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The use of gamma-irradiation and ultraviolet-irradiation in the preparation of human melanoma cells for use in autologous whole-cell vaccines.

Deacon DH, Hogan KT, Swanson EM, Chianese-Bullock KA, Denlinger CE, Czarkowski AR, Schrecengost RS, Patterson JW, Teague MW, Slingluff CL - BMC Cancer (2008)

Bottom Line: In contrast to gamma-irradiation, UV-irradiation was also capable of inducing significant levels of apoptosis.UV-irradiation, but not gamma-irradiation, was associated with the loss of tyrosinase expression.These results indicate that UV-irradiation may increase the safety of autologous melanoma vaccines, although it may do so at the expense of altering the antigenic profile of the irradiated tumor cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Surgery, University of Virginia, Charlottesville, VA 22908, USA. dmh9q@virginia.edu

ABSTRACT

Background: Human cancer vaccines incorporating autologous tumor cells carry a risk of implantation and subsequent metastasis of viable tumor cells into the patient who is being treated. Despite the fact that the melanoma cell preparations used in a recent vaccine trial (Mel37) were gamma-irradiated (200 Gy), approximately 25% of the preparations failed quality control release criteria which required that the irradiated cells incorporate 3H-thymidine at no more than 5% the level seen in the non-irradiated cells. We have, therefore, investigated ultraviolet (UV)-irradiation as a possible adjunct to, or replacement for gamma-irradiation.

Methods: Melanoma cells were gamma- and/or UV-irradiated. 3H-thymidine uptake was used to assess proliferation of the treated and untreated cells. Caspase-3 activity and DNA fragmentation were measured as indicators of apoptosis. Immunohistochemistry and Western blot analysis was used to assess antigen expression.

Results: UV-irradiation, either alone or in combination with gamma-irradiation, proved to be extremely effective in controlling the proliferation of melanoma cells. In contrast to gamma-irradiation, UV-irradiation was also capable of inducing significant levels of apoptosis. UV-irradiation, but not gamma-irradiation, was associated with the loss of tyrosinase expression. Neither form of radiation affected the expression of gp100, MART-1/MelanA, or S100.

Conclusion: These results indicate that UV-irradiation may increase the safety of autologous melanoma vaccines, although it may do so at the expense of altering the antigenic profile of the irradiated tumor cells.

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Related in: MedlinePlus

Induction of apoptosis in melanoma cell lines following gamma-irradiation and/or UV-irradiation. Melanoma cells prepared for vaccination were non-irradiated, gamma-irradiated (200 Gy), UV-irradiated (5 min), or both gamma-irradiated (200 Gy) and UV-irradiated (5 min). The cell lines were then cultured in vitro for eight hours prior to assaying (A) caspase activity or (B) DNA fragmentation.
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Figure 6: Induction of apoptosis in melanoma cell lines following gamma-irradiation and/or UV-irradiation. Melanoma cells prepared for vaccination were non-irradiated, gamma-irradiated (200 Gy), UV-irradiated (5 min), or both gamma-irradiated (200 Gy) and UV-irradiated (5 min). The cell lines were then cultured in vitro for eight hours prior to assaying (A) caspase activity or (B) DNA fragmentation.

Mentions: Eight hours following treatment with gamma-irradiation, UV-irradiation or a combination of both forms of radiation, the melanoma cell lines VMM39 and VMM86 were evaluated for apoptosis by caspase and DNA fragmentation assays (Figure 6A, 6B). Gamma-irradiation by itself led to a small increase in caspase detection with melanoma line VMM39, while UV-irradiation, either alone or in combination with gamma-irradiation led to higher levels of detectable caspase with VMM39 (Figure 6A). Similar results were obtained with melanoma line VMM86 except that gamma-irradiation alone did not result in an increase in the detectable caspase. Gamma-irradiation by itself did not increase (VMM39) or only slightly increased (VMM86) DNA fragmentation over that seen in untreated controls (Figure 6B). UV-irradiation induced DNA fragmentation in both cell lines, and the combination of both types of radiation led to the largest increases in DNA fragmentation.


The use of gamma-irradiation and ultraviolet-irradiation in the preparation of human melanoma cells for use in autologous whole-cell vaccines.

Deacon DH, Hogan KT, Swanson EM, Chianese-Bullock KA, Denlinger CE, Czarkowski AR, Schrecengost RS, Patterson JW, Teague MW, Slingluff CL - BMC Cancer (2008)

Induction of apoptosis in melanoma cell lines following gamma-irradiation and/or UV-irradiation. Melanoma cells prepared for vaccination were non-irradiated, gamma-irradiated (200 Gy), UV-irradiated (5 min), or both gamma-irradiated (200 Gy) and UV-irradiated (5 min). The cell lines were then cultured in vitro for eight hours prior to assaying (A) caspase activity or (B) DNA fragmentation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2612687&req=5

Figure 6: Induction of apoptosis in melanoma cell lines following gamma-irradiation and/or UV-irradiation. Melanoma cells prepared for vaccination were non-irradiated, gamma-irradiated (200 Gy), UV-irradiated (5 min), or both gamma-irradiated (200 Gy) and UV-irradiated (5 min). The cell lines were then cultured in vitro for eight hours prior to assaying (A) caspase activity or (B) DNA fragmentation.
Mentions: Eight hours following treatment with gamma-irradiation, UV-irradiation or a combination of both forms of radiation, the melanoma cell lines VMM39 and VMM86 were evaluated for apoptosis by caspase and DNA fragmentation assays (Figure 6A, 6B). Gamma-irradiation by itself led to a small increase in caspase detection with melanoma line VMM39, while UV-irradiation, either alone or in combination with gamma-irradiation led to higher levels of detectable caspase with VMM39 (Figure 6A). Similar results were obtained with melanoma line VMM86 except that gamma-irradiation alone did not result in an increase in the detectable caspase. Gamma-irradiation by itself did not increase (VMM39) or only slightly increased (VMM86) DNA fragmentation over that seen in untreated controls (Figure 6B). UV-irradiation induced DNA fragmentation in both cell lines, and the combination of both types of radiation led to the largest increases in DNA fragmentation.

Bottom Line: In contrast to gamma-irradiation, UV-irradiation was also capable of inducing significant levels of apoptosis.UV-irradiation, but not gamma-irradiation, was associated with the loss of tyrosinase expression.These results indicate that UV-irradiation may increase the safety of autologous melanoma vaccines, although it may do so at the expense of altering the antigenic profile of the irradiated tumor cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Surgery, University of Virginia, Charlottesville, VA 22908, USA. dmh9q@virginia.edu

ABSTRACT

Background: Human cancer vaccines incorporating autologous tumor cells carry a risk of implantation and subsequent metastasis of viable tumor cells into the patient who is being treated. Despite the fact that the melanoma cell preparations used in a recent vaccine trial (Mel37) were gamma-irradiated (200 Gy), approximately 25% of the preparations failed quality control release criteria which required that the irradiated cells incorporate 3H-thymidine at no more than 5% the level seen in the non-irradiated cells. We have, therefore, investigated ultraviolet (UV)-irradiation as a possible adjunct to, or replacement for gamma-irradiation.

Methods: Melanoma cells were gamma- and/or UV-irradiated. 3H-thymidine uptake was used to assess proliferation of the treated and untreated cells. Caspase-3 activity and DNA fragmentation were measured as indicators of apoptosis. Immunohistochemistry and Western blot analysis was used to assess antigen expression.

Results: UV-irradiation, either alone or in combination with gamma-irradiation, proved to be extremely effective in controlling the proliferation of melanoma cells. In contrast to gamma-irradiation, UV-irradiation was also capable of inducing significant levels of apoptosis. UV-irradiation, but not gamma-irradiation, was associated with the loss of tyrosinase expression. Neither form of radiation affected the expression of gp100, MART-1/MelanA, or S100.

Conclusion: These results indicate that UV-irradiation may increase the safety of autologous melanoma vaccines, although it may do so at the expense of altering the antigenic profile of the irradiated tumor cells.

Show MeSH
Related in: MedlinePlus