Oral Polio vaccination leads to oligoclonal expansion of TCRBV16+ and TCRBV13+ T cells in the colon of rhesus macaques Sabine Hörer a, ⁎, Susanne Burdak-Rothkamm b , Kristina Allers c , Thomas Schneider c , Rainer Duchmann d a Department of Neurology, City Hospital Munich, Klinikum Harlaching, Sanatoriumsplatz 2, 81545 München, Germany b Gray Cancer Institute, Mount Vernon Hospital, Northwood, UK c Medical Department I, Charité Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany d Medical Department, Hospital zum Heiligen Geist, Frankfurt/Main, Germany abstract article info Article history: Received 6 August 2008 Available online 15 August 2008 Keywords: CDR3 length analysis Intestinal immunity Macaque Mucosal immunity Oligoclonal T cells Oral Polio vaccination T cell receptor In studying immune responses towards the poliovirus, data about T cell mediated immunity in the intestine as the main portal of viral entry in disease and vaccination is lacking. We treated two macaques with oral Polio vaccine and collected duodenal and colonic biopsy specimens. RNA isolation, reverse transcription, and polymerase chain reaction were performed for fragment analysis of the complementarity determining region 3 (CDR3) of the T cell receptor beta chain variable region (TCRBV), followed by subcloning and sequencing of expanded bands. In the colon, oligoclonal expansions of TCRBV16+ or TCRBV13+ intestinal T cells with conserved motifs of the hypervariable CDR3 were found. Flow cytometric analysis of mucosal T cells revealed that activated colonic T cells were mainly CD4 + . Our results indicate that there is a local activation of oligoclonal T cells in the colon after oral Polio vaccination (OPV) which involves selected TCRBV families and may occur within the CD4 + T cell subset. © 2008 Elsevier Inc. All rights reserved. Introduction The widespread use of inactivated and attenuated oral Polio vaccine has led to eradication of poliomyelitis in large parts of the world (Goncalves et al., 2003, Slonim, 2005). However, sporadic outbreaks of the disease are still reported (Gaspar et al., 2000, Kew et al., 2002, Thorley et al., 2003). Studies about immunity towards poliovirus have often focussed on humoral immune responses, providing information about neutralizing antibody responses in various populations (Diedrich et al., 2002, Glezen et al., 1966, Hogg et al., 2002, Mastroeni et al., 1997, Winter et al., 1981) Less attention has been paid to cellular mechanisms of immunity against poliovirus, especially at the mucosal surface of the intestinal tract. T cell responses to infection with or vaccination against poliovirus have been primarily studied in systemic peripheral lymphoid organs (e.g. blood, spleen) of humans or poliovirus receptor (CD155) transgenic mice. T cell epitopes for all four capsid proteins of the virus (Kutubuddin et al., 1992a, 1992b) as well as effector human and murine T cell populations of CD4 + (Mahon et al., 1995, Simons et al., 1993) and CD8 + phenotype (Wahid et al., 2005) have been identified. In contrast to the detailed knowledge about T cell mediated immune responses towards poliovirus in systemic lymphoid tissue, there is no such evidence in terms of local T cell immunity in the intestinal tract. However, this mucosal surface represents the main site of entry in poliovirus infection. We addressed the question whether there is a local intestinal T cell response after oral Polio vaccination (OPV). We therefore obtained duodenal and colonic biopsy specimens from two healthy rhesus macaques (Macaca mulatta) after feeding oral Polio vaccine and performed a sequence analysis of the hypervariable, i.e. non-germline encoded, comple- mentarity determining region 3 (CDR3) of the T cell receptor (TCR) beta chain. As the CDR3 fragment of the TCR beta chain directly interacts with the antigenic epitope, the finding of oligoclonal T cell populations with similar CDR3 might provide evidence for a local intestinal T cell response towards OPV. In addition, we performed flow cytometric analysis of mucosal T cells of one animal to define T cell subsets activated after OPV. Materials and methods Animals Two 2-year old colony bred male macaques were imported from the Laboratory Animal Breeders and Services (Jemassee, North Carolina, USA) and caged individually at the German Primate Centre (Göttingen, Germany). Animals obtained standardised commercially available dry food (Purina Mills monkey chow; Purina, St. Louis, Missouri, USA) supplemented with fresh fruit twice a day. Animal care and handling were performed under the German Animal Protection Experimental and Molecular Pathology 85 (2008) 189–195 Abbreviations: CDR3, complementarity determining region 3; OPV, oral Polio vaccination; TCR, T cell receptor; TCRBV, T cell receptor beta chain variable region. ⁎ Corresponding author. Fax: +49 89 6210 3488. E-mail addresses: sabine.hoerer@lycos.de, hoererjs@aol.com (S. Hörer). 0014-4800/$ – see front matter © 2008 Elsevier Inc. All rights reserved. doi:10.1016/j.yexmp.2008.08.001 Contents lists available at ScienceDirect Experimental and Molecular Pathology journal homepage: www.elsevier.com/locate/yexmp