Association of vitamin D receptor genotype with leprosy type.

Host genetic factors including major histocompatibility complex (MHC) polymorphisms influence both susceptibility to leprosy per se and also to leprosy type. Non-MHC genes may play an important role, but such genes remain undefined. The influence of two non-MHC candidate genes was assessed in a case-control study of Bengali leprosy patients from Calcutta. Recent studies have implicated variation in the vitamin D receptor (VDR) gene in susceptibility to several diseases, including osteoporosis and pulmonary tuberculosis. In this population, homozygotes for the alternate alleles of the VDR polymorphism are associated, respectively, with lepromatous and tuberculoid leprosy. The NRAMP1 (natural resistance associated macrophage protein 1) gene may influence human mycobacterial disease susceptibility based on studies with the murine homologue Nramp1. However, no significant association was found between NRAMP1 and leprosy susceptibility. This study suggests that the VDR polymorphism may influence susceptibility to some diseases by affecting the type and the strength of the host immune response.

Tumor necrosis factor promoter polymorphism and susceptibility to lepromatous leprosy.

Genetically determined differences in immune responses to environmental agents may underlie susceptibility to many autoimmune and infectious diseases. Leprosy provides an example of a polarity in the type of immune response made to an infectious agent, and there is evidence that the major histocompatibility complex is genetically linked to leprosy type. It was found that HLA-DR2 is associated with both tuberculoid and lepromatous types of leprosy; however, a variant at position -308 of the promoter of the neighboring tumor necrosis factor (TNF) gene was increased in frequency in lepromatous (odds ratio = 3.0, P = .02) but not tuberculoid leprosy. Some studies have found higher serum levels of TNF in lepromatous than tuberculoid leprosy, and high TNF levels are found in malaria and leishmaniasis, which are also associated with this TNF allele. It is speculated that this association reflects genetic variability in cytokine production, which influences the immune response to and clinical outcome of leprosy.

Endemic disease, nutrition and fertility in developing countries.

PIP: Subfecundity is caused by disease and nutrition as well as by genetic, environmental, and psychological components. Sexually transmitted diseases (STDs) are caused by 21 different pathogens of which syphilis, gonorrhea, and chlamydia are the most important. Syphilis is caused by the bacterium Treponema pallidum with incidence of 10% in Thailand. 20% in Papua New Guinea, and 40% in Ethiopia. Stillbirths in infected mothers range from 66% to 80%. Gonorrhea is caused by the bacterium Neisseria gonorrhoea and its incidence was 18% in female patients in Ugandan clinic. 20% of women in Africa with cervical gonorrhea develop salpingitis. The risk of pelvic inflammatory disease is several times higher in IUD users. The bacterium Chlamydia trachomatis caused infertility in 15.4% of men in a 1991 study. Herpes simplex virus 2 infects 15-30% of sexually active adults, and the chance of fetal transmission is 40% when maternal lesions are present. Diseases other than STDs include tuberculosis (TB) whose development is aided by conditions such as malnutrition, malaria, leprosy, syphilis, and African sleeping sickness. Genital TB causes a 5-50% rate of menstrual disorders including amenorrhea and a 55-85% rate of sterility in women. Malaria is caused by Plasmodium protozoa, and the feverish state included by it can lead to oligospermia. Severe malarial anemia can lead to fetal and maternal mortality. The protozoa Trypanosoma causes African sleeping sickness that produces azoospermia and impairs the pituitary gland and ovaries. Schistosomiasis (bilharzia) and filariasis have less direct effect on fecundity but they negatively impact nutritional status. Maternal nutrition substantially impacts fetal and infant survival. During the Dutch famine of 1944-45 there was a 50% decrease in births 9 months subsequently. A 10-15% weight loss results in amenorrhea.^ieng

Skin test studies on close contacts of leprosy patients in India.

Skin-test studies with a series of tuberculins have been carried out in close contacts of multibacillary (MB) leprosy patients around three leprosy centers in India, and casual contacts of the disease around two centers. The results show that the rate of acquisition of leprosin A positivity is associated with age and the closeness of contact with MB leprosy. At the age of 15 years, the differences between the two types of contact were highly significant (p less than 0.00001). Many responses to leprosin A are directed toward the group iv species-specific, antigens of the leprosy bacillus, and the significance of positivity is discussed in relation to protective immunity from leprosy. The differences from Iran show that positivity to leprosin A is not solely the effect of the degree of contact with the disease, but must also have a genetic or environmental element, the latter being favored. The results from Miraj show that the high levels of tuberculin, scrofulin, and vaccin positivity seen in Fathimanagar, and to a lesser extent in Karigiri, are not a consequence of contact with leprosy. BCG vaccination made little difference to the leprosin A positivity of close contacts of leprosy patients, although it significantly enhanced positivity among casual contacts around Miraj (p less than 0.002). BCG vaccination significantly increased tuberculin positivity in Miraj and Karigri, and in those under 11 years of age in Fathimanagar. It made no difference to the already high level of positivity found in older persons around Fathimanagar.(ABSTRACT TRUNCATED AT 250 WORDS)

Tissue and blood T-lymphocyte subpopulations in erythema nodosum leprosum.

To study T lymphocytes in erythema nodosum leprosum (ENL), monoclonal antibodies were used to identify T-lymphocyte subpopulations in the blood and skin lesions of patients with ENL and patients with nonreactional lepromatous leprosy. The blood of nonreactional lepromatous patients had a lymphopenia and a proportionate reduction in pan T cells, helper-inducer, and suppressor-cytotoxic subsets, but a normal helper-suppressor ratio, as compared with controls. Patients with ENL did not differ significantly from the controls. In skin lesions, an admixture of helper and suppressor phenotypes among foamy histiocytes was found. The ENL tissue had more numerous cells of the helper-inducer phenotype and fewer of the suppressor-cytotoxic phenotype, as compared with nonreaction lepromatous tissues. In 22 patients with simultaneous examination of tissue and blood T-cell subsets, there was no correlation between tissue and blood helper-suppressor ratios, indicating that some sort of selection process brings lymphocytes into tissues from peripheral blood.

Comparison of S-100 and OKT6 antisera in human skin.

The monoclonal antibody OKT6 and antisera against S-100 protein have both been advocated as immunologic markers of Langerhans cells in the skin. S-100 antiserum has an advantage in its ability to stain Langerhans cells in paraffin tissues. In order to evaluate whether these antibodies stain equivalent numbers of Langerhans cells in skin, we compared the staining patterns of S-100 antiserum and OKT6 antibody on biopsy specimens from 40 patients with leprosy using immunoperoxidase techniques. Utilizing OKT6 antibody, greater numbers of positive Langerhans cells were found in the epidermis in tuberculoid leprosy, reversal reaction, and erythema nodosum leprosum than in lepromatous leprosy. However, these differences were not observed with the S-100 antiserum and, overall, fewer cells were found as compared with the OKT6 antibody. In the dermis both antibodies stained "dendritic cells" that were found encircling granulomas in tuberculoid leprosy and reversal reaction. Staining in lepromatous leprosy granulomas, in contrast to the epidermal staining pattern, revealed rare OKT6-positive cells, while S-100 cells were numerous and were more diffusely distributed throughout the granuloma. Our results indicate that antiserum to S-100 protein and OKT6 antibody stain morphologically similar cells (dendritic cells), but do not provide comparable results concerning distribution and frequency of these cells.

In situ identification of cells in human leprosy granulomas with monoclonal antibodies to interleukin 2 and its receptor.

Leprosy is a chronic granulomatous disease with an immunologic spectrum in which lepromatous leprosy patients have defective cell-mediated immune responses, in comparison to tuberculoid leprosy patients. Immunoregulatory aspects of this spectrum were investigated by using monoclonal antibodies to interleukin 2 (IL 2), IL 2 receptors (Tac), and T lymphocyte subpopulations with immunoperoxidase techniques on frozen sections of skin biopsy specimens from 10 tuberculoid and 10 lepromatous patients. A comparison of IL 2+ cells revealed markedly fewer IL 2+ cells in lepromatous specimens (lep. 0.028% +/- 0.02 vs tub. 0.46% +/- 0.28, p less than 0.001). These IL 2+ cells were large, exhibited cytoplasmic staining, and on double immunostaining were Leu-4+, Leu-3a+, Leu-2a-, Tac-, and OKT6-, consistent with the fact they are IL 2 producers. Equivalent numbers of Tac+ cells were observed in both lepromatous and tuberculoid granulomas (lep. 1.5% +/- 0.5 vs tub. 2.1% +/- 0.7, p, NS), suggesting that the responder cells are present in both conditions. The tuberculoid granuloma was highly organized, composed of a central core of mature macrophages, Leu-3a+ and Tac+ cells with a surrounding mantle of Leu-2a+, Leu-3a+, IL 2+, Tac+, and OKT6+ cells. In lepromatous granulomas, Leu-2a+, Leu-3a+, Tac+, and rare IL 2+ cells were randomly admixed with bacilli-laden macrophages. The defective cell-mediated immune responses in lepromatous leprosy appears to be associated with diminished IL 2 production and disorganization of the granuloma.

In situ characterization of T lymphocyte subsets in the reactional states of leprosy.

Using monoclonal antibodies and the immunoperoxidase technique, the numbers and distribution of T lymphocyte subsets in the tissues of reactional states of leprosy (six reversal reaction, nine erythema nodosum leprosum (ENL) and two Lucio's reaction) were determined and compared with those found in stable, non-reactional patients (six tuberculoid, two borderline lepromatous and seven lepromatous). The pattern of segregation of the suppressor/cytotoxic phenotype at the periphery of the granuloma was found in both non-reactional tuberculoid lesions and reversal reactions, but was better developed in the former. In ENL and Lucio's reaction, as well as in non-reactional lepromatous tissue, the helper/inducer and suppressor/cytotoxic phenotypes were both admixed with the aggregated histiocytes. However, the helper/suppressor ratio in ENL (2.1 +/- 0.4) was significantly larger than that in non-reactional lepromatous tissue (0.7 +/- 0.4, P less than 0.001). The immature thymocyte antigen OKT6 was found on scattered large non-lymphoid cells, most commonly in tuberculoid and reversal reaction tissues, less commonly in ENL, but only irregularly in non-reactional lepromatous tissue. The peripheral pattern of the suppressor/cytotoxic phenotype may be an immunohistological reflection of a cell-mediated immune response common to both non-reactional tuberculoid and reversal reaction patients. The reversal of the helper/suppressor ratio in ENL as compared to non-reactional lepromatous disease suggests some role for cell-mediated immunity in the pathogenesis of ENL. The OKT6 positive cell is of unknown origin and function.

Immunoperoxidase techniques applied to dermatopathology.

Immunoperoxidase techniques provide the pathologist with the capability for staining a wide range of antigens in tissue sections. More than 100 different antigens have been successfully demonstrated in fixed paraffin sections; other antigens can only be visualized in frozen sections. This latter group particularly includes lymphocyte surface antigens detectable by monoclonal antibodies. This review describes the current state of the art and provides several illustrations of the use of monoclonal antibodies for the identification of T-lymphocyte phenotypes in frozen section from cases of leprosy, mycosis fungoides, halo nevus, Kaposi's sarcoma, lichen planus and atopic dermatitis. Technical details and potential applications are discussed. The growing availability of commercial immunostaining kits makes these techniques more accessible to the surgical pathologist; indeed a whole new range of truly specific, special stains are available, as pathologists we must simply learn to use them.

In situ demonstration of T lymphocyte subsets in granulomatous inflammation: leprosy, rhinoscleroma and sarcoidosis.

T lymphocyte subpopulations in frozen tissue sections of four granulomatous conditions (five patients with tuberculoid leprosy, five with lepromatous leprosy, seven with sarcoidosis and four with rhinoscleroma) were studied using monoclonal antibodies and a modified immunoperoxidase technique. Two immunohistological patterns were observed. In tuberculoid leprosy and sarcoidosis, lymphocytes expressing the helper/inducer phenotype were present within the aggregates of mononuclear phagocytes (epithelioid cells); however, cells with the suppressor/cytotoxic phenotype were predominantly in the lymphocytic mantle surrounding each granuloma. In lepromatous leprosy and rhinoscleroma the helper/inducer T cells and suppressor/cytotoxic T cells were both diffusely distributed among the mononuclear phagocytes (histiocytes) without any discernible mantle. The segregation of the helper/inducer and suppressor/cytotoxic phenotypic subsets was associated with an epithelioid cell differentiation of mononuclear phagocytic cells, bacterial elimination and a delayed type hypersensitivity response. The intimate admixture of helper/inducer and suppressor/cytotoxic subsets was associated with undifferentiated mononuclear phagocytes, bacterial proliferation and the absence of a delayed type hypersensitivity response. Thus the different distributions of T cell subpopulations in granulomas may be associated with differences in the host's immune response in several forms of granulomatous reactions.

T lymphocyte subsets in the skin lesions of patients with leprosy.

Lymphocyte subsets in the tissues of fourteen patients with leprosy were studied using monoclonal antibodies and a modified immunoperoxidase technic. Two immunohistologic patterns were observed. In tuberculoid leprosy, helper-inducer cells were present among the aggregates of mononuclear phagocytes (epithelioid cells), but the suppressor-cytotoxic cells were predominantly in the lymphocytic mantle surrounding the epithelioid cell aggregates. In reversal reaction and lepromatous tissues, the helper-inducer and the suppressor-cytotoxic cells were both distributed among the mononuclear phagocytes (histiocytes). In tuberculoid specimens the Langerhans cells of the epidermis were increased in number as compared to lepromatous and normal tissues. The technic used appears to be of value in studying some of the cellular components of the immune response in situ.

The histopathology of the spleen from a patient with lepromatous leprosy.

The histopathology of the spleen from a young man with diffuse non-nodular lepromatous leprosy is reported. As judged by this case, other case reports, and necropsy series, involvement of the spleen in lepromatous leprosy is characterized by aggregations of large vacuolated histiocytes, containing individual bacilli and globi, in both the red and white pulp. In the white pulp the histiocytes localize about the arterioles. Findings in the present case, which may represent a comparatively early change, include numerous, small germinal centers containing nonaggregated large, vacuolated histiocytes with intracellular bacilli.

Serum and tissue lysozyme in leprosy.

Mean serum lysozyme values were found to be elevated in untreated leprosy patients. Statistically significant elevations were present in each of the three major categories of leprosy, tuberculoid, borderline, and lepromatous. Values were particularly high in patients with severe reversal reactions or Lucio's phenomenon. Prolonged sulfone therapy was associated with a fall in serum lysozyme values. With an immunoperoxidase method to localize lysozyme in leprous tissues, two distinct staining patterns were found, granular and saccular. The grandular pattern of lysozymal staining was found in epithelioid cells and in giant cells, and the intensity of staining showed a positive correlation with serum lysozyme levels. Conversely, a saccular pattern of lysozymal staining was found in lepromatous histiocytes, buth the intensity of staining was unrelated to serum lysozyme levels; the saccular structures contained dense aggregates of Mycobacterium leprae. These two patterns of staining probably represent different functional responses of monocyte-derived granuloma cells, whereas the serum levels reflect, to a varying degree, both the absolute number of such cells and the rate of secretory activity of this cell population as a whole.

Separation of M. leprae from human leproma and the development of a cytoplasmic skin test antigen from purified bacilli.

A successful method for purification of M. leprae from human leproma without subjection to heat has been developed. The "floater" phenomenon has been described which consists of bacillary tendency to float in the supernate when bacilli which are not autoclaved are separated from tissues by enzymatic digestion. A method for preparing cytoplasmic fractions from purified M. leprae has been developed for the production of a skin test antigen for leprosy. The cytoplasmic fraction of M. leprae elicited positive skin test responses in people with tuberculoid leprosy and negative responses in lepromatous leprosy. Cytoplasmic preparations from purified M. leprae had little cross-reactive relationship with the organism BCG. The small particulate fraction elicited positive reactions in PPD-S negative as well as BCG vaccinated individuals.