Purification of antigenically intact Ro ribonucleoproteins; biochemical and immunological evidence that the 52-kD protein is not a Ro protein.

Anti-Ro sera immunoprecipitate Ro ribonucleoproteins (RNPs) from human cell extracts. Ro RNPs are biochemically heterogeneous particles whose functions are unknown and whose exact composition remains controversial. In addition to 60-kD Ro and to La proteins, a 52-kD polypeptide (p52) has been proposed to be a stable component of the Ro RNPs. To confirm the immunological studies supporting this hypothesis, we have biochemically purified Ro RNPs from HeLa cells using non-denaturing conditions. Ro RNPs segregated into three distinct populations, one of which only contained hY5 RNA (RohY5 RNPs). No p52 co-purified with Ro RNPs. Despite the absence of p52, purified Ro RNPs had biochemical and immunological properties identical to those of unfractionated Ro RNPs. Many anti-Ro sera only recognize p52 in immunoblots, and are said to be monospecific anti-p52. Preincubation with purified RohY5 RNPs (free of p52) of all human anti-Ro (including so-called monospecific anti-p52) sera abolished their capacity to immunoprecipitate Ro RNPs from unfractionated HeLa cell extracts. Conversely, preincubation of anti-Ro sera with purified p52 protein specifically inhibited recognition of p52 in immunoblots, but did not interfere with immunoprecipitation of Ro RNPs. Our data demonstrate that anti-p52 antibodies do not target intact Ro RNPs, nor do they target the native 60-kD Ro protein. Contrary to previous reports, p52 protein is not a stable component of antigenically intact Ro RNPs.     

In vivo induction of CD4+ T cell responses by antigens covalently linked to synthetic microspheres does not require adjuvant

Macrophages, dendritic cells or B lymphocytes have been shownto play a major role in the presentation of soluble antigensto CD4⁺ T cells. In contrast, the capacity of these cells topresent particulate antigens such as bacterial or parasiticantigens to T cells remains controversial. To investigate thisquestion, well defined particulate antigens were prepared bycovalent linkage of proteins or peptides to 1 µm in diametersynthetic microspheres. The T cell immunogenicity of such particulateantigens was analyzed in vitro and in vivo. In vitro, a solubleprotein such as hen egg lysozyme (HEL) coupled to beads stimulateda strong proliferative T cell response of lymph node cells fromHEL-primed mice or of specific T cell hybridomas. HEL coupledto beads was presented to the specific T cell hybridomas bysplenocytes or by peritoneal macrophages, but not by lymphomaB cells. Immunization of mice with several different proteinantigens or with a synthetic peptide covalently linked to beadsinduced strong CD4⁺ T cell responses in the absence of adjuvant.The strong in vivo immunogenicity of proteins coupled to beadsdid not result from a non-specific adjuvant effect of beadssince covalent linkage of the antigen to beads was strictlyrequired to induce T cell responses in the absence of adjuvant.In vivo treatment by carrageenan showed that macrophages arerequired for the in vivo stimulation of T cell responses bythese particulate antigens. Thus, these results demonstratedthe role of phagocytic cells, especially macrophages, for invivo presentation of particulate antigens. These particulateantigens represent an interesting approach for the developmentof new vaccines, and for the in vivo analysis of the role ofvarious antigen presenting cells in T cell activation and differentiation.     

Antigen Presentation By Class I Major Histocompatibility Complex Molecules: A Context for Thinking About HLA-G

PROBLEM: There is substantial data that support the efficacy of paternal leukocyte immunization (PLI) for the treatment of alloimmune mediated miscarriage; however, there is confusion regarding the laboratory test that should be performed to determine levels of maternal anti-paternal leukocyte antibodies (MAPLA). METHOD: Popular methodologies employed include: 1) microcytotoxicity (MCX), 2) mixed lymphocyte culture (MLC), and 3) cell flow cytometry crossmatch (FCXM). Cell flow cytometry crossmatch correlates well with the more difficult MLC assay although the former proves the more sensitive study. This work compares the MCX assays with FCXM. The study group consisted of ten women who had a history of three or more spontaneous abortions (SABs). All ten had very low levels (<10%) of MAPLA as measured by FCXM. Following PLI all subjects demonstrated elevated levels (>50%) of MAPLA by FCXM. At 12 weeks gestation, sera were simultaneously measured for MAPLA by MCX and FCXM. RESULTS: Although all ten patients had very high levels of MAPLA by FCXM during pregnancy, five of ten had antibodies to HLA Class I and two of ten had antibodies to HLA Class II paternal antigens by MCX. Furthermore, all patients who were positive by MCX to paternal Class I antigens were also positive to Class I antigens not seen in either parent. Both patients who were positive by MCX to paternal Class II antigens were also positive to maternal Class II antigens. Notable is that all ten women eventually delivered healthy infants. CONCLUSION: Based on this preliminary study, the MCX assay is neither sensitive or reliable enough to determine the need and/or to monitor the effectiveness of PLI. Flow cytometry should be the modality of choice when determining the need for alloimmunotherapy and to monitor the effectiveness of treatment.     

含化复方新诺明引起过敏性休克一例

含化复方新诺明引起过敏性休克一例患者，女，３０岁，因咽峡炎于１９９２年８月７日晚１０时自行含化复方新诺明（广州白云山制药厂）两片、约７分钟后，出现全身搔痒，当即肌注苯海拉明２０ｍｇ，静推１０％葡萄糖酸钙１０ｍｌ，旋即全身泛起荨麻疹，奇痒难忍，眼睑轻度水肿，嘴唇发麻，咽部“发紧”，发音困难，烦躁，立即给予吸氧，静推地塞米松１０ｍｇ，皮下注射肾上腺素０．５ｍｇ，与此同时，患者脸色苍白、鼻尖、未梢发凉，口唇紫绀，时吸暂停，意识丧失，脉搏１４０次／分，细数，血压测不到，再次给予皮下注射肾上腺素０．５ｍｇ，静点地塞米松１５ｍｇ，并给予扩容、纠酸，约一分钟后，欲行气管切开时，患者呼吸渐恢复、继而意识清楚，紫绀减轻，血压１０／１６ｋＰａ，遂休克纠正。复方新诺明片致过敏性休克者尚属少见，该患者有青霉素过敏史，局部用药又易致过敏，考虑喉头痉挛与药物含化有关。（长治市人民医院王爱军，崔文华，郭天然）含化复方新诺明引起过敏性休克一例@王爱军，崔文华，郭天然$长治市人民医院     

Research review: DNA polymerases as molecular markers of the regenerating capacity of hepatocytes

Hepatocyte regeneration has been widely investigated, with the mitotic index and the incorporation of [³H]thymidine being used as regeneration markers. We focused on the induction of DNA replication enzymes, particularly DNA polymerases (pol) α, δ, and ε. Using rat models, we have shown that the activity of pol α in crude liver extract well represents the regenerating capacity of hepatocytes. Using pol α as an indicator, we analyzed liver regeneration in rat models under various conditions: obstructive jaundice, external or internal biliary drainage, and the obstruction of portal vein branches. It has been revealed that the ligation of the common bile duct alone induces a certain amount of hepatocyte proliferation. It was striking that external biliary drainage suppressed regeneration capacity in cholestatic rat liver after partial hepatectomy. The strong regeneration in nonligated lobes induced by portal branch ligation was similar to the liver regeneration seen after partial hepatectomy with respect to the induction of DNA polymerases. Taken together, the aspects of DNA replication, particularly the induction of DNA polymerases, may contribute to shedding new light on the regeneration of human liver. This work was supported in part by a Grant-in-Aid for General Scientific Research and for Cancer Research from the Ministry of Education, Science and Culture, Japan, and by grants from the Uehara Memorial Foundation     

Molecular oncogene markers and their significance in cutaneous malignant melanoma

Background: Oncogenes and other molecular tumor markers that predict tumor aggressiveness may allow individualization and optimization of surgical therapy of intermediate-thickness malignant melanoma. We examined the expression of selected markers, including the HLA-DR antigen, the heat shock protein-70 (HSP-70), and the c-myc oncogene in primary melanoma and regional nodes and related these findings to metastatic potential and survival. Methods: Forty patients with primary melanoma (1.5–4.0 mm) were studied, all of whom had prophylactic lymph node dissection and were followed for 18 months to 7 years. The primary tissue and nodes were examined using immunohistochemical techniques for the presence of HLA-DR antigen and HSP-70 protein and the expression of the c-myc oncogene. Results: Of 40 patients, there were 23 with lesions 1 to 2.9 mm thick and 17 with lesions 3 to 4 mm thick. Nodal metastases were present in 25 of the 40 patients who had elective node dissection. HLA-DR antibody stained the primary tumor in 10 patients (25%), but there was no correlation with survival in this group. HLA-DR antibody stained the stroma and cellular infiltrates surrounding the primary tumor in 28 of 40 patients; in this group there was a correlation of HLA-DR staining of the peritumoral stroma with improved survival overall. HLA-DR staining of the peritumoral stroma also influenced survival when patients were stratified by tumor thickness groups 1 to 2.9 mm and 3 to 4 mm and presence of nodal metastases. HSP-70 was demonstrated in the primary tumor in 25% of patients, who were also shown to have significantly improved survival when compared with those whose primary tumor did not stain with HSP-70. C-myc was expressed in the primary tumor in 25%, but showed no correlation with survival. None of these proteins correlated with or predicted the presence of nodal metastases. Conclusion: We conclude that the use of specific molecular-oncogene markers in intermediate-thickness primary melanoma may identify patients at high risk for conventional treatment failure and reduced survival who may profit from more aggressive surgery, adjuvant therapy, or both.Presented at the 48th Annual Cancer Symposium of The Society of Surgical Oncology, Boston, Massachusetts, March 23–26, 1995.     

胃肠癌患者血清及癌组织癌胚抗原的检测意义

应用放射免疫测定法,对20例胃肠癌患者进行了血清癌胚抗原(CEA)测定,并应用免疫组化法对其中16例癌组织中的CEA进行了检测.检测结果,血清CEA阳性6例,癌组织CEA阳性9例.低分化癌的血清CEA显著高于高分化癌(P<0.05).血清CEA与癌灶部位、癌组织CEA与分化程度及血清CEA均无关(P>0.05).提示血清CEA的含量与癌组织的分化程度有关,血清CEA与癌组织内的CEA无平行关系.作者并对其机制及检测CEA的意义进行讨论.     

SA、F/TPSA及PSAD在前列腺癌诊断中的作用

目的 :探讨血清前列腺特异抗原 (PSA) ,血清总PSA及游离PSA比值 (F/TPSA)及前列腺特异性抗原密度(PSAD)在前列腺癌诊断中的作用。方法 :对 5 1例前列腺癌患者及 14 5例良性前列腺增生症患者PSA、F/TPSA及PSAD值的差异进行分析、比较。结果 :前列腺癌组血清PSA及PSAD高于良性前列腺增生组 ;而F/TPSA值低于良性前列腺增生组 ,差异均有显著性。结论 :PSA >4ng·ml-1作为筛选前列腺癌的临界值存在一定缺陷 ;当PSA <10ng·ml-1,F/T值有助于鉴别前列腺癌和良性前列腺增生 ;而PSAD对于筛选前列腺活检病例亦有一定价值。     

逆转录-聚合酶链反应检测胃癌淋巴结微转移

目的 检测胃癌常规病理检查阴性的淋巴结微转移的发生及与其它临床参考指标的关系。方法 利用逆转录－聚合酶链反应（RT－PCR）方法检测68枚胃癌胃周淋巴结癌胚抗原（CEA）mRNA基因表达，同时比较RT－PCR与免疫组化（IHC）方法的检测敏感性。结果 CEAmRNA RT－PCR是一种很敏感的方法，可以检测1／10^6个转移的癌细胞；检测19例胃癌患者取材的68枚胃周淋巴结，IHC阳性率28％（19／68），RT－PCR阳性率57％（39－68），两组之间差异有非常显著性意义（P＜0.01）；RT－PCR阳性率与胃部临床参考指标密切相关，且随着病期进展而增大。结论 CEA mRNA RT-PCR是比免疫组化更敏感的方法，可以预测胃癌淋巴结微转移，能够有效地避免已有微小转移的患者被漏诊。     

Ca^＋＋对链霉素抗原—抗体反应的抑制作用

为探讨临床中使用钙制剂抢救链霉素过敏休克病人有良好效果的原因,由被动血凝试验、被动皮肤过敏试验和ELISA法证实,Ca~( )可以抑制链霉素抗原-抗体的反应。这种抑制作用是通过Ca~( )链霉素抗原结合,封闭链霉素抗原决定簇来加以实现的。