【作者】 徐美兰；

【导师】 刘辉；

【作者基本信息】 大连医科大学 ， 临床检验诊断学， 2009， 硕士

【摘要】 目的:体内抗鸡卵核抗体(anti-egg nucleus antibody)是一类针对鸡卵核抗原的特异性抗体。该抗体不仅存在于自身免疫病患者体内,还存在于胃癌患者体内。本文通过对随机人群及不同年龄组、性别组的正常人群进行抗鸡卵核抗体水平检测研究其分布特点,并将其与HLA-DQB1等位基因相关联,分析其相关机制。同时对该抗体的一般生物学特性进行分析研究,并以ICU病人作为研究对象探讨其临床意义。方法1.随机挑选大连机车医院体检人群94人(男51名、女43名、平均年龄42.7岁),用ELISA方法检测随机人群中抗鸡卵核抗体的阳性率。2.从大连医科大学附属二院正常体检人群中随机挑选,要求>70岁(老年)男女各10人、<30岁(青年)男女各10人。分别分为老年组与青年组,及男性组与女性组。研究抗鸡卵核抗体在不同性别、不同年龄人群的分布规律。3.抗鸡卵核抗体极高值的筛选: 600名体检正常人群静脉采血用ELISA检测抗鸡卵核抗体效价,每100人中筛选出高值(大于x +SD)前5人,共30人作为实验组,另选体验正常人100人作为对照组。采用SSP-PCR技术进行HLA-DQB1等位基因的扩增。探讨抗鸡卵核抗体与HLA-DQB1等位基因的关联。4.鸡卵核核酸阻断试验研究抗鸡卵核抗体是否与鸡卵核核酸相结合。5.异嗜性试验研究抗鸡卵核抗体是否具有异嗜性。6.大连机车医院收集体检正常人群20人(男9名、女11名、平均年龄49.2岁)取血分离血清备用。ELISA检测抗鸡卵核抗体、红细胞凝集法检测IgM、IgG血型抗体的效价并探讨两个指标之间的关联。7.用鸡卵核饲养昆明系幼鼠20天后取血检测抗鸡卵核抗体的效价,并做空白对照组。研究抗鸡卵核抗体产生与食物的关系,探讨该抗体的来源。8. ICU病人为大连医科大学附属二院住院患者,诊断符合入住ICU标准,并按其收住标准分为两大类:严重的中晚期器官功能衰竭(ICU慢性病组)和急性疾病(ICU急性病组)。ICU慢性病包括白血病、转移癌、糖尿病等;ICU急性病包括外伤、急性大出血等。ICU慢性病组36名(男14名、女22名、平均年龄55.4岁),ICU急性病组20名(男13名、女7名、平均年龄52.5岁),同时选取体检为正常的人群20名(男9名、女11名、平均年龄49.2岁)作对照组,对三组人群采血分离血清用于抗鸡卵核抗体与IgM、IgG血型抗体效价的检测。其中急、慢性疾病组除外了自身免疫疾病患者如SLE等。探讨抗鸡卵核抗体在危重病人体内分布特点及其临床意义。结果1.随机人群中抗鸡卵核抗体阳性率为100%。2.不同年龄组、不同性别组的抗鸡卵核抗体均无显著性差异(P>0.05)。3.抗鸡卵核抗体极高值组与对照组HLA-DQB1等位基因比较,其中HLA-DQB1*02亚型有显著性差异(P<0.05),其余各亚型均无显著性差异。在抗鸡卵核抗体极高值组中HLA-DQB1*02阳性率高达31.7%。4.鸡卵核核酸阻断试验中鸡卵核核酸阻断组与空白对照组间有显著差异(P<0.05)。5.异嗜性试验中生理盐水组与鼠肝匀浆液组间有显著差异(P<0.05)。6.体检正常人群中抗鸡卵核抗体与天然抗体——IgM、IgG血型抗体之间互无关联(P>0.05)。7.鸡卵核饲养幼鼠实验组与对照组间抗鸡卵核抗体效价水平无显著差异(P>0.05)。8. ICU慢性病组、ICU急性病组与对照组比血清中抗鸡卵核抗体效价均明显降低(P<0.05);ICU急性病组与对照组IgM血型抗体效价有显著性差异(P<0.05);ICU慢性病组与对照组IgG血型抗体效价有显著性差异(P<0.05)。结论1.抗鸡卵核抗体在随机人群中普遍存在。2.抗鸡卵核抗体参考范围与年龄、性别无关。3.抗鸡卵核抗体高效价可能与HLA-DQB1*02基因的表达有关。4.抗鸡卵核抗体不是与鸡卵核抗原的核酸类成分结合,其结合的抗原可能是鸡卵核的蛋白类物质。5.抗鸡卵核抗体未表现有异嗜性。6.抗鸡卵核抗体与血型抗体无关联,它们独立存在于人体内。7.鸡卵核饲养幼鼠试验结果不支持人体内的抗鸡卵核抗体是食物刺激产生的食物不耐受特异性抗体。8.抗鸡卵核抗体的效价在病人的危重期显著降低,提示抗鸡卵核抗体水平可能对病情评估有一定的临床意义。更多还原

【Abstract】 Objective: Anti-egg nucleus antibody is specific antibody to egg nucleus antigen in human serum. This antibody can not only be found in patients with autoimmune disease but also in patients with gastric cancer. In this study, we detected the anti-egg nucleus antibody in random people and in different years of age group、gender group, and analyzed its distribution. Furthermore, we investigated the relationships between the antibody and HLA-DQB1 allelic genes. Finally we inquired its immunological characters and explored its clinical significance in ICU patients.Methods:1. Blood samples were collected from random population in Dalian Locomotive Hospital (51 male, 43 female with average age of 42.7). We used ELISA to detect the positive rate of anti-egg nucleus antibodies in serum.2. Study on the distribution of anti-egg nucleus antibody among people of different age and gender: Subjects were selected from the people who had passed the physical examination in the second Hospital Affiliated Dalian Medical University. It included 10 males and 10 females at the age over 70 (elderly), 10 males and 10 females at the age lower than 30 (young), respectively. They were allocated to elderly group and young group and also male group and female group, respectively.3. Screening of group with the maximum value of anti-egg nucleus antibody: The titer of serum anti-egg nucleus antibodies in 600 people who had passed the physical examination were detected by ELISA. The five people with the highest value (> x +SD) in each 100 people were selected to achieve a total number of 30. 100 people who had passed the physical examination were detected as control group. In order to investigate the relationships between anti-egg nucleus antibody and HLA-DQB1 allelo- morphic genes, the allelomorphic genes of HLA-DQB1 were amplified.4. Egg nucleic acid blocking test was used to study whether the anti-egg nucleus antibody combined with egg nucleic acid.5. Heterophile test was used to study whether the anti-egg nucleus antibody has heterophile property.6. 20 normal people in physical examinations of Dalian Locomotive Hospital (9 male, 11 female with average age of 49.2) were collected. The titer of anti-egg nucleus antibody、IgM and IgG blood group antibodies in serum were detected respectively.7. The serum anti-egg nucleus antibody of Kunming young mice fed with egg nucleus for 20 days was compared with the mice of control group were fed with normal mice food. It is to study the relationships between anti-egg nucleus antibody and food to explore the source of the antibody.8. Patients came from ICU of the second Hospital Affiliated Dalian Medical Hospital. All of the patients were satisfied with the diagnosis standards of ICU. They were divided into two groups according to the hospitalization standards: Patients with severe middle and late period of organ failure group (ICU chronic diseases group) and patients with acute disease group (ICU acute diseases group). ICU chronic diseases include leukemia, metastatic carcinoma and diabetes mellitus and so on; ICU acute diseases include trauma and acute massive hemorrhage and so on. ICU chronic diseases group contained 36 patients (14 males and 22 females with average age of 55.4) and ICU acute diseases group contained 20 patients (13 males and 7 females with average age of 52.5). 20 people who had passed the physical examination were measured as control group (9 males and 11 females with average age of 49.2). Among them, patients suffered from autoimmune diseases such as SLE and gastric cancer were excluded from both of acute diseases group and chronic diseases group. The purpose of this study is to find the distribution characters of the anti-egg nucleus antibody in patients of ICU and the clinical significance. Results:1. Positive rate of anti-egg nucleus antibody in random population was 100%.2. No significant differences of anti-egg nucleus antibody were found in groups with different age and gender respectively (P>0.05).3. Significant difference in HLA-DQB1*02 was confirmed between group with maximum value of anti-egg nucleus antibody and control group (P<0.05), the positive rate of HLA-DQB1*02 was 31.7% in maximum value group. No significant differences in the other subtypes were found.4. There was a significant difference between egg nucleic acid blocking group and control group (P<0.05).5. There was a significant difference between physiological saline group and mice liver homogenized solution group (P<0.05).6. There were no significant correlations between anti-egg nucleus antibody and IgG、IgM blood group antibodies.7. There was no significant difference on the anti-egg nucleus antibody level between mice fed with egg nucleus and mice fed with normal mice food (P>0.05).8. The level of anti-egg nucleus antibody decreased significantly in both of ICU acute disease group and ICU chronic disease group, comparing with control group respectively (P<0.05); There was statistical significant different level of IgM blood group antibody between control group and ICU acute diseases group (P<0.05); Significant different level of IgG blood group antibody was found between control group and ICU chronic diseases group (P<0.05).Conclusions:1. Anti-egg nucleus antibody was ubiquitous in random population.2. Reference limit of anti-egg nucleus antibody was not related with age or gender.3. The positive rate of HLA-DQB1*02 may be related to the high expression level of anti-egg nucleus antibody.4. Anti-egg nucleus antibody was not specific antibody to the egg nucleic acid ingredients. It possible combines with egg protein components.5. Anti-egg nucleus antibody did not show heterophile. 6. Anti-egg nucleus antibody was not related with the blood group antibodies. They independently exist in human body.7. The investigations of young mice which fed with egg nucleus didn’t support that the antibody was food intolerant antibody which induced by food stimulation.8. Anti-egg nucleus antibody in critical patients significantly decree- sed. It can be used to evaluate the conditions of diseases.更多还原