A Study Of Potential Allergens In Ragweed Trilobite Pollen

Objective: Giant ragweed(Ambrosia trifida L.)can produce a large amount of pollen at flowering stage and has strong sensitization.Only two allergens,Amb t 5(unknown function)and Amb t 8(profilin,unknown sequence),have been included in the International Union of Immunological Societies(IUIS)allergen database.There are still many unknown allergens worth exploring.The main purpose of this study is to find out the potential allergens of the giant ragweed pollen and demonstrate their immunogenicity that may provide the necessary help for the individualized diagnosis and treatment of giant ragweed pollen allergy.Materials and methods: By the Next-generation sequencing and comparing with the known pollen allergens' sequences,the nucleotide sequences of potential giant ragweed pollen allergens were screened followed by PCR synthesis and cloning validation.Then,the correct genes and p ET-28a-c(+)vector were connected and expressed by E.coli expression system.Ni-affinity chromatography was used to isolate and purify the potential allergens.ELISA was used to study its immunogenicity.88 cases of giant ragweed pollen allergy and 8 cases of health were selected.Results: 9 complete fragments and 2 incomplete fragments of giant ragweed pollen potential allergens were selected and obtained.The recombinant expression and immunogenicity of the 9 complete proteins were confirmed: the ELISA positive rate of r X1 was about 13.6%,r X3 36.4%,r X4 18.2%,r X5 27.3%,r X6 5.62%,r X7 47.7%,r X8 52.3%,r X9 43.2%,and r X11 12.5%.Conclusion:All of the 11 sequences may be the potential allergens of giant ragweed pollen,and X3?X5?X7?X8?X9 may be the major allegens.