Response In Leprosy Research Articles

Sensitization in vitro of human peripheral blood mononuclear cells to phenolic glycolipid 1 of Mycobacterium leprae in liposomes

Study of primary immune responses in leprosy has been limited, since disease becomes manifest long after infection or is not detectable. To study primary immune responses, we immunized in vitro human peripheral blood mononuclear cells from unexposed individuals using phenolic glycolipid 1 (PGL-1), an important water-insoluble antigenic constituent of Mycobacterium leprae. PGL-1, encapsulated in liposomes, induced lymphoproliferation or, less frequently, suppression of lymphoproliferation in 11 -day lymphocyte cultures. The primary lymphocyte responses resembled those elicited with keyhole limpet hemocyanin (KLH). HLA-DR2 expression, associated with tuberculoid leprosy, did not influence the outcome of in vitro sensitization. The association of HLA-DR2 and tuberculoid leprosy is not explained by differential ability to generate primary lymphoproliferative responses to PGL-1 or KLH. We have extended in vitro sensitization methodology to include a water-insoluble antigen in antigen-bearing liposomes. This methodology is potentially useful for studies of immunogenetics and immunopathology, and for vaccine research.

Anti-Mycobacterium Leprae Monoclonal Antibodies Cross-React with Human Skin: An Alternative Explanation for the Immune Responses in Leprosy

A panel of 17 mouse monoclonal antibodies (MoAb) raised against Mycobacterium leprae (M. leprae) antigens was used to detect antigenic determinants in normal human skin. An indirect immunoperoxidase technique was used. Eight of the MoAb detected epidermal antigens similar to patterns well known for human sera. Five of these MoAb detected determinants in the dermis, too. These observations may indicate a certain degree of similarity between the antigenic determinants occurring in M. leprae and in the human host. We propose that such a similarity on the one hand may facilitate the survival of M. leprae in the human host when the antigens are not recognized as "non-self," a situation which seems to occur in lepromatous leprosy, when the patients' tissues are loaded with bacteria virtually without any immune response. On the other hand, M. leprae antigens which mimic host antigens may induce an auto-immune reaction against the host's own antigens, which could explain the immune reaction in tuberculoid leprosy and during a "reversal reaction" when M. leprae is not observed in the host tissues, but extensive granuloma formation occurs.

Isolation and characterization of the highly immunogenic cell wall-associated protein of Mycobacterium leprae.

In a recent study, we demonstrated that certain reactivities crucial to the immune response in leprosy are due to protein associated with the cell wall peptidoglycan "core" of Mycobacterium leprae. We now describe a primary method for the isolation of a highly immunogenic, large molecular-size, cell wall protein (CW-P) complex from M. leprae, freed of soluble proteins, bound mycolates, arabinogalactan, and much of the peptidoglycan. The complex is of apparent relative molecular size 2 x 10(6) to 20 x 10(6) Da, is distinguished by a high content of Ala, Gly, Leu, Asx, and Glx, and some peptidoglycan, and represents up to 7% of the bacterial mass. It is stable to a variety of dissociation and reductive processes and, in accord with its size, is not resolvable by polyacrylamide gel electrophoresis. The mAb to the CW-P complex also react with the heat shock 65-kDa protein of M. leprae. Conversely, antibodies that recognize internal epitopes within the polypeptide chain of the heat shock protein also react with CW-P; however, antibodies that recognize the N and C termini of the 65-kDa protein fail to react with CW-P, and some anti-CW-P mAb do not recognize any of the soluble proteins of M. leprae. Alternate methods to derive the large peptidoglycan-associated protein result in lower yield and less of the associated heat shock protein, implying that the 65-kDa protein may not be crucial to the immunogenicity of the complex. In an accompanying paper, we demonstrate that T cell clones raised to CW-P also selectively recognize soluble proteins, primarily of 7-kDa and 16-kDa size. Thus, the image of the CW-P complex of M. leprae is of a few immunoreactive polypeptides in avid association with a modicum of peptidoglycan to which the 65-kDa polypeptide may be variably attached, perhaps due to involvement in assembly of the complex.

T cell responses to fractionated Mycobacterium leprae antigens in leprosy. The lepromatous nonresponder defect can be overcome in vitro by stimulation with fractionated M. leprae components.

Protective immunity against Mycobacterium leprae is dependent on M. leprae-reactive T lymphocytes. M. lepare-directed T cell reactivity is high in the localized tuberculoid form of leprosy but specifically absent in the disseminated lepromatous type of the disease. Two important questions that are relevant for the understanding of the immune response in leprosy as well as for the design of rational immunoprophylaxis and -therapy strategies are: (a) what are the antigens that trigger T cell responses in tuberculoid patients and thus protect these individuals from developing lepromatous leprosy and (b) is it possible to restore T cell responsiveness to M. leprae in lepromatous patients by rechallenging the immune system with selected antigens that will trigger help but not suppression? We have addressed these question by directly probing the peripheral T cell repertoire of 10 tuberculoid and 18 lepromatous patients with large numbers of different M. leprae and BCG antigenic components that had been separated on the basis of their relative molecular mass (Mr) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotted onto nitrocellulose. This technique allows the identification of T cell-stimulating antigens independent of the expression of B cell epitopes by these antigens. So far T cell epitopes have only been mapped on M. leprae proteins that had previously been defined by antibodies. Our results show that: (a) tuberculoid patients' T cells responded preferentially to M. leprae and BCG antigens in the lower (i.e. less than 70 kDa) Mr range with a peak in the 10-25 kDa range; (b) 6 out of 18 lepromatous patients that did not respond to whole M. leprae responded strongly to isolated M. leprae components; antigens in the lower Mr. range were recognized by five out of these six patients and thus commonly seen by both tuberculoid and lepromatous patients' T cells; however, antigens in the higher Mr range, in particular greater than 150 kDa, were only recognized by lepromatous patients' T lymphocytes; (c) furthermore, the T and B cell repertoires in leprosy patients are skewed towards different antigenic fractions.

Studies on humoral immune response in leprosy.

Our study was undertaken to determine if the humoral immune status of leprosy patients varied with either the type of the disease or the geographic location of the disease. Another variable investigated was whether or not treatment versus non-treatment had any selected effect on the humoral immune status. Serum immunoglobulin levels were determined by single radial immunodif f usion and other immunoserologic profile testing was performed whenever possible. Immunoglobulin levels did show a difference between the different disease types of leprosy and the authors also feel that environmental and genetic factors may play a more significant role than previously thought.

Immunological Problems in Leprosy Research

This Memorandum reviews the present status of knowledge of the immunology of leprosy, with particular attention to developments since the publication of a similar review in 1970. The different types of lepromin reaction and their significance in healthy contacts and in patients with tuberculoid and lepromatous leprosy are discussed. The immunological responsiveness of patients with leprosy is also considered, with special attention to in vitro methods for evaluating this response. Part 2 of the Memorandum will cover possible mechanisms of altered immune response in leprosy (including a tentative scheme to explain the possible genesis of the lepromatous lesion); genetic, nutritional, and hormonal factors; the possibility of vaccination; attempts at immunotherapy; and areas in which further research is needed. A detailed protocol for evaluating the effect of transfer factor in leprosy will be included as an annex.

Dissociation of the Aggregates Obtained on Adding Beef-Heart Lipoid to Wassermann-Positive Sera

Summary The salt-dissociation method has proved applicable to the recovery of the Wassermann reagin, not only from syphilitic but also from leprous sera. The significance of this finding for the interpretation of the immunological response in leprosy is briefly discussed. Preliminary experiments on the preparation of purified solutions of the Wassermann reagin are also reported.