Multiple Choice
Passage
Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (Th cell) proliferation. After 7 days, cultures from both individuals were enriched for Th cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control.
![Passage Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (T<sub>h</sub> cell) proliferation. After 7 days, cultures from both individuals were enriched for T<sub>h</sub> cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control. <strong>Figure 1</strong> Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the T<sub>h</sub> cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells. Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785 -Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite? A) B) C) D)](https://d2lvgg3v3hfg70.cloudfront.net/MD0008/11ecba2d_103e_36fb_a3bf_51ceb59d9aae_MD0008_00.jpg)
Figure 1 Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the Th cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells.
Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785
-Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite?
A) ![Passage Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (T<sub>h</sub> cell) proliferation. After 7 days, cultures from both individuals were enriched for T<sub>h</sub> cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control. <strong>Figure 1</strong> Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the T<sub>h</sub> cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells. Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785 -Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite? A) B) C) D)](https://d2lvgg3v3hfg70.cloudfront.net/MD0008/11ecba2d_103e_36fc_a3bf_8114996fe950_MD0008_00.jpg)
B) ![Passage Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (T<sub>h</sub> cell) proliferation. After 7 days, cultures from both individuals were enriched for T<sub>h</sub> cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control. <strong>Figure 1</strong> Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the T<sub>h</sub> cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells. Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785 -Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite? A) B) C) D)](https://d2lvgg3v3hfg70.cloudfront.net/MD0008/11ecba2d_103e_36fd_a3bf_ed10c83ff8c4_MD0008_00.jpg)
C) ![Passage Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (T<sub>h</sub> cell) proliferation. After 7 days, cultures from both individuals were enriched for T<sub>h</sub> cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control. <strong>Figure 1</strong> Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the T<sub>h</sub> cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells. Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785 -Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite? A) B) C) D)](https://d2lvgg3v3hfg70.cloudfront.net/MD0008/11ecba2d_103e_5e0e_a3bf_035a2cab3ef9_MD0008_00.jpg)
D) ![Passage Proteins known as cancer/testis antigens (CTAs) are normally expressed exclusively in germline cells but may become reactivated in cancer cells due to chromatin remodeling (eg, gene activation due to DNA demethylation) . Because of their normally restricted expression, CTAs can elicit an immune response when expressed by cancer cells derived from other tissue types. Several CTAs are known to inhibit apoptosis or promote cell division in cancer cells, making them more resistant to traditional chemotherapy. Accordingly, cancer research instead has focused on immunotherapy treatments targeting CTAs.In a study, researchers isolated peripheral blood lymphocytes (B lymphocytes, T lymphocytes, natural killer [NK] cells) from two healthy individuals. These lymphocytes were cultured with signaling factors known to promote helper T cell (T<sub>h</sub> cell) proliferation. After 7 days, cultures from both individuals were enriched for T<sub>h</sub> cells, which expressed proteins generally involved in activating other lymphocytes. A subset of the cultures from each individual was treated with 5-Aza-CdR, a DNA-demethylating agent. Reverse transcription polymerase chain reaction (RT-PCR) was used to analyze expression of several CTAs (ie, MAGEs) in both untreated and 5-Aza-CdR-treated lymphocyte cultures, as well as in untreated melanoma (skin cancer) and breast cancer cell lines (Figure 1) . GAPDH, a protein expressed in most cell types, served as a control. <strong>Figure 1</strong> Gels showing RT-PCR results for MAGE expressionAdditional peripheral blood lymphocytes were isolated from the same healthy individuals and incubated with the T<sub>h</sub> cells that had been treated with 5-Aza-CdR. After 11 days, these cultures consisted primarily of an increased number of NK cells and cytotoxic T lymphocytes (CTLs) . Researchers found that these CTLs were able to induce apoptosis in breast cancer cell lines but did not affect healthy cells.Researchers used a similar procedure to induce CTL proliferation in lymphocytes isolated from a breast cancer patient. The CTLs were then incubated with cells obtained from a biopsy of the patient's cancerous tissue. Following this experiment, researchers found that apoptosis was induced in the patient's cancer cells. Kirkin et al. Adoptive cancer immunotherapy using DNA-demethylated T helper cells as antigen-presenting cells. Nature Communications. 2018; 9:785 -Treating DNA with the compound bisulfite converts any unmethylated cytosines into uracils. Bisulfite sequencing, which takes advantage of this mechanism, was used to compare DNA methylation of the MAGE-A1 gene in healthy cells and melanoma cells. Based on the passage, which of the following graphs most likely represents the percentage of cytosines converted to uracils in the MAGE-A1 gene after treating DNA from these cells with bisulfite? A) B) C) D)](https://d2lvgg3v3hfg70.cloudfront.net/MD0008/11ecba2d_103e_5e0f_a3bf_879de607367c_MD0008_00.jpg)
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Correct Answer:
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