Establishment and characterization of a novel genetic hybrid cell line for propagation of four pathogens.
Chromosomal DNAs were purified from human epidermoid carcinoma (HEP-2) cells and transfected into human embryonic lung (HEL) cells to establish a genetic hybrid cell line susceptible to infections by toxoplasma, rubella virus, cytomegalovirus, and herpes simplex virus. Karyotype analysis showed that the resultant hybrid cells, designated D3, had a chromosome number of 96, which was stable after passage for 100 generations. Direct microscopy and immunofluorescence showed that the D3 cells could be infected by the four pathogens with overt cytopathic effects. The toxoplasma and three viruses were purified from infected D3 cells by sucrose gradient centrifugation and used as the antigens for detection of specific IgG and/or IgM in serum samples from pregnant women with suspicious infections by the four pathogens, the results of which were consistent with those of commercial kits. These data indicate that a stable genetic hybrid cell line has been generated, which is a valuable tool for the isolation of the four intrauterine pathogens and for the preparation of antigens for serological tests.
Yan
Hua
H
Department of Microbiology and Immunology, School of Medicine, Yangzhou University, Yangzhou 225001, PR China. yzyanhua@yahoo.com.cn
Li
Guocai
G
Yan
Huishen
H
Chen
Hongju
H
Gong
Weijuan
W
Qian
Li
L
Ji
Mingchun
M
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line / Q000382:microbiology; D002875:Chromosomes / Q000737:chemistry; D003587:Cytomegalovirus / Q000254:growth & development / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000097:blood / Q000175:diagnosis; D004247:DNA / Q000235:genetics; D000078404:Epidermal Cells; D004817:Epidermis / Q000737:chemistry; D005260:Female; D006561:Herpes Simplex / Q000097:blood / Q000175:diagnosis; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007621:Karyotyping; D008168:Lung / Q000166:cytology*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000097:blood / Q000175:diagnosis; D012409:Rubella / Q000097:blood / Q000175:diagnosis; D012412:Rubella virus / Q000254:growth & development / Q000276:immunology; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000254:growth & development / Q000276:immunology; D014122:Toxoplasma / Q000254:growth & development / Q000276:immunology; D014123:Toxoplasmosis / Q000097:blood / Q000175:diagnosis; D014162:Transfection
D000913:Antibodies, Protozoan; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D004247:DNA