Development of a molecular probe to baboon interleukin-10 mRNA for in situ hybridization during experimental schistosomiasis

Ann N Y Acad Sci. 2000:916:410-6. doi: 10.1111/j.1749-6632.2000.tb05320.x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Consensus Sequence
  • Female
  • Humans
  • In Situ Hybridization
  • Interleukin-10 / genetics*
  • Lymphocytes / immunology
  • Macaca
  • Molecular Sequence Data
  • Papio
  • Polymerase Chain Reaction
  • Primate Diseases / diagnosis*
  • Primate Diseases / immunology
  • RNA, Messenger / analysis*
  • Reproducibility of Results
  • Schistosomiasis / diagnosis
  • Schistosomiasis / immunology
  • Schistosomiasis / veterinary*
  • Sequence Homology, Nucleic Acid

Substances

  • RNA, Messenger
  • Interleukin-10