A Series of Serological Tests for the Detection of Antigens and Specific Antibodies in Deep-Seated Candidosis: Experimental Aspects/Eine Gruppe von Antigen- und Antikörpertests zur Serodiagnose von tieflokalisierten Candida-Mykosen: Experimentelle Aspekte

Summary: We describe a series of six serological tests for the diagnosis of deep-seated candidosis. The array comprises two commercial tests (antigen test, Ramco Inc., and antibody test, Roche), as well as four enzyme immunoassays which have been developed in this laboratory: an antigen test for detection of Candida-proteinase, the corresponding assays for monitoring of anti-proteinase antibodies, and two assays for monitoring of IgG and IgM against heterogenous metabolic antigens of C. albicans. The highly sensitive and specific proteinase antigen-test tolerates samples with high concentration of serum proteins. Proteinase antigen was detected in 10 out of 11 normal mice after intravenous infection with C. albicans blastospores. The proteinase antigen peaked between the second and fourth day after infection. A rise in corresponding antibodies was observed in all animals. No proteinase antigen was detected in sera of healthy human individuals; anti-proteinase antibody titers in these sera amounted up to 1:8000. In related ELISAs, using metabolic fungal antigens, titer values of specific IgG and IgM amounted to 5120 and 1280, respectively. The six tests were carried out in an comparative study under diagnostic conditions, the results of which are the subject of a forthcoming communication. Zusammenfassung: Ein Satz von sechs serologischen Tests für die Diagnostik der tiefen Candida-Mykosen wird vorgestellt. Die Gruppe schließt zwei kommerziell vertriebene Testbestecke ein (Latex-Agglutinationstest zum Antigennachweis, Ramco Inc., und Hämagglutinationstest zum Antikörpernachweis, Roche). Vier weitere Enzymimmuntests wurden von uns entwickelt: Ein Antigentest zum Nachweis von sekretorischer Candida-Protease, ein entsprechender Test zum Nachweis von Antikörpem gegen Candida-Protease, und zwei Assays zum Nachweis von IgG-bzw. IgM-Antikörpem gegen heterogene metabolische Antigene von C. albicans. Der empfindliche spezifische Protease-Antigentest toleriert hohe Konzentrationen unspezifischer Serumproteine und kann deshalb auf Serumproben in geringer Verdünnung (z. B. 1:20) angewandt werden. Protease-Antigen war in 200 fach verdünnten Seren von 10 aus 11 intravenös infizierten NWNI-Mäusen nachweisbar. Die höchste Antigen-Konzentration trat zwischen dem 2. und 4. Tag nach Infektion auf; die Serum-Halbwertszeit von gereinigter Protease in der Maus betrug etwa 60 nun. Ein Anstieg korrespondierender Antikörper war in alien infizierten Tieren zu beobachten. Auch im Serum gesunder Probanden waren Antiprotease-Antikörper bis zu einem Titer von 1:8000 nachweisbar; der Protease-Antigentest fiel hingegen immer negativ aus. Die Titer von Antikörpern gegen metabolische Candida-Antigene erreichten in derselben Gruppe von Seren Werte von 1:5120 bzw. 1:1280. Die sechs Tests wurden unter diagnostischen Bedingungen verglichen; Ergebnisse dieser Studie sind Gegenstand einer weiteren Mitteilung.     

癌胚抗原及粘液组织化学在胃癌中表达的研究

对60例胃癌进行了粘液组织化学和癌胚抗原免疫组织化学研究。结果表明:胃癌发病率肠型和胃肠混合型占多数。60例胃癌中含大肠型粘液者占50例,而单纯含胃型粘液为极少数。癌胚抗原分布方式,肠型胃癌与胃肠混合型胃癌比较无显著差异性(P>0.05)。癌胚抗原的阳性率,胃肠混合型胃癌高于肠型胃癌,两者之间有非常显著差别(P<0.01)。认为选用能显示硫粘蛋白和氧乙酰基唾液酸粘蛋白的粘液组织化学方法,作为胃癌组织分型和确定胃粘膜肠化性质,对癌前病变诊断有一定价值。     

无症状性炎性前列腺炎对血清PSA、fPSA的影响

目的　探讨无症状性炎性前列腺炎(NIH分类Ⅳ型)对血清PSA、fPSA的影响。方法　对比分析36例NIH分类Ⅳ型、42 例有症状性慢性前列腺炎(NIH分类ⅢA型)患者以及22例健康对照组的血清PSA、fPSA、fPSA/tPSA之间的差异。结果　血 清PSA、fPSA、fPSA/tPSA在Ⅳ型和ⅢA型前列腺炎患者间差异无显著性(P>0.05),但与正常对照组比较差异有显著性(P< 0.01)。结论　NIH分类Ⅳ型前列腺炎可引起血清PSA、fPSA升高。对无症状、高血清PSA患者行前列腺活检前,应考虑到患 Ⅳ型前列腺炎的可能。     

CD24 expression on human keratinocytes

Abstract: CD24 or Nectadrin is a cell surface glycoprotein expressed in pre-B lymphocytes, T lymphocytes, neurons, muscle cells and carcinoma cells. Its function is not completely known, but it has been suggested that it is involved in cell adhesion and signalling. CD24 has recently been identified as the human molecule homologous to the murine heat-stable antigen (HSA). HSA is expressed by murine keratinocytes and delivers costimulatory signals in T-cell activation. Long-term cultures of normal human keratinocytes (HKC) were obtained from skin of human female breast sections and either left untreated or were treated with phorbol-12-myristate-13-acetate (PMA) at 10–100 ng/ml, calcium 0.5–2 mM or IFN-γ 100–1000 U/ml, for 24–48 h. Using RT-PCR and flow cytometry we showed that HKC express low levels of CD24 even under basal conditions, and the treatment with calcium, PMA or IFN-γ increased levels of CD24 mRNA and protein. To the best of our knowledge, this is the first report to measure CD24 expression and production by cultured HKC in basal conditions and after stimulation. Further studies are needed to determine biological and therapeutical relevance of these findings.     

Seminal Fluid and the Expression of MHC Class I Antigens in the Placenta of the Rat

PROBLEM : To determine whether seminal fluid influences the expression of MHC class I antigens on the surface of basal trophoblast cells in the placenta of the rat. METHODS : Transfer of DA × DA embryos into a WF (allogeneic) or DA (syngeneic) recipient made pseudopregnant by hormonal treatment followed by mating with a vasectomized male (seminal fluid) or by mechanical stimulation (no seminal fluid). Antigen expression was determined by electron microscopic immunocytochemistry using the appropriate gold-labeled monoclonal antibodies. RESULTS : Seminal fluid did not affect the expression of MHC class I antigens on the surface of the basal trophoblast in either allogeneic or syngeneic matings. CONCLUSIONS : The suppression of the expression of paternal class I antigens on the surface of the basal trophoblast cells in allogeneic pregnancies most likely occurs at the genome level shortly after fertilization.     

Immunobead and Density Gradient Purification of Paget Cells: In vitro Studies of Proliferation

Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramammary Paget's disease investigated whether Paget cells proliferate as other malignant cells do. Although epithelial monolayers from the involved skin were maintained for approximately 45 days, no permanent cell lines were established. The proportion of carcinoembryonic antigen (CEA)-positive cells did not increase in the long-term cultures. Herein, we report studies of whether there is a real reduction of Paget cell numbers or if this is merely a decrease in the expression of CEA by the cells. Furthermore, we investigated whether Paget cells survive longer when cultured free from any potential inhibitory keratinocytes or other epidermal cells. Skin samples were obtained from one patient with mammary Paget's disease and three with extramammary Paget's disease; epidermal cells were cultured in vitro. An enrichment of Paget cells was carried out from the cultured epidermal cells by combining an anti-epithelial membrane antigen monoclonal antibody, binding to immunobeads, and density gradient centrifugation in Nycodenz. The separated cells were re-cultured in Keratinocyte-SFM serum-free media. The proportion of CEA-positive cells did not increase in the cultures, and the purified cells did not show any increase in survival times compared to the non-purified cultured cells. These results suggest that the decrease of CEA-positive cells noted during culture results from a decline in expression of CEA in the Paget cells. Paget cells in the involved epidermis do not proliferate significantly and thus differ from many other malignant cells.     

金地鼠颊囊癌模型PCNA,BrdU及光镜观察结果的相关分析

癌变的早期诊断对提高疗效具有特殊意义，为了寻求简捷，高效，超前，可靠的诊断方法，实验以ＤＭＢＡ（二甲基苯并蒽）诱导的金地鼠颊囊致癌模型为对象，以ＰＣＮＡ（增殖细胞核抗原）ＢｒｄＶ（溴脱氧尿嘧啶）等免疫组化技术为手段；以光镜观察诊断为对照，对三种观察结果进行相关分析，发现三者间有高度显著的相关性，证实了免疫组化方法在癌变诊断中有一定的参考价值，实验还发现ＰＣＮＡ和ＢｒｄＵ检测结果比较每天敏感，简捷，