| Objective:To study the distribution of four natural antibodies (anti-egg nucleus antibody, anti-ovalbumin antibody, anti-bovine serum antibody, anti-typhoid polysaccharide antibody)and the relationships between the antibodies and age-gender, and explore the relevance between the antibodies and HLA-DQB1 alleles.These were studied by detecting the level of four natural antibodies in random people and examination normal population of different age and gender. Further, we investigated their clinical significance in antinuclear antibody positive patients.Methods:1. Eighty-six persons were selected randomly from the people who had passed the physical examination in Dalian Locomotive Hospital (36 male,50 female with average age of 28.52).The same batch of samples were detected in the same concentration ELISA plate coated with egg nuclear antigen for 5 times continuously, once a day,13 samples per time;the same sample was detected in the same concentration ELISA plate coated with egg nuclear antigen for 16 times continuously, once a day. Log units of anti-egg nucleus antibody of each sample was detected respectively, then precision and stability of the method based on ELISA, which would be used to detect anti-egg nucleus antibody, were analyzed.2. The persons who had passed the physical examination were selected randomly to be the sample in Blood Center of Dalian and Medical Center of the Second Affiliated Hospital of Dalian Medical University. It included 18, males and 19 females at the age over 70 (elderly),33 males and 41 females at the age lower than 40 (young) respectively. They were allocated to elderly group and young group and also male group and female group, respectively. The other three natural antibodies were detected with the newly established method, and investigated whether this method could be used to detect the three antibodies. The distribution of four natural antibodies among people of different age and gender was studied.3. Eighty-six persons were extracted randomly from normaland healthy people (36 male,50 female with age range 28.52±12.84),who were taken venous blood into EDTA anticoagulant tubes,and extracted their DNA.HLA-DQB1 allele was amplified by polymerase chain reaction(PCR). The level of Log units of four natural antibodies was detected by the newly established method. In order to investigate the relationships between four natural antibodies and HLA-DQB1 allele.4. The patients with antinuclear antibody positive were selected as disease group (among,35 persons from Clinical Laboratory of the First Affiliated Hospital and 23 persons from the Second One of Dalian Medical University), including 12 male and 46 female with average age 41.93. Eighty-six persons were selected randomly from the people who had passed the physical examination as control group (36 male,50 female with average age 28.52). Log units of the antibodies were detected by the previous method. The distribution and clinical significance of the antibodies were explored in the antinuclear antibody positive patients.Results:1. The same batch of samples were detected in the same concentration ELISA plate coated with egg nuclear antigen for 5 times continuously, the average value of CV (%) was 7.157 (P<0.05). The same sample was detected in the same concentration ELISA plate coated with egg nuclear antigen for 16 times continuously, the CV (%) among Log units of the antibodies was 4.8, and the general CV is 10% or less.2. The same sample was detected respectively in the ovalbumin,bovine serum and typhoid polysaccharide antigen plate for 16 times continuously, calculating Log units of corresponding antibodies,the corresponding CV% was 7.2,8.6 and 5.2. Significant difference of anti-egg nucleus antibody and anti-typhoid polysaccharide antibody was found in different age groups respectively, at the same time, Log units of the two antibodies were lower in the elderly group. No significant difference of the other two antibodies was found in different age groups respectively. There was no significant difference in different gender groups among the four antibodies.3. There was significant difference between the anti-bovine serum antibody of normal persons with HLA-DQB1*02 site and without it, and anti-typhoid polysaccharide antibody too. And the persons without HLA-DQB1*02 site have higher Log units of the two antibodies. There was statistic significance between the level of anti-bovine serum antibody of the persons with HLA-DQB1*06 site and without it, and the persons with HLA-DQB1*06 have higher Log units of the antibody. And HLA-DQB1* 03,04,05 sites were not related with Log units of four natural antibodies.4. Significant difference was confirmed between control group and disease group on the anti-bovine serum antibody, and the antibody was higher in the antinuclear antibody positive persons. There was no statistic significance between control group and disease group on the other three kinds of antibodies.Conclusions:1. The method based on ELISA, which was used to detect natural antibody, was better than the classical one, higher precision, stable and reliable.It may be used to detect natural antibody in human serum widely.2. Anti-egg nucleus antibody and anti-typhoid polysaccharide antibody in human serum were influenced by age, such phenomenon was not found on the other two antibodies. The level of the four natural antibodies was not influenced by different genders.3. HLA-DQB1*02 was a suppressor gene of the anti-bovine serum antibody and anti-typhoid polysaccharide antibody. HLA-DQB1*06 site was a priming gene of anti-bovine serum antibody. Allele HLA-DQB1*03,04,05 did not induce or inhibit the four natural antibodies.4. The level of anti-bovine serum antibody was higher in the anti-nuclear antibody positive patients.This phenomenon indicated that anti-bovine serum antibody could be used as an indicator to screen the persons with antinuclear antibody.It can be used to evaluate the conditions of diseases...
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