| Background and significance:During the flu season, pregnant women are proven to be susceptible to influenza virus. When they get infected in influenza virus, the disease procession gets rapidly and easier to become severe cases. Further more, pregnant women in the second and third trimesters have higher morbidity and mortality rates. And the patients in these two periods have higher possibilities to accompany with severe symptoms of complications. While human influenza virus gets into the human body, HA recognizes the terminal structure SA ?2-6Gal glycan. This structure is found in human airway epithelial cell. But for the avian influenza virus, the recognition is SA a2-3Gal glycan terminal structure which is found in aquatic birds of the gastrointestinal epithelial cell surface. Some research shows that the human airway epithelial cell, alveoli and bronchus contains the structure of SA a2-3Gal too, so it's possible for the avian influenza virus getting infected into the human body. Maackia Amurensis Lectin ?(MAL-?) recognizes the glycan structure of NeuAc2-3Gall-3[NeuAc2-6]GalNAc, which is similar with the human influenza virus recognizes; Sambucus Nigra lectin (SNA) recognizes the glycan structure of Siaa 2-6 Ga1?1-4Glc(NAc), which is similar with the avian influenza virus recognizes. So the simulation test of the avian influenza infecting a host cell can be made using lectin MAL-II; the simulation test of the avian influenza infecting a host cell can be made using lectin SNA. Saliva is a necessary body fluid which is covered in the whole month. In recent years, studies have shown that there are some changes of glycan structure in saliva glycoprotein when humanity is in some chronic diseases. Saliva is the first line of defense virus. It uses its glycoprotein glycan structure to close the virus'recognition to prevent the entrance of the respiratory virus. Pregnant women are susceptible to influenza virus seems due to their physiologic changes in pregnancy including respiratory, hormone, and cardiovascular systems changes. These changes can affect saliva too, so we need to study how it changes in glycoprotein.Method:Firstly, preparing saliva samples collection of non-pregnant heathy women, the first trimester of pregnancy, the second trimester of pregnancy, the third trimester of pregnancy volunteers. Then use lectin microarray to analyze and compare the changed glycan information. In this progress, focus on the expression difference of lectin MAL-? and SNA in the first trimester of pregnancy, the second trimester of pregnancy, the third trimester of pregnancy volunteers compared with non-pregnant healthy women. The changes found use saliva protein microarray as a verification. Secondly, using blotting technology, use the Cy5 labeled lectin MAL-?, SNA, chicken-origin H5N1 influenza virus, and influenza A (H1N1) virus (Influenza Vaccine) to react with the saliva sample of non-pregnant healthy women, the first trimester of pregnancy, the second trimester of pregnancy, the third trimester of pregnancy volunteers. Based on the experimental results, hepatitis B, hepatic cirrhosis, hepatic carcinoma patients are associated with susceptibility to influenza viruses analyzed and discussed. The experimental results can be discussed and analyzed for susceptibility to influenza viruses during the women in non-pregnant and in the first, second, third trimester of pregnancy.Results:1.Compared each period of pregnancy with non-pregnant women shows that some differences appear in glycan. There are some increasingly signals in the first trimester of pregnancy saliva samples in the glycan such as GalNAc?Gal?Fuca-1,6GlcNAc(core fucose), Ga1?1-3GalNAca-Ser/Thr(T-antigen)?Terminala-1,3 mannose; While the decreased signal only conclude the glycan of Ga1?1-4GlcNAc and Sia2-3Ga1?1-4GlcNAc.While compared with non-pregnant women, There are some increasingly signals in the second trimester of pregnancy saliva samples in the glycan such as (?-1,3)and(?-1,6) linked mannose, terminal GalNAc?Bisecting GlcNAc and bicentenary N-glycans?Fuca-1,6GlcNAc(core fucose), ?-Man, Fuca-1,6GlcNAc(core fucose)?Ga1?1-3GalNAca-Ser/Thr (T-antigen)?Terminala-1,3 mannose; While the decreased signals are Ga1?-1,4GlcNAc.. GlcNAc?Sia2-3Ga1?1-4Glc(NAc), aGalNAc and Gal?Fucose?Poly-LacNAc and (GlcNAc)n, ?-Gal and ?-GalNAc, ?1-4GlcNAc and LacNAc, Terminal GalNAc(especially GalNAcal-3Gal)?Non-substituteda-1,6 Man?Multivalent Sia and (GlcNAc)n, Ga1?1-4GlcNAc and Sia2-3Ga1?1-4GlcNAc? Ga1?1-3GalNAc compared with non-pregnant women. Further more, There are some increasingly signals in the third trimester of pregnancy saliva samples in the glycan such as terminal GalNAc?Bisecting GlcNAc and bicentenary N-glycans, aGalNAc,GalN Aca-Ser/Thr(Tn)and aGal?GlcNAc?Gal. While the decreased signals are Ga1?-1,4GlcNAc? Sia2-3Ga1?1-4Glc (NAc)??GalNAc T antigen/Tn antigen Poly-LacN-Ac and (GlcNAc)n, a-Man(inhibited by the presence of bisecting GlcNAc)?Multivalent Sia and (Glc-NAc)n? Ga1?1-4GlcNAc and Sia2-3Gal1?1-4GlcNAc. So we can find that the glycans in the saliva during the whole pregnancy period are different.2.Through analyzing the normalized RFI and the ratio values in the whole period of pregnancy compared with the non-pregnant women, we can find that the ratio values in the first, second, third trimester of pregnancy for the lectin MAL-II are 0.814,0.352 and 0.358. So the level of the expression of glycan SA a2-3Gal in the first trimester of pregnancy shows no significant differences compared with non-pregnant women. But they show significantly decreased signals in the second and the third trimester of pregnancy. In the same condition, the ratio values in the first, second, third trimester of pregnancy for the lectin SNA are 0.983, 0.964 and 0.924. So the level of the expression of glycan SA a2-6Gal in the first, second and third trimester of pregnancy shows no significant differences compared with non-pregnant women. The latter saliva protein microarray verifies the results.3.Lectin blotting experiments show that lectin MAL-II combined more protein in non-pregnant healthy women compared with the second and third trimester of pregnancy samples, but no significant differences with the first trimester of pregnancy samples. Lectin SNA show no significant differences in all samples. Chicken-origin H5N1 influenza virus combined more proteins in non-pregnant healthy women samples, but shows significant decreased signals in the second trimester of pregnancy samples. The results are same with lectin MAL-II. While, there is no obvious difference in H1N1 virus vaccine binding condition. So we can conclude that, the women in the second and the third trimester of pregnancy are susceptible avian influenza virus. While, the power to resist human influenza viruses is the same. |
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