Characterization of natural Epstein-Barr virus infection and replication in smooth muscle cells from a leiomyosarcoma.
Cells from a leiomyosarcoma tumor (LMS-1) from a patient with the acquired immunodeficiency syndrome (AIDS) were explanted, cultured in vitro, and studied by phase-contrast microscopy for morphologic and growth characteristics, immunostaining for cell markers, EBER in situ hybridization and polymerase chain reaction for detection of Epstein-Barr virus (EBV), and immunostaining for expression of EBV antigens. The cells exhibited very slow growth in vitro, with unusual elliptical and spindle-shaped morphology and fragmentation of the cytoplasm into long, tapering, cytoplasmic processes. Greater than 90% of cells expressed diffuse distribution of the smooth muscle isoform of actin by immunoperoxidase staining. Approximately 25% of cells expressed very bright fluorescence by immunostaining of the smooth muscle isoforms of calponin and actin. The majority of cells demonstrated a weak signal for CD21; approximately 5-10% of cells showed a strong signal that was confined to cell surfaces. The cultured cells harbored EBV, and infectious EBV continued to be detected by polymerase chain reaction and virus culture through several passages in vitro. Several EBV antigens were expressed, including latent antigen EBNA-1, immediate-early antigen BZLF1, early antigen EA-D, and late antigens, including viral capsid antigen p160, gp125, and membrane antigen gp350. Human umbilical cord lymphocytes that were transformed with virus isolated from cultured cells yielded immortalized cell lines that expressed EBV antigens similar to other EBV-transformed lymphocyte cell lines. These results confirm that EBV is capable of lytic infection of smooth muscle cells with expression of a repertoire of latent and replicative viral products and production of infectious virus. EBV infection of smooth muscle cells may contribute to the oncogenesis of leiomyosarcomas.
Jenson
H B
HB
Department of Pediatrics, The University of Texas Health Science Center at San Antonio, 78284-7811, USA. jenson@uthscsa.edu
Montalvo
E A
EA
McClain
K L
KL
Ench
Y
Y
Heard
P
P
Christy
B A
BA
Dewalt-Hagan
P J
PJ
Moyer
M P
MP
D000163:Acquired Immunodeficiency Syndrome / Q000150:complications*; D000199:Actins / Q000378:metabolism; D000328:Adult; D000956:Antigens, Viral / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002472:Cell Transformation, Viral; D004268:DNA-Binding Proteins / Q000378:metabolism; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism; D005260:Female; D015686:Gene Products, env / Q000378:metabolism; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D007890:Leiomyosarcoma / Q000473:pathology / Q000821:virology*; D008214:Lymphocytes / Q000821:virology; D008840:Microfilament Proteins; D008858:Microscopy, Phase-Contrast; D009130:Muscle, Smooth / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000378:metabolism; D017464:Receptors, Complement 3d / Q000032:analysis; D013125:Spinal Neoplasms / Q000473:pathology / Q000821:virology*; D015534:Trans-Activators / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000821:virology; D014763:Viral Matrix Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D017735:Virus Latency; D014779:Virus Replication; D054299:env Gene Products, Human Immunodeficiency Virus; D000096985:Calponins
D000199:Actins; D000956:Antigens, Viral; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D015686:Gene Products, env; D008840:Microfilament Proteins; D011498:Protein Precursors; D017464:Receptors, Complement 3d; D015534:Trans-Activators; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D054299:env Gene Products, Human Immunodeficiency Virus; D000096985:Calponins; C063676:BZLF1 protein, Herpesvirus 4, Human; C037662:EBV-associated membrane antigen, Epstein-Barr virus; C066316:gp125 envelope protein, Human immunodeficiency virus 2