Alpha 2-macroglobulin and alpha 2-macroglobulin/proteinase complexes in human seminal fluid.

OBJECTIVE: To determine the broad-spectrum proteinase inhibitor alpha 2-macroglobulin (alpha 2M) and its functional subforms, i.e., free alpha 2M and alpha 2M/proteinase complexes, in human seminal fluid by using specific enzyme immunoassay systems. SETTING: The study has been performed in the Andrology Department of the Dermatology Clinics and the Laboratory of Immunopathology of the Institute of Immunology. PATIENTS, PARTICIPATES: Routine patients attending the Andrology Department. INTERVENTIONS: The data have been obtained without particular interventions before or after collection of seminal fluid. MAIN OUTCOME MEASURES: The aim of the study was to determine whether alpha 2M or alpha 2M/proteinase complexes are present in human seminal fluid. RESULTS: The concentration of total alpha 2M in human seminal fluid ranged from 1 to greater than 1,000 ng/mL, and between 56% and 85% of the inhibitor was complexed with proteinases. CONCLUSIONS: These findings show that alpha 2M and alpha 2M/proteinase complexes have to be considered as functionally relevant biomolecules in male genital tract secretions.

Establishment of an enzyme-linked immuno-sorbent assay for urinary trypsin inhibitor by using a monoclonal antibody.

Monoclonal antibodies against inter-alpha-trypsin-inhibitor (ITI) were produced. One clone showing specificity for urinary trypsin inhibitor (UTI), a proteolytic fragment of ITI, which is excreted into urine, was selected for the establishment of an enzyme-linked immuno-sorbent assay (ELISA). The ELISA for the quantification of UTI was shown to work reproducibly in the range between 0.5 and 10 ng UTI/ml urine. Urines of several patients suffering from different lung diseases were screened for UTI using the established ELISA. Highest UTI levels were found in the urine of patients with lung empyema. A more moderate increase was observed in patients suffering from lung tuberculosis and from secondary and primary lung tumors.

Quantification of free alpha 2-macroglobulin and alpha 2-macroglobulin-protease complexes by a novel ELISA system based on streptococcal alpha 2-macroglobulin receptors.

An ELISA test system has been developed for the quantification of the two distinct forms of the proteinase inhibitor alpha 2-macroglobulin (alpha 2M): (i) free alpha 2M (functionally active), which is the electrophoretically slow form (alpha 2 MS), and (ii) the alpha 2 M-proteinase complex (functionally inactive), which is the electrophoretically fast form of alpha 2 M (alpha 2 MF). Discrimination between the two types of alpha 2 M was achieved using extracts of the two independent streptococcal strains, M1 and Sc1, which express receptors for alpha 2 MS and alpha 2 MF, respectively, in combination with a monoclonal antibody specific for alpha 2 M. The assay system described is easy and reliable and permits quantitation of alpha 2 MS and alpha 2 MF in complex biological samples such as plasma and cutaneous suction blister fluid.

Demonstration of antigen-specific T cells and histopathological alterations in mice experimentally inoculated with Borrelia burgdorferi.

Antigen-specific T-cell responses and histopathological changes were studied in mice experimentally inoculated with Borrelia burgdorferi B31. Inbred mice with different H-2 haplotypes and/or different genetic backgrounds were inoculated with B. burgdorferi organisms and tested for antigen-specific T-cell responses in vivo (delayed-type hypersensitivity [DTH]) and in vitro (T-cell proliferation). Comparable DTH responses were found after inoculation with either inactivated (in the presence of adjuvants) or viable microorganisms in all mouse strains, except BALB/c, irrespective of the H-2 haplotype (b, d, k, or s) tested and the sex of the animals. Moreover, in mice presensitized to B. burgdorferi, DTH responses could be induced only with antigen preparations derived from the corresponding strain but not with those obtained from either related spirochetes such as Treponema phagedenis and Leptospira interrogans or unrelated bacteria such as Mycobacterium tuberculosis. T cells isolated from lymph nodes or spleens of mice previously sensitized to B. burgdorferi but not those from naïve mice could be induced for antigen-specific proliferation in vitro, as revealed by [3H]thymidine incorporation. Histopathological examination of mice inoculated with viable B. burgdorferi organisms revealed significant perivascular infiltrates consisting mainly of mononuclear and a few polymorphonuclear leukocytes in different organs (brain, heart, lungs, liver, and kidneys) and the appearance of giant multinucleated cells within the spleen similar to those found in human skin specimens of patients suffering from cutaneous manifestations of Lyme disease. Our findings suggest that mice are a suitable animal model with which to study the immune response to B. burgdorferi and the pathogenesis of Lyme disease.