The vagina of women infected with Trichomonas vaginalis has numerous proteinases and antibody to trichomonad proteinases.

BACKGROUND: Patients with trichomoniasis have serum antibody to numerous T. vaginalis cysteine proteinases, indicating that the proteinases are expressed in vivo. It was important, therefore, to examine for the presence of soluble trichomonad proteinases and/or antibody to the proteinases in the vagina of infected women. METHODS: Vaginal washes (VWs) from 20 women were examined for the presence of proteinases by electrophoresis using acrylamide co-polymerised with gelatin as the indicator system. Antibody to proteinases in VWs was detected by an immunoprecipitation assay involving protein A-bearing Staphylococcus aureus first coated with anti-human immunoglobulin G (IgG) antibody, which was then added to VWs. For VWs having soluble proteinases, the bacteria were used to determine whether immune complexes between antibody and proteinases were present. VWs without soluble proteinases were incubated with the anti-human IgG treated bacteria before adding to detergent extracts of T. vaginalis. Individual isolates from the patients examined in this study were also analysed by one- and two-dimensional electrophoresis for their proteinase content. Finally, VWs were from patients without any history of other sexually transmitted diseases (STDs) as well as from individuals having numerous other STDs, including yeast, group B streptococcus, chlamydia, and syphilis. RESULTS: Approximately one-third of patients had soluble proteinases in the VWs; the remaining two-thirds (70%) of patients and normal women had no detectable proteinases in VWs. Half of the patients without soluble proteinases had IgG which, when bound to S. aureus, immunoprecipitated many proteinases from a detergent extract of T. vaginalis. All soluble proteinases and those precipitated from trichomonal extracts were inhibited by inhibitors of cysteine proteinases. Finally, patients having trichomoniasis in addition to numerous other STD agents, including yeast, group B streptococcus, chlamydia, and syphilis did not have soluble proteinases in VWs. Equally noteworthy, some patients with soluble proteinases in VWs did not have other detectable STD agents. CONCLUSIONS: Proteinases were detected in the vagina of some patients with trichomoniasis, and in most cases the proteinases were complexed with IgG, which was precipitated by S. aureus. Patients without soluble proteinases in VWs also had antibody specifically to trichomonad proteinases, again demonstrating both the expression and immunogenic nature of the proteinases in vivo. The absence of soluble proteinases in normal women and in patients having other STD agents as well as the presence of proteinases in VWs of patients without other detectable STD pathogens reinforced the idea that the proteinases were of T. vaginalis parasite origin. The findings of this study indicate that proteinases may be important to the T. vaginalis-host interrelationship.

Antibody in sera of patients infected with Trichomonas vaginalis is to trichomonad proteinases.

BACKGROUND: A recent report demonstrated the immunogenic character of the cysteine proteinases of Trichomonas vaginalis. It was of interest, therefore, to examine for the presence of serum anti-proteinase antibody among patients with trichomoniasis. METHODS: An immunoprecipitation assay was used involving protein A-bearing Staphylococcus aureus first coated with the IgG fraction of goat anti-human Ig and then mixed with individual sera of patients to bind human antibody. These antibody-coated bacteria were then added to detergent extracts of T vaginalis. Bound immune complexes on S aureus were washed and solubilised for electrophoretic analysis on acrylamide copolymerised with gelatin for detection of proteinase activity. RESULTS: Sera from patients (50/50), but none from sera of normal, uninfected women, possessed IgG to numerous trichomonad cysteine proteinases. The presence of this serum anti-proteinase antibody disappeared after drug treatment and cure of patients of the T vaginalis infection. CONCLUSIONS: The commonality of the anti-proteinase antibody in the sera of patients with trichomoniasis provided evidence for the expression of the same repertoire of parasite proteinases during infection. These observations have important implications for the in vivo relevance of the proteinases and indicate that strategies to use a specific serum antibody response for diagnosis of this infection may be possible.

Vaginal antibody of patients with trichomoniasis is to a prominent surface immunogen of Trichomonas vaginalis.

Twenty vaginal washes (VWs) and ten vaginal mucus (VM) samples from patients with trichomoniasis were examined for the presence of antibody to surface protein immunogens of Trichomonas vaginalis. Fourteen of 20 VWs (70%) and 8 of 10 VM (80%) had immunoglobulin G (IgG) antibody (Ab) that reacted in an immunoprecipitation (IP) assay with one iodinated Trichomonas vaginalis surface protein immunogen with a relative molecular mass of 230,000 daltons (230-kDa) (P230). No similar IP of any iodinated protein was observed when detergent extract was first depleted of P230 with monoclonal antibody (MAb), indicating a highly specific VW IgG response of patients to P230. VWs were also obtained from 10 patients from one to four weeks after treatment. These VWs had the same, or in one case a greater, level of IgG to P230. Under no circumstances was Ab to P230 or any other trichomonad protein detected in VWs or VM from normal, uninfected women. Flow cytofluorometry with VW Ab yielded heterogeneous fluorescent and non-fluorescent populations of trichomonads, reaffirming the restricted Ab response to one or a few epitopes on P230 in the vagina of patients. Under identical conditions, the MAb gave totally fluorescent parasite populations of some isolates, and the MAb again demonstrated variable epitope accessibility to Ab binding (Infect Immun 1987;55:1037). Finally, the MAb or VW Ab was never cytolytic for immunoreactive (fluorescent) parasites, even in the presence of complement. This study identifies the most important trichomonad surface immunogen on the basis of the vaginal Ab response, and data underscore the significance of immune evasion strategies of this sexually transmitted disease agent.

Analysis of the proteinases of representative Trichomonas vaginalis isolates.

Isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with acrylamide copolymerized with gelatin (substrate-SDS-PAGE) were combined to evaluate the proteinases of both long-term-grown and fresh isolates of Trichomonas vaginalis. This two-dimensional substrate-SDS-PAGE resolved as many as 23 distinct proteinase activities in several isolates, and proteinases had relative molecular masses between 23 and 110 kilodaltons (kDa). Isoelectric points (pI) of proteinases ranged from 5.7 to 7.0. Overall, the various representative proteinase profiles were similar among those of long-term-grown and fresh isolates, although heterogeneity existed among several cysteine proteinase activities. Pattern changes were detected in fresh isolates passaged over several weeks, showing the ability of proteinases to be differentially expressed and to undergo phase variation. The two-dimensional proteinase patterns were very reproducible for isolates analyzed over a certain period of time before expression of some proteinases varied. The heterogeneity and differential expression of certain proteinases were not coordinated with phenotypic variation of already characterized immunogens and adhesins. Data suggesting that a 43-kDa proteinase resided on the parasite surface were obtained on the basis of removal of activity following pronase or proteinase K treatment of live organisms. Finally, immunized experimental animals produced antibody to many T. vaginalis proteinases, which indicates the immunogenic nature of trichomonad proteinases.

Relatedness of structures of a major immunogen in Trichomonas vaginalis isolates.

Solubilization of live Trichomonas vaginalis organisms with detergent caused the release of cysteine proteinases in the detergent extract which were inhibitable with N-alpha-p-tosyl-L-lysine chloromethyl ketone. The detergent extracts of all isolates tested possessed similar cysteine proteinase activities. These parasite proteinases rapidly degraded a prominent immunogen whose surface disposition undergoes phenotypic variation in some isolates. The relatedness of the forms of this immunogen among all isolates tested was confirmed by identical immunoblot patterns of autolysed immunogen, and data suggest the presence of repeating units or at least equidistant sites for proteinase cleavage within the immunogen molecule.

Sore throats in adults: who sees a doctor?

In a study looking at the relationship of life stress events to health status, 500 individuals randomly selected from family practices in Hamilton, Ontario were asked to keep a health diary for three days every two weeks over a two year period. The compliance with diary keeping was remarkably high (85%), partly due to a novel method of reinforcing compliance. The present study involved reviewing these health diaries for symptoms of sore throat during the three month period January to March 1979. Over 2,700 diaries representing 8,148 person/days were reviewed; 48% recorded at least one symptom and 5.2% of all diaries recorded sore throat on at least one day. Eight to 16% of those individuals recording sore throat as a symptom contacted a health professional and/or took prescribed drugs.Although the group under study included only adults, the low medical contact rate of patients with sore throats raises questions about the effectiveness of any approach to sore throat/pharyngitis adopted in office practice.

Methodological issues in developing a scale to measure social support.

In recent years there has been considerable research and clinical interest in developing instruments to assess social supports available to individuals. There is, however, a notable deficiency of attempts to evaluate the psychometric properties of these questionnaires. The present article describes efforts made to evaluate the properties of a Social Relationship Scale (SRS) that was developed as part of a prospective study of the psychosocial influences on the health status of a population. Some descriptive scale statistics are also presented.