In vivo selective expansion of a tumour‐specific cytotoxic T‐cell clone derived from peripheral blood of a melanoma patient after vaccination with gene‐modified autologous tumour cells

Abstract

SUMMARYMelanoma-speci®c cytotoxic T lymphocytes (CTL) can be generated from peripheral blood lymphocytes (PBL) by mixed lymphocyte±tumour cell cultures. Analysis of CTL precursor frequencies in peripheral blood of melanoma patients is generally used for immunomonitoring purposes to evaluate vaccination ef®cacy. At present, it is unclear whether PBL-derived CTL generated in vitro are indicative of an anti-tumour immune response in vivo. Three tumour-speci®c human leucocyte antigen (HLA)-B/C-restricted CTL clones were derived from peripheral blood of a melanoma patient immunized with interleukin-7 (IL-7) gene-modi®ed tumour cells. CTL clones differing in their T-cell receptor-c (TCRc) rearrangement produced interferon-c, IL-4 and/or IL-10. On the basis of their unique TCRc gene rearrangements clone-speci®c primers were generated for detection of clone-speci®c DNA by polymerase chain reaction. One CTL clone (E5) of the three was found to be selectively expanded in one of seven metastases obtained at autopsy, as determined by Southern blot hybridization. However, the presence of E5 in only one of seven metastases at death indicates that the in vivo accumulation of the speci®c CTL clone was not suf®cient to contain tumour progression. Nevertheless, our data support the proposition that analysis of anti-tumour activity of PBL-derived CTLs may re¯ect an anti-tumour immune response in vivo.

Evaluating the Potential of T Cell Receptor Repertoires in Predicting the Prognosis of Resectable Non-Small Cell Lung Cancers

“…These results were consistent with previous findings that T cells infiltrating in tumor lesions comprise high numbers of clonal expansion 29 and that the proliferation of oligoclonal T cells in tumor lesions would be massively induced in response to tumor-associated antigenic peptides. 30 Our study showed the diversity of TCRs, which was calculated by Shannon diversity index 31 had no difference compared with adjacent non-tumor tissues in NSCLC ( Figure 1 ). However, a recent study that presented data from 15 lung cancer patients showed that significantly higher TCR diversity was observed in tumor tissues than in normal lung tissues, 32 the increases or decreases of the diversity of TCRs in tumors relative to paired normal tissues are inconsistent in different cancer types 33 , 34 , 35 or even same type in different studies.…”

Comparative assessment of TCRBV diversity in T lymphocytes present in blood, metastatic lesions, and DTH sites of two melanoma patients vaccinated with an IL-7 gene-modified autologous tumor cell vaccine

“…In a previous report, it was shown that vaccination with autologous melanoma cells transfected ex vivo to produce IL -7 induced, in PBL of the treated patients, both MHC-restricted as well as MHC -unrestricted lytic activity against the autologous melanoma. 13,33,34 In addition, a number of clones with antitumor specificity could be derived from PBL of one of the patients. 33 In spite of documentation of this antitumor Tcell response, no durable clinical responses were, however, observed.…”

“…13,33,34 In addition, a number of clones with antitumor specificity could be derived from PBL of one of the patients. 33 In spite of documentation of this antitumor Tcell response, no durable clinical responses were, however, observed. 13 Ex vivo sensitisation of pre -and postvaccine PBLs that identify a vaccine-specific systemic T cell response to immunisation is, however, not an unusual finding even in the absence of clinical responses.…”

“…13,33,34 The primary objective of this study was to analyse whether immunological changes occurred in the neoplastic lesions following vaccination and to measure the extent and the nature of such changes in comparison to blood and, for one of the patients, to delayed type hypersensitivity ( DTH ) skin reactions at sites of autologous tumor inoculum. Using semiquantitative polymerase chain reaction (PCR ) for BV repertoire and CDR3 length pattern analysis, we could demonstrate, in postvaccine metastases, increased frequency expression of polyclonal T-cell subsets characterised by few BVs.…”

Segregation of Effector Mechanisms in a Tumour‐Specific CD8+ T‐Cell Clone Correlates with CD30 Expression

“…Cytotoxicity assays Cytolytic activity of CTL against melanoma cells, EBV‐B cells and K562 was determined in a 6 h LDH‐release assay as described elsewhere [15]. Briefly, 5 × 10 3 target cells were cultured in triplicates with varying numbers of effector cells for 6 h at 37°C in 5% CO2.…”