title: Anti-CD3/anti-CD28 bead stimulation overcomes CD3 unresponsiveness in patients with head and neck squamous cell carcinoma.
abstract: OBJECTIVE
  To test whether T-cell CD3 responses are altered in patients with advanced-stage head and neck squamous cell carcinoma (HNSCC) and whether anti-CD3/anti-CD28 (alphaCD3/alphaCD28) bead stimulation could reverse CD3 unresponsiveness.
  
  
  METHODS
  Anti-CD3 (alphaCD3) monoclonal antibody immobilized on tissue culture plastic was used to stimulate lymph node mononuclear cells (LNMCs) and peripheral blood mononuclear cells (PBMCs) from patients with advanced-stage HNSCC. Proliferation, T-cell phenotype, and cytokines were measured during 8-day in vitro stimulation. Immune-enhancing properties of alphaCD3/ alphaCD28 beads were also tested on LNMCs and PBMCs. Cytotoxicity of bead-activated T cells (ATCs) was measured against autologous and allogeneic HNSCC.
  
  
  RESULTS
  Six patients were nonresponders to alphaCD3 stimulation defined by tritium (3H) incorporation of less than 3500 cpm, whereas 11 patients were responders with 3H incorporation of 3500 cpm or more. Responders produced higher levels of interleukin (IL)-12 and interferon gamma (IFN-gamma) after alphaCD3 stimulation than nonresponders. No phenotypic or clinical differences were identified between groups. Stimulation with alphaCD3/alphaCD28 beads enhanced IFN-gamma and IL-2 produced by both groups. Bead ATCs were generated from PBMCs of patient 11 in the responder group and lysed (+/- SD) 100% +/-1% of autologous tumor and 49% +/-1% of allogeneic tumor. Bead ATCs from LNMCs of this patient lysed 58%+/-1% of autologous tumor and 63%+/-1% of allogeneic tumor.
  
  
  CONCLUSIONS
  A subpopulation of patients with HNSCC who are nonresponders to alphaCD3 stimulation has been identified, showing reduced proliferation and IL-12 and IFN-gamma secretion. Nonresponders stimulated with alphaCD3/alphaCD28 beads reversed immune unresponsiveness and induced a type 1 cytokine response. Bead-generated ATCs from patient 11 in the responder group lysed autologous and allogeneic HNSCC in vitro, suggesting a possible effective immunotherapeutic modality in the treatment of HNSCC.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Wayne State University School of Medicine, and the Karmanos Cancer Institute, Detroit, Mich 48201, USA. Tshibuya@med.wayne.edu
authors.forename: T Y
authors.initials: TY
authors.lastname: Shibuya
authors.forename: W Z
authors.initials: WZ
authors.lastname: Wei
authors.forename: M
authors.initials: M
authors.lastname: Zormeier
authors.forename: J
authors.initials: J
authors.lastname: Ensley
authors.forename: W
authors.initials: W
authors.lastname: Sakr
authors.forename: R H
authors.initials: RH
authors.lastname: Mathog
authors.forename: R J
authors.initials: RJ
authors.lastname: Meleca
authors.forename: G H
authors.initials: GH
authors.lastname: Yoo
authors.forename: C H
authors.initials: CH
authors.lastname: June
authors.forename: B L
authors.initials: BL
authors.lastname: Levine
authors.forename: L G
authors.initials: LG
authors.lastname: Lum
mesh_terms: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D002294:Carcinoma, Squamous Cell / Q000276:immunology* / Q000628:therapy; D006258:Head and Neck Neoplasms / Q000276:immunology* / Q000628:therapy; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology; D007167:Immunotherapy; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000276:immunology; D018664:Interleukin-12 / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D017252:CD3 Complex; D018664:Interleukin-12; D007371:Interferon-gamma