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Human herpesvirus 6 (Human herpesvirus 6, HHV-6) is a class of double-stranded DNA viruses, and in 1986 by the American Cancer Center Salahuddin and other peripheral blood mononuclear cells from lymphoid hyperplasia and AIDS patients addicted to human lymphocytes isolated, and human herpesvirus 7 (HHV-7) and human cytomegalovirus (CMV) belong to the herpes virus β subfamily. That HHV-6 is the cause of infant subitum (ES), also with glioma, meningoencephalitis, AIDS, chronic fatigue syndrome, organ transplant infections, multiple sclerosis and other diseases, but its pathogenic mechanism is not yet clear. HHV-6 genome is a linear double-stranded DNA molecules, size about 160 ~ 170Kbp, of the structure can be divided into three parts, namely the sequence unique region (UL District), each of the left and right ends of a forward repeat sequence (DRL and DRR District ), and 119 open reading frame (ORF). According to the differences in gene structure and antigen can be divided into two subtypes of HHV-6A, B, GS, U1102 is a subtype of representative strains of a HHV-6A, while Z29 is a representative of the subtype B strains, and sequence homology at 96 %, HHV-6 is widely present in peripheral blood lymphocytes and saliva in healthy adults and children, the primary infection generally occurs in the 6 months to 2-year-old infant, the peak of 6 to 9 months, and subsequently establish a persistent infection in the body and were latent infection for a long time. My room was first isolated in 1994 in the country identified Nanjing local strains of HHV-6 8, and a detailed study of Nanjing strain E5 (CN5) virological, immunological, biological characteristics of the virus morphology ultrastructure, as The study has laid a good foundation. HHV-6 E5 virus antigen of this study was purified by sucrose density gradient centrifugation method of Nanjing strains 8 weeks old, female BALB / c mice, using conventional methods fusion, indirect ELISA method of screening, and by three times of limiting dilution were subcloned acquire hybridoma three steady hybridoma cell lines that secrete specific anti-HHV-6 antibody, were named JA9, JYE7, JYE8. The obtained three hybridoma strain passaged amplified injected into Pristane pre-stimulation of BALB / c mice in vivo preparation ascites, and preliminary purification. The monoclonal antibody Ig subclasses identification: the JA9 IgG1, κ subtypes, JYE7 and JYE8 IgM λ subtypes; titers were 1:0.8 X10 5,1:0.256 X10 to 5,1:0.128 X10 - 5; hybridoma chromosome count more than 3 chromosome number 95; indirect immunofluorescence assay: 3 mAb could infected with HHV-6 E5 CBMCs positive reaction, with not infected CBMCs was negative reaction; which JA9 monoclonal antibody Western-blot results further show that the and its virus protein binding with HHV-6 E5 about 75ku size. The study collected oral cancer (oral squamous cell carcinoma and precancerous lesions), brain tumors (glioma and meningioma) infant acute rash, immunosuppressants user (kidney transplantation and nephrotic syndrome) and healthy population saliva The specimens were 41, 40 parts, 36 parts, 37 parts, 40 parts of a total of 194 copies. Nested PCR and ELISA positive rate of saliva samples HHV-6, the results in patients with HHV-6 positive were higher than the healthy population and significant. This study on the basis of self-isolation and identification of HHV-6 in Nanjing strain E5, the successful preparation and preliminary identification of the three anti-human herpes virus type 6 Nanjing strain E5 monoclonal antibodies, will lay the foundation for further study of HHV-6, and may provide for the clinical diagnosis.
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