Anti-CD3/anti-CD28 bead stimulation overcomes CD3 unresponsiveness in patients with head and neck squamous cell carcinoma

Objectives: To test whether T-cell CD3 responses are altered in patients wi th advanced-stage head and neck squamous cell carcinoma (HNSCC) and whether anti-CD3/anti-CD28 (alpha CD3/alpha CD28) bead stimulation could reverse C D3 unresponsiveness. Design: Anti-CD3 (alpha CD3) monoclonal antibody immobilized on tissue cult ure plastic was used to stimulate lymph node mononuclear cells (LNMCs) and peripheral blood mononuclear cells (PBMCs) from patients with advanced-stag e HNSCC. Proliferation, T-cell phenotype, and cytokines were measured durin g 8-day in vitro stimulation. Immune-enhancing properties of alpha CD3/alph a CD28 beads were also tested on LNMCs and PBMCs. Cytotoxicity of bead-acti vated T cells (ATCs) was measured against autologous and allogeneic HNSCC. Results: Six patients were nonresponders to alpha CD3 stimulation defined b y tritium (H-3) incorporation of less than 3500 cpm, whereas 11 patients we re responders with H-3 incorporation of 3500 cpm or more. Responders produc ed higher levels of interleukin (IL)-12 and interferon gamma (IFN-gamma) af ter alpha CD3 stimulation than nonresponders. No phenotypic or clinical dif ferences were identified between groups. Stimulation with alpha CD3/alpha C D28 beads enhanced IFN-gamma and IL-2 produced by both groups. Bead ATCs we re generated from PBMCs of patient 11 in the responder group and lysed (+/- SD) 100% +/- 1% of autologous tumor and 49% +/- 1% of allogeneic tumor. Be ad 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 t o alpha CD3 stimulation has been identified, showing reduced proliferation and IL-12 and IFN-gamma secretion. Nonresponders stimulated with alpha CD3/ alpha CD28 beads reversed immune unresponsiveness and induced a type 1 cyto kine response. Bead-generated ATCs from patient 11 in the responder group l ysed autologous and allogeneic HNSCC in vitro, suggesting a possible effect ive immunotherapeutic modality in the treatment of HNSCC.