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Objective: To study the localisation of treponemes and to analyse the inflammatory infiltrate in biopsy specimens from patients with primary or secondary syphilis, or early infectious yaws., Materials and Methods: Skin biopsies originating from human lesions of primary (29x) or secondary (15x) syphilis (Rotterdam), or early yaws (18x) (West Sumatra) were studied. Different histochemical and immunohistochemical detection methods were used in this study., Results and Conclusion: The histochemical silver staining method according to Steiner revealed the presence of T. pallidum in all cases of primary syphilis studied. In 10 out of 14 cases of secondary syphilis, treponemes were demonstrated. With an immunofluorescence staining technique (IF) using anti-T. pallidum antiserum raised in rabbits (a-Tp), T. pallidum was demonstrated in 28 out of 29 cases of primary syphilis, and in 14 out of 14 studied cases of secondary syphilis. The silver staining method and IF showed identical localisations of T. pallidum (mainly in the dermal-epidermal junction zone or throughout the dermis). Using a-Tp antiserum in the indirect immunofluorescence technique, T. pertenue could be demonstrated in the dermis more often than with Steiner silver staining. However, epidermotropism of T. pertenue in yaws specimens was remarkable, compared with more mesodermotropism of T. pallidum; numbers of T. pertenue in the dermis were limited in all specimens. The dermal inflammatory infiltrate in primary and secondary syphilis was composed mainly of lymphocytes and plasma cells. In most cases more T (CD3 positive) cells than B (CD22 positive) cells were present. Regarding T cell subpopulations, in primary syphilis, T helper/inducer (CD4 positive) cells predominated in 86% of cases. In secondary syphilitic lesions, numbers of T helper/inducer cells were less frequent than or equal to T-suppressor/cytotoxic (CD8 positive) cells in 60% of cases. Remarkably, in yaws specimens the inflammatory infiltrate consisted mainly of IgG, but also IgA and IgM producing plasma cells. T or B lymphocytes were scarce, which is in sharp contrast with findings in syphilitic lesions.

Objective: To study ultrastructural aspects of infection with Treponema pertenue (Pariaman strain), originating from West Sumatra, Indonesia., Materials and Methods: Biopsy material originating from skin lesions in ten young children suffering from early infectious yaws in Indonesia, and rabbit testicular tissue inoculated with T pertenue. Human skin as well as rabbit testicular tissue was examined by means of conventional electron microscopy., Results: In human skin, treponemes were found in interepidermal spaces in 5 out of 10 specimens. In two of five positive specimens, treponemes were also seen in the dermis. In one out of five specimens from rabbit testicular tissue a profusion of treponemes was found lying in the interstitial myxomatous tissue. Microorganisms showed no adhesion to fibroblasts., Conclusion: This ultrastructural study of T pertenue demonstrated the scarcity and focal distribution of treponemes in tissue and did not reveal any morphological differences from the Gauthier strain of T pertenue. No differences from the ultrastructure of T pallidum were observed either.

The influence of storage at 4 degrees C on the survival of Treponema pallidum in donor blood, artificially infected with treponemes of the virulent Nichols strain was studied. Rabbits were inoculated in each testis with 0.5 ml of the blood-treponeme mixture, containing 5 X 10(5) microorganisms/ml, which was stored for up to 336 h. Under these circumstances, the blood-treponeme mixture appeared to be infectious for up to 96 h of storage, based on the demonstration of T. pallidum and serological findings, using FTA-ABS, TPHA and VDRL tests. The two former tests were found to be equally sensitive in detecting incubating syphilis in the rabbits, the VDRL test was significantly slower in this respect. From the absence of orchitis as well as seroconversion in rabbits, inoculated with blood-treponeme mixtures stored for 120 h or longer periods of time, it is concluded that after 120 h of storage infectivity is lost. The presently found survival time of treponemes in blood between 96 and 120 h of storage is discussed in relation with survival times found by other investigators.

The hypothesis that experimentally determined survival times of Treponema pallidum in stored donor blood could be related to the number of treponemes initially present in the treponeme-blood mixtures was investigated by inoculating rabbits with three graded doses of treponemes suspended in donor blood and stored at 4 degrees C for various periods of time. The storage periods up to which living treponemes could be detected in the testes of the inoculated rabbits as well as those storage periods up to which seroconversion occurred are related to the number of treponemes present in the treponeme-blood mixtures. Increasing numbers of treponemes present in the donor blood resulted in longer periods up to which positive results in both parameters were found. All available evidence suggests that the upper limit of seroconversion coincides with the upper limit of treponemal survival. Inoculation with 5 X 10(4) treponemes per ml of donor blood resulted in a treponemal survival time of 48 h, inoculation with 1.25 X 10(6) treponemes per ml in a treponemal survival time of 72 h and inoculation with 2.5 X 10(7) treponemes per ml donor blood in a survival time of 120 h.

The mucopolysaccharide nature of the material deposited in rabbit testes infected with Treponema pallidum was confirmed by histochemical staining with alcian blue. Differential staining of mucopolysaccharides showed the presence of sulphated mucopolysaccharides as an almost constant feature, whereas in little more than half of the orchitic testes studied variable deposits of hyaluronic acid were seen. The treponemes were almost exclusively present in the areas rich in mucopolysaccharide. A combination staining with the Warthin-Starry method and alcian blue showed treponemes in close association with pre-existing fibrils and cells contained in these fibrils. The latter findings were confirmed by electron microscopy, and the fibroblasts to which treponemes adhered displayed the characteristics of activated cells. The close parallel between the histopathological changes observed here and their descriptions in published reports shows that our specific strain still behaves the same as the original Nichols pathogenic strain of T pallidum.