Limits...
T lymphocytes derived from human cord blood provide effective antitumor immunotherapy against a human tumor.

Lee YS, Kim TS, Kim DK - BMC Cancer (2011)

Bottom Line: Although the graft-versus-tumor (GVT) effect of donor-derived T cells after allogeneic hematopoietic stem cell transplantation has been used as an effective adoptive immunotherapy, the antitumor effects of cord blood (CB) transplantation have not been well studied.NOD/SCID mice that receive neonatal CB transplants have reconstituted T cells with significant antitumor effects against human cervical and lung tumors, with a high infiltration of CD3+ T cells showing dramatic induction of apoptotic cell death.In adoptive transfer of CD3+ T cells into mice with pre-established tumors, we observed much higher antitumor effects of HPV-specific T cells by ELISPOT assays.

View Article: PubMed Central - HTML - PubMed

Affiliation: Transplantation Research Center, Samsung Biomedical Research Institute, Graduate School of Life Science and Biotechnology, CHA University, Seoul, Republic of Korea.

ABSTRACT

Background: Although the graft-versus-tumor (GVT) effect of donor-derived T cells after allogeneic hematopoietic stem cell transplantation has been used as an effective adoptive immunotherapy, the antitumor effects of cord blood (CB) transplantation have not been well studied.

Methods: We established the animal model by transplantation of CB mononuclear cells and/or tumor cells into NOD/SCID mice. The presence of CB derived T cells in NOD/SCID mice or tumor tissues were determined by flow cytometric and immunohistochemical analysis. The anti-tumor effects of CB derived T cells against tumor was determined by tumor size and weight, and by the cytotoxicity assay and ELISPOT assay of T cells.

Results: We found dramatic tumor remission following transfer of CB mononuclear cells into NOD/SCID mice with human cervical tumors with a high infiltration of CD3+ T cells in tumors. NOD/SCID mice that receive neonatal CB transplants have reconstituted T cells with significant antitumor effects against human cervical and lung tumors, with a high infiltration of CD3+ T cells showing dramatic induction of apoptotic cell death. We also confirmed that T cells showed tumor specific antigen cytotoxicity in vitro. In adoptive transfer of CD3+ T cells into mice with pre-established tumors, we observed much higher antitumor effects of HPV-specific T cells by ELISPOT assays.

Conclusions: Our results show that CB derived T lymphocytes will be useful for novel immunotherapeutic candidate cells for therapy of several tumors in clinic.

Show MeSH

Related in: MedlinePlus

Cytotoxic activity and detection of cervical tumor specific T cells. A, cervical tumor specific cytotoxic activity was shown. T cells were purified from splenocytes of NOD/SCID mice inoculated with cord blood T cells (CB-T) and Caski tumor cells. The T cells were cocultured with Caski cells, HeLa cells, or A549 cells (human lung cancer cells) as a negative control. Cytotoxicity was tested by flow cytometry with several effector-to-target ratios. B, cytotoxic activity of purified T cells against HeLa (left) and Caski (right) cells was determined for non-adoptive T cells (1st CB-T) and adoptive T cells (2nd CB-T), and human peripheral blood T lymphocytes (PBL-T) as a control. Each bar indicates the mean ± S.D. as determined from the average of quadruplicate wells (* P < 0.01). C, ELISPOT assay for interferon-gamma (INF-γ) production was conducted with E711-20 peptide (YMLDLQPETT) and E786-93 peptide (TLGIVCPI) HPV specific peptides with non-adopted T cells and adopted T cells. Each bar represents the mean ± S.D. as determined (* P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3141763&req=5

Figure 6: Cytotoxic activity and detection of cervical tumor specific T cells. A, cervical tumor specific cytotoxic activity was shown. T cells were purified from splenocytes of NOD/SCID mice inoculated with cord blood T cells (CB-T) and Caski tumor cells. The T cells were cocultured with Caski cells, HeLa cells, or A549 cells (human lung cancer cells) as a negative control. Cytotoxicity was tested by flow cytometry with several effector-to-target ratios. B, cytotoxic activity of purified T cells against HeLa (left) and Caski (right) cells was determined for non-adoptive T cells (1st CB-T) and adoptive T cells (2nd CB-T), and human peripheral blood T lymphocytes (PBL-T) as a control. Each bar indicates the mean ± S.D. as determined from the average of quadruplicate wells (* P < 0.01). C, ELISPOT assay for interferon-gamma (INF-γ) production was conducted with E711-20 peptide (YMLDLQPETT) and E786-93 peptide (TLGIVCPI) HPV specific peptides with non-adopted T cells and adopted T cells. Each bar represents the mean ± S.D. as determined (* P < 0.05).

Mentions: We assessed whether mice with both reconstituted immunity and inoculation with Caski tumor cells possessed memory T cells with specific activity against cervical tumor cells in vitro. Purified CD3+ T cells from these mice were cocultured with Caski cells, as specific target cells, and A549 human lung cancer cells, as a negative control. Cytotoxic effects were then determined by Annexin V and PI staining with flow cytometry analysis. The T cells showed significantly higher cytotoxic activity against Caski cells than against A549 control cells (Figure 6A). This finding suggests that mice with reconstituted immunity that are injected with Caski tumor cells develop memory T cells with specificity for the tumor cells. We performed an additional in vitro analysis to assess the tumor-specific cytotoxic activity of purified CD3+ T cells from spleen of mice which are either adapted or non-adapted mice followed by either Caski cell injection, HeLa cell injection, or no injection of tumor cells. We found much higher cytotoxicity in T cells from tumor adapted T cells bearing mice than T cells than non-adapted control T cells and peripheral blood (Figure 6B).


T lymphocytes derived from human cord blood provide effective antitumor immunotherapy against a human tumor.

Lee YS, Kim TS, Kim DK - BMC Cancer (2011)

Cytotoxic activity and detection of cervical tumor specific T cells. A, cervical tumor specific cytotoxic activity was shown. T cells were purified from splenocytes of NOD/SCID mice inoculated with cord blood T cells (CB-T) and Caski tumor cells. The T cells were cocultured with Caski cells, HeLa cells, or A549 cells (human lung cancer cells) as a negative control. Cytotoxicity was tested by flow cytometry with several effector-to-target ratios. B, cytotoxic activity of purified T cells against HeLa (left) and Caski (right) cells was determined for non-adoptive T cells (1st CB-T) and adoptive T cells (2nd CB-T), and human peripheral blood T lymphocytes (PBL-T) as a control. Each bar indicates the mean ± S.D. as determined from the average of quadruplicate wells (* P < 0.01). C, ELISPOT assay for interferon-gamma (INF-γ) production was conducted with E711-20 peptide (YMLDLQPETT) and E786-93 peptide (TLGIVCPI) HPV specific peptides with non-adopted T cells and adopted T cells. Each bar represents the mean ± S.D. as determined (* P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3141763&req=5

Figure 6: Cytotoxic activity and detection of cervical tumor specific T cells. A, cervical tumor specific cytotoxic activity was shown. T cells were purified from splenocytes of NOD/SCID mice inoculated with cord blood T cells (CB-T) and Caski tumor cells. The T cells were cocultured with Caski cells, HeLa cells, or A549 cells (human lung cancer cells) as a negative control. Cytotoxicity was tested by flow cytometry with several effector-to-target ratios. B, cytotoxic activity of purified T cells against HeLa (left) and Caski (right) cells was determined for non-adoptive T cells (1st CB-T) and adoptive T cells (2nd CB-T), and human peripheral blood T lymphocytes (PBL-T) as a control. Each bar indicates the mean ± S.D. as determined from the average of quadruplicate wells (* P < 0.01). C, ELISPOT assay for interferon-gamma (INF-γ) production was conducted with E711-20 peptide (YMLDLQPETT) and E786-93 peptide (TLGIVCPI) HPV specific peptides with non-adopted T cells and adopted T cells. Each bar represents the mean ± S.D. as determined (* P < 0.05).
Mentions: We assessed whether mice with both reconstituted immunity and inoculation with Caski tumor cells possessed memory T cells with specific activity against cervical tumor cells in vitro. Purified CD3+ T cells from these mice were cocultured with Caski cells, as specific target cells, and A549 human lung cancer cells, as a negative control. Cytotoxic effects were then determined by Annexin V and PI staining with flow cytometry analysis. The T cells showed significantly higher cytotoxic activity against Caski cells than against A549 control cells (Figure 6A). This finding suggests that mice with reconstituted immunity that are injected with Caski tumor cells develop memory T cells with specificity for the tumor cells. We performed an additional in vitro analysis to assess the tumor-specific cytotoxic activity of purified CD3+ T cells from spleen of mice which are either adapted or non-adapted mice followed by either Caski cell injection, HeLa cell injection, or no injection of tumor cells. We found much higher cytotoxicity in T cells from tumor adapted T cells bearing mice than T cells than non-adapted control T cells and peripheral blood (Figure 6B).

Bottom Line: Although the graft-versus-tumor (GVT) effect of donor-derived T cells after allogeneic hematopoietic stem cell transplantation has been used as an effective adoptive immunotherapy, the antitumor effects of cord blood (CB) transplantation have not been well studied.NOD/SCID mice that receive neonatal CB transplants have reconstituted T cells with significant antitumor effects against human cervical and lung tumors, with a high infiltration of CD3+ T cells showing dramatic induction of apoptotic cell death.In adoptive transfer of CD3+ T cells into mice with pre-established tumors, we observed much higher antitumor effects of HPV-specific T cells by ELISPOT assays.

View Article: PubMed Central - HTML - PubMed

Affiliation: Transplantation Research Center, Samsung Biomedical Research Institute, Graduate School of Life Science and Biotechnology, CHA University, Seoul, Republic of Korea.

ABSTRACT

Background: Although the graft-versus-tumor (GVT) effect of donor-derived T cells after allogeneic hematopoietic stem cell transplantation has been used as an effective adoptive immunotherapy, the antitumor effects of cord blood (CB) transplantation have not been well studied.

Methods: We established the animal model by transplantation of CB mononuclear cells and/or tumor cells into NOD/SCID mice. The presence of CB derived T cells in NOD/SCID mice or tumor tissues were determined by flow cytometric and immunohistochemical analysis. The anti-tumor effects of CB derived T cells against tumor was determined by tumor size and weight, and by the cytotoxicity assay and ELISPOT assay of T cells.

Results: We found dramatic tumor remission following transfer of CB mononuclear cells into NOD/SCID mice with human cervical tumors with a high infiltration of CD3+ T cells in tumors. NOD/SCID mice that receive neonatal CB transplants have reconstituted T cells with significant antitumor effects against human cervical and lung tumors, with a high infiltration of CD3+ T cells showing dramatic induction of apoptotic cell death. We also confirmed that T cells showed tumor specific antigen cytotoxicity in vitro. In adoptive transfer of CD3+ T cells into mice with pre-established tumors, we observed much higher antitumor effects of HPV-specific T cells by ELISPOT assays.

Conclusions: Our results show that CB derived T lymphocytes will be useful for novel immunotherapeutic candidate cells for therapy of several tumors in clinic.

Show MeSH
Related in: MedlinePlus