Expansion and Immunological Study of Human Tumor Infiltrating Gamma/Delta T Lymphocytes in vitro¹

Abstract

γ/δ T cells have stimulated a lot of interest because of their unique features in antigen recognition and cytotoxicities to many autologous and/or allogeneic tumor cells. We have developed a novel method to selectively expand larger amounts of human tumor–infiltrating γ/δ T lymphocytes (γ/δ TILs) ex vivo by immobilized pan– anti–TCRγ/δ monoclonal antibody in the presence of exogenous IL–2. The expanded γ/δ TILs mainly expressed CD45RO and HLA–DR molecules and did not express CD4. CD8+ γ/δ TILs accounted for 19% of γ/δ TILs. The expression of CD25 molecule on expanded γ/δ T cells was inducible and downregulated following a time course. The Vδ1 and Vδ2 subsets amount to 37 and 58%, respectively. The expanded γ/δ TILs show an IL–2–dependent proliferation, MHC class I–unrestricted and TCRγ/δ–related cytotoxicities to two MHC class I⁺ and two MHC class I⁺ allogeneic tumor cell lines in vitro.

Vaccination against the HER-2/neu oncogenic protein.

“…γδ T-cells are involved in a wide range of immune responses to infectious and non-infectious diseases, including malaria, mycobacterial infections, cancers, as well as autoimmune disorders such as multiple sclerosis (Boismenu & Havran 1998). Often, γδ T-cells clones are isolated from the tumor-infiltrating lymphocyte population of many tumors, including dysgerminoma (Zhao et al 1995), seminoma (Zhao et al 1995), renal carcinoma (Choudhary et al 1995), lung (Yu et al 1999), colorectal (Watanabe et al 1995), and melanoma (Bachelez et al 1992). The recruitment to and role of these unique cells in mediating antitumor immunity is unknown.…”

Adoptive immunotherapy of lung cancer with immobilized anti-TCRγδ antibody-expanded human γδ T Cells in peripheral blood

“…24 Briefly, each well on 24-well plate was coated with 0.5 μg anti-TCRγδ (Immunotech, Beckman Coulter, USA). Then PBMCs were transferred to the plates and cultured in RPMI 1640 medium supplemented with 5% pooled human AB plasma (Beijing Red Cross Blood Center, China) and recombinant human IL-2 (200 IU/mL, Beijing Read United Cross Pharmaceutical Co., Ltd., China).…”

“…[18][19][20][21][22][23] We have previously established the condition to expand γδ T-cells in a 2 w culture period with immobilized anti-TCRγδ antibody in vitro. 24 Such TCRγδ ligandation initiates γδ T-cell activation to display marked killing activity to both hematological and solid tumor cell lines. In particular, we are interested in antitumor activity of anti-TCRγδ antibody-expanded γδ T-cells to solid tumors, especially lung cancer.…”

Clonal diversity of the T-cell population responding to a dominant HLA-A2 epitope of HER-2/neu after active immunization in an ovarian cancer patient

“…␥␦-TCR T cells are involved in a wide range of immune responses to infectious and non-infectious diseases, including malaria, mycobacterial infections, cancers, and autoimmune disorders, such as multiple sclerosis [24]. The ␥␦-TCR T-cell clones have been isolated from tumor-infiltrating lymphocytes derived from tumors, such as dysgerminoma [25], seminoma [25], renal carcinoma [26], lung [27], colorectal [28], and melanoma [29]. Tumor-associated ␥␦-TCR T cells that have been described in colorectal cancers have a CD8 ϩ , cytolytic phenotype similar to those described in the present study [27].…”

“…The ␥␦-TCR T-cell clones have been isolated from tumor-infiltrating lymphocytes derived from tumors, such as dysgerminoma [25], seminoma [25], renal carcinoma [26], lung [27], colorectal [28], and melanoma [29]. Tumor-associated ␥␦-TCR T cells that have been described in colorectal cancers have a CD8 ϩ , cytolytic phenotype similar to those described in the present study [27]. In the present study, ␥␦-TCR T-cell clone 1D1, displayed heterogeneous expression of CD4 or CD8.…”