Tumor Infiltrating Lymphocytes in Melanoma Comprise High Numbers of T-Cell Clonotypes That Are Lost during in Vitro Culture

Straten, Per thor; Kirkin, Alexei F.; Siim, Elsebeth; Dahlstrøm, Karin; Drzewiecki, Krzysztof T.; Seremet, Tina; Zeuthen, Jesper; Becker, Jürgen C.; Guldberg, Per

doi:10.1006/clim.2000.4890

Cited by 24 publications

(15 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7,8,30 Therefore, it might result in a biased selection of CTL clones according to their proliferative potential. 33,34 In fact, we observed differences in dominant clones between cultured and non-cultured donor cells in Cases-2 and -3. The current direct single-cell analysis method is important for evaluating the in vivo dynamics of CMV-CTL more precisely.…”

Section: Discussionmentioning

confidence: 72%

Single-cell T-cell receptor-β analysis of HLA-A*2402-restricted CMV- pp65-specific cytotoxic T-cells in allogeneic hematopoietic SCT

Nakasone

Tanaka

Yamazaki

et al. 2013

Bone Marrow Transplant

View full text Add to dashboard Cite

Cellular immunity is important for the control of CMV infection after allogeneic hematopoietic cell transplantation (Allo-HCT). However, the actual in vivo dynamics of CMV-specific cytotoxic T cell (CMV-CTL) clones are still unclear. We conducted clone monitoring of tetramer(+) CMV-CTLs in HLA-A*2402-positive donor-patient pairs, using a direct single-cell analysis that enabled the simultaneous identification and quantification of CTL clones. Clone dynamics were assessed in three cases with or without CMV reactivation. In Case-1 without CMV reactivation, despite the long-term use of systemic steroid, dominant clones of Donor-1 persisted and remained dominant. The CMV-CTLs at 1 year after Allo-HCT included a high proportion of CD45RA(+)CCR7(-) effector and CD27(-)CD57(+)mature T cells. On the other hand, in Cases-2 and -3 with CMV reactivation, novel clones appeared and became dominant during the follow-up. Their CMV-CTLs included more CD27(+) immature T cells at 1 year after Allo-HCT. With regard to clonotypes, HLA-A*2402-restricted CMV-CTLs tended to select BV7 and BJ1-1 genes for complementarity-determining region 3 (CDR3) of T-cell receptor (TCR)-β. Specific amino-acid sequences of CDR3 of TCR-β were found in each case. Patterns of clone reconstitution and phenotype would be different according to CMV reactivation. In vivo clone monitoring of CMV-CTLs could provide insight into the mechanism of immunological reconstitution following Allo-HCT.

show abstract

Section: Discussionmentioning

confidence: 72%

Single-cell T-cell receptor-β analysis of HLA-A*2402-restricted CMV- pp65-specific cytotoxic T-cells in allogeneic hematopoietic SCT

Nakasone

Tanaka

Yamazaki

et al. 2013

Bone Marrow Transplant

View full text Add to dashboard Cite

show abstract

“…level (30), (b) analyzed cells might not target antigen-specific T cells without the use of specific HLA tetramers, (c) in vitro culture might induce biases in the TCR repertoire by clonal selection affected by rapid or slow proliferation and high or low sensitivity to activation-induced cell death (39,40), and (d) PCR using cDNAs derived from bulky cells might also induce biases due to intrinsic differences in the amplification efficiency of the templates (41). In this study, TCR-β CDR3 was directly sequenced using individually sorted Tax tetramer + cells without in vitro culture because we aimed to analyze the TCR repertoire of HTLV-1 Tax 301-309 peptide-specific CTLs in vivo without any technical bias in vitro.…”

Section: Discussionmentioning

confidence: 99%

Single-Cell Analysis of T-Cell Receptor Repertoire of HTLV-1 Tax-Specific Cytotoxic T Cells in Allogeneic Transplant Recipients with Adult T-Cell Leukemia/Lymphoma

et al. 2010

View full text Add to dashboard Cite

Adult T-cell leukemia (ATL) is a lymphoproliferative malignancy associated with human T-cell lymphotropic virus type 1 (HTLV-1) infection. Recently, it has been shown that allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for ATL, and that HTLV-1 Tax-specific CD8 + cytotoxic T cells (CTL) contribute to the graft-versus-ATL effect. In the present study, we, for the first time, analyzed the T-cell receptor (TCR) repertoire of isolated Tax 301-309 (SFHSLHLLF)-specific CTLs in HLA-A*2402 + ATL patients before and after allo-HSCT by single-cell reverse transcription-PCR. The Tax 301-309 -specific CTLs in bone marrow and peripheral blood showed highly restricted oligoclonal diversity. In addition, a unique conserved amino acid motif of "P-D/P-R" in TCR-β complementarity-determining region 3 in either BV7-or BV18-expressing CTLs was observed not only in all of the samples from ATL patients, but also in samples from the same patient before and after HSCT. Furthermore, the P-D/P-R motif-bearing CTL clones established from peripheral blood samples after HSCT exhibited strong killing activity against the HTLV-1-infected T cells of the patient. CTL clones were not established in vitro from samples prior to allo-HSCT. In addition, CTL clones with a strong killing activity were enriched in vivo after HSCT in the patient. Hence, Tax 301-309 -specific CTLs in ATL patients might have a preference for TCR construction and induce strong immune responses against the HTLV-1-infected T cells of patients, which contribute to the graft-versus-ATL effects after allo-HSCT. However, further analyses with a larger number of patients and more frequent sampling after allo-HSCT is required to confirm these findings. Cancer Res; 70(15);

show abstract

“…However, often the clinical relevance of the results is uncertain, because the effector cells may not be representative of the original T cell population (26). PCR-based analysis of the TCR repertoire of tumors has the advantage of providing an in situ profile of tumor-infiltrating lymphocytes without the need for their in vitro growth and expansion.…”

Section: Discussionmentioning

confidence: 99%

TCR Rearrangement in Lymphocytes Infiltrating Melanoma Metastases After Administration of Autologous Dinitrophenyl-Modified Vaccine

Manne

Mastrangelo

Sato

et al. 2002

The Journal of Immunology

View full text Add to dashboard Cite

Administration of a vaccine consisting of autologous melanoma cells modified with a hapten, dinitrophenyl (DNP), induces T cell infiltration of metastatic sites. We have reported an analysis of these infiltrating T cells, indicating that certain TCR-Vβ gene segments are greatly overexpressed. In this study, we investigate the rearrangement of the TCR-Vβ as well as the junctional diversity in T cells infiltrating melanoma metastases following treatment with DNP vaccine. In 19 of 26 control specimens, V-D-J length analysis showed the expected polyclonal patterns. In contrast, postvaccine tumors from 9 of 10 patients showed dominant peaks of V-D-J junction size in one or more Vβ families. Dominant peaks were seen most frequently in six Vβ families (Vβ7, 12, 13, 14, 16, and 23) and were never seen in seven others. Further analysis of the oligoclonal Vβ products showed dominant peaks in the J region as well. Of particular interest was the finding that Vβ and Jβ peaks were similar in inflamed metastases obtained at different times or from different sites from the same patient. Although 6 of 10 patients expressed HLA-A1, there was no common pattern of TCR rearrangements among them. Finally, the amplified PCR products from seven of these specimens were cloned and sequenced and the amino acid sequence of the complementarity-determining region 3 was deduced. In six of seven specimens, the same complementarity-determining region 3 sequence was repeated in at least two clones and in five of seven in at least three clones. Our study indicates that DNP vaccine induces the expansion of particular T cell clones that may be agents of its antitumor effects.

show abstract

Tumor Infiltrating Lymphocytes in Melanoma Comprise High Numbers of T-Cell Clonotypes That Are Lost during in Vitro Culture

Cited by 24 publications

References 32 publications

Single-cell T-cell receptor-β analysis of HLA-A*2402-restricted CMV- pp65-specific cytotoxic T-cells in allogeneic hematopoietic SCT

Single-cell T-cell receptor-β analysis of HLA-A*2402-restricted CMV- pp65-specific cytotoxic T-cells in allogeneic hematopoietic SCT

Single-Cell Analysis of T-Cell Receptor Repertoire of HTLV-1 Tax-Specific Cytotoxic T Cells in Allogeneic Transplant Recipients with Adult T-Cell Leukemia/Lymphoma

TCR Rearrangement in Lymphocytes Infiltrating Melanoma Metastases After Administration of Autologous Dinitrophenyl-Modified Vaccine

Contact Info

Product

Resources

About