ABSTRACT
Herpes simplex virus-1 (HSV) or varicella zoster virus (VZV) DNA was detected by nested polymerase chain reaction in peripheral blood mononuclear cells of patients with Meniere's disease (one of 28 patients for HSV-1, 2 of 28 patients for VZV) during acute illness (within 5 days after onset). On the other hand, neither HSV-1 DNA or VZV DNA was detected in PBMCs of 50 age- and sex-matched healthy individuals and 50 pregnant women. These findings may imply that reactivation of HSV- 1 or VZV may be associated with the development of some cases of Meniere's disease.
Subject(s)
DNA, Viral/analysis , Herpesvirus 1, Human/isolation & purification , Herpesvirus 3, Human/isolation & purification , Leukocytes, Mononuclear/virology , Meniere Disease/virology , Acute Disease , Adult , Aged , Antibodies, Viral/blood , Female , Herpesvirus 1, Human/genetics , Herpesvirus 3, Human/genetics , Humans , Male , Meniere Disease/blood , Meniere Disease/immunology , Middle Aged , Polymerase Chain ReactionABSTRACT
We previously isolated RA-A47, which is recognized as an antigen of rheumatoid arthritis (RA), from a human chondrosarcoma-derived cell line (HCS-2/8). The N-terminal 21-amino-acid sequence of RA-A47 had 81% homology to the deduced amino acid sequence of the human heat-shock protein (HSP) 47 gene, the colligin gene, and 100% homology to that of the colligin-2 gene. Moreover, as is HSP47, RA-A47 was a heat-inducible collagen-binding protein. To further characterize RA-A47, we isolated ra-a47 cDNA from HCS-2/8 cells and human periodontal ligament fibroblast (HPLF) cells. The isolated ra-a47 cDNAs from both cells were almost the same as that of colligin-2. C504 and G505 in the cDNA sequences of both cells and C598 in the cDNA of HCS-2/8 were different from the corresponding bases of colligin-2 cDNA. These differences were also observed in genomic DNA. colligin cDNA was not isolated. To show that the isolated cDNA encodes RA-A47 protein, it was expressed in Cos-7 cells. The produced protein was 47kDa and was recognized both with RA sera and antirat HSP47 antibody, indicating that it is RA-A47 and has structural similarity to HSP47. These results taken together with our previous findings show that RA-A47 is the putative colligin-2 gene product and behaves as a human HSP47. Although colligin has been considered the human HSP47 gene, failure to detect the colligin gene and its mRNA suggests that colligin does not exist in human cells and that the HSP47 gene is identical with colligin-2, which encodes RA-A47.
Subject(s)
Carrier Proteins/genetics , Heat-Shock Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , Blotting, Western , COS Cells , Carrier Proteins/isolation & purification , Carrier Proteins/metabolism , Cartilage/cytology , Cell Line , Chlorocebus aethiops , Chondrocytes/cytology , Cloning, Molecular , DNA Primers/chemistry , Fibroblasts/cytology , Genetic Vectors , Glycoproteins , HSP47 Heat-Shock Proteins , Heat-Shock Proteins/isolation & purification , Heat-Shock Proteins/metabolism , Humans , Molecular Sequence Data , Periodontal Ligament/cytology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , TransfectionABSTRACT
A novel nucleoside with an oxetanosyl-N-glycoside has been recently isolated from a culture filtrate from Bacillus megaterium and named oxetanocin A (N. Shimada, S. Hasegawa, T. Harada, T. Tomisawa, A. Fujii, and T. Takita, J. Antibiot. 39:1623-1625, 1986). In this study, we evaluated the antiherpesvirus activity of oxetanocin A and its derivatives and found that 9-(2-deoxy-2-hydroxymethyl-beta-D-erythro-oxetanosyl)guanine (OXT-G) was very potent and selective in inhibiting the replication of human cytomegalovirus (HCMV) in vitro. The median effective concentration for HCMV strain AD169 was 1.0 microgram/ml, and that for herpes simplex virus type 2 strain 186 was 3.5 micrograms/ml. The selectivity index, based on the ratio of the median inhibitory concentration for cell growth of human diploid fibroblasts to the median effective concentration for HCMV plaque formation, was more than 300. The synthesis of HCMV-induced late polypeptides such as the 150,000-molecular-weight capsid and the 68,000-molecular-weight major matrix proteins was strongly suppressed when OXT-G (5 micrograms/ml) was added to the cultures at the beginning of infection. At this concentration of OXT-G, the amount of HCMV DNA detected in the drug-treated infected cells was less than 1/10 of that detected in the infected control cells. The results suggest that the mode of action of OXT-G is inhibition of viral replication by impairing the viral DNA synthesis.