HLA markers in familial Lichen Sclerosus

Background: Lichen sclerosus (LS) has been identified with increased frequency in families,often associated with HLA markers, mainly DQ7. A genetic co‐etiology seems likely in this setting. Moreover, there is an association of LS with autoimmune disorders, such as the presence of anti‐thyroid peroxidase autoantibodies (anti‐TPO), a hallmark of autoimmune thyroid diseases. Patients and Methods: In 3 families affected by LS, we verified their HLA markers, and identified previously undiagnosed cases of LS and autoimmune disorders. 30 individuals were examined with history, skin biopsy, HLA class I and II typing by PCR‐SSP, and measurement of anti‐TPO, free thyroxine and thyroidstimulating hormones (TSH) levels. Results: There were 8 cases of LS, 50 % of them anti‐TPO+. Autoimmune disorders were found in 40 % (total) and in 87.5 % of those affected. Most common HLA markers were B*15, B*57, CW*03, CW*07, CW*18, DRB1*04, DRB1*07, DRB4*. The three latter have been previously associated with LS. Conclusion: New cases of LS and autoimmune disorders can be detected in first degree relatives of patients with LS. The presence of anti‐TPO antibodies strongly suggests autoimmune thyroiditis. There is intra‐familial association between the haplotype HLA‐B*15 ‐DRB1*04 ‐DRB4* and anti‐TPO,emphasizing their link with thyroiditis. New familial approaches might help to make clear the pathogenesis of LS and its association with autoimmune diseases.     

A Comparison of Surface and Cytoplasmic Antigens of Aspergillus fumigatus in an Enzyme-Linked Immunosorbent Assay (ELISA)

Summary: Cytoplasmic antigens of Aspergillus fumigatus were prepared by exposing the mycelium to a lytic enzyme system from Trichodenna hanianum to yield protoplasts. Following physical rupture, the watersoluble constituents gave multiple peaks with homologous antiserum on 2-dimensional immunoelectrophoresis. The same system was used to identify reactive terminal sugar residues on antigen molecules by the incorporation of lectins of appropriate specificities in an intermediate gel. On affinity chromatography on Conmnavalin A (ConA) - linked Sepharose, the bulk of the applied cytoplasmic material eluted in the void volume. The elution profile obtained on column chromatography of the extract on wheat germ agglutinin (WGA)-linked Sepharose showed two major components, one of which was elutable only with N-acetyl-D-glucosamine. Cytoplasmic antigens and antigens derived from the mycelial wall by detergent extraction showed comparable reactivity towards homologous antisera when tested in an en-zymelinked immunosorbent assay (ELISA). In the same system, the wallassociated antigens proved far superior in the detection of specific IgG in sera from patients with aspergillus-related diseases. Zusammenfassung: Die zytoplasmatischen Antigene von Aspergillus fumigatus wurden gewonnen, in dem das Myzel einem lytischen Enzymsystem von Trichoderma harzianum ausgesetzt wurde, um so Protoplasten zu gewinnen. Nach Aufbrechen der Protoplasten ergaben die wasser-1öslichen Bestandteile zahlreiche Peaks mit homologem Antiserum bei der 2-dimensionalen Immunelektrophorese. Das gleiche System wurde benutzt, um reaktive terminale Zuckerreste an Antigenmolekülen zu identifizieren, indem Lectine mit entsprechender Spezifität in ein intermediäres Gel eingebracht wnrden. Bei der Affinitätschromatographie über Conavalin A (ConA) beschickter Sepharose, wurde der Hauptteil des aufgetragenen zytoplasmatischen Materials ausgewaschen. Dagegen ergab das Elutionsprofil mit Säulenchromatographie bei der die Sepharose mit Weizenkeimagglutinim (WGA) beschickt war, zwei Hauptkomponenten von denen eine nur mit N-acetyl-D-glucosamin eluiert werden konnte. Zytoplasmatische Antigene und Antigene von der Zellwand der Hyphen, die durch Extraktion mit Detergentien gewonnen worden waren, zeigten eine vergleichbare Reaktivität gegenüber homologen Antisera, wenn sie mit einem Enzymimmunosorbenttest (ELISA) untersucht wurden. Mit dem gleichen System zeigte sich, daß die Zellwandantigene für die Entdeckung von spezifischem IgG von Patienten mit Krankheiten, bei denen Aspergillen eine Role spielen, welt überlegen waren.     

Purification and Characterization of Deep Candidosis Related Antigens/Reindarstellung und Charakterisierung von Antigenen mit Beziehung zu tiefen Candidosen

Summary: Deep-candidosis related antigens were purified by means of bioaffinity chromatography on sepharose anti-Candida rabbit IgG immunosorbents. Specific immunoglobulins obtained from rabbits with chronic systemic candidosis were fixed on CNBr-Sepharose 4B and constituted our first affinity chromatography column. Rabbits were immunized with the corresponding purified antigens and a second affinity chromatography column was constituted with these hyperimmune anti-Candida rabbit IgG. The corresponding purified antigens showed 20 precipitation peaks in crossed Immunoelectrophoresis. Enzyme activities using API-ZYM and a proteinase were also detected in the purified antigens. Acute and chronic infections in mice and rabbits were done and confirmed previous works. Possible applications of these purified antigens to specific serodiagnosis of systemic candidosis are being evaluated. Zusammenfassung: Antigene mit Beziehung zu tiefen Candidosen wurden mit Hilfe der Bioaffinitätschromatographie auf Sepharose unter Verwendung von anti-Candida Kaninchen-IgG-Immunsorbentien gereinigt. Spezifische Immunglobuline von Kaninchen mit chronischer systemischer Candidose wurden an CNBr-Sepharose 4B fixiert und für die erste Säule bei der Affinitäts-Chromatographie verwendet. Kaninchen wurden sodann mit den gereinigten Antigenen immunisiert und eine zweite Affinitäts-Chromatographiesäule wurde mit dem so gewonnenen anti-Candida-Kaninchen-Hyperinunun-IgG beschickt. Die hiermit gewonnenen und gereinigten Antigene zeigten in der zweidimensionalen Immunelektrophorese 20 Präzipitationsgipfel. Einige Enzymaktivitäten wurden mit Hilfe von API-ZYM und zusätzlich wurde eine Proteinase ebenfalls unter den gereinigten Antigenen entdeckt. An akuten und chronischen Candida-Infektionen von Mäusen und Kaninchen konnten die Ergebnisse bestätigt werden. Die mögliche An wendbarkeit der gereinigten Antigene für eine spezifische Serodiagnostik systemischer Candidosen wird diskutiert.     

The effect of sulconazole on the antigenic composition of two dermatophyte species

Trichophyton rubrum and Trichophyton interdigitale have been grown in liquid culture in the presence of sulconazole. The antigenic activity of detergent extracts of intact organisms was analysed following SDS-PAGE and the probing of Western blots with homologous antisera raised in rabbits and with sera from patients with dermatophyte infections. Differences in protein-band patterns were noted; some bands present in control samples were absent in azole-treated samples and vice versa. These differences were reflected in antigenic band patterns, especially among components of approximate molecular weight of 30-40, 50-60 and 92-100 kDa.     

75. Immunologie des malignen Melanoms

Zusammenfassung Melanomzellen tragen membrangebundene Antigene, die humorale und celluläre Immunantworten auslösen. Diese Antigene scheinen aber nicht tumorspezifisch zu sein, da Absorption mit fetalen Antigenen die Aktivität menschlicher Antiseren löscht. Das Phänomen der Spontanregression zeigt jedoch, obwohl es meist nur partiell auftritt, daß wirksame Tumorabwehr existiert. Gleichzeitige Tumorprogression findet statt, weil Tumorzellpopulationen heterogen sind und in ihrer Antigenexpression stark variieren. Nicht erkennbare Zellen unterlaufen die Abwehr und verursachen Metastasen.     

Coexistence of Free Antigens, Free Antibodies and Immune Complexes in Sera from Patients with Suspected Deep-seated Candidosis

Two commercially available latex agglutination tests: Cand-TEC(TM) from RAMCO Laboratories, Inc., Houston, Texas, USA, for the detection of a heat labile Candida antigen, and LA-Candida Antigen Detection System from Immuno-Mycologics (IMMY), Inc., Norman, Oklahoma, USA, for the detection of Candida mannan antigen, and Own LAT, a self-prepared latex agglutination test for the detection of Candida mannan antigen were tested on 467 sera of patients at risk for deep-seated Candidosis. Separate, parallel investigations were made to demonstrate anti-Candida antibodies (3 methods), circulating immune complexes (not antigen-specific, 2 methods), and rheumatoid factor (2 methods). Specificity and sensitivity of the mannan antigen detection systems were studied. Conditions of specific and non-specific mannan binding to antisera were studied in a defined mannan-antiserum system. The potential and the limitations of the Candida mannan antigen and protein antigen detection systems using latex particle agglutination are evaluated.     

Antigenic Relationship Between Pythium insidiosum de Cock et al. 1987 and its Synonym Pythium destruens Shipton 1987

Antigens and rabbit-antisera from holotypes of Pythium insidiosum and P. destruens were prepared to elucidate their antigenic relationship. The antigens and rabbit-antisera of P. insidiosum as well as P. destruens used as a reference system showed that both shared three precipitin bands in common. The antigen and rabbit-antisera of P. destruens and P. insidiosum used as a reference system against other strains isolated from humans and animals with pythiosis, also showed three precipitin bands in common. When we used sera taken from horses with proven pythiosis against antigens of P. insidiosum and P. destruens, six common bands were observed. We concluded that the etiologic agent of pythiosis is a single species P. insidiosum, and could be identified by serologic methods.     

狼疮性肾炎中ANCN及其靶抗原的临床研究

目的探讨狼疮性肾炎(LN)患者血清中抗中性粒细胞胞浆抗体(ANCN)及其靶抗原的表达及临床意义。方法收集103例LN病人血清,分别用间接免疫荧光法(IIF)及免疫酶联吸附实验(ELISA)法检测ANCA的阳性率及其靶抗原的表达率。结果活动期LN的ANCA阳性率及靶抗原的表达率均高于稳定期LN,且以组蛋白酶G(CG)及乳铁蛋白(LF)为主,活动期LN中抗dsDNA、低补体血症、血沉增快及肾功能下降的发生率均明显高于稳定期。结论LN患者血清中ANCN阳性及靶抗原的表达与LN的活动有关,肾脏损害以ANCA阳性者为重。     

Identification of Candida albicans Antigens Recognized by Sera of Patients with Candidiasis/Identifizierung von Candida albicans Antigenen, mit denen Seren von Candidiasis-Patienten reagieren

Summary: Antigens of Candida albicans derived from both growth phases employing five different media and from five different strains using one medium were analysed by gel-electrophoresis and immunoblot-technique. No growth phase or growth-medium dependent differences were noted when commercially available rabbit anti-Candida or human antisera were employed. The SDS-PAGE patterns of the five strains analysed were quite similar. However, one strain isolated from a patient with proven systemic candidiasis showed an additional 60 kd membrane protein which was recognized only by human but not by rabbit antisera. The immunoblot-patterns of patient and blood donor sera were compared. A striking result was that patients sera specifically recognized a 43 kd soluble antigen, whereas blood donor sera did not. Interestingly, early sera of patients with systemic candidiasis showed an isolated IgM-response towards an 80 kd antigen of the soluble fraction. Our results suggest that tests based on these two antigens may facilitate the diagnosis of systemic candidiasis. Zusammenfassung: Wir verglichen durch Gel Elektrophorese und Immunoblot Antigen Prepa-rationen, die von fünf verschiedenen Candida albicans Stämmen sowie unter Verwendung von fünf verschiedenen Anzuchtmedien gewonnen worden waren. Es konnten im Immunoblot keine von der Wachstumsphase des Organismus abhängige Unterschiede festgestellt werden. Die Protein-Bandenmuster der SDS-Gelelektrophorese waren ebenfalls vergleichbar. Bei der Analyse unterschiedlicher Stämme zeigte sich jedoch, daß ein Isolat aus einer Blutkultur ein zusätzliches Antigen in der Membranfunktion aufwies, das nur von menschlichen, aber nicht von Kaninchen-Antiseren erkannt wurde. Weiterhin wurden die Immunoblot-Muster von Patienten und Blutspenderseren verglichen. Dabei zeigte sich, daß Seren von infizierten Patienten mit einem 43kd Antigen reagierten, während Blutspenderseren diese Reaktivität nicht zeigten. Ein weiterer interessanter Befund war, daß Patienten mit systemischer Candidainfektion in der Frühphase IgM-Antikörperzeigten, die gegen ein 80 kd Antigen gerichtet waren. Unsere Ergebnisse lassen vermuten, daß durch Verwendung dieser Antigene in einem ELISA-Test eine Verbesserung der Diagnostik systemischer Candidainfektionen erreicht werden kann.     

Synthetic oligonucleotides: Useful molecules? A review.

Specific inhibition of mammalian genes is possible through the use of antisense oligonucleotides (AS ODNs) or ribozymes. These strategies have led to a better understanding of several cellular and molecular mechanisms, among which cancer development. Recently, these strategies have been applied also for therapeutical purposes in diseases such as AIDS and cancer. In some of these therapeutical trials the antisense strategy is combined with gene transfer technology: the AS ODN or the ribozyme are expressed within the cell by the use of adenoviral or retroviral vectors. However, many difficulties have still to be overcome before ODNs and ribozymes can be used routinely in the clinic.