HN proteins of human parainfluenza type 4A virus expressed in cell lines transfected with a cloned cDNA have an ability to induce interferon in mouse spleen cells.
Primary monkey kidney cells infected with human parainfluenza type 4A virus (HPIV-4A) were treated with various concentrations of formaldehyde. Formaldehyde (0.275%) treatment completely blocked virus production. However, when mouse spleen cells were cocultured with the fixed virus-infected cells, interferon was produced in the culture fluid. On the other hand, when mouse spleen cells were incubated with the fixed virus-infected cells in the presence of anti-HPIV-4A antiserum or a mixture of anti-HN protein monoclonal antibodies, interferon activity could scarcely be detected in the culture fluid. These findings indicated that the fixed virus-infected cells had an ability to induce interferon in mouse spleen cells and that the HN protein was related to interferon induction. Subsequently, a recombinant plasmid was constructed by inserting the cDNA of the HN gene of HPIV-4A into a pcDL-SR alpha expression vector. Mouse spleen cells produced interferon when cocultured with COS7 cells transfected with the recombinant plasmid, but did not when cocultured with COS7 cells transfected with the vector alone. Furthermore, we established HeLa cells constitutively expressing HPIV-4A HN (HeLa-4aHN cells) or F protein (HeLa-4aF cells). Type I (alpha/beta) interferon was detected in culture fluids of mouse spleen cells with HeLa-4aHN cells, but was not detected in those with HeLa-4aF cells. Therefore, it was concluded that the HN glycoproteins on the cell surface were sufficient for interferon induction to occur.
Ito
Y
Y
Department of Microbiology, Mie University School of Medicine, Japan.
Bando
H
H
Komada
H
H
Tsurudome
M
M
Nishio
M
M
Kawano
M
M
Matsumura
H
H
Kusagawa
S
S
Yuasa
T
T
Ohta
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005557:Formaldehyde / Q000494:pharmacology; D006168:Guinea Pigs; D015707:HN Protein / Q000235:genetics / Q000502:physiology*; D000882:Haplorhini; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000096:biosynthesis*; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D011994:Recombinant Proteins; D018108:Respirovirus / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D013154:Spleen / Q000166:cytology / Q000276:immunology*; D014162:Transfection
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D015707:HN Protein; D011994:Recombinant Proteins; D005557:Formaldehyde; D007372:Interferons