Development of a molecular probe to baboon interleukin-10 mRNA for in situ hybridization during experimental schistosomiasis.
A nucleic acid probe complementary to baboon interleukin 10 (IL-10) mRNA was developed for in situ hybridization. Highly conserved IL-10 protein sequences from several mammals were aligned to design oligonucleotide primers flanking a 270-bp sequence of the target cDNA. RNA was isolated from stimulated peripheral blood mononuclear cells (PBMC). IL-10 cDNA was reverse-transcribed from the total PBMC RNA and amplified with the polymerase chain reaction (PCR). Cloning and sequencing of the PCR product confirmed it to be of baboon IL-10 origin, with 97.8% identity to human and 100% identity to macaque mRNA sequences. The baboon IL-10 DNA probe hybridized in Southern blots to a 7.9-Kbp or 8.6-Kbp band after digestion of genomic baboon DNA with Bam H1 or Eco R1, respectively. Preliminary results with an antisense riboprobe derived from this sequence showed the presence of IL-10 mRNA in sections of granulomatous tissues.
Chou
T M
TM
Department of Cellular Biology, University of Georgia, Athens, Georgia 30602, USA. wchou@fdah.com
Stich
R W
RW
Marsala
C
C
Scott
M
M
Brown
C C
CC
Rawlings
C A
CA
Damian
R T
RT
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D005260:Female; D006801:Humans; D017403:In Situ Hybridization; D016753:Interleukin-10 / Q000235:genetics*; D008214:Lymphocytes / Q000276:immunology; D008251:Macaca; D008969:Molecular Sequence Data; D010215:Papio; D016133:Polymerase Chain Reaction; D018419:Primate Diseases / Q000175:diagnosis* / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D012552:Schistosomiasis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D012689:Sequence Homology, Nucleic Acid
D012333:RNA, Messenger; D016753:Interleukin-10