Generation, maintenance and reactivity of ovine T-lymphocyte clones derived from sheep immunized with pili from Bacteroides nodosus.
Peripheral blood leucocytes (PBL) from sheep immunized with pilus protein purified from Bacteroides nodosus serogroup A were cultivated in vitro and cloned in the presence of the specific antigen and autologous antigen-presenting cells (APC). The efficiency of cloning was enhanced by high proliferative responses to pili during the initial week of cultivation, and the provision of recombinant human interleukin-2 (rec-IL-2). After three passages at weekly intervals, bulk cultures of PBL and cloned T-lymphocytes were greater than 99% CD4+, CD8-, sIg-, i.e. the characteristic phenotype of helper T-lymphocytes. Cloned T-lymphocytes were devoid of allo-reactivity, and were restricted by class II antigens of the major histocompatibility complex (MHC). Both bulk PBL and cloned T-lymphocytes exhibited similar patterns of reactivity against pili from different serogroups of B. nodosus and the T-lymphocytes reacted to three of six peptides synthesized from the amino-acid sequence of pilus from serogroup A. Although clones of T-lymphocytes could retain antigen specificity for up to 2 months of cultivation, several attempts to recover clones with specific reactivity after storage in liquid nitrogen were unsuccessful.
Emery
D L
DL
CSIRO, Division of Animal Health, McMaster Laboratory, Australia.
Rothel
J S
JS
Kirkpatrick
A
A
Maclaren
J A
JA
D000818:Animals; D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D001439:Bacteroides / Q000276:immunology*; D002455:Cell Division; D002999:Clone Cells; D000939:Epitopes; D010861:Fimbriae, Bacterial / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007376:Interleukin-2 / Q000494:pharmacology; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010641:Phenotype; D011994:Recombinant Proteins / Q000494:pharmacology; D012756:Sheep / Q000235:genetics / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D010455:Peptides; D011994:Recombinant Proteins