| Objective:To explore the possible effects of environmental factors(including automobile exhaust and smoking)on humoral immunity based on ELISA.HPLC/MS was used to investigate the components of automobile exhaust and cigarette smoke-aqueous solution.The samples of smokers and non-smokers were collected in the questionnaire,and the automobile exhaust antibody,smoke liquid antibody,inhaled and food allergens and antinuclear antibodies were tested,to investigate the correlation between automobile exhaust antibody and smoke liquid antibody and autoantibody.The subjects of COPD,lung cancer and health examination were randomly selected for the detection of automobile exhaust antibody and smoke liquid antibody,and the relationship between HLA-DQB1 gene difference and antibodies was studied at the molecular level.Methods:1.Questionnaire survey of physical examination department of people's Hospital of Lushunkou District on 24 male subjects of smokers and non-smokers respectively,the average age of smokers was 40.08 ± 10.80 years old,the average age of non-smokers was 33.63 ± 11.40 years old.Automobile exhaust and cigarette smoke-aqueous solution were prepared by liquid collision method,coated with automobile exhaust and cigarette liquid antigen enzyme-linked strip,and the Ig Gantibodies of automobile exhaust and cigarette liquid were detected by ELISA.2.Kunming mice(6 weeks old,about 20 g)were selected as animal experiments to study the antigenicity of automobile exhaust and smoke-related substances,and automobile exhaust and smoke solution antigenthe were injected repeatedly by intraperitoneal injection.After three times of immunization,the whole blood of the animals was collected and the serum was separated for the detection of automobile exhaust and smoke liquid Ig G antibody.3.Antinuclear antibodies and specific antibodies to inhaled and food allergens in the serum of smokers were detected by western blot to study the relationship between smoking and autoantibodies.4.The components of cigarette smoke-water solution produced by liquid collision method of 1,10 and 20 cigarettes were analyzed by HPLC/MS.5.Cotinine in the serum of 24 male smokers and 24 male non-smokers were detected by CGI.The serum was diluted 10 times and 100 times with normal saline and the degree of cotinine in the serum of the subjects was initially graded.6.The whole blood and serum of 85 subjects in the laboratory of the Second Affiliated Hospital of Dalian Medical University were randomly selected,including 30 patients with lung cancer(average age 71.90 ± 4.21),25 patients with COPD(average age 71.52 ± 10.00)and 30 healthy physical examination controls(average age 72.17 ±8.15).The serum was tested for the automobile exhaust and smoke liquid Ig G antibody,at the same time,PCR-SSP was used to detect the frequency distribution of HLA-DQB1 alleles in the whole blood of 85 subjects,and to analyze the association between HLA-DQB1 gene polymorphism and Ig G antibodies.Results:1.Enzyme-linked immunosorbent test revealed the presence of tobacco liquid antibodies and automobile exhaust Ig G antibodies in the serum(p < 0.01),and there was a significant correlation between the two antibodies(r = 0.321;p = 0.026(p < 0.05)).2.Smokers and non-smokers,patients with lung cancer,COPD and healthy physical examination showed no significant difference between the subjects in thedetection of automobile exhaust antibodies and smoke liquid Ig G antibodies.3.The composition of cigarette smoke-aqueous solution was analyzed by HPLC/MS,and it was determined that the composition of cigarette smoke-aqueous solution contained a large number of alkaloids,mainly including nicotine,cotinine,nornicotine,anabasine and anatabine.4.The CGI was used to qualitatively detect the cotinine in the serum of 48 subjects,and the results of 24 non-smokers were all negative,while the results of 24 smokers were all positive.After the serum of 24 smokers was diluted by 10 times with normal saline,10 of them were negative,while the serum of 24 smokers was diluted by100 times with normal saline,and the results were negative.5.There was no significant correlation between the levels of smoke liquid and automobile exhaust Ig G antibodies and the polymorphisms of HLA-DQB1 gene,anti-nuclear antibody,inhalation and food-allergen-specific antibodies.Conclusions:1.Serum may contain Ig G antibodies against automobile exhaust and cigarette smoke-aqueous solution associated antigens.2.Automobile exhaust and smoke liquid Ig G antibodies may have specific effects on the body's humoral immunity.3.There was no statistical relationship between the level of automobile exhaust and smoke liquid Ig G antibody and the polymorphism of HLA-DQB1 gene,and it had nothing to do with the incidence of lung cancer and COPD.There may be a special mechanism for its influence on humoral immunity and pathogenesis. |
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