 2 (IL12RB2), and interferon gamma receptor 1 (IFNGR1) perform important roles in the host defense against intracellular pathogens such as Mycobacteria. Several mutations within their genes have been confirmed as associated with increased susceptibility to mycobacterial infection. However, the association between mutations of the IL12RB1, IL12RB2, and IFNGR1 encoding genes and lepromatous leprosy has not been studied. This study screened for polymorphisms within IL12RB1, IL12RB2, and IFNGR1 encoding genes in the Korean populations using polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) DNA sequencing assay, and an association study was performed using the missense mutations of 705 A/G (Q214R), 1196 G/C (G378R), 1637 G/A (A525T), and 1664 C/T (P534S) of the IL12RB1, 83 G/A (V14M), and 1443 T/C (L467P) for the IFNGR1 encoding genes. There were no differences in the genotype and allele frequencies of IL12RB1 and IFNGR1 genes between 93 lepromatous leprosy patients and 94 control subjects. In conclusion, missense mutations of 705 A/G (Q214R), 1196 G/C (G378R), 1637 G/A (A525T), 1664 C/T (P534S) of the IL12RB1, 83 G/A (V14 M), and 1443 T/C (L467P) of the IFNGR1 encoding genes have no association with the susceptibility to lepromatous leprosy in the Korean population.—Authors’ Abstract

Boshoff, H. I., Reed, M. B., Barry, C. E., and Mizrahi, V. DnaE2 Polymerase Contributes to In Vivo Survival and the Emergence of Drug Resistance in Mycobacterium tuberculosis. Cell. 113(2) (2003) 183–193.

The presence of multiple copies of the major replicative DNA polymerase (DnaE) in some organisms, including important pathogens and symbionts, has remained an unresolved enigma. We postulated that one copy might participate in error-prone DNA repair synthesis. We found that UV irradiation of Mycobacterium tuberculosis results in increased mutation frequency in the surviving fraction. We identified dnaE2 as a gene that is upregulated in vitro by several DNA damaging agents, as well as during infection of mice. Loss of this protein reduces both survival of the bacillus after UV irradiation and the virulence of the organism in mice. Our data suggest that DnaE2, and not a member of the Y family of error-prone DNA polymerases, is the primary mediator of survival through inducible mutagenesis and can contribute directly to the emergence of drug resistance in vivo. These results may indicate a potential new target for therapeutic intervention.—Authors’ Abstract

Brennan, P. J. Structure, function, and biogenesis of the cell wall of Mycobacterium tuberculosis. Tuberculosis (Edinb). 83(1–3) (2003) 91–97.

Much of the early structural definition of the cell wall of Mycobacterium spp. was initiated in the 1960s and 1970s. There was a long period of inactivity, but more recent developments in NMR and mass spectral analysis and definition of the M. tuberculosis genome have resulted in a thorough understanding, not only of the structure of the mycobacterial cell wall and its lipids but also the basic genetics and biosynthesis. Our understanding nowadays of cell-wall architecture amounts to a massive “core” comprised of peptidoglycan covalently attached via a linker unit (L-Rha-D-GlcNAc-P) to a linear galactofuran, in turn attached to several strands of a highly branched arabinofuran, in turn attached to mycolic acids. The mycolic acids are oriented perpendicular to the plane of the membrane and provide a truly special lipid barrier responsible for many of the physiological and disease-inducing aspects of M. tuberculosis. Intercalated within this lipid environment are the lipids that have intrigued researchers for over five decades: the phthiocerol dimycocerosate, cord factor/dimycolyltrehalose, the sulfolipids, the phosphatidylinositol mannosides, etc. Knowledge of their roles in “signaling” events, in pathogenesis, and in the immune response is now emerging, sometimes piecemeal and sometimes in an organized fashion. Some of the more intriguing observations are those demonstrating that mycolic acids are recognized by CD1-restricted T-cells, that antigen 85, one of the most powerful protective antigens of M. tuberculosis, is a mycolyltransferase, and that lipoarabinomannan (LAM), when “capped” with short mannose oligosaccharides, is involved in phagocytosis of M. tuberculosis. Definition of the genome of M. tuberculosis has greatly aided efforts to define the biosynthetic pathways for all of these exotic molecules: the mycolic acids, the mycocerosates, phthiocerol, LAM, and the polyprenyl phosphates. For example, we know that synthesis of the entire core is initiated on a decaprenyl-P with synthesis of the linker unit, and then there is concomitant extension of the galactan and arabinan chains while this intermediate is transported through the cytoplasmic membrane. The final steps in these events, the attachment of mycolic acids and ligation to peptidoglycan, await definition and will prove to be excellent targets for a new generation of anti-tuberculosis drugs.—Author’s Abstract

Ehlers, S., Holscher, C., Scheu, S., Tertilt, C., Hehlgans, T., Suwinski, J., Endres, R., and Pfeffer, K. The Lymphotoxin beta Receptor Is Critically Involved in Controlling Infections with the Intracellular Pathogens Mycobacterium tuberculosis and Listeria monocytogenes. J. Immunol. 170(10) (2003) 5210–5218.

Containment of intracellularly viable microorganisms requires an intricate cooperation between macrophages and T cells, the most potent mediators known to date being IFN-gamma and TNF. To identify novel mechanisms involved in combating intracellular infections, experiments were performed in mice with selective defects in the lymphotoxin (LT)/LTbetaR pathway. When mice deficient in LTalpha or LTbeta were challenged intranasally with Mycobacterium tuberculosis, they showed a significant increase in bacterial loads in lungs and livers compared with wild-type mice, suggesting a role for LTalphabeta heterotrimers in resistance to infection. Indeed, mice deficient in the receptor for LTalpha(1)beta(2) heterotrimers (LTbetaR-knockout (KO) mice) also had significantly higher numbers of M. tuberculosis in infected lungs and exhibited widespread pulmonary necrosis already by day 35 after intranasal infection. Furthermore, LTbetaR-KO mice were dramatically more susceptible than wild-type mice to i.p. infection with Listeria monocytogenes. Compared with wild-type mice, LTbetaR-KO mice had similar transcript levels of TNF and IFN-gamma and recruited similar numbers of CD3(+) T cells inside granulomatous lesions in M. tuberculosis-infected lungs. Flow cytometry revealed that the LTbetaR is expressed on pulmonary macrophages obtained after digestion of M. tuberculosis-infected lungs. LTbetaR-KO mice showed delayed expression of inducible NO synthase protein in granuloma macrophages, implicating deficient macrophage activation as the most likely cause for enhanced susceptibility of these mice to intracellular infections. Since LIGHT-KO mice proved to be equally resistant to M. tuberculosis infection as wild-type mice, these data demonstrate that signaling of LTalpha(1)beta(2) heterotrimers via the LTbetaR is an essential prerequisite for containment of intracellular pathogens.—Authors’ Abstract

Fenhalls, G., Squires, G. R., Stevens-Muller, L., Bezuidenhout, J., Amphlett, G., Duncan, K., and Lukey, P. T. Associations between Toll-like Receptors and IL-4 in the Lungs of Patients with Tuberculosis. Am. J. Respir. Cell. Mol. Biol. 29(1) (2003) 28–38.

Toll-like receptors (TLRs) are implicated in the intracellular killing of Mycobacterium tuberculosis and their expression is modulated by interleukin-4 (IL-4) in vitro. Our aim was to examine the expression of TLRs at the site of pathology in tuberculous lung granulomas and to explore the effect of the immune response on TLR expression. Immunohistochemistry was performed on lung granulomas from nine patients with tuberculosis undergoing lobectomy for haemoptysis. All nine patients expressed all of the TLRs studied (TLRs1–5 and 9), whereas only five out of the nine patients had any granulomas positive for IL-4. Statistical analysis of TLR and cytokine staining patterns in 183 individual granulomas from the nine patients revealed significant associations between pairs of receptors and IL-4. A positive association between TLR2 and TLR4 (p <0.0001) and a negative association between TLR2 and IL-4 (p <0.0001) was observed. The associations between TLRs 1, 5 and 9 were significantly different in IL-4 negative compared to IL-4 positive patients. In conclusion, TLRs are expressed by various cell types in the human tuberculous lung and their expression patterns are reflected by differences in the immune response.—Authors’ Abstract

Fratti, R. A., Chua, J., Vergne, I., and Deretic, V. Mycobacterium tuberculosis glycosylated phosphatidylinositol causes phagosome maturation arrest. Proc. Natl. Acad. Sci. U. S. A. 100(9) (2003) 5437– 5442.

The tubercle bacillus parasitizes macrophages by inhibiting phagosome maturation into the phagolysosome. This phenomenon underlies the tuberculosis pandemic involving 2 billion people. We report here how Mycobacterium tuberculosis causes phagosome maturation arrest. A glycosylated M. tuberculosis phosphatidylinositol [mannose-capped lipoarabinomannan (ManLAM)] interfered with the phagosomal acquisition of the lysosomal cargo and syntaxin 6 from the trans-Golgi network. ManLAM specifically inhibited the pathway dependent on phosphatidylinositol 3-kinase activity and phosphatidylinositol 3-phosphate-binding effectors. These findings identify ManLAM as the M. tuberculosis product responsible for the inhibition of phagosomal maturation.—Authors’ Abstract

Hsiao, P. F., Tzen, C. Y., Chen, H. C., and Su, H. Y. Polymerase chain reaction based detection of Mycobacterium tuberculosis in tissues showing granulomatous inflammation without demonstrable acid-fast bacilli. Int. J. Dermatol. 42(4) (2003) 281–286.

BACKGROUND: Cutaneous tuberculosis is especially difficult to distinguish from other granulomatous dermatoses. We used polymerase chain reaction (PCR) to evaluate the incidence of cutaneous tuberculosis and atypical mycobacterial infection in formalin-fixed, paraffin-embedded tissues with unspecified granulomatous inflammation and negative results for acid-fast bacilli (AFB), and analyzed the pattern of cutaneous tuberculosis in this group of patients. METHODS: A total of 38 specimens which had been collected from 36 patients and fulfillled the criteria for tissues described above were used in this study. Two different primer pairs targeting the gene encoding for 16S ribosomal RNA (common to all mycobacteria) and the insertion sequence IS6110 (specific for M. tuberculosis complex) were used in the PCR assays. The clinical characteristics, histopathologic findings, and culture results of the patients were also analyzed. RESULTS: Four specimens were excluded from the analysis due to the lack of internal control testing. Of the remaining 34 specimens, 22 were PCR positive for the 16S rRNA gene. Among them, 18 specimens were PCR positive for both the 16S rRNA gene and IS6110. Cutaneous tuberculosis could be diagnosed in these 18 cases (56.2%). Out of the 18 cases, there were 8 women and 10 men. The age range was 15–77 years (mean: 44.2 years). After reviewing their clinical presentation, 11 cases were considered as tuberculosis verrucosa cutis, 6 cases as lupus vulgaris, and 1 case as erythema induratum. The remaining 4 cases (12.5%) positive only for 16S rRNA gene were considered as possible atypical mycobacteria infection. CONCLUSIONS: These results show that in paucibacillary form of cutaneous tuberculosis with unclassical clinical and histological presentation, this PCR system provides rapid and sensitive detection of M. tuberculosis DNA in formalin-fixed, paraffin-embedded specimens. Cutaneous tuberculosis represents a significant proportion in specimens showing granulomatous inflammation. In areas like Taiwan, where prevalence of pulmonary tuberculosis is still high, tuberculosis verrucosa cutis and lupus vulgaris are common forms of cutaneous tuberculosis and are seen more frequently than atypical mycobacterial infection.—Authors’ Abstract

Jung, Y. J., Ryan, L., LaCourse, R., and North, R. J. Increased interleukin-10 expression is not responsible for failure of T helper 1 immunity to resolve airborne Mycobacterium tuberculosis infection in mice. Immunology 109(2) (2003) 295–299.

With a view to determining whether failure of mice to resolve Mycobacterium tuberculosis (Mtb) infection is a consequence of downregulation of T helper 1 (Th1) immunity by interleukin (IL)-10, mice deleted of the gene for IL-10 were compared with wild-type (WT) mice in terms of their ability to make IL-10 mRNA, generate Th1-mediated immunity [as measured by synthesis of mRNA for interferon-gamma (IFN-gamma)], IL-12p40 and inducible nitric oxide synthase (iNOS), and to control lung infection. It was found that the response of WT mice to infection included a substantial and sustained increase in IL-10 mRNA synthesis in the lungs. A Th1 response in the lungs of WT and IL-10–/– mice was evidenced by a large and sustained increase in the synthesis of mRNA for IFN-gamma, IL-12p40 and iNOS, with somewhat higher levels of these mRNA species being made in the lungs of IL-10–/– mice, particularly at an early stage of infection. However, IL-10–/– mice were no more capable than WT mice at combating infection.—Authors’ Abstract

Kaul, D., Coffey, M. J., Phare, S. M., and Kazanjian, P. H. Capacity of neutrophils and monocytes from human immunodeficiency virus-infected patients and healthy controls to inhibit growth of Mycobacterium bovis. J. Lab. Clin. Med. 141(5) (2003) 330–334.

We compared the differences in growth inhibition of Mycobacterium bovis by monocytes and neutrophils from human immunodeficiency virus (HIV)-infected persons (N = 12; mean CD4 count = 451/mm(3)) and healthy controls (N = 6). Phagocytes from all HIV-infected patients were incubated with or without exogenous granulocyte-macrophate colony-stimulating factor (GMCSF; 500–1000 U/mL). In two of the HIV-infected patients, phagocytes were incubated with or without interleukin (IL)-2 or IL-8 (500–1000 U/mL). Compared with that in HIV-infected patients, the reduction of M. bovis growth at 24 hours was 81% greater among monocytes and 69% greater among neutrophils from healthy controls (p = .03 and .04, respectively). Among HIV-infected patients, we noted greater mycobacterial reduction in monocytes (49%, p = .04) and neutrophils (42%, p = .05) from the early-stage patients (mean CD4 count = 760/mm(3)) compared with that in late-stage patients (mean CD4 count = 172/mm(3)). Incubation with GM-CSF, IL-2, or IL-8 did not augment mycobactericidal activity. These findings suggest that the capacity of neutrophils and monocytes from HIV-infected patients to inhibit the growth of M. bovis is impaired, and this impairment is more pronounced in later stages of HIV infection.—Authors’ Abstract

Koguchi, Y., Kawakami, K., Uezu, K., Fukushima, K., Kon, S., Maeda, M., Nakamoto, A., Owan, I., Kuba, M., Kudeken, N., Kadota, J. I., Mukae, H. Kohno, S., Uede, T., and Saito, A. High plasma osteopontin level and its relationship with IL-12-mediated Th1 response in tuberculosis. Am. J. Respir. Crit. Care Med. 167(10) (2003) 1355–1359.

Osteopontin (OPN, also known as Eta-1), a noncollagenous matrix protein produced by macrophages and T lymphocytes, is expressed in granulomatous lesions caused by Mycobacterium tuberculosis infection. In the present study, we compared plasma concentrations of OPN in patients with active pulmonary tuberculosis with those of healthy control subjects and patients with sarcoidosis, another disease associated with granuloma formation. Plasma OPN levels were significantly higher in patients with tuberculosis (N = 48) than control subjects (N = 34) and patients with sarcoidosis (N = 20). OPN levels correlated well with severity of pulmonary tuberculosis, as indicated by the size of lung lesions on chest X-ray films. Furthermore, chemotherapy resulted in a significant fall in plasma OPN levels. In patients with tuberculosis, plasma OPN concentrations correlated significantly with those of interleukin (IL)-12. In vitro experiments showed that OPN production by peripheral blood mononuclear cells infected with M. bovis BCG preceded the synthesis of IL-12 and interferon (IFN)-gamma, and that neutralizing anti-OPN monoclonal antibody significantly reduced the production of IL-12 and IFN-gamma. Our results suggest that OPN may be involved in the pathological process associated with active pulmonary tuberculosis by inducing IL-12-mediated Th1 responses.—Authors’ Abstract

Munoz, S., Hernandez-Pando, R., Abraham, S. N., and Enciso, J. A. Mast cell activation by Mycobacterium tuberculosis: mediator release and role of CD48. J. Immunol. 170(11) (2003) 5590–5596.

Mast cells (MC) are abundant in the lung and other peripheral tissue, where they participate in inflammatory processes against bacterial infections. Like other effector cells of the innate immune system, MC interact directly with a wide variety of infectious agents. This interaction results in MC activation and inflammatory mediator release. We demonstrated that MC interact with Mycobacterium tuberculosis, triggering the release of several prestored reagents, such as histamine and beta-hexosaminidase, and de novo synthesized cytokines, such as TNF-alpha and IL-6. A number of M. tuberculosis Ags, ESAT-6, MTSA-10, and MPT-63, have been implicated in MC activation and mediator release. A MC plasmalemmal protein, CD48, was implicated in interactions with mycobacteria because CD48 appeared to aggregate in the MC membrane at sites of bacterial binding and because Abs to CD48 inhibited the MC histamine response to mycobacteria. Cumulatively, these findings suggest that MC, even in the absence of opsonins, can directly recognize M. tuberculosis and its Ags and have the potential to play an active role in mediating the host’s innate response to M. tuberculosis infection.—Authors’ Abstract

Natarajan, K., Latchumanan, V. K., Singh, B., Singh, S., and Sharma, P. Down-regulation of T helper 1 responses to mycobacterial antigens due to maturation of dendritic cells by 10-kDa Mycobacterium tuberculosis secretory antigen. J. Infect. Dis. 187(6) (2003) 914–928.

Interactions of 10-kDa Mycobacterium tuberculosis secretory antigen (MTSA) with dendritic cells (DCs) were investigated to elucidate the role of secretory antigens in regulating immune responses to M. tuberculosis early in the course of infection. MTSA induced the maturation of different DC subsets. The cytokine profiles of these DCs were characteristic to each DC subset. Of interest, coculture of