Proteomic analysis of Glossina pallidipes salivary gland hypertrophy virus virions for immune intervention in tsetse fly colonies.
Many species of tsetse flies (Diptera: Glossinidae) can be infected by a virus that causes salivary gland hypertrophy (SGH). The genomes of viruses isolated from Glossina pallidipes (GpSGHV) and Musca domestica (MdSGHV) have recently been sequenced. Tsetse flies with SGH have reduced fecundity and fertility which cause a serious problem for mass rearing in the frame of sterile insect technique (SIT) programmes to control and eradicate tsetse populations in the wild. A potential intervention strategy to mitigate viral infections in fly colonies is neutralizing of the GpSGHV infection with specific antibodies against virion proteins. Two major GpSGHV virion proteins of about 130 and 50&emsp14;kDa, respectively, were identified by Western analysis using a polyclonal rabbit antibody raised against whole GpSHGV virions. The proteome of GpSGHV, containing the antigens responsible for the immune-response, was investigated by liquid chromatography tandem mass spectrometry and 61 virion proteins were identified by comparison with the genome sequence. Specific antibodies were produced in rabbits against seven candidate proteins, including the ORF10/C-terminal fragment, ORF47 and ORF96 as well as proteins involved in peroral infectivity PIF-1 (ORF102), PIF-2 (ORF53), PIF-3 (ORF76) and P74 (ORF1). Antiserum against ORF10 specifically reacted to the 130&emsp14;kDa protein in a Western blot analysis and to the envelope protein of GpSGHV, detected by using immunogold-electron microscopy. This result suggests that immune intervention of viral infections in colonies of G. pallidipes is a realistic option.
Kariithi
Henry M
HM
Laboratory of Virology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
Ince
Ikbal Agah
IA
Boeren
Sjef
S
Vervoort
Jacques
J
Bergoin
Max
M
van Oers
Monique M
MM
Abd-Alla
Adly M M
AM
Vlak
Just M
JM
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D015153:Blotting, Western; D004267:DNA Viruses / Q000737:chemistry* / Q000302:isolation & purification; D023061:Gene Order; D005814:Genes, Viral; D013058:Mass Spectrometry; D008970:Molecular Weight; D020543:Proteome / Q000032:analysis*; D011817:Rabbits; D012469:Salivary Glands / Q000821:virology*; D014370:Tsetse Flies / Q000821:virology*; D014764:Viral Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D014771:Virion / Q000737:chemistry* / Q000302:isolation & purification
D000914:Antibodies, Viral; D000956:Antigens, Viral; D020543:Proteome; D014764:Viral Proteins